Methods for preventing or treating coronavirus infections

Abstract

The present invention is in the field of medical treatment and relates to a method for the treatment of coronavirus infections. In particular, the present invention relates to a method for the prophylactic and / or therapeutic treatment of coronavirus infections by mucosal administration of antibodies against coronaviruses.

Description

[Technical Field]

[0001] The present invention is in the field of medical treatment and relates to methods for the treatment of β-coronavirus infections in animals and humans, including severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2).

[0002] In particular, the present invention relates to a method for the prophylactic and / or therapeutic treatment of SARS-CoV-2 by transmucosal administration, in particular intranasal and / or oral inhalation, of antibodies against β-coronaviruses, in particular SARS-CoV-2. [Background technology]

[0003] The SARS-CoV-2 virus causes the disease COVID-19 in humans and is spreading rapidly, having a significant impact on human society. The SARS-CoV-2 virus and its variants of concern are expected to continue to cause high rates of infection and severe illness and death. As of January 2023, the ongoing COVID-19 pandemic caused by SARS-CoV-2 has resulted in more than 6.7 million deaths worldwide since 2019.

[0004] In recent years, other coronavirus outbreaks have caused severe illness and death. While scientists are developing therapeutic antibodies and vaccines against SARS-CoV-2, the risk of future emergence of novel SARS-CoV-2 variants and novel coronaviruses remains. These novel coronaviruses could cause morbidity and mortality and could spark pandemics. Identifying broad-spectrum protective therapies that can combat current and future coronavirus outbreaks remains crucial.

[0005] In addition to SARS-CoV-2, four other betacoronaviruses are known to cause disease in humans: betacoronaviruses HCoV-OC43 (human coronavirus OC43), HCoV-HKU1 (human coronavirus HKU1), SARS-CoV (severe acute respiratory syndrome coronavirus), and MERS-CoV (Middle East respiratory syndrome coronavirus). Spillover events, in which humans become infected with coronaviruses circulating in animal reservoirs, are common. Recently, three betacoronaviruses, including SARS-CoV-2, have crossed over from animals to humans, causing severe epidemics.

[0006] Furthermore, two coronaviruses previously associated only with animal infections have recently been detected in humans presenting with influenza-like symptoms.

[0007] The name coronavirus comes from its crown-like appearance. Coronaviruses are a large group of viruses that have spike proteins on their surface that resemble the spikes of a crown. Antibodies elicited by natural infection or by the COVID-19 vaccines used for herd immunity in 2020–2023 primarily target the variable receptor-binding domain (RBD) of these spike proteins.

[0008] Spike protein structure Coronavirus infection is a multistep process involving the enzymatic cleavage and rearrangement of the surface spike protein, which contains an S1 subunit that contains the receptor-binding domain (RBD) and an S2 subunit that is involved in the fusion of viral and (host) cell membranes, thereby facilitating cell entry.

[0009] The SARS-CoV-2 viral spike protein facilitates viral entry primarily by binding to the angiotensin-converting enzyme 2 (ACE2) receptor on host cells.

[0010] The SARS-CoV-2 spike contains two cleavage sites: a furin cleavage site at the interface of the S1 and S2 subunits, and an S2' site that is highly conserved among different coronaviruses.

[0011] S1 subunit The SARS-CoV-2 spike protein uses the RBD on the S1 subunit to engage with the ACE2 receptor on target cells.

[0012] The S1 subunit is more accessible and remains the primary target of many neutralizing antibodies. However, the S1 subunit is more genetically variable than the S2 subunit, especially when subjected to antibody selection pressure. This tendency toward genetic variability can lead to viral variants with major changes occurring in the S1 subunit. Changes in the receptor-binding domain in SARS-CoV-2 variants of concern dramatically reduce protection from antibodies elicited by previous infection and / or vaccines, often resulting in reinfection.

[0013] S2 subunit The viral spike component, which is important for infection, also contains the structurally complex S2 subunit, which contains dynamic elements important for fusion with host cells. Upon receptor binding, the S1 subunit is discarded, and the membrane enzyme transmembrane serine protease 2 (TMPRSS2) or endosomal cathepsins cleave the S2 region.

[0014] This cleavage leads to insertion of the fusion peptide into the cell membrane, ultimately resulting in viral fusion.

[0015] These S2 subunit elements are less susceptible to genetic variability than the RBD, which has so far been able to retain or even increase its ability to bind ACE2 despite various mutations.

[0016] The S2 domain site presents an inaccessible target for novel therapeutics that protect against a broader range of coronaviruses.

[0017] Stem Helix The stem helices at the base of the viral spike protein are even less accessible than the elements on the S2 subunit, but have the advantage of being historically better conserved in amino acid sequence.

[0018] Treatment and Prevention There are limited viral treatment options for SARS-CoV-2, in the form of small molecule or antiviral antibody drugs. Few interventions exist to prevent SARS-CoV-2 infection; most are non-pharmaceutical, such as social distancing and wearing masks. These methods are non-specific, and efficacy is largely dependent on adherence. Therefore, there is a need for different treatments, especially those that simultaneously target multiple SARS-CoV-2 variants of concern, such as broadly neutralizing antibodies (bnAbs).

[0019] Although treatment of SARS-CoV-2 infection with one, two, or more antibodies is feasible, treatments involving a single antibody or a combination of antibodies would be more cost-effective. The currently predominant SARS-CoV-2 omicron subvariant BA.5 is resistant to most monoclonal antibody therapeutics. Betacoronaviruses, including SARS-CoV-2, also infect a range of animal species known to frequently come into contact with humans, thus increasing the risk of animal transmission and novel outbreaks caused by SARS-CoV-2 variants or viruses that have not previously infected humans and for which no treatment exists.

[0020] Therefore, there is an urgent need for the development of therapeutic mAbs that broadly target β-coronaviruses and novel SARS-CoV-2 variants of concern that are not neutralized by antibodies elicited by immunity and / or natural infection. Summary of the Invention [Problem to be solved by the invention]

[0021] Identifying broad-spectrum protective therapies that can combat the novel coronavirus remains crucial, and the inventors have surprisingly developed a novel array of broad-spectrum protective antibodies, as disclosed herein. [Means for solving the problem]

[0022] The present inventors have discovered that antibodies having the complementarity determining regions (CDRs) disclosed herein can be advantageously used in the treatment of coronavirus infections, particularly in the prophylactic treatment of coronavirus infections.

[0023] The inventors have surprisingly established that mucosal administration of antibodies having the CDRs disclosed herein is a more potent route of prophylactic administration compared to intraperitoneal administration when treating coronavirus infections.

[0024] The inventors have surprisingly established that intranasal administration of antibodies having the CDRs disclosed herein is a more potent route of prophylactic administration compared to intraperitoneal administration when treating coronavirus infection.

[0025] Significantly, the transmucosal administration method of the treatment of the present invention is effective against SARS-Cov-2 and variants of concern and does not require the use of additional antibodies.

[0026] The transmucosal treatment method of the present invention is effective at low dosages and prevents weight loss.

[0027] Complementarity-Determining Regions (CDRs) Preferably, the CDR regions are according to Kabat et al. (1991), as described in Sequences of Proteins of Immunological Interest.

[0028] In a preferred embodiment, the present invention relates to a heavy chain variable dopant antibody comprising a heavy chain CDR1 region comprising any one of SEQ ID NOs: 006 to 011, 015 to 020, 026 to 031, 046 to 051, 054 to 059, or 064 to 069; a heavy chain CDR2 region comprising any one of SEQ ID NOs: 0082 to 087, 093 to 098, 114 to 119, 136 to 141, 158 to 163, or 191 to 196; and a heavy chain CDR3 region comprising any one of SEQ ID NOs: 202 to 207, 218 to 223, 237 to 242, 254 to 259, 280 to 285, or 291 to 296. The present invention provides an antibody comprising a light chain variable domain including a main, a light chain CDR1 region comprising any one of SEQ ID NOs: 301 to 318, 324 to 329, 345 to 350, 367 to 372, 378 to 383, and 400 to 405, a light chain CDR2 region comprising any one of SEQ ID NOs: 430 to 435, 441 to 446, 452 to 457, 463 to 468, 479 to 484, and 502 to 507, and a light chain CDR3 region comprising any one of SEQ ID NOs: 513 to 518, 524 to 529, 545 to 550, 584 to 589, 595 to 600, and 617 to 622.

[0029] In a preferred embodiment, the present invention provides a heavy chain variable domain comprising a heavy chain CDR1 region comprising any one of SEQ ID NOs: 006 to 011, 015 to 020, 026 to 031, 046 to 051, 054 to 059, and 064 to 069; a heavy chain CDR2 region comprising any one of SEQ ID NOs: 082 to 087, 093 to 098, 114 to 119, 136 to 141, 158 to 163, and 191 to 196; and a heavy chain CDR3 region comprising any one of SEQ ID NOs: 202 to 207, 218 to 223, 237 to 242, 254 to 259, 280 to 285, and 291 to 296; Provided is an antibody comprising a light chain variable domain including a light chain CDR1 region comprising any one of SEQ ID NOs: 313 to 318, 324 to 329, 345 to 350, 367 to 372, 378 to 383, and 400 to 405; a light chain CDR2 region comprising any one of SEQ ID NOs: 430 to 435, 441 to 446, 452 to 457, 463 to 468, 479 to 484, and 502 to 507; and a light chain CDR3 region comprising any one of SEQ ID NOs: 513 to 518, 524 to 529, 545 to 550, 584 to 589, 595 to 600, and 617 to 622, wherein the antibody is administered to a mucosal epithelium.

[0030] In a preferred embodiment, the present invention provides a method for treating a coronavirus infection in an individual, the method comprising administering to an individual a heavy chain variable dopant comprising a heavy chain CDR1 region comprising any one of SEQ ID NOs: 006 to 011, 015 to 020, 026 to 031, 046 to 051, 054 to 059, or 064 to 069, a heavy chain CDR2 region comprising any one of SEQ ID NOs: 082 to 087, 093 to 098, 114 to 119, 136 to 141, 158 to 163, or 191 to 196, and a heavy chain CDR3 region comprising any one of SEQ ID NOs: 202 to 207, 218 to 223, 237 to 242, 254 to 259, 280 to 285, or 291 to 296. The method includes a step of administering to an individual in need thereof an antibody comprising a light chain variable domain including a main, a light chain CDR1 region comprising any one of SEQ ID NOs: 313 to 318, 324 to 329, 345 to 350, 367 to 372, 378 to 383, and 400 to 405, a light chain CDR2 region comprising any one of SEQ ID NOs: 430 to 435, 441 to 446, 452 to 457, 463 to 468, 479 to 484, and 502 to 507, and a light chain CDR3 region comprising any one of SEQ ID NOs: 513 to 518, 524 to 529, 545 to 550, 584 to 589, 595 to 600, and 617 to 622, wherein the antibody is administered to the mucosal epithelium.

[0031] In a preferred embodiment, the present invention provides a heavy chain variable domain comprising: a heavy chain CDR1 region comprising any one of SEQ ID NOs: 006 to 011, 015 to 020, 026 to 031, 046 to 051, 054 to 059, or 064 to 069; a heavy chain CDR2 region comprising any one of SEQ ID NOs: 082 to 087, 093 to 098, 114 to 119, 136 to 141, 158 to 163, or 191 to 196; and a heavy chain CDR3 region comprising any one of SEQ ID NOs: 202 to 207, 218 to 223, 237 to 242, 254 to 259, 280 to 285, or 291 to 296; Provided is a composition for mucosal application comprising an antibody comprising a light chain variable domain including a light chain CDR1 region comprising any one of SEQ ID NOs: 313 to 318, 324 to 329, 345 to 350, 367 to 372, 378 to 383, and 400 to 405; a light chain CDR2 region comprising any one of SEQ ID NOs: 430 to 435, 441 to 446, 452 to 457, 463 to 468, 479 to 484, and 502 to 507; and a light chain CDR3 region comprising any one of SEQ ID NOs: 513 to 518, 524 to 529, 545 to 550, 584 to 589, 595 to 600, and 617 to 622.

[0032] In a preferred embodiment, the present invention provides an antibody for use in a method for treating a coronavirus infection in an individual, the antibody comprising a heavy chain CDR1 region comprising any one of SEQ ID NOs: 006 to 011, 015 to 020, 026 to 031, 046 to 051, 054 to 059, or 064 to 069; a heavy chain CDR2 region comprising any one of SEQ ID NOs: 082 to 087, 093 to 098, 114 to 119, 136 to 141, 158 to 163, or 191 to 196; and a heavy chain CDR2 region comprising any one of SEQ ID NOs: 202 to 207, 218 to 223, 237 to 242, 254 to 259, 280 to 285, or 291 to 296. and a light chain variable domain comprising a light chain CDR1 region comprising any one of SEQ ID NOs: 313 to 318, 324 to 329, 345 to 350, 367 to 372, 378 to 383, and 400 to 405; a light chain CDR2 region comprising any one of SEQ ID NOs: 430 to 435, 441 to 446, 452 to 457, 463 to 468, 479 to 484, and 502 to 507; and a light chain CDR3 region comprising any one of SEQ ID NOs: 513 to 518, 524 to 529, 545 to 550, 584 to 589, 595 to 600, and 617 to 622, and the antibody is administered to a mucosal epithelium.

[0033] In a preferred embodiment, the invention provides an antibody or a method or composition comprising an antibody, wherein the antibody comprises a heavy chain variable domain comprising a heavy chain CDR1 region comprising SEQ ID NO: 046, a heavy chain CDR2 region comprising SEQ ID NO: 114, and a heavy chain CDR3 region comprising SEQ ID NO: 254, and a light chain variable domain comprising a light chain CDR1 region comprising SEQ ID NO: 367, a light chain CDR2 region comprising SEQ ID NO: 430, and a light chain CDR3 region comprising SEQ ID NO: 545.

[0034] In a preferred embodiment, the invention provides an antibody or a method or composition comprising an antibody, wherein the antibody comprises a heavy chain variable domain comprising a heavy chain CDR1 region consisting of SEQ ID NO: 046, a heavy chain CDR2 region consisting of SEQ ID NO: 114, and a heavy chain CDR3 region consisting of SEQ ID NO: 254, and a light chain variable domain comprising a light chain CDR1 region consisting of SEQ ID NO: 367, a light chain CDR2 region consisting of SEQ ID NO: 430, and a light chain CDR3 region consisting of SEQ ID NO: 545.

[0035] In a preferred embodiment, disclosed herein is an antibody for use in a method for the prophylactic treatment of a coronavirus infection in an individual, the antibody comprising a heavy chain CDR1 region comprising any one of SEQ ID NOs: 006-011, 015-020, 026-031, 046-051, 054-059, or 064-069; a heavy chain CDR2 region comprising any one of SEQ ID NOs: 082-087, 093-098, 114-119, 136-141, 158-163, or 191-196; and a heavy chain CDR2 region comprising any one of SEQ ID NOs: 202-207, 218-223, 237-242, 254-259, 280-285, or 291-296. The antibody comprises a heavy chain variable domain including a heavy chain CDR3 region; a light chain CDR1 region including any one of SEQ ID NOs: 313 to 318, 324 to 329, 345 to 350, 367 to 372, 378 to 383, and 400 to 405; a light chain CDR2 region including any one of SEQ ID NOs: 430 to 435, 441 to 446, 452 to 457, 463 to 468, 479 to 484, and 502 to 507; and a light chain CDR3 region including any one of SEQ ID NOs: 513 to 518, 524 to 529, 545 to 550, 584 to 589, 595 to 600, and 617 to 622, and 0.1 mg to 20 mg of the antibody is administered to the mucosa.

[0036] In a preferred embodiment, disclosed herein is an antibody for use in a method for the prophylactic treatment of a coronavirus infection in an individual, the antibody comprising a heavy chain CDR1 region comprising any one of SEQ ID NOs: 006-011, 015-020, 026-031, 046-051, 054-059, or 064-069; a heavy chain CDR2 region comprising any one of SEQ ID NOs: 082-087, 093-098, 114-119, 136-141, 158-163, or 191-196; and a heavy chain CDR2 region comprising any one of SEQ ID NOs: 202-207, 218-223, 237-242, 254-259, 280-285, or 291-296. The antibody comprises a heavy chain variable domain including a heavy chain CDR3 region; a light chain CDR1 region including any one of SEQ ID NOs: 313 to 318, 324 to 329, 345 to 350, 367 to 372, 378 to 383, and 400 to 405; a light chain CDR2 region including any one of SEQ ID NOs: 430 to 435, 441 to 446, 452 to 457, 463 to 468, 479 to 484, and 502 to 507; and a light chain CDR3 region including any one of SEQ ID NOs: 513 to 518, 524 to 529, 545 to 550, 584 to 589, 595 to 600, and 617 to 622, and 0.1 mg to 20 mg of the antibody is administered intranasally.

[0037] In a preferred embodiment, disclosed herein is an antibody for use in a method for the prophylactic treatment of a coronavirus infection in an individual, the antibody comprising a heavy chain CDR1 region comprising any one of SEQ ID NOs: 006-011, 015-020, 026-031, 046-051, 054-059, 064-069, a heavy chain CDR2 region comprising any one of SEQ ID NOs: 082-087, 093-098, 114-119, 136-141, 158-163, 191-196, and a heavy chain CDR3 region comprising any one of SEQ ID NOs: 202-207, 218-223, 237-242, 254-259, 280-285, 291-296. The antibody comprises a light chain variable domain comprising a heavy chain variable domain including a DR3 region; a light chain CDR1 region including any one of SEQ ID NOs: 313 to 318, 324 to 329, 345 to 350, 367 to 372, 378 to 383, and 400 to 405; a light chain CDR2 region including any one of SEQ ID NOs: 430 to 435, 441 to 446, 452 to 457, 463 to 468, 479 to 484, and 502 to 507; and a light chain CDR3 region including any one of SEQ ID NOs: 513 to 518, 524 to 529, 545 to 550, 584 to 589, 595 to 600, and 617 to 622, and 0.1 mg to 20 mg of the antibody is administered by oral inhalation.

[0038] In a preferred embodiment, disclosed herein is a composition comprising an antibody for use in a method for preventing or treating coronavirus infection, wherein the antibody comprises a heavy chain variable domain comprising: a heavy chain CDR1 region comprising any one of SEQ ID NOs: 006-011, 015-020, 026-031, 046-051, 054-059, or 064-069; a heavy chain CDR2 region comprising any one of SEQ ID NOs: 082-087, 093-098, 114-119, 136-141, 158-163, or 191-196; and a heavy chain CDR3 region comprising any one of SEQ ID NOs: 202-207, 218-223, 237-242, 254-259, 280-285, or 291-296. the antibody comprises a light chain variable domain comprising: a light chain CDR1 region comprising any one of SEQ ID NOs: 313 to 318, 324 to 329, 345 to 350, 367 to 372, 378 to 383, and 400 to 405; a light chain CDR2 region comprising any one of SEQ ID NOs: 430 to 435, 441 to 446, 452 to 457, 463 to 468, 479 to 484, and 502 to 507; and a light chain CDR3 region comprising any one of SEQ ID NOs: 513 to 518, 524 to 529, 545 to 550, 584 to 589, 595 to 600, and 617 to 622; the administration route includes at least one of pulmonary administration, nasal administration, and oropharyngeal administration; and the nominal dose of the antibody is 0.1 mg to 20 mg.

[0039] Preferably, the antibody disclosed herein comprises a heavy chain variable domain comprising a heavy chain CDR1 region comprising any one of SEQ ID NOs: 006 to 011, 015 to 020, 026 to 031, 046 to 051, 054 to 059, and 064 to 069; a heavy chain CDR2 region comprising any one of SEQ ID NOs: 082 to 087, 093 to 098, 114 to 119, 136 to 141, 158 to 163, and 191 to 196; and a heavy chain CDR3 region comprising any one of SEQ ID NOs: 202 to 207, 218 to 223, 237 to 242, 254 to 259, 280 to 285, and 291 to 296.

[0040] Preferably, the antibody disclosed herein comprises a light chain variable domain comprising: a light chain CDR1 region comprising any one of SEQ ID NOs: 313 to 318, 324 to 329, 345 to 350, 367 to 372, 378 to 383, and 400 to 405; a light chain CDR2 region comprising any one of SEQ ID NOs: 430 to 435, 441 to 446, 452 to 457, 463 to 468, 479 to 484, and 502 to 507; and a light chain CDR3 region comprising any one of SEQ ID NOs: 513 to 518, 524 to 529, 545 to 550, 584 to 589, 595 to 600, and 617 to 622.

[0041] Preferably, in the method for treatment of the present invention, the antibody comprises a heavy chain variable domain comprising a heavy chain CDR1 region comprising any one of SEQ ID NOs: 006 to 011, 015 to 020, 026 to 031, 046 to 051, 054 to 059, and 064 to 069; a heavy chain CDR2 region comprising any one of SEQ ID NOs: 082 to 087, 093 to 098, 114 to 119, 136 to 141, 158 to 163, and 191 to 196; and a heavy chain CDR3 region comprising any one of SEQ ID NOs: 202 to 207, 218 to 223, 237 to 242, 254 to 259, 280 to 285, and 291 to 296.

[0042] Preferably, in the method for treatment of the present invention, the antibody comprises a light chain variable domain comprising a light chain CDR1 region comprising any one of SEQ ID NOs: 313 to 318, 324 to 329, 345 to 350, 367 to 372, 378 to 383, and 400 to 405; a light chain CDR2 region comprising any one of SEQ ID NOs: 430 to 435, 441 to 446, 452 to 457, 463 to 468, 479 to 484, and 502 to 507; and a light chain CDR3 region comprising any one of SEQ ID NOs: 513 to 518, 524 to 529, 545 to 550, 584 to 589, 595 to 600, and 617 to 622.

[0043] In an alternative preferred embodiment, the present invention provides a heavy chain variable fragment comprising a heavy chain CDR1 region comprising any one of SEQ ID NOs: 006 to 011, 015 to 020, 026 to 031, 047 to 051, 054 to 059, or 064 to 069, a heavy chain CDR2 region comprising any one of SEQ ID NOs: 082 to 087, 093 to 098, 115 to 119, 136 to 141, 158 to 163, or 191 to 196, and a heavy chain CDR3 region comprising any one of SEQ ID NOs: 202 to 207, 218 to 223, 237 to 242, 255 to 259, 280 to 285, or 291 to 296. and a light chain CDR3 region comprising any one of SEQ ID NOs: 513 to 518, 524 to 529, 546 to 550, 584 to 589, 595 to 600, and 617 to 622.

[0044] Embodiments with more than one CDR Preferably, the antibodies disclosed herein comprise a heavy chain variable domain comprising a heavy chain CDR2 region comprising any one of SEQ ID NOs: 082-087, 093-098, 114-119, 136-141, 158-163, and 191-196.

[0045] Route of administration In a preferred embodiment, the invention provides an antibody disclosed herein or a method or composition comprising an antibody disclosed herein, wherein the antibody is administered by transmucosal administration.

[0046] In a preferred embodiment, the invention provides an antibody disclosed herein, or a method or composition comprising an antibody disclosed herein, wherein the antibody is administered by intravenous administration.

[0047] In a preferred embodiment, the invention provides an antibody disclosed herein or a method or composition comprising an antibody disclosed herein, wherein the antibody is administered by at least one of oral inhalation, nasal administration, intraocular administration, vaginal administration, rectal administration, and oropharyngeal administration.

[0048] In a preferred embodiment, the antibodies disclosed herein are administered intranasally.

[0049] Such antibodies are useful for treating a coronavirus infection in an individual. Preferably, the method for treating a coronavirus infection is a method for prophylactic and / or therapeutic treatment of a coronavirus infection. Preferably, the antibody is provided to an individual infected with a coronavirus. Preferably, the antibody is provided to an individual prophylactically. Preferably, the antibody is provided to an individual after infection but before the onset of symptoms.

[0050] In a preferred embodiment, the present disclosure provides a method of treating a coronavirus infection in an individual, the method comprising administering to an individual a heavy chain variable domain comprising: a heavy chain CDR1 region comprising any one of SEQ ID NOs: 006-011, 015-020, 026-031, 046-051, 054-059, or 064-069; a heavy chain CDR2 region comprising any one of SEQ ID NOs: 082-087, 093-098, 114-119, 136-141, 158-163, or 191-196; and a heavy chain CDR3 region comprising any one of SEQ ID NOs: 202-207, 218-223, 237-242, 254-259, 280-285, or 291-296. and a light chain CDR3 region comprising any one of SEQ ID NOs: 513 to 518, 524 to 529, 545 to 550, 584 to 589, 595 to 600, and 617 to 622.

[0051] In a preferred embodiment, the individual is infected with or at risk for coronavirus infection.

[0052] In a preferred embodiment, the present disclosure provides an antibody disclosed herein for use in the manufacture of a medicament for use in the treatment of a coronavirus infection.

[0053] section Each of the following numbered sections represents a preferred embodiment of the present invention and is part of this specification. Verse 1: 1. A method for treating a coronavirus infection in an individual, comprising administering to an individual a heavy chain variable domain comprising: a heavy chain CDR1 region comprising any one of SEQ ID NOs: 006-011, 015-020, 026-031, 046-051, 054-059, or 064-069; a heavy chain CDR2 region comprising any one of SEQ ID NOs: 082-087, 093-098, 114-119, 136-141, 158-163, or 191-196; and a heavy chain CDR3 region comprising any one of SEQ ID NOs: 202-207, 218-223, 237-242, 254-259, 280-285, or 291-296; a light chain CDR1 region comprising any one of SEQ ID NOs: 3 to 318, 324 to 329, 345 to 350, 367 to 372, 378 to 383, and 400 to 405; a light chain CDR2 region comprising any one of SEQ ID NOs: 430 to 435, 441 to 446, 452 to 457, 463 to 468, 479 to 484, and 502 to 507; and a light chain CDR3 region comprising any one of SEQ ID NOs: 513 to 518, 524 to 529, 545 to 550, 584 to 589, 595 to 600, and 617 to 622, wherein the antibody is administered mucosally to an individual in need thereof. Verse 2: a heavy chain variable domain comprising a heavy chain CDR1 region comprising any one of SEQ ID NOs: 006 to 011, 015 to 020, 026 to 031, 046 to 051, 054 to 059, and 064 to 069, a heavy chain CDR2 region comprising any one of SEQ ID NOs: 082 to 087, 093 to 098, 114 to 119, 136 to 141, 158 to 163, and 191 to 196, and a heavy chain CDR3 region comprising any one of SEQ ID NOs: 202 to 207, 218 to 223, 237 to 242, 254 to 259, 280 to 285, and 291 to 296; A composition for mucosal application comprising an antibody comprising a light chain variable domain comprising: a light chain CDR1 region comprising any one of SEQ ID NOs: 430 to 435, 441 to 446, 452 to 457, 463 to 468, 479 to 484, and 502 to 507; and a light chain CDR3 region comprising any one of SEQ ID NOs: 513 to 518, 524 to 529, 545 to 550, 584 to 589, 595 to 600, and 617 to 622. Verse 3: 1. An antibody for use in a method for preventing or treating a coronavirus infection in an individual, the antibody comprising: a heavy chain CDR1 region comprising any one of SEQ ID NOs: 006 to 011, 015 to 020, 026 to 031, 046 to 051, 054 to 059, or 064 to 069; a heavy chain CDR2 region comprising any one of SEQ ID NOs: 082 to 087, 093 to 098, 114 to 119, 136 to 141, 158 to 163, or 191 to 196; and a heavy chain CDR3 region comprising any one of SEQ ID NOs: 202 to 207, 218 to 223, 237 to 242, 254 to 259, 280 to 285, or 291 to 296. an antibody for mucosal administration, the antibody comprising a light chain variable domain including a heavy chain variable domain containing an R3 region; a light chain CDR1 region comprising any one of SEQ ID NOs: 313 to 318, 324 to 329, 345 to 350, 367 to 372, 378 to 383, and 400 to 405; a light chain CDR2 region comprising any one of SEQ ID NOs: 430 to 435, 441 to 446, 452 to 457, 463 to 468, 479 to 484, and 502 to 507; and a light chain CDR3 region comprising any one of SEQ ID NOs: 513 to 518, 524 to 529, 545 to 550, 584 to 589, 595 to 600, and 617 to 622. Verse 4: The method, composition, or antibody according to any preceding clause, wherein the antibody comprises a heavy chain variable domain comprising a heavy chain CDR1 region comprising SEQ ID NO: 046, a heavy chain CDR2 region comprising SEQ ID NO: 114, and a heavy chain CDR3 region comprising SEQ ID NO: 254; a light chain variable domain comprising a light chain CDR1 region comprising SEQ ID NO: 367, a light chain CDR2 region comprising SEQ ID NO: 430, and a light chain CDR3 region comprising SEQ ID NO: 545. Verse 5: The method, composition, or antibody according to any preceding clause, wherein the method for treatment of coronavirus virus infection is a method for prophylactic and / or therapeutic treatment of coronavirus, preferably the method for treatment is for prophylactic treatment of SARS-COV-2 infection. Verse 6: The method, composition, or antibody according to any preceding clause, wherein the antibody comprises a heavy chain variable domain having the sequence of SEQ ID NO: 629 with at most 15, preferably 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, amino acid insertions, deletions, or substitutions that are not in the heavy chain CDRs. Verse 7: The method, composition, or antibody according to any preceding clause, wherein the antibody comprises a light chain variable domain having the sequence of SEQ ID NO: 630 with at most 15, preferably 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, amino acid insertions, deletions, or substitutions that are not in the light chain CDRs. Verse 8: The method, composition, or antibody according to any preceding clause, wherein the antibody is an IgG antibody, preferably an IgG1 antibody. Verse 9: The method, composition, or antibody according to any preceding clause, wherein the antibody is provided prophylactically to the individual. Verse 10: The method, composition, or antibody according to any preceding clause, wherein the route of administration comprises at least one of oral inhalation, nasal administration, intraocular administration, and oropharyngeal administration. Verse 11: The method, composition, or antibody according to any preceding clause, wherein the antibody is administered at least once or at least twice monthly. Verse 12: The method, composition, or antibody according to any preceding clause, wherein the antibody is administered to the individual in a dosage of between 0.01 mg and 20 mg. Verse 13: 16. A composition according to any preceding clause comprising a single dose unit of 0.01 mg to 20 mg, preferably 0.1 mg to 15 mg, or preferably 0.5 mg to 10 mg of the antibody, wherein the antibody is defined in any one of clauses 1 to 12. Verse 14: A composition according to any preceding clause that is self-administered. Verse 15: A pharmaceutical delivery device comprising a composition according to any preceding clause.

[0054] Dosage Preferably, 0.01 mg to 20 mg of antibody is administered to an individual, and more preferably, 0.1 mg to 20 mg of antibody is administered to an individual.

[0055] The fixed dose, or nominal dose, of the antibody is preferably 0.01 mg to 17 mg, preferably 0.011 mg to 16 mg, preferably 0.012 mg to 15 mg, preferably 0.013 mg to 14 mg, preferably 0.014 mg to 13 mg, preferably 0.015 mg to 12 mg, preferably 0.016 mg to 11 mg, preferably 0.017 mg to 10 mg, preferably 0.018 mg to 9 mg, preferably 0.020 mg to 8 mg, preferably 0.023 mg to 7 mg, preferably 0.025 mg to 6 mg, preferably 0.030 mg to 5 mg, preferably 0.040 mg to 4 mg, preferably 0.050 mg to 3 mg, preferably 0.075 mg to 2 mg, or preferably 0.10 mg to 1 mg.

[0056] The fixed dose, or nominal dose, of the antibody is preferably 0.010 mg to 5.0 mg, preferably 0.020 mg to 4.5 mg, preferably 0.030 mg to 4.0 mg, preferably 0.040 mg to 3.5 mg, preferably 0.050 mg to 3.0 mg, preferably 0.060 mg to 2.5 mg, preferably 0.070 mg to 2.0 mg, preferably 0.080 mg to 1.5 mg, preferably 0.090 mg to 1.0 mg, or preferably 0.100 mg to 0.5 mg.

[0057] The fixed dose, or nominal dose, of the antibody is preferably 0.010 mg to 15.0 mg, preferably 0.020 mg to 14.5 mg, preferably 0.030 mg to 4.0 mg, preferably 0.040 mg to 13.5 mg, preferably 0.050 mg to 13.0 mg, preferably 0.060 mg to 12.5 mg, preferably 0.070 mg to 12.0 mg, preferably 0.080 mg to 11.5 mg, preferably 0.090 mg to 11.0 mg, or preferably 0.100 mg to 10.5 mg.

[0058] The fixed dose, or nominal dose, of antibody to be delivered to a human subject is preferably 0.1 μg to 10 μg, preferably 0.2 μg to 9 μg, preferably 0.3 μg to 8 μg, preferably 0.4 μg to 7 μg, preferably 0.5 μg to 6 μg, preferably 0.6 μg to 5 μg, preferably 0.7 μg to 4 μg, preferably 0.8 μg to 3 μg, preferably 0.9 μg to 2 μg, or preferably about 1 μg.

[0059] The fixed dose, or nominal dose, of antibody to be delivered to a human subject is preferably 1 mg to 11 mg, preferably 2 mg to 12 mg, preferably 3 mg to 13 mg, preferably 4 mg to 14 mg, preferably 5 mg to 15 mg, preferably 6 mg to 16 mg, preferably 7 mg to 17 mg, preferably 8 mg to 18 mg, preferably 9 mg to 19 mg, or preferably 10 mg to 20 mg.

[0060] The fixed dose, or nominal dose, of antibody to be delivered to a human subject is preferably 1 mg to 20 mg, preferably 2 mg to 19 mg, preferably 3 mg to 18 mg, preferably 4 mg to 17 mg, preferably 5 mg to 16 mg, preferably 6 mg to 15 mg, preferably 7 mg to 14 mg, preferably 8 mg to 13 mg, preferably 9 mg to 12 mg, or preferably 10 mg to 11 mg.

[0061] The fixed dose, or nominal dose, of antibody to be delivered to a human subject is preferably 11 mg to 20 mg, preferably 10 mg to 19 mg, preferably 9 mg to 18 mg, preferably 8 mg to 17 mg, preferably 7 mg to 16 mg, preferably 6 mg to 15 mg, preferably 5 mg to 14 mg, preferably 4 mg to 13 mg, preferably 3 mg to 12 mg, preferably 2 mg to 11 mg, or preferably 1 mg to 10 mg.

[0062] In preferred embodiments, the present disclosure further provides compositions formulated for mucosal administration comprising an antibody disclosed herein in a single dosage unit of between 0.1 mg and 20 mg, preferably between 5 mg and 15 mg, or preferably between 7.5 mg and 12.5 mg.

[0063] In preferred embodiments, the present disclosure further provides compositions formulated for intranasal administration comprising an antibody disclosed herein in a single dosage unit of between 0.1 mg and 20 mg, preferably between 5 mg and 15 mg, or preferably between 7.5 mg and 12.5 mg.

[0064] In preferred embodiments, the present disclosure further provides compositions formulated for oral inhalation comprising an antibody disclosed herein in a single dosage unit of between 0.1 mg and 20 mg, preferably between 5 mg and 15 mg, or preferably between 7.5 mg and 12.5 mg.

[0065] Timing and Spacing Preferably, the antibody is administered at least once or at least twice per month.

[0066] Preferably, the antibody is administered at least once or at least twice per week.

[0067] The fixed dose, or nominal dose, of antibody per week is 0.010 mg to 17 mg, preferably 0.011 mg to 16 mg, preferably 0.012 mg to 15 mg, preferably 0.013 mg to 14 mg, preferably 0.014 mg to 13 mg, preferably 0.015 mg to 12 mg, preferably 0.016 mg to 11 mg, preferably 0.017 mg to 10 mg, preferably 0.018 mg to 9 mg, preferably 0.020 mg to 8 mg, preferably 0.023 mg to 7 mg, preferably 0.025 mg to 6 mg, preferably 0.030 mg to 5 mg, preferably 0.040 mg to 4 mg, preferably 0.050 mg to 3 mg, preferably 0.075 mg to 2 mg, or preferably 0.10 mg to 1 mg.

[0068] The fixed dose, or nominal dose, of antibody per week is 0.010 mg to 5.0 mg, preferably 0.020 mg to 4.5 mg, preferably 0.030 mg to 4.0 mg, preferably 0.040 mg to 3.5 mg, preferably 0.050 mg to 3.0 mg, preferably 0.060 mg to 2.5 mg, preferably 0.070 mg to 2.0 mg, preferably 0.080 mg to 1.5 mg, preferably 0.090 mg to 1.0 mg, or preferably 0.100 mg to 0.5 mg.

[0069] Preferably, the antibody is administered at least once or at least twice per day. Preferably, the antibody is administered daily. Preferably, the antibody is administered twice daily.

[0070] Preferably, the daily fixed dose, or nominal dose, is between 0.001 mg and 17 mg, preferably between 0.011 mg and 16 mg, preferably between 0.012 mg and 15 mg, preferably between 0.013 mg and 14 mg, preferably between 0.014 mg and 13 mg, preferably between 0.015 mg and 12 mg, preferably between 0.016 mg and 11 mg, preferably between 0.017 mg and 10 mg, preferably between 0.018 mg and 9 mg, preferably between 0.020 mg and 8 mg, preferably between 0.023 mg and 7 mg, preferably between 0.025 mg and 6 mg, preferably between 0.030 mg and 5 mg, preferably between 0.040 mg and 4 mg, preferably between 0.050 mg and 3 mg, preferably between 0.075 mg and 2 mg, or preferably between 0.10 mg and 1 mg.

[0071] Framework Area Preferably, the antibody comprises a heavy chain variable domain having the sequence of SEQ ID NO: 629 and / or a light chain variable domain having the sequence of SEQ ID NO: 630.

[0072] Preferably, the heavy chain variable domain of the antibody further comprises a heavy chain framework region FR1 of SEQ ID NO: 631, a heavy chain framework region FR2 of SEQ ID NO: 632, a heavy chain framework region FR3 of SEQ ID NO: 633, and / or a heavy chain framework region FR4 of SEQ ID NO: 634.

[0073] Preferably, the light chain variable domain further comprises a light chain framework region RF1 of SEQ ID NO: 635, a light chain framework region FR2 of SEQ ID NO: 636, a light chain framework region FR3 of SEQ ID NO: 637, and / or a light chain framework region FR4 of SEQ ID NO: 638.

[0074] Combination Compositions In a preferred embodiment, the inventors disclose a composition comprising a first binding fragment as disclosed herein and a second binding fragment, wherein the first fragment is selected from the group consisting of SEQ ID NOs: 006-011, 015-020, 026-031, 046-051, 054-059, 064-069, 082-087, 093-098, 114-119, 136-141, 158-163, 191-196, 202-207, 218-223, 237-240, 241-242, 251-252, 261-262, 263-264, 270-272, 275-276, 280-282, 283-284, 285-286, 287-288, 290-300, 301-302, 303-304, 305-306, 307-308, 309-409, 410-411, 412-413, 414-415, 416-417, 418-419, 420-421, 422-423, 424-425, 426-427, 428-429, 430-431, 432-433, 434-435, 436-437, 438-440, 440-441, 442-443, 444-445, 446-447, 448-449, 450-451, More preferably, the first fragment comprises a sequence comprising any one of SEQ ID NOs: 046, 114, 254, 367, 430, 545, 550, 567, 570, 571, 572, 573, 574, 575, 576, 577, 578, 579, 584, 589, 595, 600, and 617 to 622. More preferably, the first fragment comprises a sequence comprising any one or more of SEQ ID NOs: 046, 114, 254, 367, 430, and 545. The second binding fragment comprises a sequence comprising any one of SEQ ID NOs: 639, 640, 641, 642, 643, or SEQ ID NO: 644. Preferably, the fragment is selected from any one of the group comprising a full-length antibody, a Fab, a modified Fab, a Fab', a modified Fab', a F(ab')2, an Fv, a single domain antibody, a scFv, a scFv-Fc, a bivalent, trivalent, or tetravalent antibody, a Bis-scFv, a diabody, a triabody, a tetrabody, and epitope-binding fragments thereof.

[0075] In a preferred embodiment, the inventors disclose a composition comprising a first binding fragment as disclosed herein and a second binding fragment, wherein the first fragment is selected from the group consisting of SEQ ID NOs: 006-011, 015-020, 026-031, 046-051, 054-059, 064-069, 082-087, 093-098, 114-119, 136-141, 158-163, 191-196, 202-207, 218-223, 237-240, 241-242, 251-252, 261-262, 263-264, 270-272, 275-276, 280-282, 283-284, 285-286, 287-288, 290-300, 301-302, 303-304, 305-306, 307-308, 309-409, 410-411, 412-413, 414-415, 416-417, 418-419, 420-421, 422-423, 424-425, 426-427, 428-429, 430-431, 432-433, 434-435, 436-437, 438-440, 440-441, 442-443, 444-445, 446-447, 448-449, 450-451, More preferably, the first fragment comprises a sequence comprising any one of SEQ ID NOs: 046, 114, 254, 367, 430, 441, 442, 254 to 259, 280 to 285, 291 to 296, 313 to 318, 324 to 329, 345 to 350, 367 to 372, 378 to 383, 400 to 405, 430 to 435, 441 to 446, 452 to 457, 463 to 468, 479 to 484, 502 to 507, 513 to 518, 524 to 529, 545 to 550, 584 to 589, 595 to 600, and 617 to 622. More preferably, the first fragment comprises any one or more of SEQ ID NOs: 046, 114, 254, 367, 430, or 545. The second binding fragment comprises a sequence comprising any one of SEQ ID NOs: 645, 646, 647, 648, 649, or SEQ ID NO: 650. Preferably, the fragment is selected from any one of the group comprising a full-length antibody, a Fab, a modified Fab, a Fab', a modified Fab', a F(ab')2, an Fv, a single domain antibody, a scFv, a scFv-Fc, a bivalent, trivalent, or tetravalent antibody, a Bis-scFv, a diabody, a triabody, a tetrabody, and epitope-binding fragments thereof.

[0076] In a preferred embodiment, the inventors disclose a composition comprising a first binding fragment as disclosed herein and a second binding fragment, wherein the first fragment is selected from the group consisting of SEQ ID NOs: 006-011, 015-020, 026-031, 046-051, 054-059, 064-069, 082-087, 093-098, 114-119, 136-141, 158-163, 191-196, 202-207, 218-223, 237-240, 241-242, 251-252, 261-262, 263-264, 270-272, 275-276, 280-282, 283-284, 285-286, 287-288, 290-300, 301-302, 303-304, 305-306, 307-308, 309-409, 410-411, 412-413, 414-415, 416-417, 418-419, 420-421, 422-423, 424-425, 426-427, 428-429, 430-431, 432-433, 434-435, 436-437, 438-440, 440-441, 442-443, 444-445, 446-447, 448-449, 450-451, More preferably, the first fragment comprises a sequence comprising any one of SEQ ID NOs: 046, 114, 254, 367, 430, 545, 550, 567, 570, 571, 572, 573, 574, 575, 576, 577, 578, 579, 584, 589, 595, 600, and 617 to 622. More preferably, the first fragment comprises a sequence comprising any one or more of SEQ ID NOs: 046, 114, 254, 367, 430, and 545. The second binding fragment comprises a sequence comprising any one of SEQ ID NOs: 651, 652, 653, 654, 655, or SEQ ID NO: 656. Preferably, the fragment is selected from any one of the group comprising a full-length antibody, a Fab, a modified Fab, a Fab', a modified Fab', a F(ab')2, an Fv, a single domain antibody, a scFv, a scFv-Fc, a bivalent, trivalent, or tetravalent antibody, a Bis-scFv, a diabody, a triabody, a tetrabody, and epitope-binding fragments thereof.

[0077] In a preferred embodiment, the inventors disclose a composition comprising a first binding fragment as disclosed herein and a second binding fragment, wherein the first fragment is selected from the group consisting of SEQ ID NOs: 006-011, 015-020, 026-031, 046-051, 054-059, 064-069, 082-087, 093-098, 114-119, 136-141, 158-163, 191-196, 202-207, 218-223, 237-240, 241-242, 251-252, 261-262, 263-264, 270-272, 275-276, 280-282, 283-284, 285-286, 287-288, 290-300, 301-302, 303-304, 305-306, 307-308, 309-409, 410-411, 412-413, 414-415, 416-417, 418-419, 420-421, 422-423, 424-425, 426-427, 428-429, 430-431, 432-433, 434-435, 436-437, 438-440, 440-441, 442-443, 444-445, 446-447, 448-449, 450-451, More preferably, the first fragment comprises a sequence comprising any one of SEQ ID NOs: 046, 114, 254, 367, 430, and 545. The second binding fragment comprises a sequence comprising any one of SEQ ID NOs: 657, 658, 659, 660, 661, or SEQ ID NO: 662. Preferably, the fragment is selected from any one of the group comprising a full-length antibody, a Fab, a modified Fab, a Fab', a modified Fab', a F(ab')2, an Fv, a single domain antibody, a scFv, a scFv-Fc, a bivalent, trivalent, or tetravalent antibody, a Bis-scFv, a diabody, a triabody, a tetrabody, and epitope-binding fragments thereof.

[0078] bispecific antibody In a preferred embodiment, the inventors disclose a bispecific antibody, which is capable of binding to the stem helix of a coronavirus. In a preferred embodiment, the inventors disclose a bispecific antibody, which is capable of binding to the stem helix of SARS-CoV-2.

[0079] In a preferred embodiment, the inventors disclose an anti-coronavirus bispecific antibody or antigen-binding fragment thereof having a first Fab capable of binding to the stem helix of a coronavirus and a second Fab capable of binding to the fusion peptide of a coronavirus. In a preferred embodiment, the inventors disclose an anti-SARS-CoV-2 bispecific antibody or antigen-binding fragment thereof having a first Fab capable of binding to the stem helix of SARS-CoV-2 and a second Fab capable of binding to the fusion peptide of SARS-CoV-2.

[0080] In a preferred embodiment, the inventors disclose an anti-SARS-CoV-2 bispecific antibody or antigen-binding fragment thereof having a first Fab capable of binding to the stem helix of SARS-CoV-2 and a second Fab capable of binding to the fusion peptide of SARS-CoV-2, wherein the first Fab comprises a sequence comprising any one or more of the preferred SEQ ID NOs: 046, 114, 254, 367, 430, or 545, or the first Fab comprises a sequence comprising any one or more of the preferred SEQ ID NOs: 651, 652, 653, 654, 655, 656, 657, 658, 659, 660, 661, 662, 663, 664, 665, 666, 667, 668, 669, 670, 671, 672, 673, 674, 675, 676, 677, 678, 679, 680, 681, 682, 683, 684, 685, 686, 687, 688, 689, 690, 700, 701, 702, 703, 704, 705, 706, 707, 708, 709, 710, 711, 712, 713, 714, 715, 716, 717, 718, 719, 720, 721, 722, 723, 724, 725, 726, 727, 728, 729, 730, 731, 732 Alternatively, the first Fab comprises a sequence comprising one or more of preferred SEQ ID NOs: 657, 658, 659, 660, 661-662, and the second Fab comprises a sequence comprising one or more of preferred SEQ ID NOs: 639, 640, 641, 642, 643-644, or the second Fab comprises a sequence comprising one or more of preferred SEQ ID NOs: 645, 646, 647, 648, 649-650.

[0081] In a preferred embodiment, the inventors disclose an anti-SARS-CoV-2 bispecific antibody or antigen-binding fragment thereof, having a first Fab capable of binding to the stem helix of SARS-CoV-2 and a second Fab capable of binding to the fusion peptide of SARS-CoV-2, wherein the first Fab comprises a sequence comprising any one or more of preferred SEQ ID NOs: 046, 114, 254, 367, 430, or 545, and the second Fab comprises a sequence comprising any one or more of preferred SEQ ID NOs: 639, 640, 641, 642, 643-644. Preferably, the fragment is selected from any one of the group comprising a full-length antibody, a Fab, a modified Fab, a Fab', a modified Fab', a F(ab'), an Fv, a single-domain antibody, a scFv, a scFv-Fc, a bivalent, trivalent, or tetravalent antibody, a Bis-scFv, a diabody, a triabody, a tetrabody, and epitope-binding fragments thereof.

[0082] In a preferred embodiment, the inventors disclose an anti-SARS-CoV-2 bispecific antibody or antigen-binding fragment thereof, having a first Fab capable of binding to the stem helix of SARS-CoV-2 and a second Fab capable of binding to the fusion peptide of SARS-CoV-2, wherein the first Fab comprises a sequence comprising any one or more of preferred SEQ ID NOs: 046, 114, 254, 367, 430, or 545, and the second Fab comprises a sequence comprising any one or more of preferred SEQ ID NOs: 645, 646, 647, 648, 649-650. Preferably, the fragment is selected from any one of the group comprising a full-length antibody, a Fab, a modified Fab, a Fab', a modified Fab', a F(ab'), an Fv, a single-domain antibody, a scFv, a scFv-Fc, a bivalent, trivalent, or tetravalent antibody, a Bis-scFv, a diabody, a triabody, a tetrabody, and epitope-binding fragments thereof.

[0083] In a preferred embodiment, the inventors disclose an anti-SARS-CoV-2 bispecific antibody or antigen-binding fragment thereof having a first Fab capable of binding to the stem helix of SARS-CoV-2 and a second Fab also capable of binding to the stem helix of SARS-CoV-2, wherein the first Fab comprises a sequence comprising any one or more of preferred SEQ ID NOs: 046, 114, 254, 367, 430, or 545, and the second Fab comprises a sequence comprising any one or more of preferred SEQ ID NOs: 651, 652, 653, 654, 655-656. Preferably, the fragment is selected from any one of the group comprising a full-length antibody, a Fab, a modified Fab, a Fab', a modified Fab', a F(ab'), an Fv, a single-domain antibody, a scFv, a scFv-Fc, a bivalent, trivalent, or tetravalent antibody, a Bis-scFv, a diabody, a triabody, a tetrabody, and epitope-binding fragments thereof.

[0084] In a preferred embodiment, the inventors disclose an anti-SARS-CoV-2 bispecific antibody or antigen-binding fragment thereof having a first Fab capable of binding to the stem helix of SARS-CoV-2 and a second Fab also capable of binding to the stem helix of SARS-CoV-2, wherein the first Fab comprises a sequence comprising any one or more of preferred SEQ ID NOs: 046, 114, 254, 367, 430, or 545, and the second Fab comprises a sequence comprising any one or more of preferred SEQ ID NOs: 657, 658, 659, 660, 661-662. Preferably, the fragment is selected from any one of the group comprising a full-length antibody, a Fab, a modified Fab, a Fab', a modified Fab', a F(ab'), an Fv, a single-domain antibody, a scFv, a scFv-Fc, a bivalent, trivalent, or tetravalent antibody, a Bis-scFv, a diabody, a triabody, a tetrabody, and epitope-binding fragments thereof.

[0085] In a preferred embodiment, we disclose a bispecific antibody comprising a first Fab and a second Fab, wherein the first Fab comprises any one or more of the sequences set forth in preferred SEQ ID NOs: 046, 114 or 254, and the second Fab comprises any one or more of the sequences set forth in preferred SEQ ID NOs: 660, 661 or 662.

[0086] In a preferred embodiment, we disclose a bispecific antibody comprising a first Fab and a second Fab, wherein the first Fab comprises any one or more of the sequences set forth in preferred SEQ ID NOs: 046, 114 or 254, and the second Fab comprises any one or more of the sequences set forth in preferred SEQ ID NOs: 642, 643 or 644.

[0087] In a preferred embodiment, we disclose a bispecific antibody comprising a first Fab and a second Fab, wherein the first Fab comprises any one or more of the sequences set forth in preferred SEQ ID NOs: 046, 114 or 254, and the second Fab comprises any one or more of the sequences set forth in preferred SEQ ID NOs: 648, 649 or 650.

[0088] In a preferred embodiment, we disclose a bispecific antibody comprising a first Fab and a second Fab, wherein the first Fab comprises any one or more of the sequences set forth in preferred SEQ ID NOs: 046, 114 or 254, and the second Fab comprises any one or more of the sequences set forth in preferred SEQ ID NOs: 654, 655 or 656.

[0089] In a preferred embodiment, we disclose a bispecific antibody comprising a first Fab and a second Fab, wherein the first Fab comprises any one or more of the sequences set forth in preferred SEQ ID NOs: 657, 658 or 659, and the second Fab comprises any one or more of the sequences set forth in preferred SEQ ID NOs: 367, 430 or 545.

[0090] In a preferred embodiment, we disclose a bispecific antibody comprising a first Fab and a second Fab, wherein the first Fab comprises any one or more of the sequences set forth in preferred SEQ ID NOs: 639, 640 or 641, and the second Fab comprises any one or more of the sequences set forth in preferred SEQ ID NOs: 367, 430 or 545.

[0091] In a preferred embodiment, we disclose a bispecific antibody comprising a first Fab and a second Fab, wherein the first Fab comprises any one or more of the sequences set forth in preferred SEQ ID NOs: 645, 646 or 647, and the second Fab comprises any one or more of the sequences set forth in preferred SEQ ID NOs: 367, 430 or 545.

[0092] In a preferred embodiment, we disclose a bispecific antibody comprising a first Fab and a second Fab, wherein the first Fab comprises any one or more of the sequences set forth in preferred SEQ ID NOs: 651, 652 or 653, and the second Fab comprises any one or more of the sequences set forth in preferred SEQ ID NOs: 367, 430 or 545.

[0093] The present invention also provides methods for the construction, expression, and purification of bispecific antibodies having the function of binding to the stem helix of SARS-CoV-2, and the use of bispecific antibodies in the field of medicine, particularly in the prevention and / or treatment of SARS-CoV-2 infection.

[0094] section Each of the following numbered sections represents a preferred embodiment of the present invention and is part of this specification. Verse 26: An anti-coronavirus, preferably anti-SARS-CoV-2 bispecific antibody or antigen-binding fragment thereof, having a first Fab capable of binding to the stem helix of a coronavirus, preferably SARS-CoV-2, and a second Fab capable of binding to the fusion peptide of a coronavirus, preferably SARS-CoV-2, wherein the first Fab comprises a sequence comprising preferably any one or more of SEQ ID NOs: 046, 114, 254, 367, 430, or 545, and the second Fab comprises a sequence comprising preferably any one or more of SEQ ID NOs: 639, 640, 641, 642, 643-644. Verse 27: 27. The bispecific antibody or antigen-binding fragment thereof according to clause 26, comprising a first Fab capable of binding to the stem helix of a coronavirus, preferably SARS-CoV-2, and a second Fab capable of binding to the fusion peptide of a coronavirus, preferably SARS-CoV-2, wherein the first Fab comprises a sequence comprising preferably any one or more of SEQ ID NOs: 046, 114, 254, 367, 430, and / or 545, and the second Fab comprises a sequence comprising preferably any one or more of SEQ ID NOs: 639, 640, 641, 642, 643, and / or 644. Verse 28: a first Fab that specifically binds to the stem helix of a coronavirus, preferably SARS-CoV-2, and a second Fab that specifically binds to the fusion peptide of a coronavirus, preferably SARS-CoV-2; a.) a first Fab comprises a heavy chain variable region comprising, as CDRs, the heavy chain CDR1 region of SEQ ID NO: 046, the heavy chain CDR2 region of SEQ ID NO: 114, and the heavy chain CDR3 region of SEQ ID NO: 254, and a light chain variable region comprising, as CDRs, the light chain CDR1 region of SEQ ID NO: 367, the light chain CDR2 region of SEQ ID NO: 430, and the light chain CDR3 region of SEQ ID NO: 545; b.) the second Fab comprises a heavy chain variable region comprising, as CDRs, the heavy chain CDR1 region of SEQ ID NO: 639, the heavy chain CDR2 region of SEQ ID NO: 640, and the heavy chain CDR3 region of SEQ ID NO: 641, and a light chain variable region comprising, as CDRs, the light chain CDR1 region of SEQ ID NO: 642, the light chain CDR2 region of SEQ ID NO: 643, and the light chain CDR3 region of SEQ ID NO: 644; 28. The bispecific antibody or antigen-binding fragment thereof according to clause 26 or 27, Verse 29: a) a light chain comprising a VL-CL domain and a heavy chain comprising a VH-CH1-CH2-CH3 domain of an antibody or antigen-binding fragment thereof capable of binding to the stem helix of a coronavirus, preferably SARS-CoV-2; and b) a light chain comprising the VL-CL domains and a heavy chain comprising the VH-CH1-CH2-CH3 domains of an antibody or antigen-binding fragment thereof capable of binding to the fusion peptide of a coronavirus, preferably SARS-CoV-2; Including, the constant domains CL and CH1 from an antibody or antigen-binding fragment thereof capable of binding to the stem helix of a coronavirus, preferably SARS-CoV-2, are replaced by each other; 29. The bispecific antibody or antigen-binding fragment thereof according to any one of clauses 26 to 28. Verse 30: the VH domain of the antibody or antigen-binding fragment thereof capable of binding to the stem helix of a coronavirus, preferably SARS-CoV-2, comprises, as CDRs, a heavy chain CDR1 region of SEQ ID NO: 046, a heavy chain CDR2 region of SEQ ID NO: 114, and a heavy chain CDR3 region of SEQ ID NO: 254; and the VL domain of the antibody or antigen-binding fragment thereof capable of binding to the stem helix of a coronavirus, preferably SARS-CoV-2, comprises, as CDRs, a light chain CDR1 region of SEQ ID NO: 367, a light chain CDR2 region of SEQ ID NO: 430, and a light chain CDR3 region of SEQ ID NO: 545; The VH domain of the antibody or antigen-binding fragment thereof capable of binding to the fusion peptide of coronavirus, preferably SARS-CoV-2, comprises, as CDRs, the heavy chain CDR1 region of SEQ ID NO: 639, the heavy chain CDR2 region of SEQ ID NO: 640, and the heavy chain CDR3 region of SEQ ID NO: 641; and the VL domain of the antibody or antigen-binding fragment thereof capable of binding to the fusion peptide of coronavirus, preferably SARS-CoV-2, comprises, as CDRs, the light chain CDR1 region of SEQ ID NO: 642, the light chain CDR2 region of SEQ ID NO: 643, and the light chain CDR3 region of SEQ ID NO: 644. 29. A bispecific antibody or antigen-binding fragment thereof according to clause 29. Verse 31: a) a light chain comprising a VL-CL domain and a heavy chain comprising a VH-CH1-CH2-CH3 domain of an antibody capable of binding to the stem helix of a coronavirus, preferably SARS-CoV-2, and b) a light chain comprising VL-CL domains and a heavy chain comprising VH-CH1-CH2-CH3 domains of an antibody capable of binding to the fusion peptide of a coronavirus, preferably SARS-CoV-2 Including, the constant domains CL and CH1 from an antibody capable of binding to the fusion peptide of a coronavirus, preferably SARS-CoV-2, are replaced by each other; 29. The bispecific antibody or antigen-binding fragment thereof according to any one of clauses 26 to 28. Verse 32: the VH domain of the antibody or antigen-binding fragment thereof capable of binding to the stem helix of a coronavirus, preferably SARS-CoV-2, comprises, as CDRs, a heavy chain CDR1 region of SEQ ID NO: 046, a heavy chain CDR2 region of SEQ ID NO: 114, and a heavy chain CDR3 region of SEQ ID NO: 254; and the VL domain of the antibody capable of binding to the stem helix of a coronavirus, preferably SARS-CoV-2, comprises, as CDRs, a light chain CDR1 region of SEQ ID NO: 367, a light chain CDR2 region of SEQ ID NO: 430, and a light chain CDR3 region of SEQ ID NO: 545; The VH domain of the antibody capable of binding to the fusion peptide of coronavirus, preferably SARS-CoV-2, comprises as CDRs the heavy chain CDR1 region of SEQ ID NO: 639, the heavy chain CDR2 region of SEQ ID NO: 640, and the heavy chain CDR3 region of SEQ ID NO: 641, and the VL domain of the antibody capable of binding to the fusion peptide of coronavirus, preferably SARS-CoV-2, comprises as CDRs the light chain CDR1 region of SEQ ID NO: 642, the light chain CDR2 region of SEQ ID NO: 643, and the light chain CDR3 region of SEQ ID NO: 644; 32. A bispecific antibody or antigen-binding fragment thereof according to clause 31. Verse 33: a) a light chain comprising a VL-CL domain and a heavy chain comprising a VH-CH1-CH2-CH3 domain of an antibody capable of binding to the stem helix of a coronavirus, preferably SARS-CoV-2, and b) a light chain comprising VL-CL domains and a heavy chain comprising VH-CH1-CH2-CH3 domains of an antibody capable of binding to the fusion peptide of a coronavirus, preferably SARS-CoV-2 Including, the domains VL-CL and VH-CH1 from an antibody that specifically binds to the fusion peptide of a coronavirus, preferably SARS-CoV-2, are replaced by each other; 29. The bispecific antibody or antigen-binding fragment thereof according to any one of clauses 26 to 28. Verse 34: 34. The bispecific antibody or antigen-binding fragment thereof according to any one of clauses 26 to 33, wherein the first Fab comprises a heavy chain variable region having an amino acid sequence that is at least 90% identical, at least 91% identical, at least 92% identical, at least 93% identical, at least 94% identical, at least 95% identical, at least 96% identical, at least 97% identical, at least 98% identical, or at least 99% identical to the amino acid sequence set forth in SEQ ID NO: 629. Verse 35: 34. The bispecific antibody or antigen-binding fragment thereof according to any one of clauses 26 to 33, wherein the second Fab comprises a heavy chain variable region having an amino acid sequence that is at least 90% identical, at least 91% identical, at least 92% identical, at least 93% identical, at least 94% identical, at least 95% identical, at least 96% identical, at least 97% identical, at least 98% identical, or at least 99% identical to the amino acid sequence set forth in SEQ ID NO: 663. Verse 36: 34. The bispecific antibody or antigen-binding fragment thereof according to any one of clauses 26 to 33, wherein the first Fab comprises a light chain variable region having an amino acid sequence that is at least 90% identical, at least 91% identical, at least 92% identical, at least 93% identical, at least 94% identical, at least 95% identical, at least 96% identical, at least 97% identical, at least 98% identical, or at least 99% identical to the amino acid sequence set forth in SEQ ID NO: 630. Verse 37: 34. The bispecific antibody or antigen-binding fragment thereof according to any one of clauses 26 to 33, wherein the second Fab comprises a light chain variable region having an amino acid sequence that is at least 90% identical, at least 91% identical, at least 92% identical, at least 93% identical, at least 94% identical, at least 95% identical, at least 96% identical, at least 97% identical, at least 98% identical, or at least 99% identical to the amino acid sequence set forth in SEQ ID NO: 664. Verse 38: 34. The bispecific antibody or antigen-binding fragment thereof according to any one of clauses 26 to 33, selected from any one of the group comprising a full length antibody, a Fab, a modified Fab, a Fab', a modified Fab', a F(ab')2, an Fv, a single domain antibody, a scFv, a scFv-Fc, a bivalent, trivalent or tetravalent antibody, a Bis-scFv, a diabody, a triabody, a tetrabody, and epitope-binding fragments thereof. Verse 39: 39. A method for the production of a bispecific antibody or antigen-binding fragment thereof according to any one of clauses 26 to 38, comprising: a) culturing a host cell comprising an expression vector comprising a polynucleotide encoding the bispecific antibody or antibody fragment according to any one of the preceding clauses under conditions that allow the production of said bispecific antibody or antigen-binding fragment thereof, and b) isolating the bispecific antibody or antigen-binding fragment thereof. A method comprising: Verse 40: 39. A nucleic acid molecule comprising a nucleic acid sequence encoding the bispecific antibody or antigen-binding fragment thereof, the heavy chain variable region and / or the light chain variable region of the bispecific antibody or antigen-binding fragment thereof according to any one of clauses 26 to 38, which is preferably an isolated nucleic acid molecule. Verse 41: a bispecific antibody or antigen-binding fragment thereof according to any one of clauses 26 to 38 and clause 40, comprising a nucleic acid sequence encoding the heavy chain variable region and / or the light chain variable region of the bispecific antibody or antigen-binding fragment thereof, The nucleic acid sequence is (i) a first nucleic acid sequence encoding the amino acid sequence set forth in SEQ ID NO: 046, or a first nucleic acid sequence encoding an amino acid sequence that differs from SEQ ID NO: 046 by at most one or two amino acids; (ii) a second nucleic acid sequence encoding the amino acid sequence set forth in SEQ ID NO: 114, or a second nucleic acid sequence encoding an amino acid sequence that differs from SEQ ID NO: 114 by at most 1, 2, 3, 4, or 5 amino acids; (iii) a third nucleic acid sequence encoding the amino acid sequence set forth in SEQ ID NO: 254, or a third nucleic acid sequence encoding an amino acid sequence that differs from SEQ ID NO: 254 by at most 1, 2, or 3 amino acids; (iv) a fourth nucleic acid sequence encoding the amino acid sequence set forth in SEQ ID NO: 367, or an amino acid sequence that differs from SEQ ID NO: 367 by at most 1, 2, 3, or 4 amino acids; (v) a fifth nucleic acid sequence encoding the amino acid sequence set forth in SEQ ID NO: 430, or an amino acid sequence that differs from SEQ ID NO: 430 by at most one or two amino acids; (vi) a sixth nucleic acid sequence encoding the amino acid sequence set forth in SEQ ID NO: 545, or an amino acid sequence that differs from SEQ ID NO: 545 by at most 1, 2, 3, or 4 amino acids; (vii) a seventh nucleic acid sequence encoding the amino acid sequence set forth in SEQ ID NO: 639, or an amino acid sequence that differs from SEQ ID NO: 639 by at most one or two amino acids; (viii) an eighth nucleic acid sequence encoding the amino acid sequence set forth in SEQ ID NO: 640, or an eighth nucleic acid sequence encoding an amino acid sequence that differs from SEQ ID NO: 640 by at most 1, 2, 3, 4, or 5 amino acids; (ix) a ninth nucleic acid sequence encoding the amino acid sequence set forth in SEQ ID NO: 641, or an amino acid sequence that differs from SEQ ID NO: 641 by at most 1, 2, 3, or 4 amino acids; (x) a tenth nucleic acid sequence encoding the amino acid sequence set forth in SEQ ID NO: 642, or an amino acid sequence that differs from SEQ ID NO: 642 by at most 1, 2, 3, or 4 amino acids; (xi) an eleventh nucleic acid sequence encoding the amino acid sequence set forth in SEQ ID NO: 643, or an eleventh nucleic acid sequence encoding an amino acid sequence differing from SEQ ID NO: 643 by at most one or two amino acids; and (xii) a twelfth nucleic acid sequence encoding the amino acid sequence set forth in SEQ ID NO: 644, or an amino acid sequence that differs from SEQ ID NO: 644 by at most 1, 2, or 3 amino acids; A nucleic acid molecule comprising at least one of the following: Verse 42: 39. The bispecific antibody or antigen-binding fragment thereof according to any one of clauses 26 to 38 for use in a method of treating a coronavirus infection in a subject. Verse 43: 39. A composition comprising a bispecific antibody or antigen-binding fragment thereof according to any one of clauses 26 to 38. Verse 44: 44. A composition according to clause 43 further comprising a second medicament for simultaneous, separate or sequential administration. Verse 45: 45. The composition according to clause 44, wherein the second medicament comprises a second antibody or antigen-binding fragment thereof. Verse 46: 45. The composition according to clause 44, wherein the second medicament comprises a bispecific antibody or a bispecific antigen-binding fragment thereof. Verse 47: 44. The composition according to clause 43, further comprising a pharmaceutically acceptable excipient or carrier. Verse 48: 44. A composition according to clause 43 for use in preventing and / or treating a coronavirus infection in a subject, preferably a SARS-CoV-1 or SARS-CoV-2 infection in a subject, comprising an effective amount of the bispecific antibody or antigen-binding fragment thereof, nucleic acid molecule, or vector, and optionally a pharmaceutically acceptable excipient or carrier. Verse 49: 44. The composition of clause 43 for use as a medicament. Verse 50: 44. An inhalation device comprising the composition of paragraph 43. Verse 51: 39. The bispecific antibody or antigen-binding fragment thereof according to any one of clauses 26 to 38, which is bivalent. Verse 52: 39. The bispecific antibody or antigen-binding fragment thereof according to any one of clauses 26 to 38, capable of binding to at least one stem helix and / or fusion peptide of an α-coronavirus, a β-coronavirus, a γ-coronavirus, and a δ-coronavirus, preferably at least an α-coronavirus and / or a β-coronavirus.

[0095] section Each of the following numbered sections represents a preferred embodiment of the present invention and is part of this specification. Verse 53: An anti-coronavirus, preferably anti-SARS-CoV-2 bispecific antibody or antigen-binding fragment thereof, comprising a first Fab capable of binding to the stem helix of a coronavirus, preferably SARS-CoV-2, and a second Fab capable of binding to the fusion peptide of a coronavirus, preferably SARS-CoV-2, wherein the first Fab comprises a sequence comprising preferably any one or more of SEQ ID NOs: 046, 114, 254, 367, 430, or 545, and the second Fab comprises a sequence comprising preferably any one or more of SEQ ID NOs: 645, 646, 647, 648, 649 to 650. Verse 54: 54. The bispecific antibody or antigen-binding fragment thereof according to clause 53, comprising a first Fab capable of binding to the stem helix of a coronavirus, preferably SARS-CoV-2, and a second Fab capable of binding to the fusion peptide of a coronavirus, preferably SARS-CoV-2, wherein the first Fab comprises a sequence comprising preferably any one or more of SEQ ID NOs: 046, 114, 254, 367, 430, and / or 545, and the second Fab comprises a sequence comprising preferably any one or more of SEQ ID NOs: 645, 646, 647, 648, 649, and / or 650. Verse 55: a first Fab that specifically binds to the stem helix of a coronavirus, preferably SARS-CoV-2, and a second Fab that specifically binds to the fusion peptide of a coronavirus, preferably SARS-CoV-2; a.) a first Fab comprises a heavy chain variable region comprising, as CDRs, the heavy chain CDR1 region of SEQ ID NO: 046, the heavy chain CDR2 region of SEQ ID NO: 114, and the heavy chain CDR3 region of SEQ ID NO: 254, and a light chain variable region comprising, as CDRs, the light chain CDR1 region of SEQ ID NO: 367, the light chain CDR2 region of SEQ ID NO: 430, and the light chain CDR3 region of SEQ ID NO: 545; b.) the second Fab comprises a heavy chain variable region comprising, as CDRs, the heavy chain CDR1 region of SEQ ID NO: 645, the heavy chain CDR2 region of SEQ ID NO: 646, and the heavy chain CDR3 region of SEQ ID NO: 647, and a light chain variable region comprising, as CDRs, the light chain CDR1 region of SEQ ID NO: 648, the light chain CDR2 region of SEQ ID NO: 649, and the light chain CDR3 region of SEQ ID NO: 650; 55. The bispecific antibody or antigen-binding fragment thereof according to clause 53 or 54, Verse 56: a) a light chain comprising a VL-CL domain and a heavy chain comprising a VH-CH1-CH2-CH3 domain of an antibody or antigen-binding fragment thereof capable of binding to the stem helix of a coronavirus, preferably SARS-CoV-2; and b) a light chain comprising the VL-CL domains and a heavy chain comprising the VH-CH1-CH2-CH3 domains of an antibody or antigen-binding fragment thereof capable of binding to the fusion peptide of a coronavirus, preferably SARS-CoV-2; Including, 56. The bispecific antibody or antigen-binding fragment thereof according to any one of clauses 53 to 55, wherein the constant domains CL and CH1 from an antibody or antigen-binding fragment thereof capable of binding to the stem helix of a coronavirus, preferably SARS-CoV-2, are replaced by each other. Verse 57: the VH domain of the antibody or antigen-binding fragment thereof capable of binding to the stem helix of a coronavirus, preferably SARS-CoV-2, comprises, as CDRs, a heavy chain CDR1 region of SEQ ID NO: 046, a heavy chain CDR2 region of SEQ ID NO: 114, and a heavy chain CDR3 region of SEQ ID NO: 254; and the VL domain of the antibody or antigen-binding fragment thereof capable of binding to the stem helix of a coronavirus, preferably SARS-CoV-2, comprises, as CDRs, a light chain CDR1 region of SEQ ID NO: 367, a light chain CDR2 region of SEQ ID NO: 430, and a light chain CDR3 region of SEQ ID NO: 545; The VH domain of the antibody or antigen-binding fragment thereof capable of binding to the fusion peptide of coronavirus, preferably SARS-CoV-2, comprises, as CDRs, the heavy chain CDR1 region of SEQ ID NO: 645, the heavy chain CDR2 region of SEQ ID NO: 646, and the heavy chain CDR3 region of SEQ ID NO: 647, and the VL domain of the antibody or antigen-binding fragment thereof capable of binding to the fusion peptide of coronavirus, preferably SARS-CoV-2, comprises, as CDRs, the light chain CDR1 region of SEQ ID NO: 648, the light chain CDR2 region of SEQ ID NO: 649, and the light chain CDR3 region of SEQ ID NO: 650. 57. A bispecific antibody or antigen-binding fragment thereof according to clause 56. Verse 58: a) a light chain comprising a VL-CL domain and a heavy chain comprising a VH-CH1-CH2-CH3 domain of an antibody capable of binding to the stem helix of a coronavirus, preferably SARS-CoV-2, and b) a light chain comprising VL-CL domains and a heavy chain comprising VH-CH1-CH2-CH3 domains of an antibody capable of binding to the fusion peptide of a coronavirus, preferably SARS-CoV-2 Including, the constant domains CL and CH1 from an antibody capable of binding to the fusion peptide of a coronavirus, preferably SARS-CoV-2, are replaced by each other; 56. The bispecific antibody or antigen-binding fragment thereof according to any one of clauses 53 to 55. Verse 59: the VH domain of the antibody or antigen-binding fragment thereof capable of binding to the stem helix of a coronavirus, preferably SARS-CoV-2, comprises, as CDRs, a heavy chain CDR1 region of SEQ ID NO: 046, a heavy chain CDR2 region of SEQ ID NO: 114, and a heavy chain CDR3 region of SEQ ID NO: 254; and the VL domain of the antibody capable of binding to the stem helix of a coronavirus, preferably SARS-CoV-2, comprises, as CDRs, a light chain CDR1 region of SEQ ID NO: 367, a light chain CDR2 region of SEQ ID NO: 430, and a light chain CDR3 region of SEQ ID NO: 545; The VH domain of the antibody capable of binding to the fusion peptide of coronavirus, preferably SARS-CoV-2, comprises as CDRs the heavy chain CDR1 region of SEQ ID NO: 645, the heavy chain CDR2 region of SEQ ID NO: 646, and the heavy chain CDR3 region of SEQ ID NO: 647, and the VL domain of the antibody capable of binding to the fusion peptide of coronavirus, preferably SARS-CoV-2, comprises as CDRs the light chain CDR1 region of SEQ ID NO: 648, the light chain CDR2 region of SEQ ID NO: 649, and the light chain CDR3 region of SEQ ID NO: 650. 59. A bispecific antibody or antigen-binding fragment thereof according to clause 58. Verse 60: a) a light chain comprising a VL-CL domain and a heavy chain comprising a VH-CH1-CH2-CH3 domain of an antibody capable of binding to the stem helix of a coronavirus, preferably SARS-CoV-2, and b) a light chain comprising VL-CL domains and a heavy chain comprising VH-CH1-CH2-CH3 domains of an antibody capable of binding to the fusion peptide of a coronavirus, preferably SARS-CoV-2 Including, the domains VL-CL and VH-CH1 from an antibody that specifically binds to the fusion peptide of a coronavirus, preferably SARS-CoV-2, are replaced by each other; 56. The bispecific antibody or antigen-binding fragment thereof according to any one of clauses 53 to 55. Verse 61: 61. The bispecific antibody or antigen-binding fragment thereof according to any one of clauses 53 to 60, wherein the first Fab comprises a heavy chain variable region having an amino acid sequence that is at least 90% identical, at least 91% identical, at least 92% identical, at least 93% identical, at least 94% identical, at least 95% identical, at least 96% identical, at least 97% identical, at least 98% identical, or at least 99% identical to the amino acid sequence set forth in SEQ ID NO: 629. Verse 62: 61. The bispecific antibody or antigen-binding fragment thereof according to any one of clauses 53 to 60, wherein the second Fab comprises a heavy chain variable region having an amino acid sequence that is at least 90% identical, at least 91% identical, at least 92% identical, at least 93% identical, at least 94% identical, at least 95% identical, at least 96% identical, at least 97% identical, at least 98% identical, or at least 99% identical to the amino acid sequence set forth in SEQ ID NO: 665. Verse 63: 61. The bispecific antibody or antigen-binding fragment thereof according to any one of clauses 53 to 60, wherein the first Fab comprises a light chain variable region having an amino acid sequence that is at least 90% identical, at least 91% identical, at least 92% identical, at least 93% identical, at least 94% identical, at least 95% identical, at least 96% identical, at least 97% identical, at least 98% identical, or at least 99% identical to the amino acid sequence set forth in SEQ ID NO: 630. Verse 64: 61. The bispecific antibody or antigen-binding fragment thereof according to any one of clauses 53 to 60, wherein the second Fab comprises a light chain variable region having an amino acid sequence that is at least 90% identical, at least 91% identical, at least 92% identical, at least 93% identical, at least 94% identical, at least 95% identical, at least 96% identical, at least 97% identical, at least 98% identical, or at least 99% identical to the amino acid sequence set forth in SEQ ID NO: 666. Verse 65: 61. The bispecific antibody or antigen-binding fragment thereof according to any one of clauses 53 to 60, selected from any one of the group comprising a full length antibody, a Fab, a modified Fab, a Fab', a modified Fab', a F(ab')2, an Fv, a single domain antibody, a scFv, a scFv-Fc, a bivalent, trivalent or tetravalent antibody, a Bis-scFv, a diabody, a triabody, a tetrabody, and epitope-binding fragments thereof. Verse 66: 66. A method for the production of a bispecific antibody or antigen-binding fragment thereof according to any one of clauses 53 to 65, comprising: a) culturing a host cell comprising an expression vector comprising a polynucleotide encoding the bispecific antibody or antibody fragment according to any one of the preceding clauses under conditions that allow the production of said bispecific antibody or antigen-binding fragment thereof, and b) isolating the bispecific antibody or antigen-binding fragment thereof. A method comprising: Verse 67: 66. A nucleic acid molecule comprising a nucleic acid sequence encoding the bispecific antibody or antigen-binding fragment thereof, the heavy chain variable region and / or the light chain variable region of the bispecific antibody or antigen-binding fragment thereof according to any one of clauses 53 to 65, which is preferably an isolated nucleic acid molecule. Verse 68: 68. A bispecific antibody or antigen-binding fragment thereof according to any one of clauses 53 to 65 and clause 67, a nucleic acid molecule comprising a nucleic acid sequence encoding the heavy chain variable region and / or the light chain variable region of the bispecific antibody or antigen-binding fragment thereof, The nucleic acid sequence is (i) a first nucleic acid sequence encoding the amino acid sequence set forth in SEQ ID NO: 046, or a first nucleic acid sequence encoding an amino acid sequence that differs from SEQ ID NO: 046 by at most one or two amino acids; (ii) a second nucleic acid sequence encoding the amino acid sequence set forth in SEQ ID NO: 114, or a second nucleic acid sequence encoding an amino acid sequence that differs from SEQ ID NO: 114 by at most 1, 2, 3, 4, or 5 amino acids; (iii) a third nucleic acid sequence encoding the amino acid sequence set forth in SEQ ID NO: 254, or a third nucleic acid sequence encoding an amino acid sequence that differs from SEQ ID NO: 254 by at most 1, 2, or 3 amino acids; (iv) a fourth nucleic acid sequence encoding the amino acid sequence set forth in SEQ ID NO: 367, or an amino acid sequence that differs from SEQ ID NO: 367 by at most 1, 2, 3, or 4 amino acids; (v) a fifth nucleic acid sequence encoding the amino acid sequence set forth in SEQ ID NO: 430, or an amino acid sequence that differs from SEQ ID NO: 430 by at most one or two amino acids; (vi) a sixth nucleic acid sequence encoding the amino acid sequence set forth in SEQ ID NO: 545, or an amino acid sequence that differs from SEQ ID NO: 545 by at most 1, 2, 3, or 4 amino acids; (vii) a seventh nucleic acid sequence encoding the amino acid sequence set forth in SEQ ID NO: 645, or an amino acid sequence that differs from SEQ ID NO: 645 by at most one or two amino acids; (viii) an eighth nucleic acid sequence encoding the amino acid sequence set forth in SEQ ID NO: 646, or an eighth nucleic acid sequence encoding an amino acid sequence that differs from SEQ ID NO: 646 by at most 1, 2, 3, 4, or 5 amino acids; (ix) a ninth nucleic acid sequence encoding the amino acid sequence set forth in SEQ ID NO: 647, or an amino acid sequence that differs from SEQ ID NO: 647 by at most 1, 2, 3, 4, or 5 amino acids; (x) a tenth nucleic acid sequence encoding the amino acid sequence set forth in SEQ ID NO: 648, or an amino acid sequence that differs from SEQ ID NO: 648 by at most 1, 2, or 3 amino acids; (xi) an eleventh nucleic acid sequence encoding the amino acid sequence set forth in SEQ ID NO: 649, or an eleventh nucleic acid sequence encoding an amino acid sequence that differs from SEQ ID NO: 649 by at most one or two amino acids; and (xii) a twelfth nucleic acid sequence encoding the amino acid sequence set forth in SEQ ID NO: 650, or an amino acid sequence that differs from SEQ ID NO: 650 by at most 1, 2, or 3 amino acids. A nucleic acid molecule comprising at least one of the following: Verse 69: 66. The bispecific antibody or antigen-binding fragment thereof according to any one of clauses 53 to 65 for use in a method of treating a coronavirus infection in a subject. Verse 70: 66. A composition comprising a bispecific antibody and / or antigen-binding fragment thereof according to any one of clauses 53 to 65. Verse 71: 71. A composition according to paragraph 70 further comprising a second medicament for simultaneous, separate or sequential administration. Verse 72: 72. The composition according to clause 71, wherein the second medicament comprises a second antibody or antigen-binding fragment thereof. Verse 73: 72. The composition according to clause 71, wherein the second medicament comprises a bispecific antibody or a bispecific antigen-binding fragment thereof. Verse 74: 71. The composition according to clause 70, further comprising a pharmaceutically acceptable excipient or carrier. Verse 75: 71. A composition according to clause 70 for use in preventing and / or treating a coronavirus infection in a subject, preferably a SARS-CoV-1 or SARS-CoV-2 infection in a subject, comprising an effective amount of the bispecific antibody or antigen-binding fragment thereof, nucleic acid molecule, or vector, and optionally a pharmaceutically acceptable excipient or carrier. Verse 76: 71. The composition of clause 70 for use as a medicament. Verse 77: An inhalation device comprising the composition of clause 70. Verse 78: 66. The bispecific antibody or antigen-binding fragment thereof according to any one of clauses 53 to 65, which is bivalent. Verse 79: 66. The bispecific antibody or antigen-binding fragment thereof according to any one of clauses 53 to 65, capable of binding to at least one stem helix and / or fusion peptide of an α-coronavirus, a β-coronavirus, a γ-coronavirus, and a δ-coronavirus, preferably at least an α-coronavirus and / or a β-coronavirus. [Brief explanation of the drawings]

[0096] [Figure 1] Survival after lethal challenge - Kaplan-Meier survival curve for CV3-25 intraperitoneal prophylactic treatment group. Animals (n=10 per group) were treated at dose-adjusted intervals with the control antibody CV3-25 on day -1. Animals were infected with 103.5 TCID50 of SARS-CoV-2 delta on day 0. A vehicle control group (PBS) was included. The lines are slightly shifted on the y-axis to improve visual representation. [Figure 2]Survival after lethal challenge - Kaplan-Meier survival curves for CV3-25 intranasal prophylactic treatment groups. Animals (n=10 per group, except for n=8 at 15 mg / kg) were treated at dose titration with control antibody CV3-25 on day -1. Animals were infected with 103.5 TCID50 of SARS-CoV-2 delta on day 0. A vehicle control group (PBS) was included. Lines are slightly shifted on the y-axis to improve visual representation. [Figure 3] Body Weight Change - % Body Weight Change relative to Day 0 for the CV3-25 intraperitoneal prophylactic treatment group. Animals (n=10 per group) were treated intranasally with titrated doses of the control antibody CV3-25 on Day -1. Animals were infected with 103.5 TCI D50 of SARS-CoV-2 delta on Day 0. A vehicle control group (PBS) was included. Bars represent the 95% CI of the mean. If mice died or were euthanized during study follow-up, the last observed body weight was carried forward. [Figure 4] Body Weight Change - % Body Weight Change relative to Day 0 for the CV3-25 intranasal prophylactic treatment groups. Animals (n=10 per group, except for n=8 in the 15 mg / kg group) were treated intranasally with titrated doses of the control antibody CV3-25 on Day -1. Animals were infected with 103.5 TCID50 of SARS-CoV-2 delta on Day 0. A vehicle control group (PBS) was included. Bars represent the 95% CI of the mean. If mice died or were euthanized during study follow-up, the last observed body weight was carried forward. [Figure 5A] Survival and body weight change for intraperitoneal administration - antibodies according to the invention Survival and body weight change for intraperitoneal administration of antibodies according to the invention in a pre-exposure efficacy study in K18-hACE2 mice challenged with lethal SARS-CoV-2 delta. Animals (n=10 per group) were treated intraperitoneally with antibody dose adjustment or vehicle control. Kaplan-Meier survival curves for treatment groups are shown. Lines are slightly shifted on the Y-axis to improve visual presentation. [Figure 5B]Survival and body weight change for intraperitoneal administration - antibodies according to the invention Survival and body weight change for intraperitoneal administration of antibodies according to the invention in a pre-exposure efficacy study in K18-hACE2 mice challenged with lethal SARS-CoV-2 delta. Animals (n=10 per group) were treated intraperitoneally with antibody dose adjustment or vehicle control. Weight change (%) relative to day 0 is shown. Error bars represent the 95% confidence interval (CI) of the mean. If mice died or were euthanized during study follow-up, the last observed body weight was carried forward. Asterisks indicate significant differences compared to the vehicle control group. [Figure 6A] Survival and weight change for intranasal administration - antibodies according to the invention Survival and weight change for intranasal administration of antibodies according to the invention in a pre-exposure efficacy study in K18-hACE2 mice challenged with lethal SARS-CoV-2 delta. Animals (n=10 per group) were treated intranasally with antibody dose adjustment or vehicle control. Kaplan-Meier survival curves for treatment groups are shown. Lines are slightly shifted on the Y-axis to improve visual presentation. [Figure 6B] Survival and weight change for intranasal administration - antibodies according to the invention Survival and weight change for intranasal administration of antibodies according to the invention in a pre-exposure efficacy study in K18-hACE2 mice challenged with lethal SARS-CoV-2 delta. Animals (n=10 per group) were treated intranasally with antibody titration or vehicle control. Weight change (%) relative to day 0 is shown. Error bars represent the 95% confidence interval (CI) of the mean. If mice died or were euthanized during study follow-up, the last observed weight was carried forward. Asterisks indicate significant differences compared to the vehicle control group. [Figure 7] Linear epitope mapping to the spike helix domains of alpha- and beta-coronaviruses. Amino acids highlighted in gray were identified as epitopes to which antibodies bind on the spike protein. Target sequences for the seven strains tested are aligned by Clustal. * may include multiple smaller portions of the epitope. DETAILED DESCRIPTION OF THE INVENTION

[0097] For purposes of clarity and brevity, features may be described herein as part of the same or separate embodiments; however, it will be recognized that the present disclosure includes embodiments having all or any combination of the described features. References to "methods for treatment," "method of treatment," "antibody for use," and "use of an antibody in the manufacture of a medicament" may be used interchangeably, and embodiments disclosed with respect to any one of these aspects also apply with respect to the other aspects. In other words, they all refer to medical treatments involving the antibodies disclosed herein.

[0098] Preventive measures As used herein, the term "prophylactic treatment" includes reference to treatment to prevent an individual from contracting a coronavirus, or to prevent symptoms after a coronavirus infection, or to prevent severe symptoms after a known coronavirus infection, with or without symptoms, or to prevent hospitalization and death after a coronavirus infection.

[0099] Prevention of infection is preferably achieved by administration of the antibodies disclosed herein prior to exposure to coronavirus, i.e., pre-exposure prophylaxis.

[0100] Thus, "prophylactically" preferably means prior to viral exposure, although it can also include administration after infection, e.g., to reduce viral replication or spread or increase viral clearance, i.e., post-exposure prophylaxis.

[0101] An infected individual may be asymptomatic, or alternatively, an infected individual may be symptomatic.

[0102] Preferably, post-exposure prophylaxis involves administration of the antibodies disclosed herein after exposure to the coronavirus to prevent symptomatic disease.

[0103] Preferably, post-exposure prophylaxis involves administration of the antibodies disclosed herein after exposure to the coronavirus to prevent severe disease, particularly hospitalization.

[0104] In the treatment methods of the present invention, prophylactic treatment involves administering an antibody to coronavirus at a time when the individual is not infected with coronavirus. Preferably, the antibody binds to a conserved epitope of the spike protein of the coronavirus virion. In some embodiments, the individual in need thereof is not (yet) infected with coronavirus.

[0105] Therapeutic treatment As used herein, the term "therapeutic treatment" includes reference to treating viral infections (including coronavirus diseases) after the viral infection has occurred. Viral infection involves the entry of the virus into the body and / or replication of the virus within the body and / or the spread of the virus to previously uninfected cells, tissues, or locations within the body. Viral infections may cause one or more diseases, but may also remain latent, i.e., remain in the body without causing symptoms or disease.

[0106] coronavirus As used herein, the term "coronavirus" includes reference to positive-sense, single-stranded RNA viruses belonging to the Coronaviridae family.

[0107] Preferably, the antibodies disclosed herein are capable of specifically binding to the stem helix of coronaviruses, particularly SARS-CoV-2.

[0108] Preferably, the antibodies disclosed herein are capable of specifically binding to the stem helix of a SARS-CoV-2 variant of concern.

[0109] Preferably, the antibodies disclosed herein are capable of neutralizing coronaviruses, particularly SARS-CoV-2.

[0110] Preferably, the antibodies disclosed herein are capable of neutralizing at least one or more, preferably two or more, preferably three or more, preferably four or more, and even more preferably five or more coronavirus subtypes.

[0111] As used herein, the phrase "capable of" means that the ability of the antibodies or antigen-binding fragments thereof described herein is sufficient to bind to at least a portion of an antigen, such as the stem helix of a coronavirus, particularly SARS-CoV-2, and / or a fusion protein of a coronavirus, particularly SARS-CoV-2. The binding affinity or binding ability of the antibodies and / or antigen-binding fragments thereof described herein can be measured using any method known to one of skill in the art. As used herein, the phrase "capable of" includes, but is not limited to, the phrases "specifically bind" or "specifically binds to."

[0112] coronavirus infection As used herein, the term "coronavirus infection" includes reference to the pathological or non-pathological, preferably pathological, entry and retention of any type of coronavirus in a human host. The infecting virus may replicate within the host, its cells, or the cells of its microbiota. The infecting virus may or may not cause disease, e.g., COVID-19. The infection may or may not be detectable by methods known in the art for detecting viral infection. An infected individual may or may not be aware of the infection. Typical, but not exclusive, sites in the human body where SARS-CoV-2, for example, may be located in an infected individual are the respiratory system and / or its cells, and the cardiovascular system and / or its cells. As used herein, the term "coronavirus infection" further includes reference to the entry and retention of any type of coronavirus moiety capable of initiating viral replication in a human host. The term "coronavirus infection" encompasses symptoms or disease subsequent to infection, e.g., COVID-19. The term "coronavirus infection" encompasses "SARS-CoV-2 infection."

[0113] epitope As used herein, the term "epitope" includes reference to a moiety capable of binding to an antibody disclosed herein with sufficiently high affinity to form a detectable antigen-antibody complex.

[0114] individual As used herein, the term "individual" includes reference to a mammal or human subject to or at risk of contracting a viral infection. Infection can occur in any system, tissue, or cell belonging to the host, including the host's microbiota. Coronavirus infection and coronavirus disease can occur in individuals of all age groups and genders. Nevertheless, preferably, the individual is a human, particularly an elderly human, e.g., at least 60, 65, 70, 75, 80, or at least 85 years of age, or a human who is at increased risk of infection due to their occupation or living environment. Preferably, the individual is at risk of contracting a serious disease upon infection, e.g., COVID-19. In preferred embodiments, the individual has an underlying condition, such as (i) a respiratory disease, such as asthma, COPD, chronic bronchitis, and emphysema; (ii) a cardiovascular disease, such as cardiac arrhythmia or an individual who has undergone cardiac surgery; (iii) diabetes; (iv) renal failure; and / or (v) a disease affecting the immune system, e.g., an immunocompromised individual or an increased risk of viral infection due to their occupation.

[0115] As used herein, the term "individual" includes reference to a mammal, such as, but not limited to, a human, that will benefit from a specified therapy; for example, the term "individual" can encompass a non-human primate (NHP).

[0116] Preferably, the individual is a mammal, more preferably a human.

[0117] Administering and Administration As used herein, the terms "administering" and "administration" include reference to providing one or more drugs and, optionally, one or more adjuvants, for the purpose of treating, curing, reducing, or preventing a disease or its symptoms in an individual, or promoting the individual's sense of well-being. Preferred methods of administering the antibodies disclosed herein include transmucosal administration, preferably intranasal administration, and oral inhalation.

[0118] Individuals who need it As used herein, the phrase "individual in need thereof" includes reference to a mammal, such as a human, that will benefit from the identified therapy. The treatment methods of the present invention may be used prophylactically and do not require the presentation of symptoms or signs of coronavirus infection. Individuals particularly in need of the methods or antibodies for use of the present invention are those at high risk of coronavirus infection, at high risk of developing severe symptoms (disease), such as COVID-19, and / or at high risk of dying from COVID-19. Those skilled in the art are familiar with risk factors for high risk of coronavirus infection, high risk of developing severe symptoms of coronavirus infection, and high risk of dying from coronavirus infection.

[0119] antibody The term "antibody," as used herein, includes reference to an intact immunoglobulin, including a monoclonal antibody, e.g., a chimeric, humanized, or human monoclonal antibody, or a binding molecule that comprises an antigen-binding domain (e.g., heavy chain CDRs 1-3 of the variable domain) of an antibody disclosed herein, or an antibody that competes with an antibody disclosed herein for specific binding to an immunoglobulin binding partner.

[0120] In other words, functional fragments of antibodies are also encompassed by the term "antibody." Antibodies are generally Y-shaped proteins. In antibodies, a constant domain and a variable domain are generally present. The variable domain facilitates antigen binding. Antibodies generally comprise two heavy chains and two light chains. Both the heavy and light chains are partly constant and partly variable. Antibodies occur in several classes, namely IgA, IgD, IgE, IgG, and IgM. Preferably, the antibodies of the present invention are of the IgG class, particularly the IgG1 class. Some classes are further subdivided into subclasses or isotypes. For example, the IgG class is subdivided into subclasses IgG1, IgG2, IgG3, and IgG4. Preferably, the antibodies of the present invention are of the IgG class, preferably the IgG1 class. Antigen-binding regions or antigen-binding fragments of antibodies are encompassed by the term "antibody" and are therefore part of the present invention, and may include, for example, Fab, F(ab'), F(ab')2, dAb, Fv, Fd, CDR fragments, diabodies, triabodies, tetrabodies, single-chain antibodies (scFv, scFv-Fc), bivalent single-chain antibodies, single-chain phage antibodies, (poly)peptides comprising at least a fragment of an immunoglobulin sufficient to confer specific antigen-binding properties to the (poly)peptide, and others. Such fragments may be produced synthetically or by enzymatic or chemical cleavage of the original immunoglobulin, or may be genetically engineered by recombinant DNA techniques. Methods for producing antibodies and antigen-binding fragments are well known to those skilled in the art. Antibodies may be conjugated or unconjugated. Antibodies may be conjugated, linked, or otherwise physically or functionally associated with effector moieties or tags, such as, for example, enzymes, liposomes, radioactive substances, fluorescent dyes, toxic substances, among others. Antibodies may be stabilized, multimerized, humanized, or otherwise engineered.

[0121] The antibody may be neutralizing. This includes reference to the inhibition of virus, e.g., as measured by an in vitro neutralization assay of virus entry into host cells and / or virus replication. Neutralization can be achieved, for example, by inhibiting viral attachment or adhesion to the cell surface, or by inhibiting fusion of the virus with the cell membrane after the virus has attached to the target cell, or by inhibiting viral egress from the cell. Neutralization does not specify the method of neutralization. Preferably, the antibody is cross-neutralizing. This includes reference to the ability of the antibody of the present invention to bind to different sets of molecules, preferably different sets of molecules of different subtypes belonging to the Coronaviridae family.

[0122] As used herein, the terms "domain" or "region" in reference to a portion of an antibody, bispecific antibody, or antigen-binding fragment thereof, may be used interchangeably herein.

[0123] As used herein, the phrases "antigen-binding fragment," "antigen-binding fragment," "antigen fragment," "fragment of an antibody," or "antigen-binding molecule" refer to a protein, polypeptide, or molecular complex that comprises or consists of at least one CDR, alone or in combination with one or more additional complementarity-determining regions (CDRs) and / or framework regions (FRs) described herein, that is capable of binding to the stem helix of a coronavirus, particularly SARS-CoV-2, and / or a portion of an antigen comprising the stem helix of a coronavirus, particularly SARS-CoV-2 and its variants of concern. As used herein, the phrases "antigen-binding fragment," "antigen-binding fragment," "antigen fragment," "fragment of an antibody," or "antigen-binding molecule" can be used interchangeably. For example, the phrase "antigen-binding fragment" as used herein is used interchangeably with the phrase "antibody fragment" as used herein.

[0124] Antibodies contain complementarity-determining regions (CDRs) located in the variable domains of the heavy and light chains. CDRs contribute significantly to the antigen-binding site. Three CDRs can be identified: CDR1, CDR2, and CDR3. Because each CDR can be located on either the light or heavy chain, for each antigen receptor, there are generally six CDRs that collectively contact the antigen: light chain CDR1, light chain CDR2, light chain CDR3, heavy chain CDR1, heavy chain CDR2, and heavy chain CDR3. CDR3-type CDRs are the most variable. CDRs can be specific for linear, discontinuous, or conformational epitopes of proteins or protein fragments, whether present on the protein in its native conformation or, in some cases, on denatured or inactivated proteins. Epitopes can also consist of or include post-translational modifications of proteins. Of particular interest are antibodies of the present invention that contain CDRs that recognize coronavirus antigens, such as spike proteins, and in particular the stem helices of spike proteins. The antibodies disclosed herein bind to conserved epitopes in the stem helices at the base of the spike protein of the SARS-CoV-2 virion. The epitopes of the antibodies disclosed herein use the CDR loops of the light and heavy chains.

[0125] The antibodies disclosed herein can be used in isolated or non-isolated form.

[0126] Preferably, the compositions of the present invention comprise a single anti-coronavirus antibody disclosed herein.

[0127] Preferably, the antibodies of the invention disclosed herein are capable of cross-neutralizing coronaviruses, particularly SARS-CoV-2.

[0128] Furthermore, the antibodies disclosed herein can be used alone or in mixtures comprising the antibodies (or variants, fragments, or bispecifics thereof) disclosed herein and / or with other antibodies that bind to coronavirus and have a coronavirus inhibitory effect. In other words, the antibodies disclosed herein can be used in combination, e.g., as pharmaceutical compositions or co-administered compositions comprising two or more antibodies that specifically bind to coronavirus. For example, antibodies with different but complementary activities can be combined in a single therapy to achieve a desired therapeutic or prophylactic effect. Alternatively, antibodies with the same activity can be combined in a single therapy to achieve a desired prophylactic or therapeutic effect. Optionally, the mixture further comprises at least one other therapeutic agent.

[0129] Preferably, the antibodies disclosed herein are bispecific antibodies and / or antigen-binding fragments thereof, and the bispecific antibodies are capable of binding to the stem helix of a coronavirus and / or the fusion peptide of a coronavirus. The phrase "capable of binding" encompasses "specifically binds to," in other words, the antibodies, bispecific antibodies, and / or antigen-binding fragments thereof disclosed herein that are capable of binding to an epitope may also bind to other epitopes.

[0130] Preferably, the coronaviruses described herein include SARS-CoV-2 and variants.

[0131] As used herein, the phrases "replace each other," "interchanged," "swap positions with each other," or "replace one another" include reference to when at least a portion of a heavy chain domain, e.g., CH1 and / or VH, of an antibody or antigen-binding fragment described herein is replaced by at least a portion of the light chain domain with which it is paired, e.g., CL and / or VL, and at the same time, when at least a portion of the light chain domain is replaced by at least a portion of the heavy chain domain. The phrases "replace each other," "interchanged," "swap positions with each other," or "replace one another" are used interchangeably herein.

[0132] Preferably, the antibodies disclosed herein are human antibodies.

[0133] Framework Area Antibodies also contain framework regions, generally four framework regions (FR1, FR2, FR3, and FR4), in each of the variable heavy and light domains. The CDRs are located between the framework regions (preferably in the order FR1-CDR1-FR2-CDR2-FR3-CDR3-FR4), and together the CDRs and framework regions define a heavy chain variable domain (or region) and a light chain variable domain (or region), which are also called antigen-binding regions or domains.

[0134] IgG antibodies As used herein, the term "IgG antibody" includes reference to an antibody that contains two antigen-binding sites. IgG is the most common antibody in human serum. The heavy chain of an IgG antibody is of the γ type, which can be subdivided into γ1, γ2, γ3, and γ4. Preferably, the antibodies described herein are γ1 heavy chains. The light chain of an IgG antibody is of the λ or κ type. Preferably, the antibodies described herein are λ light chains.

[0135] Preferably, the antibody is an IgG antibody, preferably an IgG1 antibody.

[0136] IgA antibody As used herein, the term "IgA antibody" includes reference to antibodies containing two to four antigen-binding sites. IgA is the most abundant antibody in mucosal secretions. IgA antibodies are produced in monomeric, dimeric, or secretory forms, each with two or four antigen-binding sites, and can exist as IgA1 or IgA2 isotypes. The heavy chain of an IgA antibody is of the α type, which can be subdivided into α1 and α2. The light chain of an IgA antibody is of the λ or κ type.

[0137] Preferably, the antibody is an IgA antibody.

[0138] IgM antibody As used herein, the term "IgM antibody" includes reference to antibodies containing 10 to 12 antigen-binding sites. IgM antibodies can be produced in pentameric or hexamer forms. The heavy chains of IgM antibodies are of the μ type. The light chains of IgM antibodies are of the λ or κ type.

[0139] Preferably, the antibody is an IgM antibody.

[0140] Anti-coronavirus antibodies As used herein, the term "anti-coronavirus antibody" includes reference to the antibodies disclosed herein, i.e., antibodies that can be used in the treatment methods of the present invention. Preferably, the anti-coronavirus antibodies of the present invention can treat various subtypes of coronavirus.

[0141] For the avoidance of doubt, the term "anti-coronavirus antibody" can be used interchangeably with "coronavirus antibody."

[0142] composition In preferred embodiments, the antibodies disclosed herein are the only active ingredient in the composition administered for treatment, eg, the antibody is provided in the composition as the only active ingredient.

[0143] In a preferred embodiment, the active ingredient comprises an antibody disclosed herein, e.g., the antibody is provided as a composition as the sole active ingredient in the composition administered for treatment or in combination with another antibody.

[0144] In a preferred embodiment, a first antibody disclosed herein is provided as a composition in combination with a second antibody, wherein the first antibody is selected from the group consisting of SEQ ID NOs: 001-006, 012-017, 023-028, 033-038, 042-047, 050-055, 077-082, 088-093, 099-104, 110-115, 121-126, 132-137, 197-202, 208-213, 219-224, 230-232, 232-234, 234-236, 236-238, 240-242, 242-246, 244-248, 246-248, 248-249, 250-251, 252-253, 254-255, 256-257, 258-259, 260-261, 262-263, 264-265, 266-267, 268-269, 270-271, 272-273, 274-275, 276-277, 278-279, 280-281, 282-283, 284-285, 286-287, 288-289, 290-291, 300-302, 304-305, 306-307, 308-309, 310-311, 312-313, 314-315, 316 The CDR sequence includes any one of 0 to 235, 241 to 246, 252 to 257, 303 to 308, 314 to 319, 325 to 330, 336 to 341, 347 to 352, 358 to 363, 421 to 426, 432 to 437, 443 to 448, 454 to 459, 465 to 470, 475 to 480, 514 to 519, 525 to 530, 536 to 541, 547 to 552, 558 to 563, and 569 to 574. More preferably, the first antibody comprises CDR sequences comprising any one or more of SEQ ID NOs: 046, 114, 254, 367, 430, or 545, and the second antibody comprises CDR sequences comprising any one or more of SEQ ID NOs: 639, 640, 641, 642, 643, or SEQ ID NO: 644. Preferably, the composition is administered for prophylactic and / or therapeutic treatment.

[0145] In a preferred embodiment, a first antibody disclosed herein is provided as a composition in combination with a second antibody, wherein the first antibody is selected from the group consisting of SEQ ID NOs: 001-006, 012-017, 023-028, 033-038, 042-047, 050-055, 077-082, 088-093, 099-104, 110-115, 121-126, 132-137, 197-202, 208-213, 219-224, 230-232, 232-234, 234-236, 236-238, 240-242, 242-246, 244-248, 246-248, 248-249, 250-251, 252-253, 254-255, 256-257, 258-259, 260-261, 262-263, 264-265, 266-267, 268-269, 270-271, 272-273, 274-275, 276-277, 278-279, 280-281, 282-283, 284-285, 286-287, 288-289, 290-291, 300-302, 304-305, 306-307, 308-309, 310-311, 312-313, 314-315, 316 The CDR sequence includes any one of 0 to 235, 241 to 246, 252 to 257, 303 to 308, 314 to 319, 325 to 330, 336 to 341, 347 to 352, 358 to 363, 421 to 426, 432 to 437, 443 to 448, 454 to 459, 465 to 470, 475 to 480, 514 to 519, 525 to 530, 536 to 541, 547 to 552, 558 to 563, and 569 to 574. More preferably, the first antibody comprises CDR sequences comprising any one or more of SEQ ID NOs: 046, 114, 254, 367, 430, or 545, and the second antibody comprises CDR sequences comprising any one or more of SEQ ID NOs: 645, 646, 647, 648, 649, or SEQ ID NO: 650. Preferably, the composition is administered for prophylactic and / or therapeutic treatment.

[0146] In a preferred embodiment, a first antibody disclosed herein is provided as a composition in combination with a second antibody, wherein the first antibody is selected from the group consisting of SEQ ID NOs: 001-006, 012-017, 023-028, 033-038, 042-047, 050-055, 077-082, 088-093, 099-104, 110-115, 121-126, 132-137, 197-202, 208-213, 219-224, 230-232, 232-234, 234-236, 236-238, 240-242, 242-246, 244-248, 246-248, 248-249, 250-251, 252-253, 254-255, 256-257, 258-259, 260-261, 262-263, 264-265, 266-267, 268-269, 270-271, 272-273, 274-275, 276-277, 278-279, 280-281, 282-283, 284-285, 286-287, 288-289, 290-291, 300-302, 304-305, 306-307, 308-309, 310-311, 312-313, 314-315, 316 The CDR sequence includes any one of 0 to 235, 241 to 246, 252 to 257, 303 to 308, 314 to 319, 325 to 330, 336 to 341, 347 to 352, 358 to 363, 421 to 426, 432 to 437, 443 to 448, 454 to 459, 465 to 470, 475 to 480, 514 to 519, 525 to 530, 536 to 541, 547 to 552, 558 to 563, and 569 to 574. More preferably, the first antibody comprises CDR sequences comprising any one or more of SEQ ID NOs: 046, 114, 254, 367, 430, or 545, and the second antibody comprises CDR sequences comprising any one or more of SEQ ID NOs: 651, 652, 653, 654, 655, or SEQ ID NO: 656. Preferably, the composition is administered for prophylactic and / or therapeutic treatment.

[0147] In a preferred embodiment, a first antibody disclosed herein is provided as a composition in combination with a second antibody, wherein the first antibody is selected from the group consisting of SEQ ID NOs: 001-006, 012-017, 023-028, 033-038, 042-047, 050-055, 077-082, 088-093, 099-104, 110-115, 121-126, 132-137, 197-202, 208-213, 219-224, 230-232, 232-234, 234-236, 236-238, 240-242, 242-246, 244-248, 246-248, 248-249, 250-251, 252-253, 254-255, 256-257, 258-259, 260-261, 262-263, 264-265, 266-267, 268-269, 270-271, 272-273, 274-275, 276-277, 278-279, 280-281, 282-283, 284-285, 286-287, 288-289, 290-291, 300-302, 304-305, 306-307, 308-309, 310-311, 312-313, 314-315, 316 The CDR sequence includes any one of 0 to 235, 241 to 246, 252 to 257, 303 to 308, 314 to 319, 325 to 330, 336 to 341, 347 to 352, 358 to 363, 421 to 426, 432 to 437, 443 to 448, 454 to 459, 465 to 470, 475 to 480, 514 to 519, 525 to 530, 536 to 541, 547 to 552, 558 to 563, and 569 to 574. More preferably, the first antibody comprises CDR sequences comprising any one or more of SEQ ID NOs: 046, 114, 254, 367, 430, or 545, and the second antibody comprises CDR sequences comprising any one or more of SEQ ID NOs: 657, 658, 659, 660, 661, or SEQ ID NO: 662. Preferably, the composition is administered for prophylactic and / or therapeutic treatment.

[0148] Preferably, the compositions of the present invention are water-based compositions, such as aqueous liquids. Preferably, the compositions of the present invention further comprise one or more salts, such as sodium chloride. The compositions of the present invention may further comprise one or more buffering agents, such as sodium acetate. The compositions of the present invention may further comprise one or more carbohydrates, such as sucrose, or other active ingredients, such as other antibodies, neuraminidase inhibitors, endonuclease inhibitors, and adjuvants, such as oils, cytokines, emulsifiers, or combinations thereof. The pH of the compositions of the present invention may be between 4 and 8, and more preferably, the pH is around 5.5. Preferably, the pH is around 7.4.

[0149] Composition for mucous membranes As used herein, the term "mucosal composition" refers to any pharmaceutical preparation that is topically applied to a mucosal surface to retard or resist the effects of mucosal shedding or removal.

[0150] The term "mucosal composition" is used in reference to the route of administration in which the antibodies disclosed herein are provided to the nasal cavity, oropharyngeal region, or upper respiratory tract, preferably through the nasal passages, as part of the prophylactic and / or therapeutic treatments disclosed herein.

[0151] Transmucosal Mucous membranes line the internal cavities of the human body, cover internal organs, and comprise membranes composed of one or more layers of epithelial cells and loose connective tissue, which may produce mucus. As used herein, the term "transmucosal," also referred to as "mucosal administration," refers to a route of administration in which a drug is provided prophylactically and / or therapeutically to a mucous membrane, such as those found in the nose, mouth, lungs, vagina, rectum, or stomach. Mucous membranes lubricate these organs and cavities, protecting them from abrasive particles and fluids, as well as invasive pathogens.

[0152] Preferably, the antibodies of the invention are administered transmucosally.

[0153] Intranasal As used herein, the term "intranasal," also referred to as "nasal administration," includes reference to a route of administration in which a drug is preferably provided to the upper and / or lower respiratory tract through the nostrils as part of the prophylactic and / or therapeutic treatments disclosed herein. Preferably, administration provides the drug to the nasal cavity. The back of the nasal cavity is also referred to as the pharynx. Nasal administration preferably provides delivery of the antibodies disclosed herein to the mucous membranes lining the nasal cavity. Nasal administration can be performed, for example, using a nasal spray or nasal drops. In some embodiments, the drug is delivered to the nasal cavity via the oral route. For example, RetroNose uses a breath-activated pressurized metered-dose inhaler (pMDI) to administer the drug through the oral cavity during the nasal exhalation phase. This method allows drug particles to enter the nasal cavity through the pharynx.

[0154] Preferably, the antibodies of the invention are administered intranasally.

[0155] In nasal administration, a drug is provided to the upper respiratory tract through the nostrils as part of the preventive and / or therapeutic treatments disclosed herein.Preferably, the administration provides the drug to the nasal cavity.Since the drug delivered locally in this way can continue to have a local or systemic effect, nasal administration can be a form of local or systemic administration.In the present case with the antibody disclosed herein, nasal administration is preferably a form of local administration.

[0156] The intranasal administration disclosed herein may be carried out using a medicament in liquid form, preferably in the form of drops or nasal spray. The aqueous liquid may contain an adjuvant. These adjuvants may be, for example, salts, oils, cytokines, emulsifiers, buffers, carbohydrates, and combinations thereof. Nasal administration may also be carried out using a medicament in solid form, such as a powder.

[0157] Oral inhalation As used herein, the term "oral inhalation," also referred to as "mouth inhalation," includes reference to a route of administration in which a drug is provided through the mouth to the upper and / or lower respiratory tract, e.g., the lungs, as part of the prophylactic and / or therapeutic treatment of the invention. Oral inhalation may apply, for example, to drugs in powder form as well as drugs in the form of liquid droplets or aerosols.

[0158] Preferably, the antibodies of the invention are administered by oral inhalation.

[0159] In oral inhalation, a drug is provided through the mouth to the respiratory tract, preferably the lower respiratory tract, e.g., the lungs, as part of the prophylactic and / or therapeutic treatment of the invention. As further described herein, oral inhalation also includes nasal drug delivery (also referred to as nasal drug delivery via the oral route).

[0160] Oral inhalation may be applied to, for example, drugs in powder form and drugs in droplet or aerosol form. Oral inhalation may involve the use of an inhaler. The inhaler may be involved in achieving a determined dose. Drugs administered by oral inhalation may reach the lungs, but may also be partially excreted through exhaled air.

[0161] The oral inhalation administration disclosed herein may be carried out using a medicament containing an aerosol in powder (solid) or liquid form. Powdered aerosols containing particles smaller than 3 μm in diameter will primarily reach the respiratory portion of the lungs and therefore will be better absorbed than larger particles. The medicament may contain adjuvants. These adjuvants may be, for example, salts, oils, cytokines, emulsifiers, buffers, carbohydrates, and combinations thereof.

[0162] Oral pharyngeal administration As used herein, the term "oropharyngeal administration," also referred to as delivery to the portion of the pharynx located between the soft palate and the hyoid bone, includes reference to routes of administration in which a drug is provided via the oral or nasal passages as part of a prophylactic and / or therapeutic treatment. Oropharyngeal administration may be used, for example, for drugs in powder form as well as for drugs in the form of liquid drops or aerosols.

[0163] Preferably, the antibodies of the invention are administered by oral pharyngeal administration.

[0164] in front As used herein, the term "prior" includes reference to administration of the antibody before an individual is exposed to or infected with coronavirus.

[0165] Preferably, the disclosed antibodies are administered to an individual up to 24 hours before exposure to a coronavirus, for example, 0-24 hours before the individual is exposed to said coronavirus.

[0166] Preferably, the disclosed antibodies are administered to an individual up to 48 hours before exposure to a coronavirus, for example, 0-48 hours before the individual is exposed to said coronavirus.

[0167] In a preferred embodiment, the antibody is administered two or more days prior to exposure to coronavirus. In a preferred embodiment, the antibody is administered three or more days prior to exposure to coronavirus. In a preferred embodiment, the antibody is administered four or more days prior to exposure to coronavirus. In a preferred embodiment, the antibody is administered five or more days prior to exposure to coronavirus. In a preferred embodiment, the antibody is administered six or more days prior to exposure to coronavirus. In a preferred embodiment, the antibody is administered seven or more days prior to exposure to coronavirus.

[0168] Dosage As used herein, the term "dosage" refers to an amount of antibody given over a particular period of time (e.g., over the course of 24 hours, 12 hours, 30 minutes, etc.). A dose refers to a single administration episode (e.g., taking two or more tablets, receiving two or more nasal doses), whether the dose is a unit dosage form or a multiple unit dosage form combined together. A dosage includes reference to pharmaceutical dosage forms in which the medication is packaged for administration, e.g., as a single unit dose or multiple unit doses. A dosage may be administered, for example, as one or more drops of an antibody-containing composition (e.g., a nasal spray) or one or more sprays of an antibody-containing composition (e.g., a nasal spray).

[0169] Preferably, suitable dosages of the antibodies disclosed herein, for example, when the dosage is for intranasal administration, include doses of 0.01 mg to 20 mg, preferably 0.1 mg to 15 mg, more preferably about 0.5 mg and 10 mg, or about 1 mg. Such dosages are also referred to as "fixed dosages" or "nominal dosages," as opposed to dosages based on patient weight. Fixed dosages have the advantage that the medicament can be packaged, for example, in a single unit dose. A single dose of the antibodies according to the present invention can provide protection from coronavirus infection for several days and can be provided "on demand" or "as needed." For example, an individual can receive the antibody before leaving home or before coming into contact with other individuals.

[0170] To provide long-lasting protection, the antibody may be administered periodically. For example, the antibody is administered once a month, or at least once a month. In a preferred embodiment, the antibody is administered once a week, or at least once a week, for example, twice a week. In a preferred embodiment, the antibody is administered once a day, or at least once a day. As will be apparent to those skilled in the art, less antibody may be administered if the antibody is administered more frequently (e.g., daily). In a preferred embodiment, 0.01 mg to 20 mg of antibody per week is administered (e.g., once or twice a week or daily). In an exemplary embodiment, 0.1 mg to 25 mg (i.e., 0.7 mg to 175 mg per week) is administered daily. In an exemplary embodiment, 0.5 mg to 3.5 mg (i.e., 3.5 mg to 24.5 mg per week) is administered daily.

[0171] The present invention also provides compositions comprising an antibody disclosed herein, preferably in a single dosage unit of between 0.1 mg and 20 mg, preferably between 0.5 mg and 15 mg, or preferably between 1 mg and 12.5 mg, formulated for intranasal administration and / or oral inhalation.

[0172] Use in methods for treatment The present invention particularly relates to the antibodies disclosed herein for use in methods for the treatment of coronavirus infection in an individual. More specifically, the antibodies can be used in methods for the prophylactic and / or therapeutic treatment of coronavirus infection in an individual.

[0173] Complementarity-Determining Regions (CDRs) Preferably, the CDR regions are identified according to Kabat et al. (1991) as described in Sequences of Proteins of Immunological Interest.

[0174] In a preferred embodiment, the binding interaction between the binding molecule, preferably an antibody, and the stem helix is ​​mediated exclusively by the variable sequences of the light and heavy chains.

[0175] The antibodies disclosed herein can specifically bind to coronavirus in attenuated or inactivated form or in live and / or infectious form.

[0176] The antibodies disclosed herein can also specifically bind to one or more fragments of a coronavirus.

[0177] The antibodies disclosed herein preferably comprise a heavy chain CDR1 sequence comprising any one of the following SEQ ID NOs: 006 to 011, 015 to 020, 026 to 031, 046 to 051, 054 to 059, and 064 to 069. A CDR comprising the amino acid sequence of SEQ ID NO: 046 is particularly preferred.

[0178] The antibodies disclosed herein preferably comprise a heavy chain CDR2 sequence comprising any one of the following SEQ ID NOs: 082 to 087, 093 to 098, 114 to 119, 136 to 141, 158 to 163, and 191 to 196. A CDR comprising the amino acid sequence of SEQ ID NO: 114 is particularly preferred.

[0179] The antibodies disclosed herein preferably comprise a heavy chain CDR3 sequence comprising any one of the following SEQ ID NOs: 202 to 207, 218 to 223, 237 to 242, 254 to 259, 280 to 285, and 291 to 296. A CDR comprising the amino acid sequence of SEQ ID NO: 254 is particularly preferred.

[0180] The antibodies disclosed herein preferably comprise a light chain CDR1 sequence comprising any one of the following SEQ ID NOs: 313 to 318, 324 to 329, 345 to 350, 367 to 372, 378 to 383, and 400 to 405. A CDR comprising the amino acid sequence of SEQ ID NO: 367 is particularly preferred.

[0181] The antibodies disclosed herein preferably comprise a light chain CDR2 sequence comprising any one of the following SEQ ID NOs: 430 to 435, 441 to 446, 452 to 457, 463 to 468, 479 to 484, and 502 to 507. A CDR comprising the amino acid sequence of SEQ ID NO: 430 is particularly preferred.

[0182] The antibodies disclosed herein preferably comprise a light chain CDR3 sequence comprising any one of the following SEQ ID NOs: 513 to 518, 524 to 529, 545 to 550, 584 to 589, 595 to 600, and 617 to 622. A CDR comprising the amino acid sequence of SEQ ID NO: 545 is particularly preferred.

[0183] The antibodies disclosed herein comprise a heavy chain variable domain further comprising at least the heavy chain framework region FR1 of SEQ ID NO:631.

[0184] The antibodies disclosed herein comprise a heavy chain variable domain that further comprises at least the heavy chain framework region FR2 of SEQ ID NO:632.

[0185] The antibodies disclosed herein comprise a heavy chain variable domain that further comprises at least the heavy chain framework region FR3 of SEQ ID NO:633.

[0186] The antibodies disclosed herein comprise a heavy chain variable domain further comprising at least the heavy chain framework region FR4 of SEQ ID NO:634.

[0187] In a preferred embodiment, the heavy chain variable domain of the antibody comprises a heavy chain framework region FR1 of SEQ ID NO: 631, a heavy chain framework region FR2 of SEQ ID NO: 632, a heavy chain framework region FR3 of SEQ ID NO: 633, and / or a heavy chain framework region FR4 of SEQ ID NO: 634, preferably all of the heavy chain framework regions FR1 to FR4.

[0188] The antibodies disclosed herein comprise a heavy chain variable domain that further comprises at least the light chain framework region FR1 of SEQ ID NO:635.

[0189] The antibodies disclosed herein comprise a heavy chain variable domain that further comprises at least the light chain framework region FR2 of SEQ ID NO:636.

[0190] The antibodies disclosed herein comprise a heavy chain variable domain that further comprises at least the light chain framework region FR3 of SEQ ID NO:637.

[0191] The antibodies disclosed herein comprise a heavy chain variable domain that further comprises at least the light chain framework region FR4 of SEQ ID NO:638.

[0192] In a preferred embodiment, the light chain variable domain of the antibody comprises a light chain framework region FR1 of SEQ ID NO: 635, a light chain framework region FR2 of SEQ ID NO: 636, a light chain framework region FR3 of SEQ ID NO: 637, and / or a light chain framework region FR4 of SEQ ID NO: 638, preferably all of the light chain framework regions FR1 to FR4.

[0193] Most preferably, the antibodies disclosed herein comprise a heavy chain variable domain having the sequence of SEQ ID NO:629 and / or a light chain variable domain having the sequence of SEQ ID NO:630.

[0194] In the treatment methods or compositions of the present invention, the heavy chain variable domain (VH) of the antibody is preferably set forth as SEQ ID NO: 629. In the treatment methods or compositions of the present invention, the light chain variable domain (VL) of the antibody is preferably set forth as SEQ ID NO: 630. In the treatment methods or compositions of the present invention, the antibody may be an antibody disclosed herein or a binding molecule comprising an antigen-binding domain thereof, such as a variable domain (V).

[0195] Preferably, the antibodies disclosed herein comprise a heavy chain variable domain having the sequence of SEQ ID NO: 629 with at most 15, preferably 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, more preferably 0, 1, 2, 3, 4, or 5 amino acid insertions, deletions, or substitutions.

[0196] Preferably, the antibodies disclosed herein comprise a light chain variable domain having the sequence of SEQ ID NO: 630 with at most 15, preferably 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, more preferably 0, 1, 2, 3, 4, or 5 amino acid insertions, deletions, or substitutions.

[0197] Preferably, said amino acid insertions, deletions or substitutions for the heavy and / or light chain variable domains are not within the CDRs.

[0198] array CDR and framework regions as defined herein are based on the Kabat numbering scheme set forth in Sequences of Proteins of Immunological Interest (Kabat et al., 1991).

[0199] Heavy chain CDR1 region (SEQ ID NO: 001 to SEQ ID NO: 076) Sequence number 001 DTFS Sequence number 002 DTFSS Sequence number 003 DTFSSH Sequence number 004 DTFSSHY Sequence number 005 DTFSSHYM Sequence number 006 DTFSSHYMH Sequence number 007 DTFSSHYMHW Sequence number 008 DTFSSHYMHWV Sequence number 009 DTFSSHYMHWVR Sequence number 010 DTFSSHYMHWVRQ Sequence number 011 DTFSSHYMHWVRQA Sequence number 012 FSSH Sequence number 013 FSSHY Sequence number 014 FSSHYM Sequence number 015 FSSHYMH Sequence number 016 FSSHYMHW Sequence number 017 FSSHYMHWV Sequence number 018 FSSHYMHWVR Sequence number 019 FSSHYMHWVRQ Sequence number 020 FSSHYMHWVRQA Sequence number 021 GDTFS Sequence number 022 GDTFSS Sequence number 023 GDTFSSH SEQ ID NO: 024 GDTFSSHY Sequence number 025 GDTFSSHYM SEQ ID NO: 026 GDTFSSHYMH Sequence number 027 GDTFSSHYMHW Sequence number 028 GDTFSSHYMHWV Sequence number 029 GDTFSSHYMHWVR Sequence number 030 GDTFSSHYMHWVRQ Sequence number 031 GDTFSSHYMHWVRQA Sequence number 032 HWVR SEQ ID NO. 033 HWVRQ Sequence number 034 HWVRQA SEQ ID NO. 035 HYMH Sequence number 036 HYMHW SEQ ID NO: 037 HYMHWV SEQ ID NO. 038 HYMHWVR Sequence number 039 HYMHWVRQ Sequence number 040 HYMHWVRQA SEQ ID NO: 041 MHWV Sequence number 042 MHWVR SEQ ID NO: 043 MHWVRQ Sequence number 044 MHWVRQA SEQ ID NO. 045 SHYM SEQ ID NO. 046 SHYMH Sequence number 047 SHYMHW SEQ ID NO: 048 SHYMHWV SEQ ID NO. 049 SHYMHWVR Sequence number 050 SHYMHWVRQ SEQ ID NO. 051 SHYMHWVRQA SEQ ID NO. 052 SSHY Sequence number 053 SSHYM Sequence number 054 SSHYMH Sequence number 055 SSHYMHW Sequence number 056 SSHYMHWV Sequence number 057 SSHYMHWVR Sequence number 058 SSHYMHWVRQ Sequence number 059 SSHYMHWVRQA SEQ ID NO. 060 TFSS Sequence number 061 TFSSH SEQ ID NO. 062 TFSSHY SEQ ID NO. 063 TFSSHYM SEQ ID NO: 064 TFSSHYMH Sequence number 065 TFSSHYMHW Sequence number 066 TFSSHYMHWV Sequence number 067 TFSSHYMHWVR SEQ ID NO: 068 TFSSHYMHWVRQ Sequence number 069 TFSSHYMHWVRQA Sequence number 070 WVRQ Sequence number 071 WVRQA Sequence number 072 YMHW Sequence number 073 YMHWV Sequence number 074 YMHWVR Sequence number 075 YMHWVRQ Sequence number 076 YMHWVRQA Heavy chain CDR2 region (SEQ ID NO: 077 to SEQ ID NO: 196) Sequence number 077 EWMGIINPSGSGTAYG Sequence number 078 EWMGIINPSGSGTAYGQ Sequence number 079 EWMGIINPSGSGTAYGQK Sequence number 080 EWMGIINPSGSGTAYGQKF Sequence number 081 EWMGIINPSGSGTAYGQKFQ Sequence number 082 EWMGIINPSGSGTAYGQKFQG Sequence number 083 EWMGIINPSGSGTAYGQKFQGR Sequence number 084 EWMGIINPSGSGTAYGQKFQGRL Sequence number 085 EWMGIINPSGSGTAYGQKFQGRLT Sequence number 086 EWMGIINPSGSGTAYGQKFQGRLTM Sequence number 087 EWMGIINPSGSGTAYGQKFQGRLTMT Sequence number 088 GIINPSGSGTAYG Sequence number 089 GIINPSGSGTAYGQ Sequence number 090 GIINPSGSGTAYGQK Sequence number 091 GIINPSGSGTAYGQKF Sequence number 092 GIINPSGSGTAYGQKFQ Sequence number 093 GIINPSGSGTAYGQKFQG Sequence number 094 GIINPSGSGTAYGQKFQGR Sequence number 095 GIINPSGSGTAYGQKFQGRL Sequence number 096 GIINPSGSGTAYGQKFQGRLT Sequence number 097 GIINPSGSGTAYGQKFQGRLTM Sequence number 098 GIINPSGSGTAYGQKFQGRLTMT Sequence number 099 GSGTAYGQ Sequence number 100 GSGTAYGQK Sequence number 101 GSGTAYGQKF SEQ ID NO: 102 GSGTAYGQKFQ SEQ ID NO: 103 GSGTAYGQKFQG SEQ ID NO: 104 GSGTAYGQKFQGR SEQ ID NO: 105 GSGTAYGQKFQGRL SEQ ID NO: 106 GSGTAYGQKFQGRLT SEQ ID NO: 107 GSGTAYGQKFQGRLTM SEQ ID NO: 108 GSGTAYGQKFQGRLTMT SEQ ID NO: 109 IINPSGSGTAYG SEQ ID NO: 110 IINPSGSGTAYGQ Sequence number 111 IINPSGSGTAYGQK Sequence number 112 IINPSGSGTAYGQKF Sequence number 113 IINPSGSGTAYGQKFQ SEQ ID NO: 114 IINPSGSGTAYGQKFQG SEQ ID NO: 115 IINPSGSGTAYGQKFQGR SEQ ID NO: 116 IINPSGSGTAYGQKFQGRL SEQ ID NO: 117 IINPSGSGTAYGQKFQGRLT SEQ ID NO: 118 IINPSGSGTAYGQKFQGRLTM SEQ ID NO: 119 IINPSGSGTAYGQKFQGRLTMT SEQ ID NO: 120 INPSGSGTAYG SEQ ID NO: 121 INPSGSGTAYGQ SEQ ID NO: 122 INPSGSGTAYGQK SEQ ID NO: 123 INPSGSGTAYGQKF SEQ ID NO: 124 INPSGSGTAYGQKFQ SEQ ID NO: 125 INPSGSGTAYGQKFQG SEQ ID NO: 126 INPSGSGTAYGQKFQGR SEQ ID NO: 127 INPSGSGTAYGQKFQGRL SEQ ID NO: 128 INPSGSGTAYGQKFQGRLT SEQ ID NO: 129 INPSGSGTAYGQKFQGRLTM SEQ ID NO: 130 INPSGSGTAYGQKFQGRLTMT SEQ ID NO: 131 MGIINPSGSGTAYG SEQ ID NO: 132 MGIINPSGSGTAYGQ Sequence number 133 MGIINPSGSGTAYGQK Sequence number 134 MGIINPSGSGTAYGQKF Sequence number 135 MGIINPSGSGTAYGQKFQ SEQ ID NO: 136 MGIINPSGSGTAYGQKFQG Sequence number 137 MGIINPSGSGTAYGQKFQGR SEQ ID NO: 138 MGIINPSGSGTAYGQKFQGRL SEQ ID NO: 139 MGIINPSGSGTAYGQKFQGRLT SEQ ID NO: 140 MGIINPSGSGTAYGQKFQGRLTM SEQ ID NO: 141 MGIINPSGSGTAYGQKFQGRLTMT SEQ ID NO: 142 NPSGSGTAYG SEQ ID NO: 143 NPSGSGTAYGQ SEQ ID NO: 144 NPSGSGTAYGQK SEQ ID NO: 145 NPSGSGTAYGQKF SEQ ID NO: 146 NPSGSGTAYGQKFQ SEQ ID NO: 147 NPSGSGTAYGQKFQG SEQ ID NO: 148 NPSGSGTAYGQKFQGR SEQ ID NO: 149 NPSGSGTAYGQKFQGRL SEQ ID NO: 150 NPSGSGTAYGQKFQGRLT SEQ ID NO: 151 NPSGSGTAYGQKFQGRLTM SEQ ID NO: 152 NPSGSGTAYGQKFQGRLTMT SEQ ID NO: 153 PEWMGIINPSGSGTAYG SEQ ID NO: 154 PEWMGIINPSGSGTAYGQ Sequence number 155 PEWMGIINPSGSGTAYGQK Sequence number 156 PEWMGIINPSGSGTAYGQKF Sequence number 157 PEWMGIINPSGSGTAYGQKFQ Sequence number 158 PEWMGIINPSGSGTAYGQKFQG Sequence number 159 PEWMGIINPSGSGTAYGQKFQGR Sequence number 160 PEWMGIINPSGSGTAYGQKFQGRL SEQ ID NO: 161 PEWMGIINPSGSGTAYGQKFQGRLT SEQ ID NO: 162 PEWMGIINPSGSGTAYGQKFQGRLTM SEQ ID NO: 163 PEWMGIINPSGSGTAYGQKFQGRLTMT SEQ ID NO: 164 PSGSGTAYG SEQ ID NO: 165 PSGSGTAYGQ SEQ ID NO: 166 PSGSGTAYGQK SEQ ID NO: 167 PSGSGTAYGQKF SEQ ID NO: 168 PSGSGTAYGQKFQ SEQ ID NO: 169 PSGSGTAYGQKFQG SEQ ID NO: 170 PSGSGTAYGQKFQGR SEQ ID NO: 171 PSGSGTAYGQKFQGRL SEQ ID NO: 172 PSGSGTAYGQKFQGRLT SEQ ID NO: 173 PSGSGTAYGQKFQGRLTM SEQ ID NO: 174 PSGSGTAYGQKFQGRLTMT SEQ ID NO: 175 SGSGTAYG SEQ ID NO: 176 SGSGTAYGQ Sequence number 177 SGSGTAYGQK Sequence number 178 SGSGTAYGQKF Sequence number 179 SGSGTAYGQKFQ SEQ ID NO: 180 SGSGTAYGQKFQG SEQ ID NO: 181 SGSGTAYGQKFQGR SEQ ID NO: 182 SGSGTAYGQKFQGRL SEQ ID NO: 183 SGSGTAYGQKFQGRLT Sequence number 184 SGSGTAYGQKFQGRLTM SEQ ID NO: 185 SGSGTAYGQKFQGRLTMT Sequence number 186 WMGIINPSGSGTAYG Sequence number 187 WMGIINPSGSGTAYGQ Sequence number 188 WMGIINPSGSGTAYGQK Sequence number 189 WMGIINPSGSGTAYGQKF Sequence number 190 WMGIINPSGSGTAYGQKFQ Sequence number 191 WMGIINPSGSGTAYGQKFQG Sequence number 192 WMGIINPSGSGTAYGQKFQGR Sequence number 193 WMGIINPSGSGTAYGQKFQGRL Sequence number 194 WMGIINPSGSGTAYGQKFQGRLT Sequence number 195 WMGIINPSGSGTAYGQKFQGRLTM Sequence number 196 WMGIINPSGSGTAYGQKFQGRLTMT Heavy chain CDR3 region (SEQ ID NO: 197 to SEQ ID NO: 296) SEQ ID NO: 197 CGGGSG SEQ ID NO: 198 CGGGSGG SEQ ID NO: 199 CGGGSGGL Sequence number 200 CGGGSGGLF SEQ ID NO: 201 CGGGSGGLFA SEQ ID NO: 202 CGGGSGGLFAY Sequence number 203 CGGGSGGLFAYW Sequence number 204 CGGGSGGLFAYWG SEQ ID NO: 205 CGGGSGGLFAYWGQ SEQ ID NO: 206 CGGGSGGLFAYWGQG SEQ ID NO: 207 CGGGSGGLFAYWGQGT Sequence number 208 FAYW Sequence number 209 FAYWG SEQ ID NO: 210 FAYWGQ Sequence number 211 FAYWGQG SEQ ID NO: 212 FAYWGQGT SEQ ID NO: 213 GGGSG SEQ ID NO: 214 GGGSGG SEQ ID NO: 215 GGGSGGL SEQ ID NO: 216 GGGSGGLF SEQ ID NO: 217 GGGSGGLFA SEQ ID NO: 218 GGGSGGLFAY SEQ ID NO: 219 GGGSGGLFAYW SEQ ID NO: 220 GGGSGGLFAYWG SEQ ID NO: 221 GGGSGGLFAYWGQ SEQ ID NO: 222 GGGSGGLFAYWGQG SEQ ID NO: 223 GGGSGGLFAYWGQGT SEQ ID NO: 224 GGLF SEQ ID NO: 225 GGLFA SEQ ID NO: 226 GGLFAY Sequence number 227 GGLFAYW SEQ ID NO: 228 GGLFAYWG SEQ ID NO: 229 GGLFAYWGQ SEQ ID NO: 230 GGLFAYWGQG SEQ ID NO: 231 GGLFAYWGQGT SEQ ID NO: 232 GGSG SEQ ID NO: 233 GGSGG SEQ ID NO: 234 GGSGGL SEQ ID NO: 235 GGSGGLF SEQ ID NO: 236 GGSGGLFA SEQ ID NO: 237 GGSGGLFAY SEQ ID NO: 238 GGSGGLFAYW Sequence number 239 GGSGGLFAYWG SEQ ID NO: 240 GGSGGLFAYWGQ SEQ ID NO: 241 GGSGGLFAYWGQG SEQ ID NO: 242 GGSGGLFAYWGQGT SEQ ID NO: 243 GLFA SEQ ID NO: 244 GLFAY Sequence number 245 GLFAYW Sequence number 246 GLFAYWG SEQ ID NO: 247 GLFAYWGQ SEQ ID NO: 248 GLFAYWGQG SEQ ID NO: 249 GLFAYWGQGT SEQ ID NO: 250 GSGG SEQ ID NO: 251 GSGGL SEQ ID NO: 252 GSGGLF SEQ ID NO: 253 GSGGLFA SEQ ID NO: 254 GSGGLFAY Sequence number 255 GSGGLFAYW Sequence number 256 GSGGLFAYWG SEQ ID NO: 257 GSGGLFAYWGQ SEQ ID NO: 258 GSGGLFAYWGQG SEQ ID NO: 259 GSGGLFAYWGQGT Sequence number 260 LFAY Sequence number 261 LFAYW Sequence number 262 LFAYWG SEQ ID NO: 263 LFAYWGQ Sequence number 264 LFAYWGQG Sequence number 265 LFAYWGQGT SEQ ID NO: 266 SGGL Sequence number 267 SGGLF SEQ ID NO: 268 SGGLFA Sequence number 269 SGGLFAY Sequence number 270 SGGLFAYW Sequence number 271 SGGLFAYWG SEQ ID NO: 272 SGGLFAYWGQ SEQ ID NO: 273 SGGLFAYWGQG SEQ ID NO: 274 SGGLFAYWGQGT SEQ ID NO: 275 YCGGGSG SEQ ID NO: 276 YCGGGSGG SEQ ID NO: 277 YCGGGSGGL SEQ ID NO: 278 YCGGGSGGLF SEQ ID NO: 279 YCGGGSGGLFA SEQ ID NO: 280 YCGGGSGGLFAY Sequence number 281 YCGGGSGGLFAYW Sequence number 282 YCGGGSGGLFAYWG SEQ ID NO: 283 YCGGGSGGLFAYWGQ Sequence number 284 YCGGGSGGLFAYWGQG Sequence number 285 YCGGGSGGLFAYWGQGT SEQ ID NO: 286 YYCGGGSG SEQ ID NO: 287 YYCGGGSGG SEQ ID NO: 288 YYCGGGSGGL SEQ ID NO: 289 YYCGGGSGGLF SEQ ID NO: 290 YYCGGGSGGLFA SEQ ID NO: 291 YYCGGGSGGLFAY Sequence number 292 YYCGGGSGGLFAYW Sequence number 293 YYCGGGSGGLFAYWG SEQ ID NO: 294 YYCGGGSGGLFAYWGQ Sequence number 295 YYCGGGSGGLFAYWGQG SEQ ID NO: 296 YYCGGGSGGLFAYWGQGT Light chain CDR1 region (SEQ ID NO: 297 to SEQ ID NO: 414) Sequence number 297 ASQIVR SEQ ID NO: 298 ASQIVRS Sequence number 299 ASQIVRSN Sequence number 300 ASQIVRSNY SEQ ID NO: 301 ASQIVRSNYL SEQ ID NO: 302 ASQIVRSNYLA Sequence number 303 ASQIVRSNYLAW SEQ ID NO: 304 ASQIVRSNYLAWY Sequence number 305 ASQIVRSNYLAWYQ Sequence number 306 ASQIVRSNYLAWYQQ Sequence number 307 ASQIVRSNYLAWYQQK SEQ ID NO: 308 ATLSCRASQIVR SEQ ID NO: 309 ATLSCRASQIVRS SEQ ID NO: 310 ATLSCRASQIVRSN SEQ ID NO: 311 ATLSCRASQIVRSNY SEQ ID NO: 312 ATLSCRASQIVRSNYL SEQ ID NO: 313 ATLSCRASQIVRSNYLA SEQ ID NO: 314 ATLSCRASQIVRSNYLAW SEQ ID NO: 315 ATLSCRASQIVRSNYLAWY SEQ ID NO: 316 ATLSCRASQIVRSNYLAWYQ SEQ ID NO: 317 ATLSCRASQIVRSNYLAWYQQ SEQ ID NO: 318 ATLSCRASQIVRSNYLAWYQQK SEQ ID NO: 319 CRASQIVR SEQ ID NO: 320 CRASQIVRS SEQ ID NO: 321 CRASQIVRSN SEQ ID NO: 322 CRASQIVRSNY SEQ ID NO: 323 CRASQIVRSNYL SEQ ID NO: 324 CRASQIVRSNYLA SEQ ID NO: 325 CRASQIVRSNYLAW SEQ ID NO: 326 CRASQIVRSNYLAWY SEQ ID NO: 327 CRASQIVRSNYLAWYQ SEQ ID NO: 328 CRASQIVRSNYLAWYQQ SEQ ID NO: 329 CRASQIVRSNYLAWYQQK SEQ ID NO: 330 IVRS SEQ ID NO: 331 IVRSN Sequence number 332 IVRSNY SEQ ID NO: 333 IVRSNYL SEQ ID NO: 334 IVRSNYLA Sequence number 335 IVRSNYLAW SEQ ID NO: 336 IVRSNYLAWY Sequence number 337 IVRSNYLAWYQ Sequence number 338 IVRSNYLAWYQQ Sequence number 339 IVRSNYLAWYQQK SEQ ID NO: 340 LSCRASQIVR SEQ ID NO: 341 LSCRASQIVRS SEQ ID NO: 342 LSCRASQIVRSN SEQ ID NO: 343 LSCRASQIVRSNY SEQ ID NO: 344 LSCRASQIVRSNYL SEQ ID NO: 345 LSCRASQIVRSNYLA SEQ ID NO: 346 LSCRASQIVRSNYLAW SEQ ID NO: 347 LSCRASQIVRSNYLAWY SEQ ID NO: 348 LSCRASQIVRSNYLAWYQ SEQ ID NO: 349 LSCRASQIVRSNYLAWYQQ SEQ ID NO: 350 LSCRASQIVRSNYLAWYQQK Sequence number 351 QIVR SEQ ID NO: 352 QIVRS Sequence number 353 QIVRSN SEQ ID NO: 354 QIVRSNY Sequence number 355 QIVRSNYL SEQ ID NO: 356 QIVRSNYLA Sequence number 357 QIVRSNYLAW SEQ ID NO:358 QIVRSNYLAWY Sequence number 359 QIVRSNYLAWYQ Sequence number 360 QIVRSNYLAWYQQ SEQ ID NO: 361 QIVRSNYLAWYQQK SEQ ID NO: 362 RASQIVR SEQ ID NO: 363 RASQIVRS SEQ ID NO. 364 RASQIVRSN SEQ ID NO: 365 RASQIVRSNY SEQ ID NO: 366 RASQIVRSNYL SEQ ID NO: 367 RASQIVRSNYLA SEQ ID NO: 368 RASQIVRSNYLAW SEQ ID NO: 369 RASQIVRSNYLAWY SEQ ID NO: 370 RASQIVRSNYLAWYQ SEQ ID NO: 371 RASQIVRSNYLAWYQQ SEQ ID NO: 372 RASQIVRSNYLAWYQQK SEQ ID NO: 373 SCRASQIVR SEQ ID NO: 374 SCRASQIVRS SEQ ID NO: 375 SCRASQIVRSN SEQ ID NO: 376 SCRASQIVRSNY SEQ ID NO: 377 SCRASQIVRSNYL SEQ ID NO: 378 SCRASQIVRSNYLA SEQ ID NO: 379 SCRASQIVRSNYLAW SEQ ID NO: 380 SCRASQIVRSNYLAWY SEQ ID NO: 381 SCRASQIVRSNYLAWYQ SEQ ID NO: 382 SCRASQIVRSNYLAWYQQ SEQ ID NO: 383 SCRASQIVRSNYLAWYQQK Sequence number 384 SQIVR SEQ ID NO: 385 SQIVRS Sequence number 386 SQIVRSN SEQ ID NO: 387 SQIVRSNY SEQ ID NO: 388 SQIVRSNYL SEQ ID NO: 389 SQIVRSNYLA SEQ ID NO: 390 SQIVRSNYLAW SEQ ID NO: 391 SQIVRSNYLAWY SEQ ID NO: 392 SQIVRSNYLAWYQ SEQ ID NO: 393 SQIVRSNYLAWYQQ Sequence number 394 SQIVRSNYLAWYQQK SEQ ID NO: 395 TLSCRASQIVR SEQ ID NO: 396 TLSCRASQIVRS SEQ ID NO: 397 TLSCRASQIVRSN SEQ ID NO: 398 TLSCRASQIVRSNY SEQ ID NO: 399 TLSCRASQIVRSNYL SEQ ID NO: 400 TLSCRASQIVRSNYLA SEQ ID NO:401 TLSCRASQIVRSNYLAW SEQ ID NO:402 TLSCRASQIVRSNYLAWY SEQ ID NO:403 TLSCRASQIVRSNYLAWYQ SEQ ID NO: 404 TLSCRASQIVRSNYLAWYQQ SEQ ID NO: 405 TLSCRASQIVRSNYLAWYQQK Sequence number 406 VRSN Sequence number 407 VRSNY Sequence number 408 VRSNYL Sequence number 409 VRSNYLA Sequence number 410 VRSNYLAW SEQ ID NO: 411 VRSNYLAWY SEQ ID NO: 412 VRSNYLAWYQ SEQ ID NO: 413 VRSNYLAWYQQ Sequence number 414 VRSNYLAWYQQK Light chain CDR2 region (SEQ ID NOs: 415 to 507) SEQ ID NO: 415 ASSR Sequence number 416 ASSRA SEQ ID NO: 417 ASSRAT SEQ ID NO: 418 ASSRATG SEQ ID NO: 419 ASSRATGT Sequence number 420 ASSRATGTP SEQ ID NO: 421 ASSRATGTPD SEQ ID NO: 422 ASSRATGTPDR SEQ ID NO: 423 ATGT SEQ ID NO: 424 ATGTP SEQ ID NO: 425 ATGTPD SEQ ID NO: 426 ATGTPDR SEQ ID NO: 427 GASS SEQ ID NO: 428 GASSR Sequence number 429 GASSRA Sequence number 430 GASSRAT SEQ ID NO: 431 GASSRATG SEQ ID NO: 432 GASSRATGT SEQ ID NO: 433 GASSRATGTP SEQ ID NO: 434 GASSRATGTPD SEQ ID NO: 435 GASSRATGTPDR SEQ ID NO: 436 IYGA SEQ ID NO: 437 IYGAS SEQ ID NO: 438 IYGASS SEQ ID NO: 439 IYGASSR SEQ ID NO. 440 IYGASSRA SEQ ID NO: 441 IYGASSRAT SEQ ID NO: 442 IYGASSRATG SEQ ID NO: 443 IYGASSRATGT SEQ ID NO: 444 IYGASSRATGTP SEQ ID NO: 445 IYGASSRATGTPD SEQ ID NO: 446 IYGASSRATGTPDR SEQ ID NO: 447 LIYGA SEQ ID NO: 448 LIYGAS SEQ ID NO: 449 LIYGASS Sequence number 450 LIYGASSR Sequence number 451 LIYGASSRA Sequence number 452 LIYGASSRAT Sequence number 453 LIYGASSRATG SEQ ID NO: 454 LIYGASSRATGT Sequence number 455 LIYGASSRATGTP SEQ ID NO: 456 LIYGASSRATGTPD SEQ ID NO: 457 LIYGASSRATGTPDR SEQ ID NO: 458 LLIYGA SEQ ID NO: 459 LLIYGAS Sequence number 460 LLIYGASS SEQ ID NO: 461 LLIYGASSR Sequence number 462 LLIYGASSRA Sequence number 463 LLIYGASSRAT Sequence number 464 LLIYGASSRATG Sequence number 465 LLIYGASSRATGT Sequence number 466 LLIYGASSRATGTP SEQ ID NO: 467 LLIYGASSRATGTPD Sequence number 468 LLIYGASSRATGTPDR SEQ ID NO: 469 RATG SEQ ID NO: 470 RATGT SEQ ID NO: 471 RATGTP SEQ ID NO: 472 RATGTPD SEQ ID NO: 473 RATGTPDR SEQ ID NO: 474 RLLIYGA SEQ ID NO: 475 RLLIYGAS SEQ ID NO: 476 RLLIYGASS SEQ ID NO: 477 RLLIYGASSR SEQ ID NO: 478 RLLIYGASSRA Sequence number 479 RLLIYGASSRAT Sequence number 480 RLLIYGASSRATG Sequence number 481 RLLIYGASSRATGT Sequence number 482 RLLIYGASSRATGTP SEQ ID NO: 483 RLLIYGASSRATGTPD SEQ ID NO: 484 RLLIYGASSRATGTPDR Sequence number 485 SRAT SEQ ID NO: 486 SRATG Sequence number 487 SRATGT SEQ ID NO: 488 SRATGTP SEQ ID NO: 489 SRATGTPD SEQ ID NO: 490 SRATGTPDR Sequence number 491 SSRA SEQ ID NO: 492 SSRAT SEQ ID NO: 493 SSRATG SEQ ID NO:494 SSRATGT Sequence number 495 SSRATGTP SEQ ID NO:496 SSRATGTPD SEQ ID NO: 497 SSRATGTPDR SEQ ID NO: 498 YGAS Sequence number 499 YGASS Sequence number 500 YGASSR Sequence number 501 YGASSRA Sequence number 502 YGASSRAT Sequence number 503 YGASSRATG SEQ ID NO:504 YGASSRATGT Sequence number 505 YGASSRATGTP SEQ ID NO:506 YGASSRATGTPD Sequence number 507 YGASSRATGTPDR Light chain CDR3 region (SEQ ID NO: 508 to SEQ ID NO: 622) SEQ ID NO:508 AVYYCLQYDSS SEQ ID NO:509 AVYYCLQYDSSP SEQ ID NO: 510 AVYYCLQYDSSPP SEQ ID NO:511 AVYYCLQYDSSPPT SEQ ID NO:512 AVYYCLQYDSSPPTY SEQ ID NO:513 AVYYCLQYDSSPPTYI SEQ ID NO:514 AVYYCLQYDSSPPTYIF SEQ ID NO:515 AVYYCLQYDSSPPTYIFG SEQ ID NO:516 AVYYCLQYDSSPPTYIFGQ SEQ ID NO:517 AVYYCLQYDSSPPTYIFGQG SEQ ID NO: 518 AVYYCLQYDSSPPTYIFGQGT SEQ ID NO:519 CLQYDSS SEQ ID NO: 520 CLQYDSSP SEQ ID NO:521 CLQYDSSPP SEQ ID NO:522 CLQYDSSPPT SEQ ID NO:523 CLQYDSSPPTY SEQ ID NO:524 CLQYDSSPPTYI SEQ ID NO:525 CLQYDSSPPTYIF SEQ ID NO:526 CLQYDSSPPTYIFG SEQ ID NO:527 CLQYDSSPPTYIFGQ SEQ ID NO: 528 CLQYDSSPPTYIFGQG SEQ ID NO: 529 CLQYDSSPPTYIFGQGT SEQ ID NO: 530 DSSP SEQ ID NO:531 DSSPP SEQ ID NO: 532 DSSPPT SEQ ID NO:533 DSSPPTY SEQ ID NO:534 DSSPPTYI SEQ ID NO:535 DSSPPTYIF SEQ ID NO:536 DSSPPTYIFG SEQ ID NO:537 DSSPPTYIFGQ SEQ ID NO:538 DSSPPTYIFGQG SEQ ID NO:539 DSSPPTYIFGQGT SEQ ID NO:540 LQYDSS SEQ ID NO:541 LQYDSSP SEQ ID NO:542 LQYDSSPP SEQ ID NO:543 LQYDSSPPT SEQ ID NO:544 LQYDSSPPTY SEQ ID NO:545 LQYDSSPPTYI SEQ ID NO:546 LQYDSSPPTYIF SEQ ID NO:547 LQYDSSPPTYIFG SEQ ID NO:548 LQYDSSPPTYIFGQ SEQ ID NO:549 LQYDSSPPTYIFGQG Sequence number 550 LQYDSSPPTYIFGQGT SEQ ID NO:551 QYDSS SEQ ID NO:552 QYDSSP SEQ ID NO:553 QYDSSPP SEQ ID NO:554 QYDSSPPT SEQ ID NO:555 QYDSSPPTY SEQ ID NO:556 QYDSSPPTYI SEQ ID NO:557 QYDSSPPTYIF SEQ ID NO:558 QYDSSPPTYIFG Sequence number 559 QYDSSPPTYIFGQ SEQ ID NO: 560 QYDSSPPTYIFGQG SEQ ID NO:561 QYDSSPPTYIFGQGT SEQ ID NO:562 SPPT SEQ ID NO:563 SPPTY SEQ ID NO:564 SPPTYI SEQ ID NO:565 SPPTYIF SEQ ID NO:566 SPPTYIFG SEQ ID NO:567 SPPTYIFGQ SEQ ID NO:568 SPPTYIFGQG SEQ ID NO:569 SPPTYIFGQGT SEQ ID NO:570 SSPP SEQ ID NO:571 SSPPT SEQ ID NO:572 SSPPTY SEQ ID NO:573 SSPPTYI SEQ ID NO:574 SSPPTYIF SEQ ID NO:575 SSPPTYIFG SEQ ID NO:576 SSPPTYIFGQ Sequence number 577 SSPPTYIFGQG SEQ ID NO: 578 SSPPTYIFGQGT Sequence number 579 VYYCLQYDSS SEQ ID NO:580 VYYCLQYDSSP SEQ ID NO:581 VYYCLQYDSSPP SEQ ID NO:582 VYYCLQYDSSPPT SEQ ID NO:583 VYYCLQYDSSPPTY SEQ ID NO:584 VYYCLQYDSSPPTYI Sequence number 585 VYYCLQYDSSPPTYIF Sequence number 586 VYYCLQYDSSPPTYIFG Sequence number 587 VYYCLQYDSSPPTYIFGQ Sequence number 588 VYYCLQYDSSPPTYIFGQG Sequence number 589 VYYCLQYDSSPPTYIFGQGT SEQ ID NO:590 YCLQYDSS SEQ ID NO:591 YCLQYDSSP SEQ ID NO:592 YCLQYDSSPP SEQ ID NO:593 YCLQYDSSPPT SEQ ID NO:594 YCLQYDSSPPTY SEQ ID NO: 595 YCLQYDSSPPTYI SEQ ID NO:596 YCLQYDSSPPTYIF SEQ ID NO:597 YCLQYDSSPPTYIFG SEQ ID NO:598 YCLQYDSSPPTYIFGQ Sequence number 599 YCLQYDSSPPTYIFGQG Sequence number 600 YCLQYDSSPPTYIFGQGT Sequence number 601 YDSS SEQ ID NO: 602 YDSSP SEQ ID NO: 603 YDSSPP SEQ ID NO: 604 YDSSPPT SEQ ID NO: 605 YDSSPPTY SEQ ID NO: 606 YDSSPPTYI Sequence number 607 YDSSPPTYIF SEQ ID NO: 608 YDSSPPTYIFG Sequence number 609 YDSSPPTYIFGQ SEQ ID NO: 610 YDSSPPTYIFGQG SEQ ID NO: 611 YDSSPPTYIFGQGT SEQ ID NO: 612 YYCLQYDSS SEQ ID NO: 613 YYCLQYDSSP SEQ ID NO: 614 YYCLQYDSSPP SEQ ID NO: 615 YYCLQYDSSPPT SEQ ID NO: 616 YYCLQYDSSPPTY SEQ ID NO: 617 YYCLQYDSSPPTYI SEQ ID NO: 618 YYCLQYDSSPPTYIF SEQ ID NO: 619 YYCLQYDSSPPTYIFG SEQ ID NO: 620 YYCLQYDSSPPTYIFGQ SEQ ID NO: 621 YYCLQYDSSPPTYIFGQG SEQ ID NO: 622 YYCLQYDSSPPTYIFGQGT Heavy Chain Variable Domain (SEQ ID NO: 629) QVQLVQSGAEVKKPAASVKVSCKASGDTFSSHYMHWVRQAPGQGPEWMGIINPSGSGTAYGQKFQGRLTMTRDTSTSTVYMELSSLTSDDTAVYYCGGGSGGLFAYWGQGTLVTVSS Light Chain Variable Domain (SEQ ID NO: 630) EIVLTQSPGTLSLSPGERATLSCRASQIVRSNYLAWYQQKPGQAPRLLIYGASSRATGTPDRFSGGGSGTDFTLTINRLEPEDFAVYYCLQYDSSPPTYIFGQGTKLEIK Heavy chain FR1 region (SEQ ID NO: 631) Sequence number 631 QVQLVQSGAEVKKPAASVKVSCKASGDTFS Heavy chain FR2 region (SEQ ID NO: 632) Sequence number 632 WVRQAPGQGPEWMG Heavy chain FR3 region (SEQ ID NO: 633) SEQ ID NO: 633 RLTMTRDTSTSTVYMELSSLTSDDTAVYYCGG Heavy chain FR4 region (SEQ ID NO: 634) Sequence number 634 WGQGTLVTVSS Light chain FR1 region (SEQ ID NO: 635) SEQ ID NO: 635 EIVLTQSPGTLSLSPGERATLSC Light chain FR2 region (SEQ ID NO: 636) SEQ ID NO: 636 WYQQKPGQAPRLLIY Light chain FR3 region (SEQ ID NO: 637) SEQ ID NO: 637 GTPDRFSGGGSGTDFTLTINRLEPEDFAVYYC Light chain FR4 region (SEQ ID NO: 638) Sequence number 638 FGQGTKLEIK Heavy chain CDR1 region (SEQ ID NO: 639) SEQ ID NO: 639 DYRIH Heavy chain CDR2 region (SEQ ID NO: 640) Sequence number 640 RMNPKSGDTNFAQKFQG Heavy chain CDR3 region (SEQ ID NO: 641) SEQ ID NO: 641 LLIVGGFDPLDDFEV Light chain CDR1 region (SEQ ID NO: 642) SEQ ID NO: 642 SGTSSDVGGYNFVS Light chain CDR2 region (SEQ ID NO: 643) SEQ ID NO: 643 EVTKRPS Light chain CDR3 region (SEQ ID NO: 644) SEQ ID NO: 644 SSYGGTNNLL Heavy chain CDR1 region (SEQ ID NO: 645) SEQ ID NO: 645 GYAMH Heavy chain CDR2 region (SEQ ID NO: 646) SEQ ID NO: 646 VISRDARNKYYADSVKG Heavy chain CDR3 region (SEQ ID NO: 647) SEQ ID NO: 647 LIIPGITEPGSPDALDI Light chain CDR1 region (SEQ ID NO: 648) Sequence number 648 RASQDISKWLA Light chain CDR2 region (SEQ ID NO: 649) SEQ ID NO: 649 AASSLQS Light chain CDR3 region (SEQ ID NO: 650) Sequence number 650 QQASSFPWSIT Heavy chain CDR1 region (SEQ ID NO: 651) SEQ ID NO: 651 YFYLH Heavy chain CDR2 region (SEQ ID NO: 652) SEQ ID NO: 652 IINPRGDGTRYAQKFQG Heavy chain CDR3 region (SEQ ID NO: 653) SEQ ID NO: 653 GADHGAFDI Light chain CDR1 region (SEQ ID NO: 654) SEQ ID NO: 654 RASQSVRRNYFA Light chain CDR2 region (SEQ ID NO: 655) SEQ ID NO. 655 DASTRAT Light chain CDR3 region (SEQ ID NO: 656) SEQ ID NO: 656 QQYDSSPPMYI Heavy chain CDR1 region (SEQ ID NO: 657) SEQ ID NO: 657 SYYMH Heavy chain CDR2 region (SEQ ID NO: 658) Sequence number 658 LITPSGDDTYYAQRFQG Heavy chain CDR3 region (SEQ ID NO: 659) SEQ ID NO: 659 MSRAGGFDV Light chain CDR1 region (SEQ ID NO: 660) SEQ ID NO: 660 RASQSITGRYLA Light chain CDR2 region (SEQ ID NO: 661) SEQ ID NO: 661 GESSRVT Light chain CDR3 region (SEQ ID NO: 662) Sequence number 662 QHFASSPPTYT Heavy Chain Variable Domain (SEQ ID NO: 663) QMQLMQSGAEVKKPGASVTVSCKASGDTFSDYRIHWVRQAPGQGLEWMGRMNPKSGDTNFAQKFQGRVTMTRDMSINTAYMTLSGLTFDDTALYYCASLLIVGGDPLDDFEVWGQGTMVTISS Light Chain Variable Domain (SEQ ID NO: 664) QSALTQPPSASGSPGQSVTISCSGTSSDVGGYNFVSWYQHHPGKAPKILIYEVTKRPSGVPDRFSGSKSGNTASLTVSGLQAEDEADYYCSSYGGTNNLLFGGGTKLTVL Heavy Chain Variable Domain (SEQ ID NO: 665) QVQLVESGGGVVQPGRSLRLSCAASGLTFSGYAMHWVRQAPGKGLEWVAVISRDARNKYYADSVKGRFTISRDNSKKTVYLEMNSLRVEDTAVYYCAILIIPGITEPGSPDALDIWGQGTMVSVSS Light Chain Variable Domain (SEQ ID NO: 666) DIQMTQSPSSMSASVGDRVTITCRASQDISKWLAWYQQRPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQASSFPWSITFGQGTRLEIR [Example]

[0200] The monoclonal antibody CV3-25 specifically binds to the stem helix of SARS-CoV-2, a binding region shared with the antibodies of the present invention, and has demonstrated in vitro protection against the original Wuhan strain of SARS-CoV-2. When administered mucosally at low dosages, CV3-25 provides a lower level of protection compared to its systemic administration. In contrast, when administered mucosally, the antibodies of the present invention provide a greater level of protection, as demonstrated by the in vivo studies presented below.

[0201] Example 1 CV3-25 The objective of this study was to evaluate the pre-exposure efficacy of monoclonal antibody CV3-25 after systemic intraperitoneal administration in a SARS-CoV-2 delta K18 hACE2 Tg mouse model.

[0202] On day -1 of the study, animals (n=10 per group) were treated intraperitoneally with a range of doses of test CV3-25 (15-0.2 mg / kg) or PBS control as vehicle. On day 0 of the study, animals were administered 10 3.5 TCID 50 were loaded with the SARS-CoV-2 delta variant.

[0203] Materials and Methods The monoclonal antibody CV3-25 was dissolved and diluted in phosphate buffered saline (PBS) to prepare a CV3-25 dilution.

[0204] Dilutions of CV3-25 were made at various concentrations so that administration of 200 μL resulted in doses of CV3-25 ranging from 15 mg / kg, 5 mg / kg, 1.7 mg / kg, 0.5 mg / kg to 0.2 mg / kg relative to the mean body weight of the dosing group.

[0205] Antibodies were stored at -80°C ± 10°C, diluents were stored at 4°C until use, and the temperature of the storage units was monitored.

[0206] The virus strain tested was SARS-CoV-2 delta variant (Aus / VIC / 18440 / 2021 (B.1.617.2)), which had been passaged three times in Vero-hSLAM cells.

[0207] animal The animal species used was the SARS-CoV-2 delta K18 hACE2 Tg (B6.Cg-Tg(K18-ACE2)2Prlmn / JArc; Jax™ Stock Number: 034860) mouse model, weighing approximately 17-26 g on day -9. Sixty mice, 11-13 weeks old on the day of test material administration, were used, with 83.3% female. Ten animals were randomly assigned to six treatment groups based on their body weight on day -9 to create groups with similar average body weights. Males comprised 20% of the animals per group, except for the 15 mg / kg group, which was all female. All mice were housed in individually ventilated cages (IVCs) with corncob bedding, tissue or shredded paper as nesting material, wooden chew blocks, accessible food pellets and acidified water for animal nutrition, and red plastic tunnels. Each cage system housed 3-5 mice per cage. All mice were moved to new cages every 14 days. Food and water were checked daily and replenished as needed.

[0208] Study design The dose levels of CV3-25 administered in this example were based on the dose range proven to have prophylactic activity. A total of 60 mice, 11-13 weeks old, were transported to the animal facility and assigned to six experimental groups according to Table 1 (see below). Mice were allowed more than 3 days for acclimatization.

[0209] [Table 1]

[0210] SARS-CoV-2 Delta K18 hACE2 mice (20% males per group, except for the 15 mg / kg group, which were all female) were treated via the intraperitoneal route with antibody at doses ranging from 15 mg / kg to 0.2 mg / kg, based on the mean group weight on Day -9, adjusted for a mean weight gain of 4% (based on the subset of animals weighed on Day -1). On Day 0, all mice received a lethal dose (10 3.5 TCID 50 ) and monitored for weight loss and mortality until the study was terminated on day 11.

[0211] Antibody administration Upon arrival, the test antibodies were stored at -80°C ± 10°C. The appropriate dose according to the treatment schedule (Table 1) was prescribed according to the mean group weight on day -9, adjusted for a mean weight gain of 4% (based on the subset of animals weighed on day -1).

[0212] Prior to dosing, the material was drawn up into a 1 mL syringe with a 26 G needle, briefly warmed to room temperature, and then administered to each mouse. Mice received the indicated dose by intraperitoneal administration of 200 μL of antibody solution into the peritoneal cavity. For administration, mice were grasped by the neck and held upside down at a -45° angle (200 μL per mouse).

[0213] Virus administration The virus material was stored at -80°C ± 10°C and thawed before administration. 3.5 TCID 50 The material was diluted in sterile PBS equivalent to 50 μL / mL. Animals were anesthetized with isoflurane (4% v / v in 2 L / min O) as needed. Each animal received approximately 10 3.5 TCID 50 Approximately 50 μL (25 μL in each nostril) of virus equivalent to 100 μg of IgG was given by intranasal inoculation. Unused material was frozen at -20°C to -80°C for back titration.

[0214] Laboratory analysis The administered virus dose was verified by back titrating the inoculum and titrating replicate samples on Vero cells.

[0215] Clinical monitoring General health observations were performed on each animal at least once daily (during normal work procedures) from the day of arrival until the end of the study. Each animal was weighed daily, beginning on the day of infection (day 0).

[0216] Final Inspection Mice were euthanized by cervical dislocation at the end of the study on day 11. No complete necropsy was performed.

[0217] Data analysis and statistical methods Survival fraction, survival time, and body weight change (area under the curve) at day 11 were compared with the control group using Fisher's exact test, log-rank, and Welch's t-test, respectively. All groups were compared with the vehicle (PBS solution) control group. P values ​​were adjusted according to Bonferroni (for two comparisons with vehicle), following a stepwise approach (starting with the highest antibody dose and conditionally testing lower doses if the previous step was statistically significant).

[0218] Statistical analysis was performed using R, and statistical significance was set at α = 0.05.

[0219] survival rate Prophylactic treatment with test antibody at 0.5 mg / kg or greater provided a statistically significant increase in survival compared to the control group (see Figure 1). Animals treated with test antibody at 0.5 mg / kg or greater had an 80% survival rate, and animals treated with 1.7 mg / kg or greater had a 100% survival rate, while the control group had a 20% survival rate at day 11. The median survival time in the control group was 6.5 days.

[0220] Prophylactic treatment with 0.5 mg / kg or greater of the test antibody resulted in a significant improvement in survival compared to the control group.

[0221] body weight Body weight changes were analyzed using area under the curve (AUC) analysis. If a mouse died or was euthanized during the study, the last observed body weight was carried forward. The body weight per mouse on day 0 was used as the baseline, and body weight changes were determined relative to the baseline, with the net AUC defined as the sum of the areas above and below the baseline using the percentage change per day. Prophylactic treatment with 0.5 mg / kg or higher of the test antibody resulted in a significant reduction in body weight loss compared to the control group.

[0222] conclusion In this lethal SARS-CoV-2 DeltaK18 hACE2 Tg mouse model, prophylactic intraperitoneal administration of the test antibody at 0.5 mg / kg or higher provided a statistically significant improvement in survival and reduced weight loss compared to the control group, whereas 8 of 10 animals in the control group did not survive.

[0223] Example 2 CV3-25 The objective of this study was to evaluate the pre-exposure efficacy of monoclonal antibody CV3-25 after intranasal administration in the SARS-CoV-2 delta K18 hACE2 Tg mouse model.

[0224] On day -1 of the study, animals (n=10 per group) were treated with a range of intranasal doses of test CV3-25 (15-0.2 mg / kg) or PBS control as vehicle. On day 0 of the study, animals were treated with 10 3.5 TCID 50 were loaded with the SARS-CoV-2 delta variant.

[0225] Materials and Methods The monoclonal antibody CV3-25 was dissolved and diluted in phosphate buffered saline (PBS) to prepare a CV3-25 dilution.

[0226] Dilutions of CV3-25 were made at various concentrations so that administration of 50 μL resulted in doses of CV3-25 ranging from 15 mg / kg, 5 mg / kg, 1.7 mg / kg, 0.5 mg / kg to 0.2 mg / kg relative to the mean body weight of the dosing group.

[0227] The formulations were stored at -80°C ± 10°C and the temperature of the storage units was monitored.

[0228] The virus strain tested was SARS-CoV-2 delta variant (Aus / VIC / 18440 / 2021 (B.1.617.2)), which had been passaged three times in Vero-hSLAM cells.

[0229] animal The animal species used was the SARS-CoV-2 delta K18 hACE2 Tg (B6.Cg-Tg(K18-ACE2)2Prlmn / JArc; Jax™ Stock Number: 034860) mouse model, weighing approximately 17-26 g on day -9. Sixty mice, 11-13 weeks old on the day of test material administration, were used, with 83.3% female. Ten animals were randomly assigned to six treatment groups based on their body weight on day -9 to create groups with similar average body weights. Males comprised 20% of the animals per group, except for the 15 mg / kg group, which was all female. All mice were housed in individually ventilated cages (IVCs) with corncob bedding, tissue or shredded paper as nesting material, wooden chew blocks, accessible food pellets and acidified water for animal nutrition, and red plastic tunnels. Each cage system housed 3-5 mice per cage. All mice were moved to new cages every 14 days. Food and water were checked daily and replenished as needed.

[0230] Study design The dose levels of CV3-25 administered in this example were extrapolated from the dose range proven to have prophylactic activity intravenously. Upon arrival, a total of 60 mice, aged 11-13 weeks, were transported to the animal facility and assigned to six experimental groups according to Table 2 (see below). Mice were allowed more than 3 days for acclimatization.

[0231] [Table 2]

[0232] Female SARS-CoV-2 Delta K18 hACE2 mice (20% males per group, except for the 15 mg / kg group, which was all female) were treated via the intranasal route with antibody at doses ranging from 15 mg / kg to 0.2 mg / kg, based on the mean group weight on Day -9, adjusted for a mean weight gain of 4% (based on the subset of animals weighed on Day -1). On Day 0, all mice received a lethal dose (10 3.5 TCID 50 ) and monitored for weight loss and mortality until the study was terminated on day 11.

[0233] Antibody administration Upon arrival, the test antibodies were stored at -80°C ± 10°C. The appropriate dose according to the treatment schedule (Table 2) was prescribed according to the mean group weight on day -9, adjusted for a mean weight gain of 4% (based on the subset of animals weighed on day -1).

[0234] Immediately prior to dosing, the material was loaded into a syringe, briefly warmed to room temperature, and then administered to each mouse. Mice were anesthetized with isoflurane (4% v / v in 2 L / min O2) for 2 minutes. The mouse was then grasped by the neck and held at a +45° angle. 50 μL of compound or vehicle control was then slowly administered into each mouse's nostril (25 μL in each nostril). To ensure compound delivery to the lower respiratory tract, the mouse was held for an additional 5-10 seconds before being returned to the nest box.

[0235] Virus administration The virus material was stored at -80°C ± 10°C and thawed before administration. 3.5 TCID 50 The material was diluted in cold PBS equivalent to 50 μL / mL. Animals were anesthetized with isoflurane (4% v / v in 2 L / min O) as needed. Approximately 10 μL of the material was added to each animal using a pipette tip. 3.5 TCID 50 Approximately 50 μL (25 μL in each nostril) of virus equivalent to 100 μg of IgG was given by intranasal inoculation. Unused material was frozen at -20°C to -80°C for back titration.

[0236] Laboratory analysis The inoculum was returned to the laboratory and replicate samples were titrated on Vero cells to verify the administered virus dose.

[0237] Clinical monitoring General health observations were performed on each animal at least once daily (during normal work procedures) from the day of arrival until the end of the study. Each animal was weighed daily, beginning one day before infection (day -1).

[0238] Final Inspection Mice were euthanized by cervical dislocation at the end of the study on day 11. No complete necropsy was performed.

[0239] Data analysis and statistical methods Two animals in the CV3-25 15 mg / kg group failed to dosing (lost part of their antibody volume by sneezing), and these two animals were excluded from statistical analysis.

[0240] Survival fraction, survival time, and body weight change (area under the curve) at day 11 were compared with the control group using Fisher's exact test, log-rank, and Welch's t-test, respectively. All groups were compared with the vehicle (PBS solution) control group. P values ​​were adjusted according to Bonferroni (for two comparisons with vehicle), following a stepwise approach (starting with the highest antibody dose and conditionally testing lower doses if the previous step was statistically significant).

[0241] Statistical analysis was performed using R, and statistical significance was set at α = 0.05.

[0242] survival rate Prophylactic treatment with test antibody at 1.7 mg / kg or greater provided a statistically significant increase in survival compared to the control group (see Figure 2). The median survival time in the control group was 6.0 days.

[0243] Prophylactic treatment with test antibody at 1.7 mg / kg or higher resulted in a significant improvement in survival compared to the control group.

[0244] body weight Body weight changes were analyzed using area under the curve (AUC) analysis. If a mouse died or was euthanized during the study, the last observed body weight was carried forward. The weight per mouse on day 0 was used as the baseline, and body weight changes were determined relative to the baseline, with the net AUC defined as the sum of the areas above and below the baseline using the percentage change per day. Prophylactic treatment with 0.2 g / kg or higher of the test antibody resulted in a significant reduction in body weight loss compared to the control group.

[0245] conclusion In this lethal SARS-CoV-2 DeltaK18 hACE2 Tg mouse model, prophylactic intranasal administration of the test antibody at doses of 1.7 mg / kg or higher provided a statistically significant improvement in survival and reduced weight loss compared to controls, which exhibited a 10% survival rate and a median survival time of 6 days.

[0246] Example 3 Antibodies according to the present invention The aim of this study was to evaluate the pre-exposure efficacy of a monoclonal antibody according to the invention having heavy chain CDR1 as SEQ ID NO: 046, heavy chain CDR2 as SEQ ID NO: 114, heavy chain CDR3 as SEQ ID NO: 254, light chain CDR1 as SEQ ID NO: 367, light chain CDR2 as SEQ ID NO: 430, and light chain CDR3 as SEQ ID NO: 545 after systemic intraperitoneal administration in a SARS-CoV-2 Delta K18 hACE2 Tg mouse model.

[0247] On day -1 of the study, animals (n=10 per group) were treated intraperitoneally with a range of doses of the test antibodies listed above (10-0.2 mg / kg) or a PBS control as vehicle. On day 0 of the study, animals were treated with 10 3.5 TCID 50 were loaded with the SARS-CoV-2 delta variant.

[0248] Materials and Methods The monoclonal test antibodies described above were buffered with phosphate buffered saline (PBS) and diluted to a final concentration for administration (10-0.2 mg / kg in 200 μL), thereby preparing the test antibody dilutions.

[0249] Test antibody dilutions were made at various concentrations such that administration of 200 μL relative to the average body weight per cage for the dosing group resulted in test antibody doses ranging from 10 mg / kg, 5 mg / kg, 1.7 mg / kg, 0.5 mg / kg to 0.2 mg / kg.

[0250] Antibodies were stored at -80°C ± 10°C, diluents were stored at 4°C until use, and the temperature of the storage units was monitored.

[0251] The virus strain tested was SARS-CoV-2 delta variant (Aus / VIC / 18440 / 2021 (B.1.617.2)), which had been passaged three times in Vero-hSLAM cells.

[0252] animal The animal species used was the SARS-CoV-2 delta K18 hACE2 Tg (B6.Cg-Tg(K18-ACE2)2Prlmn / JArc; Jax™ Stock Number: 034860) mouse model. Sixty mice were used, with 10 animals assigned to six treatment groups. All mice were housed in individually ventilated cages (IVCs) with corncob bedding, tissue or shredded paper as nesting material, wooden chewing blocks, accessible food pellets, acidified water for animal nutrition, and a red plastic tunnel. Each cage system housed 3–5 mice per cage. All mice were transferred to new cages every 14 days. Food and water were inspected daily and replenished as needed. Each animal was weighed daily, starting on the day of infection (day 0).

[0253] Study design The dose levels of the test antibodies applied in this example were based on the maximum possible dosage for the stock concentration of the material (4 mg / mL), and on literature and previous experience with an anti-S2 antibody (CV3-25) that demonstrated protective IP at doses of 0.5 mg / kg or higher. A total of 60 mice were assigned to six experimental groups according to Table 3 (see below). Mice were allowed more than three days for acclimatization.

[0254] [Table 3]

[0255] Female SARS-CoV-2 Delta K18 hACE2 mice were treated with antibody at doses ranging from 10 mg / kg to 0.2 mg / kg, based on average group weight, via intraperitoneal administration. On day 0, all mice received a lethal dose (10 3.5 TCID 50 ) and monitored for weight loss and mortality until the study was terminated on day 14.

[0256] Antibody administration Upon arrival, the test antibodies were stored at −80° C.±10° C. The appropriate dose according to the treatment schedule (Table 3) was formulated using the average body weight per cage and group on day −5.

[0257] Prior to dosing, the material was loaded into a 1 mL syringe with a 26G needle and then administered to each mouse. Mice in the treatment group received the indicated dose via intraperitoneal administration of 200 μL of antibody solution into the peritoneal cavity. For administration (200 μL per mouse), mice were grasped by the neck and held upside down at a -45° angle. Mice in the vehicle group, which received PBS, were anesthetized with isoflurane (4% v / v in 2 L / min O2) for 2 minutes. Mice were then grasped by the neck and held at a +45° angle. 50 μL of compound or vehicle control was then slowly administered into each mouse's nostril (25 μL in each nostril). To ensure delivery of the compound to the lower respiratory tract, mice were held for an additional 5–10 seconds before being returned to the nest box.

[0258] Virus administration The virus material was stored at -80°C ± 10°C and kept at room temperature for 10 minutes before administration. 6.3 TCID 50 / mL. Upon thawing, the titer was approximately 10 3.5 TCID 50 The material was diluted in sterile PBS equivalent to 50 μL / mL. Animals were anesthetized with isoflurane (4% v / v in 2 L / min O) as needed. Each animal received approximately 10 3.5 TCID 50 Approximately 50 μL (25 μL in each nostril) of virus equivalent to 100 μg of IgG was given by intranasal inoculation. Unused material was frozen at -20°C to -80°C for back titration.

[0259] Laboratory analysis The actual dose of virus administered was verified by back titrating the inoculum and titrating replicate samples on Vero cells.

[0260] Final Inspection Mice were euthanized by cervical dislocation at the end of the study on day 14. No complete necropsy was performed.

[0261] Data analysis and statistical methods Survival fraction, survival time, and body weight change (area under the curve) at day 14 were compared with the control group using Fisher's exact test, log-rank, and Welch's t-test, respectively. All groups were compared with the vehicle (PBS solution) control group. P values ​​were adjusted according to Bonferroni (for two comparisons with vehicle), following a stepwise approach (starting with the highest antibody dose and conditionally testing lower doses if the previous step was statistically significant).

[0262] Statistical analysis was performed using R, and statistical significance was set at α = 0.05.

[0263] Viability - i.p. Prophylactic treatment with the test antibody provides a statistically significant increase in survival compared to the control group.

[0264] The prophylactic treatment test antibody resulted in a significant improvement in survival compared to the control group.

[0265] body weight Body weight changes were analyzed using area under the curve (AUC) analysis. If a mouse died or was euthanized during the study, the last observed weight was carried forward. The weight per mouse on day 0 was used as the baseline, and body weight changes were determined relative to the baseline, with the net AUC defined as the sum of the areas above and below the baseline using the percentage change per day. Prophylactic treatment with the test antibody resulted in a significant reduction in weight loss compared to the control group.

[0266] conclusion In this lethal SARS-CoV-2 DeltaK18 hACE2 Tg mouse model, prophylactic intraperitoneal administration of the test antibody significantly improved survival and reduced weight loss compared to vehicle, while control animals died.

[0267] Example 4 Antibody IP according to the present invention The aim of this study was to evaluate the pre-exposure efficacy of a monoclonal antibody according to the invention having heavy chain CDR1 as SEQ ID NO: 046, heavy chain CDR2 as SEQ ID NO: 114, heavy chain CDR3 as SEQ ID NO: 254, light chain CDR1 as SEQ ID NO: 367, light chain CDR2 as SEQ ID NO: 430, and light chain CDR3 as SEQ ID NO: 545 after systemic intraperitoneal administration in a SARS-CoV-2 Delta K18 hACE2 Tg mouse model.

[0268] On day -1 of the study, animals (n=10 per group) were treated intraperitoneally with a range of doses of the test antibodies listed above (10-0.2 mg / kg) or a PBS control as a vehicle control. On day 0 of the study, animals were treated with 10 3.5 TCID 50 were loaded with the SARS-CoV-2 delta variant.

[0269] Materials and Methods The monoclonal test antibody described above, buffered in phosphate buffered saline (PBS), was diluted to a final concentration for administration (10-0.2 mg / kg in 200 μL), thereby preparing the test antibody dilution.

[0270] Test antibody dilutions were made at various concentrations such that administration of 200 μL relative to the average body weight per cage resulted in test antibody doses ranging from 10 mg / kg, 5 mg / kg, 1.7 mg / kg, 0.5 mg / kg to 0.2 mg / kg.

[0271] Antibodies were stored at -80°C ± 10°C, diluents were stored at 4°C until use, and the temperature of the storage units was monitored.

[0272] The virus strain tested was SARS-CoV-2 delta variant (Aus / VIC / 18440 / 2021 (B.1.617.2)), which had been passaged three times in Vero-hSLAM cells.

[0273] animal The animal species used was the SARS-CoV-2 delta K18 hACE2 Tg (B6.Cg-Tg(K18-ACE2)2Prlmn / JArc; Jax™ stock number: 034860) mouse model, with body weights ranging from 17.1 to 23.4 g on the day of viral challenge. Sixty 12- to 13-week-old mice, all female, were used. Ten animals were assigned to six treatment groups. All mice were housed in individually ventilated cages (IVCs) with corncob bedding, tissue or shredded paper as nesting material, wooden chewing blocks, accessible food pellets, acidified water for animal nutrition, and a red plastic tunnel. Each cage system housed 3 to 5 mice per cage. All mice were transferred to new cages every 14 days. Food and water were inspected daily and replenished as needed. Each animal was weighed daily, starting on the day of infection (day 0).

[0274] Study design The dose levels of the test antibodies applied in this example were based on the maximum possible dosage for the stock concentration of the material (4 mg / mL), and on literature and previous experience with an anti-S2 antibody (CV3-25) that demonstrated protective IP at doses of 0.5 mg / kg or higher. A total of 60 mice were assigned to six experimental groups according to Table 4 (see below). Mice were allowed more than three days for acclimatization.

[0275] [Table 4]

[0276] Female SARS-CoV-2 Delta K18 hACE2 mice were treated with antibody at doses ranging from 10 mg / kg to 0.2 mg / kg, based on average group weight, via intraperitoneal administration. On day 0, all mice received a lethal dose (10 3.5 TCID 50 ) and monitored for weight loss and mortality until the study was terminated on day 14.

[0277] Antibody administration Upon arrival, the test antibodies were stored at -80° C.±10° C. Appropriate doses according to the treatment schedule (Table 4) were formulated using the average body weight per cage.

[0278] Prior to dosing, the material was loaded into a 1 mL syringe with a 26G needle and then administered to each mouse. Mice in the treatment group received the indicated dose via intraperitoneal administration of 200 μL of antibody solution into the peritoneal cavity. For administration (200 μL per mouse), mice were grasped by the neck and held upside down at a -45° angle. Mice in the vehicle group, which received PBS, were anesthetized with isoflurane (4% v / v in 2 L / min O2) for 2 minutes. Mice were then grasped by the neck and held at a +45° angle. 50 μL of compound or vehicle control was then slowly administered into each mouse's nostril (25 μL in each nostril). To ensure delivery of the compound to the lower respiratory tract, mice were held for an additional 5–10 seconds before being returned to the nest box.

[0279] Virus administration The virus material was stored at -80°C ± 10°C and kept at room temperature for 10 minutes before administration. 6.3 TCID 50 / mL. 3.5 TCID 50 The material was diluted in sterile PBS equivalent to 50 μL / mL. Animals were anesthetized with isoflurane (4% v / v in 2 L / min O) as needed. Each animal received approximately 10 3.5 TCID 50 Approximately 50 μL (25 μL in each nostril) of virus equivalent to 100 μg of IgG was given by intranasal inoculation. Unused material was frozen at -20°C to -80°C for back titration.

[0280] Laboratory analysis The actual dose of virus administered was verified by back titrating the inoculum and titrating replicate samples on Vero cells.

[0281] Final Inspection Mice were euthanized by cervical dislocation at the end of the study on day 14. No complete necropsy was performed.

[0282] Data analysis and statistical methods Survival fraction, survival time, and body weight change (area under the curve) at day 14 were compared with the control group using Fisher's exact test, log-rank, and Welch's t-test, respectively. All groups were compared with the vehicle (PBS solution) control group. P values ​​were adjusted according to Bonferroni (for two comparisons with vehicle control), following a stepwise approach (starting with the highest antibody dose and conditionally testing lower doses if the previous step was statistically significant).

[0283] Statistical analysis was performed using R, and statistical significance was set at α = 0.05.

[0284] Viability - i.p. Prophylactic treatment with 0.5 mg / kg or higher of the test antibody provided a statistically significant increase in survival compared to the vehicle control group (see Figure 5A). The survival rate in the control group at day 6 was 0%. The median survival time in the control group was 6 days.

[0285] Survival times in the test antibody-treated groups were compared with the control group using the log-rank test. Prophylactic treatment with 0.2 mg / kg or higher of the test antibody resulted in a significant improvement in survival times.

[0286] body weight Body weight changes were analyzed using area under the curve (AUC) analysis. If a mouse died or was euthanized during the study, the last observed body weight was carried forward. The body weight per mouse on day 0 was used as the baseline, and body weight change was determined relative to the baseline, with the net AUC defined as the sum of the areas above and below the baseline using the percentage change per day. Prophylactic treatment with 0.5 mg / kg or higher of the test antibody resulted in a significant reduction in body weight loss compared to the control group (Figure 5B).

[0287] conclusion In this lethal SARS-CoV-2 DeltaK18 hACE2 Tg mouse model, prophylactic intraperitoneal administration of test antibodies at 0.5 mg / kg or higher provided significantly improved survival and reduced weight loss compared to the vehicle control group, while none of the animals in the control group survived (Figure 5).

[0288] Example 5 Antibodies according to the present invention The aim of this study was to evaluate the pre-exposure efficacy of a monoclonal antibody according to the invention having heavy chain CDR1 as SEQ ID NO: 046, heavy chain CDR2 as SEQ ID NO: 114, heavy chain CDR3 as SEQ ID NO: 254, light chain CDR1 as SEQ ID NO: 367, light chain CDR2 as SEQ ID NO: 430, and light chain CDR3 as SEQ ID NO: 545 after intranasal administration in a SARS-CoV-2 Delta K18 hACE2 Tg mouse model.

[0289] On day -1 of the study, animals (n=10 per group) were treated with a range of intranasal doses of the test antibodies described above (10-0.2 mg / kg) or a PBS control. On day 0 of the study, animals were treated with 10 3.5 TCID 50 were loaded with the SARS-CoV-2 delta variant.

[0290] Materials and Methods The monoclonal antibodies according to the invention were buffered in phosphate buffered saline (PBS) and diluted to a final concentration for administration (10-0.2 mg / kg in 50 μL).

[0291] Monoclonal dilutions were made at various concentrations such that administration of 50 μL relative to the average body weight per cage for the dosing group on day -5 resulted in test antibody doses ranging from 10 mg / kg, 5 mg / kg, 1.7 mg / kg, 0.5 mg / kg to 0.2 mg / kg.

[0292] Antibodies were stored at -80°C ± 10°C, diluents were stored at 4°C until use, and the temperature of the storage units was monitored.

[0293] The virus strain tested was SARS-CoV-2 delta variant (Aus / VIC / 18440 / 2021 (B.1.617.2)), which had been passaged three times in Vero-hSLAM cells.

[0294] animal The animal species used was the SARS-CoV-2 Delta K18 hACE2 Tg mouse model. Sixty mice were used, with 10 animals assigned to each of six treatment groups. All mice were housed in individually ventilated cages (IVCs) with corncob bedding, tissue or shredded paper as nesting material, wooden chewing blocks, easily accessible food pellets, acidified water for animal nutrition, and a red plastic tunnel. Each cage system housed 3–5 mice per cage. All mice were transferred to new cages every 14 days. Food and water were inspected daily and replenished as needed. Each animal was weighed daily, beginning on the day of infection (day 0).

[0295] Study design The dose levels of the monoclonal antibodies applied in this example were based on the maximum possible dosage for the stock concentration of the material (4 mg / mL) and on literature and previous experience with an anti-S2 antibody (CV3-25) that showed protective IN at doses of 1.7 mg / kg or higher. A total of 60 mice were transported to the animal facility and assigned to six experimental groups according to Table 5 (see below). Mice were allowed more than 3 days for acclimatization.

[0296] [Table 5]

[0297] Female SARS-CoV-2 DeltaK18 hACE2 mice were treated with antibody at doses ranging from 10 mg / kg to 0.2 mg / kg, based on group mean mouse weight per cage, via the intranasal route. On day 0, all mice received a lethal dose (10 3.5 TCID 50) and monitored for mortality and weight loss until the study was terminated on day 14.

[0298] Antibody administration Upon arrival, the test antibodies were stored at −80° C.±10° C. The appropriate dose according to the treatment schedule (Table 5) was formulated using the average body weight per cage and group on day −5.

[0299] Immediately prior to dosing, the material was drawn into a 1 mL syringe equipped with a 26G needle and then administered to each mouse. Mice were anesthetized with isoflurane (4% v / v in 2 L / min O2) for 2 minutes. The mice were then grasped by the neck and held at a +45° angle. 50 μL of compound or vehicle control was then slowly administered into each mouse's nostril (25 μL in each nostril). To ensure compound delivery to the lower respiratory tract, the mice were held for an additional 5-10 seconds before being returned to the nest box.

[0300] Virus administration The virus material was stored at -80°C ± 10°C and thawed before administration. 3.5 TCID 50 The material was diluted in cold PBS equivalent to 50 μL / mL. Animals were anesthetized with isoflurane (4% v / v in 2 L / min O) as needed. Each animal received approximately 10 3.5 TCID 50 Approximately 50 μL (25 μL in each nostril) of virus equivalent to 100 μg of IgG was given by intranasal inoculation. Unused material was frozen at -20°C to -80°C for back titration.

[0301] Laboratory analysis The administered virus dose was verified by back titrating the inoculum and titrating replicate samples on Vero cells.

[0302] Final Inspection Mice were euthanized by cervical dislocation at the end of the study on day 14. No complete necropsy was performed.

[0303] Data analysis and statistical methods Survival fraction, survival time, and body weight change (area under the curve) at day 14 were compared with corresponding control groups using Fisher's exact test, log-rank, and Welch's t-test, respectively. All groups were compared to the vehicle (PBS solution) control group. P values ​​were adjusted according to Bonferroni (for two comparisons with vehicle), following a stepwise approach (starting with the highest antibody dose and conditionally testing lower doses if the previous step was statistically significant).

[0304] Statistical analysis was performed using R, and statistical significance was set at α = 0.05.

[0305] Survival rate – intranasal Prophylactic treatment with the test antibody provides statistically significant protection compared to vehicle.

[0306] Survival times in the test antibody-treated group were compared with the control group using the log-rank test. Prophylactic treatment with the test antibody resulted in a significant improvement in survival times.

[0307] body weight Body weight changes were analyzed using area under the curve (AUC) analysis. If a mouse died or was euthanized during the study, the last observed weight was carried forward. The weight per mouse on day 0 was used as the baseline, and body weight changes were determined relative to the baseline, with the net AUC defined as the sum of the areas above and below the baseline using the percentage change per day. Prophylactic treatment with the test antibody resulted in a significant reduction in weight loss compared to the control group.

[0308] conclusion In this lethal SARS-CoV-2 DeltaK18 hACE2 Tg mouse model, prophylactic intranasal administration of the test antibody provided significantly improved survival and reduced weight loss compared to vehicle, while control animals died by day 6.

[0309] Example 6 Antibodies according to the present invention The aim of this study was to evaluate the pre-exposure efficacy of a monoclonal antibody according to the invention having heavy chain CDR1 as SEQ ID NO: 046, heavy chain CDR2 as SEQ ID NO: 114, heavy chain CDR3 as SEQ ID NO: 254, light chain CDR1 as SEQ ID NO: 367, light chain CDR2 as SEQ ID NO: 430, and light chain CDR3 as SEQ ID NO: 545 after intranasal administration in a SARS-CoV-2 Delta K18 hACE2 Tg mouse model.

[0310] On day -1 of the study, animals (n=10 per group) were treated with a range of intranasal doses of the test antibodies described above (10-0.2 mg / kg) or a PBS control. On day 0 of the study, animals were treated with 10 3.5 TCID 50 were loaded with the SARS-CoV-2 delta variant.

[0311] Materials and Methods The monoclonal antibodies according to the invention were buffered in phosphate buffered saline (PBS) and diluted to a final concentration for administration (10-0.2 mg / kg in 50 μL).

[0312] Monoclonal dilutions were made at various concentrations such that administration of 50 μL per cage relative to average body weight resulted in test antibody doses ranging from 10 mg / kg, 5 mg / kg, 1.7 mg / kg, 0.5 mg / kg to 0.2 mg / kg.

[0313] Antibodies were stored at -80°C ± 10°C, diluents were stored at 4°C until use, and the temperature of the storage units was monitored.

[0314] The virus strain tested was SARS-CoV-2 delta variant (Aus / VIC / 18440 / 2021 (B.1.617.2)), which had been passaged three times in Vero-hSLAM cells.

[0315] animal The SARS-CoV-2 Delta K18 hACE2 Tg mouse model was used, with body weights ranging from 17.1 to 23.4 g on the day of test material administration. Sixty 12- to 13-week-old mice, all female, were used. Ten animals were assigned to six treatment groups based on the creation of groups with similar mean body weights and body weight variability. All mice were housed in individually ventilated cages (IVCs) with corncob bedding, tissue or shredded paper as nesting material, wooden chew blocks, accessible food pellets, acidified water for animal nutrition, and a red plastic tunnel. Each cage system housed three to five mice per cage. All mice were transferred to new cages every 14 days. Food and water were inspected daily and replenished as needed. Each animal was weighed daily, beginning on the day of infection (day 0).

[0316] Study design The dose levels of the monoclonal antibodies applied in this example were based on the maximum possible dosage for the stock concentration of the material (4 mg / mL) and on literature and previous experience with an anti-S2 antibody (CV3-25) that demonstrated protective IN at doses of 1.7 mg / kg or higher. A total of 60 12- to 13-week-old mice were transported to the animal facility and assigned to six experimental groups according to Table 6 (see below). Mice were allowed more than 3 days for acclimatization.

[0317] [Table 6]

[0318] Female SARS-CoV-2 DeltaK18 hACE2 mice were treated with antibody at doses ranging from 10 mg / kg to 0.2 mg / kg, based on group mean mouse weight per cage, via the intranasal route. On day 0, all mice received a lethal dose (10 3.5 TCID 50 ) and monitored for mortality and weight loss until the study was terminated on day 14.

[0319] Antibody administration Upon arrival, the test antibodies were stored at -80° C.±10° C. Appropriate doses according to the treatment schedule (Table 6) were formulated using the average body weight per cage.

[0320] Immediately prior to dosing, the material was drawn into a 1 mL syringe equipped with a 26G needle and then administered to each mouse. Mice were anesthetized with isoflurane (4% v / v in 2 L / min O2) for 2 minutes. The mice were then grasped by the neck and held at a +45° angle. 50 μL of compound or vehicle control was then slowly administered into each mouse's nostril (25 μL in each nostril). To ensure compound delivery to the lower respiratory tract, the mice were held for an additional 5-10 seconds before being returned to the nest box.

[0321] Virus administration The virus material was stored at -80°C ± 10°C and thawed before administration. 3.5 TCID 50 The material was diluted in cold PBS equivalent to 50 μL / mL. Animals were anesthetized with isoflurane (4% v / v in 2 L / min O) as needed. Each animal received approximately 10 3.5 TCID 50 Approximately 50 μL (25 μL in each nostril) of virus, equivalent to 100 μL of nasal inoculation, was given by intranasal inoculation.

[0322] Laboratory analysis The administered virus dose was verified by back titrating the inoculum and titrating replicate samples on Vero cells.

[0323] Final Inspection Mice were euthanized by cervical dislocation at the end of the study on day 14. No complete necropsy was performed.

[0324] Data analysis and statistical methods Survival fraction, survival time, and body weight change (area under the curve) at day 14 were compared with the control group using Fisher's exact test, log-rank, and Welch's t-test, respectively. All groups were compared with the vehicle (PBS solution) control group. P values ​​were adjusted according to Bonferroni (for two comparisons with vehicle), following a stepwise approach (starting with the highest antibody dose and conditionally testing lower doses if the previous step was statistically significant).

[0325] Statistical analysis was performed using R, and statistical significance was set at α = 0.05.

[0326] Survival rate – intranasal Prophylactic treatment with 0.5 mg / kg or higher of the test antibody provided a statistically significant increase in survival compared to vehicle (see Figure 6A). The survival rate at day 7 in the control group was 0%. The median survival time in the control group was 6 days.

[0327] Prophylactic treatment with 0.2 mg / kg or greater of the test antibody resulted in a significant improvement in survival.

[0328] body weight Body weight changes were analyzed using area under the curve (AUC) analysis. If a mouse died or was euthanized during the study, the last observed body weight was carried forward. The weight per mouse on day 0 was used as the baseline, and body weight change was determined relative to the baseline, with the net AUC defined as the sum of the areas above and below the baseline using the percentage change per day. Prophylactic treatment with test antibody at 0.2 mg / kg or higher resulted in a significant reduction in body weight loss compared to the control group (Figure 6B).

[0329] conclusion In this lethal SARS-CoV-2 DeltaK18 hACE2 Tg mouse model, prophylactic intranasal administration of test antibody at 0.5 mg / kg or higher provided a significant improvement in survival and reduced weight loss compared to vehicle, while control animals did not survive to day 6 (see Figure 6).

[0330] Example 7 Antibodies according to the present invention The objective of this study is to evaluate the pre- and post-exposure efficacy of a monoclonal antibody according to the invention having heavy chain CDR1 as SEQ ID NO: 046, heavy chain CDR2 as SEQ ID NO: 114, heavy chain CDR3 as SEQ ID NO: 254, light chain CDR1 as SEQ ID NO: 367, light chain CDR2 as SEQ ID NO: 430, and light chain CDR3 as SEQ ID NO: 545 after intranasal administration in an NHP model in which each subject has been pre-screened for the absence of serum antibodies against the SARS-CoV-2 virus.

[0331] NHP subjects are treated with an intranasal dose of a composition comprising the antibody described above. The fixed, or nominal, dose of antibody administered to each NHP subject is 0.01 mg to 10 mg (e.g., 0.01 to 2.0 mg ("low dose") or 2.0 to 10 mg ("high dose")) or a control (e.g., vehicle only) on day -1 of the study.

[0332] Materials and Methods Monoclonal antibodies according to the invention are provided in sodium acetate buffer (e.g., 20 mM sodium acetate, 75 mM sodium chloride, 5% sucrose, pH 5.5) and diluted to a final concentration for administration ranging from a nominal dose of 0.01 mg to 2.0 mg or 2.0 to 10 mg, administered in 50 μL to 500 μL per nostril.

[0333] NHP subjects assigned to the control group (sodium acetate buffer) will receive vehicle (e.g., 20 mM sodium acetate, 75 mM sodium chloride, 5% sucrose, pH 5.5), administered as 50 µL–500 µL per nostril.

[0334] All NHPs will receive an intranasal challenge with SARS-CoV-2 on day 0.

[0335] Samples will be collected during the study, and sample types will include nasal sampling (e.g., nasal swabs, washes, or scrapes), serum and / or bronchoalveolar lavage (BAL) samples. Sampling periods will encompass the duration of the study within the time frame of Day -1 to the end of the study (e.g., a 7- to 21-day follow-up period).

[0336] Samples may be analyzed for virus quantification (quantitative or RT-PCR and / or TCID 50 The antibody may be analyzed for specificity (e.g., by PCR) and / or the antibody titer may be analyzed by ELISA, HPLC, or similar methods.

[0337] animal NHP subjects are used at a weight of 3-12 kg (e.g., 5 kg per animal) at the start of the study. Five to twelve animals are assigned to each treatment group.

[0338] Study design The dose levels of monoclonal antibodies applied in this example are based on maximum doses and lower doses based on allometric calculations derived from mouse studies.

[0339] [Table 7]

[0340] NHP subjects will receive intranasal administration of a composition containing an antibody at a dose (nominal dose) of 0.01 mg to 2.0 mg or 2.0 mg to 10 mg. On day 0, all NHP subjects will be challenged with a dose of SARS-CoV-2 and will be observed for clinical symptoms (e.g., weight loss and / or temperature) and virological measures until the end of the study (e.g., 7 to 21 days).

[0341] Antibody administration Test antibody or vehicle alone is administered (eg, by pipette or spray) to each nostril according to the treatment schedule (Table 7), using a volume of 50 μL to 500 μL per nostril.

[0342] Virus administration The viral material is thawed prior to administration. Once thawed, the material is diluted and each animal receives 50 μL to 500 μL of viral solution per nostril. NHP subjects are administered 10 2 ~10 8 TCID 50 of SARS-CoV-2 (e.g., 10 5 TCID 50 They are infected with SARS-CoV-2.

[0343] Data analysis and statistical methods Clinical symptoms (eg, weight loss and temperature) and virological measures are used to compare treated and control groups.

[0344] Results - Clinical symptoms Prophylactic treatment with low and high dose antibody administration provides a reduction in clinical symptom measures compared to control groups.

[0345] Results – Virological Measurements Prophylactic treatment with low and high doses of antibody administration provides a reduction in virological measurements compared to the control group, with all control NHPs showing a viral load in samples following SARS-CoV-2 challenge.

[0346] conclusion In this SARS-CoV-2 NHP model, prophylactic intranasal administration of 2.0-10 mg (high dose) and 0.01-2.0 mg (low dose) of a test antibody according to the invention provides a reduction in virological measures and / or a reduction in clinical symptoms compared to control NHPs.

[0347] Example 8 Compositions comprising antibodies according to the present invention The objective of this study is to evaluate the pre- and post-exposure efficacy of a composition comprising a first monoclonal antibody according to the invention ("first antibody") having a heavy chain CDR1 as SEQ ID NO: 046, a heavy chain CDR2 as SEQ ID NO: 114, a heavy chain CDR3 as SEQ ID NO: 254, a light chain CDR1 as SEQ ID NO: 367, a light chain CDR2 as SEQ ID NO: 430, and a light chain CDR3 as SEQ ID NO: 545, and a second monoclonal antibody according to the invention ("second antibody") having a heavy chain CDR1 as SEQ ID NO: 639, a heavy chain CDR2 as SEQ ID NO: 640, a heavy chain CDR3 as SEQ ID NO: 641, a light chain CDR1 as SEQ ID NO: 642, a light chain CDR2 as SEQ ID NO: 643, and a light chain CDR3 as SEQ ID NO: 644, following intranasal administration in an NHP model in which each subject has been pre-screened for the absence of serum antibodies to the SARS-CoV-2 virus.

[0348] NHP subjects are treated with an intranasal dose of a composition containing the above-described antibodies (either a single antibody or a combination of both antibodies). The fixed, or nominal, dose of antibody administered to each NHP subject is 0.003 mg to 5 mg or a control (e.g., vehicle only) on day -1 of the study.

[0349] Materials and Methods Monoclonal antibodies according to the invention are provided in sodium acetate buffer (e.g., 20 mM sodium acetate, 75 mM sodium chloride, 5% sucrose, pH 5.5) and diluted to a final concentration for administration in the range of nominal doses of 0.003 mg to 5 mg, administered in 50 μL to 500 μL per nostril.

[0350] NHP subjects assigned to the control group (sodium acetate buffer) will receive vehicle (e.g., 20 mM sodium acetate, 75 mM sodium chloride, 5% sucrose, pH 5.5), administered as 50 µL–500 µL per nostril.

[0351] All NHPs will receive an intranasal challenge with SARS-CoV-2 on day 0.

[0352] Samples will be collected during the study, and sample types will include nasal sampling (e.g., nasal swabs, washes, or scrapes), serum and / or bronchoalveolar lavage (BAL) samples. Sampling periods will encompass the duration of the study within the time frame of Day -1 to the end of the study (e.g., a 7- to 21-day follow-up period).

[0353] Samples may be analyzed for virus quantification (quantitative or RT-PCR and / or TCID 50 The antibody is analyzed for antibody titer (by ELISA, HPLC, or similar methods).

[0354] animal NHP subjects are used at a weight of 3-12 kg (e.g., 5 kg per animal) at the start of the study. Four to 12 animals are assigned to each treatment group.

[0355] Study design The dose levels of the monoclonal antibodies and combinations applied in this example are based on maximum dosages and on allometric calculations derived from mouse studies.

[0356] [Table 8]

[0357] NHP subjects will receive intranasal administration of a composition containing either a single antibody at a nominal dose of 0.003-5 mg (treatment group 1 or 2). Another NHP subject will receive intranasal administration of a composition containing the first and second antibodies at a nominal dose of 0.003-5 mg (treatment group 3). On day 0, all NHP subjects will be challenged with a dose of SARS-CoV-2 and will be observed for clinical symptoms (e.g., weight loss and / or temperature) and virological measures until the end of the study (e.g., 7-21 days).

[0358] Antibody administration According to the treatment schedule (Table 8), antibody compositions (treatment groups 1-3) or vehicle alone are administered (e.g., by pipette or spray) to each nostril using volumes of 50 μL to 500 μL per nostril.

[0359] Virus administration The viral material is thawed prior to administration. Once thawed, the material is diluted and each animal receives 50 μL to 500 μL of viral solution per nostril. NHP subjects are administered 10 2 ~10 8 TCID 50 of SARS-CoV-2 (e.g., 10 5 TCID 50 They are infected with SARS-CoV-2.

[0360] Data analysis and statistical methods Clinical symptoms (e.g., weight loss and temperature) and virological measures are used to compare treatment groups (e.g., treatment groups 1-3) with the control group and / or between treatment groups (e.g., treatment group 1 versus treatment group 3).

[0361] Results - Clinical symptoms Prophylactic treatment with treatments (shown in Table 8) provides a reduction in clinical symptom measures compared to the control group or between treatment groups.

[0362] Results – Virological Measurements Prophylactic treatment with treatments (shown in Table 8) provides a reduction in virological measures compared to the control group or between treatment groups.

[0363] conclusion In this SARS-CoV-2 NHP model, prophylactic intranasal administration of a treatment according to the invention (shown in Table 8) provides a reduction in clinical symptoms and / or a reduction in virological measures compared to control NHPs and in comparisons between treatments.

[0364] Example 9 Bispecific antibodies according to the present invention The purpose of this intranasal mouse study was to evaluate the pre- and post-exposure efficacy of bispecific monoclonal antibodies according to the invention, which have a first Fab capable of binding to the stem helix of SARS-CoV-2 and a second Fab capable of binding to the fusion peptide of SARS-CoV-2, compared to parental antibodies. The first parent antibody ("Parent mAb 1") has the following CDRs represented by SEQ ID NOs: 046, 114, 254, 367, 430, and 545. The second parent antibody ("Parent mAb 2") has the following CDRs represented by SEQ ID NOs: 645, 646, 647, 648, 649, and 650. Bispecific antibodies according to the invention have a first Fab comprising a sequence comprising any one or more of SEQ ID NOs: 046, 114, 254, 367, 430, or 545, and a second Fab comprising a sequence comprising any one or more of SEQ ID NOs: 645, 646, 647, 648, 649-650.

[0365] Animals are treated on day -1 of the study with intranasal nominal doses (0.001 mg / kg to 15 mg / kg) containing either parental mAb 1 or parental mAb 2, or the bispecific antibodies described above, or vehicle control.

[0366] Materials and Methods The parent monoclonal antibodies and bispecific antibodies according to the invention are provided in sodium acetate buffer (e.g., 20 mM sodium acetate, 75 mM sodium chloride, 5% sucrose, pH 5.5) and diluted to a final concentration for administration ranging from 0.001 mg / kg to 15 mg / kg, administered in 25 μL to 50 μL per nostril.

[0367] Mice assigned to the control group (sodium acetate buffer) receive vehicle (e.g., 20 mM sodium acetate, 75 mM sodium chloride, 5% sucrose, pH 5.5), administered as 25 µL–50 µL per nostril.

[0368] All mice will receive an intranasal challenge with SARS-CoV-2 delta on day 0.

[0369] animal SARS-CoV-2 Delta K18 hACE2 Tg mice are used at the start of the study, weighing approximately 15 g to 22 g (e.g., 20 g per animal). Six to ten animals are assigned to each treatment group.

[0370] Study design The dose levels of the monoclonal and bispecific antibodies applied in this example are based on maximum dosages and on previous results of efficacy studies in mice using the parental antibody alone. Starting at a maximum dose of 15 mg / kg, a 3- or 4-fold dilution curve is applied for each parental or bispecific antibody.

[0371] [Table 9]

[0372] Mice are treated with compositions containing either the parental or bispecific antibodies at doses ranging from 0.001 to 15 mg / kg via the intranasal route. On day 0, all mice are challenged with a dose of SARS-CoV-2 and monitored for survival and body weight until the end of the study (e.g., 10-15 days).

[0373] Antibody administration According to the treatment schedule (Table 9), the parental monoclonal or bispecific antibody or vehicle alone will be administered (e.g., by pipette or spray) to each nostril using a volume of 25 μL to 50 μL per nostril.

[0374] Virus administration The viral material was thawed prior to administration. Once thawed, the material was diluted and each animal received 25 μL to 50 μL of virus per nostril. Mice were 10 2 ~10 8 TCID 50 of SARS-CoV-2 (e.g., 10 3.5 TCID 50 They are infected with SARS-CoV-2.

[0375] Laboratory analysis The inoculum is returned to the laboratory and replicate samples are titrated on Vero cells to verify the administered virus dose.

[0376] Clinical monitoring General health observations will be performed on each animal at least once daily (during normal work procedures) from the day of arrival until the end of the study. Each animal will be weighed daily beginning one day before infection (day -1).

[0377] Data analysis and statistical methods Survival fraction, survival time, and weight change (area under the curve) are compared with corresponding control groups using Fisher's exact test, log-rank, and Welch's t-test, respectively. All groups are compared to the vehicle control group. P values ​​are adjusted according to Bonferroni (for triplicate comparisons with vehicle), followed by a stepwise approach within the antibody (starting with the highest dose and conditionally testing lower doses if the previous step is statistically significant).

[0378] Combination Index A survival dose-response curve was fitted for each of the treatments (parental mAb 1, parental mAb 2, and bispecific) and the ED 50 The efficacy of the bispecific antibody is then compared to the parental antibodies by calculating a combination index.

[0379] survival rate Prophylactic treatment with parental mAb 1, parental mAb 2, or the bispecific antibody (shown in Table 9) provided statistically significant protection against mortality compared to the control group and a significant improvement in survival compared to the control group.

[0380] body weight Weight change is analyzed using area under the curve (AUC) analysis. If a mouse dies or is euthanized during the study, the last observed weight is carried forward. The weight per mouse on day 0 is used as the baseline, and weight change is determined relative to the baseline, with the net AUC defined as the sum of the areas above and below the baseline using the percentage change per day.

[0381] Prophylactic treatment with the parental or bispecific antibodies (shown in Table 9) provides a statistically significant reduction in weight loss compared to the control group.

[0382] conclusion In this SARS-CoV-2 delta mouse model, prophylactic intranasal administration of bispecific antibodies according to the invention (shown in Table 9) provides a significant improvement in survival and reduced weight loss compared to the control group. Combination index results suggest at least comparable, additive, or possibly synergistic effects of the bispecific antibodies compared to the parental antibodies.

[0383] Example 10 Compositions Comprising Antibody Combinations According to the Invention The purpose of this intranasal mouse study is to evaluate the pre- and post-exposure efficacy of a composition comprising a first antibody of the invention capable of binding to the stem helix of SARS-CoV-2 and a second antibody capable of binding to the fusion peptide of SARS-CoV-2. The first antibody ("first mAb") has the following CDRs set forth in SEQ ID NOs: 046, 114, 254, 367, 430, or 545. The second antibody ("second mAb") has the following CDRs set forth in SEQ ID NOs: 639, 640, 641, 642, 643, and 644.

[0384] Animals are treated on day -1 of the study with either a nominal intranasal dose (0.001 mg / kg to 15 mg / kg) of the first mAb or the second mAb, or a composition comprising the first and second antibodies described above, or a vehicle control.

[0385] Materials and Methods The first and second mAbs are provided in sodium acetate buffer (e.g., 20 mM sodium acetate, 75 mM sodium chloride, 5% sucrose, pH 5.5) and diluted to a final concentration for administration ranging from 0.001 mg / kg to 15 mg / kg, administered as 25 μL to 50 μL per nostril.

[0386] Mice assigned to the control group (sodium acetate buffer) receive vehicle (e.g., 20 mM sodium acetate, 75 mM sodium chloride, 5% sucrose, pH 5.5), administered as 25 µL–50 µL per nostril.

[0387] All mice will receive an intranasal challenge with SARS-CoV-2 delta on day 0.

[0388] animal SARS-CoV-2 Delta K18 hACE2 Tg mice are used at the start of the study, weighing approximately 15 g to 22 g (e.g., 20 g per animal). Six to ten animals are assigned to each treatment group.

[0389] Study design The dose levels for the first or second mAb, or compositions comprising the first and second antibodies described above, in this example are based on maximum dosages and on previous results of efficacy studies in mice using the parent antibody alone, with a 3- or 4-fold dilution curve applied for each parent antibody or bispecific antibody, starting at a maximum dose of 15 mg / kg or less.

[0390] [Table 10]

[0391] Mice are treated via intranasal administration with a composition containing either the first or second antibody or a composition containing the combination at a dose ranging from 0.001 to 15 mg / kg. On day 0, all mice are challenged with a dose of SARS-CoV-2 and monitored for survival and body weight until the end of the study (e.g., 10-15 days).

[0392] Antibody administration According to the treatment schedule (Table 10), a composition containing the first or second antibody or combination, or vehicle alone, is administered (e.g., by pipette or spray) to each nostril using a volume of 25 μL to 50 μL per nostril.

[0393] Virus administration The viral material was thawed prior to administration. Once thawed, the material was diluted and each animal received 25 μL to 50 μL of viral solution per nostril. Mice were 10 2 ~10 8 TCID 50 of SARS-CoV-2 (e.g., 10 3.5 TCID 50 They are infected with SARS-CoV-2.

[0394] Laboratory analysis The inoculum is returned to the laboratory and replicate samples are titrated on Vero cells to verify the administered virus dose.

[0395] Clinical monitoring General health observations will be performed on each animal at least once daily (during normal work procedures) from the day of arrival until the end of the study. Each animal will be weighed daily beginning one day before infection (day -1).

[0396] Data analysis and statistical methods Survival fraction, survival time, and weight change (area under the curve) are compared with corresponding control groups using Fisher's exact test, log-rank, and Welch's t-test, respectively. All groups are compared to the vehicle control group. P values ​​are adjusted according to Bonferroni (for triplicate comparisons with vehicle), followed by a stepwise approach within the antibody (starting with the highest dose and conditionally testing lower doses if the previous step is statistically significant).

[0397] Combination Index A survival dose-response curve was fitted for each of the treatments (compositions containing the first or second antibody or combination) and the ED 50 The effectiveness of the combination is then compared to the first and second antibodies by calculating a combination index.

[0398] survival rate Prophylactic treatment with a composition comprising the first or second antibody or combination (as shown in Table 10) compared to the control group provides statistically significant protection against mortality and significant improvement in survival compared to the control group.

[0399] body weight Weight change is analyzed using area under the curve (AUC) analysis. If a mouse dies or is euthanized during the study, the last observed weight is carried forward. The weight per mouse on day 0 is used as the baseline, and weight change is determined relative to the baseline, with the net AUC defined as the sum of the areas above and below the baseline using the percentage change per day.

[0400] Prophylactic treatment with a composition containing the combination (shown in Table 10) provides a statistically significant reduction in weight loss compared to the control group.

[0401] conclusion In this SARS-CoV-2 delta mouse model, prophylactic intranasal administration of a composition comprising a combination according to the invention (shown in Table 10) provides a significant improvement in survival and reduced weight loss compared to the control group. The combination index results suggest at least comparable, additive, or possibly synergistic effects for the combination compared to the first or second antibody administered alone.

[0402] Example 11 ELISA binding assessment of antibodies according to the invention The purpose of this study was to evaluate antibody binding to various coronaspike antigens of various alpha- and beta-coronaviruses.

[0403] Binding was tested by enzyme-linked immunosorbent assay (ELISA). Briefly, spike antigens were used to coat the surface of microwells of an ELISA plate. Decreasing concentrations of antibodies were then added in duplicate to the coated, pre-blocked microwell surface, allowing them to attach to the coated antigen. Detection antibodies were added, labeled with horseradish peroxidase (HRP), and a colorimetric signal was generated upon addition of substrate. The readout was optical density, reflecting binding. Antibodies were assayed in duplicate against several alpha- and beta-coronavirus spike antigens, including, but not limited to, SARS-CoV-2 delta, Omicron XBB.1.5, and human coronavirus NL63. Antibodies were tested in a range of 10.0–0.000002 nM relative to other anti-S1 antibodies. The half-maximal effective concentration (EC) was used for each antibody on each antigen as an estimate of binding affinity. 50 ) was calculated.

[0404] [Table 11]

[0405] ELISA binding assays (Table 12) show that the antibodies according to the invention can effectively bind to all spike antigens tested from SARS-CoV-2 delta and betacoronaviruses, including Omicron XBB.1.5. The alphacoronavirus NL63 did not bind to the antibodies. The anti-S1 antibody tested bound only to SARS-CoV-2 delta and showed no breadth among the alphacoronaviruses NL63, as expected from the literature. The isotype control antibody did not show any binding to coronavirus antigens.

[0406] Example 12 Affinity Binding by Mesoscale Discovery (MSD) The goal of this study was to evaluate the relative binding affinity for spike antigens in a multiplexed method with extremely high specificity and low sample input. Therefore, antibodies were tested in Mesoscale Discovery (MSD), a multiplexed assay using electrochemiluminescent labels conjugated to detection antibodies. In this assay, up to 10 trimeric viral antigens are individually printed onto spots in each well of a plate, allowing for multiplexed assay readout. Antibodies according to the present invention bind to the viral antigens through Fab-mediated recognition, followed by the addition of an anti-human IgG sulfotag detection antibody, which recognizes the human IgG Fab. Addition of a read buffer containing substrate and passage of current through the plate electrodes initiates an electrochemical (current), buffer-substrate, and luminescent (light) cascade, resulting in light emission. The intensity of the emitted light is measured for each spot, revealing the level of analyte binding. Antibodies were assayed in duplicate in separate assays against spike antigens from SARS-CoV-2, SARS-CoV-1, MERS, hCoV NL63, hCoV HKU, hCoV OC43, and hCoV 229E (V-Plex COVID-19 Coronavirus Panel 3 (IgG) Kit cat# K15399U-2) (Table 13), as well as a set of Omicron variants (V-Plex SARS-CoV-2 Panel 34 (IgG) Kit cat# K15690U-2) (Table 14). Antibodies were tested at a dilution range starting at 555 ng / ml for Panel 3 and 10 ng / ml for the Omicron Panel 24 Kit. Light emission from the MSD sulfo-tag antibodies was quantified on an MSD Discovery WorkBench. Using the calibration curve, antibody concentrations were calculated by fitting raw electrochemiluminescence unit (eCLU) data from the calibrators to a logistic regression for curve fitting (sigmoidal, 4PL, X = concentration). Model asymptote was constrained to the lower limit of detection (LLOQ) and upper limit of detection (ULOQ). The concentration (ng / ml) corresponding to the midpoint of the dynamic range (i.e., the range between the lower and upper limits of quantitation (LLOQ and ULOQ)) was reported.

[0407] [Table 12]

[0408] MSD binding arrays (see Table 13) showed that the antibodies could effectively bind to all spike antigens from betacoronaviruses, including SARS-CoV-2, MERS, HKU1, and OC43. The antibodies did not bind to alphacoronaviruses, including 229E and NL63, or the SARS-CoV-2 receptor-binding domain (RBD) of the S1 portion of the spike. The anti-S1 antibodies tested here bound to the RBD domain and wild-type SARS-CoV-2 at low concentrations, as expected from the literature, and did not show breadth among other coronaviruses. While the antibodies of the present invention bound to all betacoronavirus spike antigens tested, the anti-RBD antibodies bound only to the SARS-CoV-2 spike and RBD antigens. An isotype control antibody showed no binding to any coronavirus antigens.

[0409] [Table 13]

[0410] MSD binding arrays showed that antibodies according to the invention can effectively bind to all omicron variants tested in this assay (Table 14). The anti-S1 control antibody bound to the SARS-CoV-2 spike at lower concentrations than the antibodies according to the invention, but only bound early omicron variants at very high concentrations and did not bind later omicron variants such as BQ and XBB. This suggests that antibodies according to the invention exhibit greater breadth across the omicron variants tested than anti-RBD antibodies. The isotype control antibody showed no binding to coronavirus antigens at all.

[0411] Example 13 Live virus neutralization The purpose of this study was to evaluate the ability of antibodies to neutralize live coronaviruses. Antibodies were tested for functional activity in live virus microneutralization assays against MERS, SARS-CoV-1, and SARS-CoV-2 Wuhan. Briefly, a dilution series of antibodies was pre-incubated with the corresponding virus and then added in quadruplicate to the respective cell lines (Vero: MERS and SARS-CoV-1, SARS-CoV-2: Vero E6 cells). After incubation, cells were fixed and stained with anti-nucleocapsid antibodies, and an enzyme-tagged detection antibody was added. The colored precipitate, which signals the presence of nucleocapsid, was read on an Immunospot analyzer and the 50% inhibitory concentration (IC) was determined by the Zielinska method (REF: https: / / doi.org / 10.1186 / 1743-422X-2-84). 50 ) reported. Antibodies were tested for neutralizing activity in the range of 0.025 to 500 μg / ml.

[0412] [Table 14]

[0413] Overall, the antibodies were able to neutralize the tested viruses SARS-CoV-1, SARS-CoV-2, and MERS at various concentrations (see Table 5), with IC for MERS-CoV. 50 The concentrations were minimal, suggesting that the binding efficacy observed in the binding assays resulted in neutralizing activity for the antibodies according to the invention. 50 A titer is required.

[0414] Example 14 Pseudovirion Neutralization The purpose of this study was to evaluate the ability of antibodies to neutralize various pseudotyped virus particles. Advantages over live virus assays include the ability to use pseudotyped viruses in laboratories with lower biosafety levels, the ability to produce and test novel variants of concern more rapidly, and broader testing across variants of concern or interest. Briefly, pseudovirions were produced by cotransfecting HEK293T cells with a virus-expressing plasmid and the pHIV-1NL43 ΔEnv-NanoLuc reporter virus plasmid. A dilution series of antibodies was then preincubated with the corresponding pseudovirus and added to HEK293T cells expressing ACE2, the entry receptor for SARS-CoV-2, SARS-CoV-1, and NL63. For hCoV 229E, diluted antibodies mixed with the virus were added to Huh7 cells expressing the aminopeptidase N (APN) receptor, which facilitates hCoV 229E entry into host cells. After incubation, cells were washed and lysed, and luciferase activity in the cell lysates was measured by reading relative optical units (RLU) using the Nano-Glo Luciferase Assay System and the GloMax System. The 50% inhibitory concentration (IC) was determined as the antibody concentration at which infectivity was inhibited by 50% using a four-parameter logistic regression (4PL) curve fit. 50 The bispecific antibodies were tested for neutralizing activity in the range of 0.0042 to 250 μg / ml against SARS-CoV-1, SARS-CoV-2 Wuhan, Delta, Omicron BA4, Omicron BQ.1.1, Omicron XBB.1, hCoV NL63, and hCoV 229E.

[0415] [Table 15]

[0416] Overall, the antibodies were able to neutralize the tested betaviruses SARS-CoV-1 and SARS-CoV-2, including variants of concern, at various concentrations (see Table 16), with an overall trend of higher IC for omicron variants. 50 Concentration, lowest IC for SARS-CoV-2 Wuhan 50 As expected based on the lack of binding in the ELISA and MSD assays, the antibodies did not neutralize the alphacoronaviruses hCoV NL63 and 229E.

[0417] Example 15 Epitope mapping The goal of this study was to precisely determine antibody-bound amino acid sequences in spike proteins across various alpha- and beta-coronaviruses. The method involves mapping linear epitopes using a library of overlapping synthetic peptides derived from spike proteins. Briefly, linear epitopes of sequences of interest were synthesized directly on a solid-state chip, generating a library of linear mimetics to aid in the identification of the correct amino acid sequence for target antibodies. The peptide library consisted of overlapping 15-amino acid fragments, with a 14-amino acid overlap. Antibody binding to each of the synthesized peptide chips was tested by enzyme-linked immunosorbent assay (ELISA) assay via incubation of the peptide array with an antibody solution. After washing, the peptide array was incubated with an antibody-peroxidase conjugate, substrate was added, and the reaction was stopped with hydrogen peroxide. The color development was measured and the intensity was reported.

[0418] The antibodies bound to peptide arrays of alpha- and beta-coronaviruses in specific epitopes of 10-20 amino acids (see Figure 7). This method allowed for high confidence in the definition of the epitopes, although there was some variability in the outer regions of the epitopes for each strain, indicated by asterisks.

Claims

1. 1. A method for treating a coronavirus infection in an individual, comprising administering to an individual a heavy chain variable domain comprising: a heavy chain CDR1 region comprising any one of SEQ ID NOs: 006-011, 015-020, 026-031, 046-051, 054-059, 064-069; a heavy chain CDR2 region comprising any one of SEQ ID NOs: 082-087, 093-098, 114-119, 136-141, 158-163, 191-196; and a heavy chain CDR3 region comprising any one of SEQ ID NOs: 202-207, 218-223, 237-242, 254-259, 280-285, 291-296; 1. A method comprising the step of administering to an individual in need thereof an antibody comprising a light chain variable domain comprising a light chain CDR1 region comprising any one of SEQ ID NOs: 430-435, 441-446, 452-457, 463-468, 479-484, 502-507, and a light chain CDR3 region comprising any one of SEQ ID NOs: 513-518, 524-529, 545-550, 584-589, 595-600, 617-622, wherein the antibody is administered mucosally.

2. a heavy chain variable domain comprising a heavy chain CDR1 region comprising any one of SEQ ID NOs: 006-011, 015-020, 026-031, 046-051, 054-059, and 064-069, a heavy chain CDR2 region comprising any one of SEQ ID NOs: 082-087, 093-098, 114-119, 136-141, 158-163, and 191-196, and a heavy chain CDR3 region comprising any one of SEQ ID NOs: 202-207, 218-223, 237-242, 254-259, 280-285, and 291-296; and SEQ ID NO:

313. A composition for mucosal application comprising an antibody comprising a light chain variable domain comprising: a light chain CDR1 region comprising any one of SEQ ID NOs: 318, 324, 329, 345, 350, 367, 372, 378, 383, and 400, 405; a light chain CDR2 region comprising any one of SEQ ID NOs: 430, 435, 441, 446, 452, 457, 463, 468, 479, 484, and 502, 507; and a light chain CDR3 region comprising any one of SEQ ID NOs: 513, 518, 524, 529, 545, 550, 584, 589, 595, 600, and 617, 622.

3. 1. An antibody for use in a method for preventing or treating a coronavirus infection in an individual, the antibody comprising a heavy chain CDR1 region comprising any one of SEQ ID NOs: 006-011, 015-020, 026-031, 046-051, 054-059, 064-069, a heavy chain CDR2 region comprising any one of SEQ ID NOs: 082-087, 093-098, 114-119, 136-141, 158-163, 191-196, and a heavy chain CDR3 region comprising any one of SEQ ID NOs: 202-207, 218-223, 237-242, 254-259, 280-285, 291-296. a light chain CDR1 region comprising any one of SEQ ID NOs: 313-318, 324-329, 345-350, 367-372, 378-383, and 400-405; a light chain CDR2 region comprising any one of SEQ ID NOs: 430-435, 441-446, 452-457, 463-468, 479-484, and 502-507; and a light chain CDR3 region comprising any one of SEQ ID NOs: 513-518, 524-529, 545-550, 584-589, 595-600, and 617-622, wherein the antibody is administered to a mucosa.

4. 4. The method, composition, or antibody of any one of claims 1 to 3, wherein the antibody comprises a heavy chain variable domain comprising a heavy chain CDR1 region comprising SEQ ID NO: 046, a heavy chain CDR2 region comprising SEQ ID NO: 114, and a heavy chain CDR3 region comprising SEQ ID NO: 254, a light chain variable domain comprising a light chain CDR1 region comprising SEQ ID NO: 367, a light chain CDR2 region comprising SEQ ID NO: 430, and a light chain CDR3 region comprising SEQ ID NO:

545.

5. 5. The method, composition or antibody according to any one of claims 1 to 4, wherein said method for the treatment of coronavirus virus infection is a method for prophylactic and / or therapeutic treatment of coronavirus, preferably said method for treatment is for prophylactic treatment of SARS-COV-2 infection.

6. 6. The method, composition or antibody of any one of claims 1 to 5, wherein the antibody comprises a heavy chain variable domain having the sequence of SEQ ID NO: 629 with at most 15, preferably 0, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10, amino acid insertions, deletions or substitutions not present in the heavy chain CDRs.

7. 7. The method, composition or antibody of any one of claims 1 to 6, wherein the antibody comprises a light chain variable domain having the sequence of SEQ ID NO: 630 with at most 15, preferably 0, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10, amino acid insertions, deletions or substitutions not present in the light chain CDRs.

8. 8. The method, composition or antibody of claim 1, wherein the antibody is an IgG antibody, preferably an IgG1 antibody.

9. 9. The method, composition, or antibody of any one of claims 1 to 8, wherein the antibody is provided to the individual prophylactically.

10. 10. The method, composition, or antibody of any one of claims 1 to 9, wherein the route of administration comprises at least one of oral inhalation, nasal administration, intraocular administration, and oropharyngeal administration.

11. 11. The method, composition, or antibody of any one of claims 1 to 10, wherein the antibody is administered at least once or at least twice monthly.

12. 12. The method, composition or antibody of any one of claims 1 to 11, wherein the antibody is administered to the individual in a dosage of 0.01 mg to 20 mg.

13. 13. The composition of any one of claims 1 to 12, comprising a single dose unit of 0.01 mg to 20 mg, preferably 0.1 mg to 15 mg, or preferably 0.5 mg to 10 mg of antibody, wherein the antibody is defined in any one of claims 1 to 12.

14. 14. The composition of any one of claims 1 to 13, which is self-administered.

15. 15. A pharmaceutical delivery device comprising the composition of any one of claims 1 to 14.

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