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, six other coronaviruses are known to cause disease in humans: the 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 jumped 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 domains (RBDs) present on 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 and an S2 subunit that is involved in the fusion of viral and cellular 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 human 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 the 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 helix at the base of the viral spike protein is even less accessible than the element on the S2 subunit, but has the advantage of being better conserved in amino acid sequence so far.

[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 provides a heavy chain variable domain comprising a heavy chain CDR1 region comprising any one of SEQ ID NOs: 004 to 009, 015 to 020, 029 to 034, 039 to 044, 047 to 052, and 065 to 070; a heavy chain CDR2 region comprising any one of SEQ ID NOs: 082 to 087, 103 to 108, 125 to 130, 136 to 141, 147 to 152, and 191 to 196; and a heavy chain CDR3 region comprising any one of SEQ ID NOs: 210 to 215, 221 to 226, 245 to 250, 256 to 261, 286 to 291, and 297 to 302. The present invention provides antibodies comprising a light chain variable domain including a light chain CDR1 region comprising any one of SEQ ID NOs: 319 to 324, 330 to 335, 350 to 355, 372 to 377, 383 to 388, and 415 to 420; a light chain CDR2 region comprising any one of SEQ ID NOs: 432 to 437, 443 to 448, 454 to 459, 465 to 470, 476 to 481, and 508 to 513; and a light chain CDR3 region comprising any one of SEQ ID NOs: 529 to 534, 540 to 545, 573 to 578, 601 to 606, 612 to 617, and 623 to 628.

[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: 004 to 009, 015 to 020, 029 to 034, 039 to 044, 047 to 052, and 065 to 070; a heavy chain CDR2 region comprising any one of SEQ ID NOs: 082 to 087, 103 to 108, 125 to 130, 136 to 141, 147 to 152, and 191 to 196; and a heavy chain CDR3 region comprising any one of SEQ ID NOs: 210 to 215, 221 to 226, 245 to 250, 256 to 261, 286 to 291, and 297 to 302; Provided is an antibody comprising a light chain variable domain including a light chain CDR1 region comprising any one of SEQ ID NOs: 319 to 324, 330 to 335, 350 to 355, 372 to 377, 383 to 388, and 415 to 420; a light chain CDR2 region comprising any one of SEQ ID NOs: 432 to 437, 443 to 448, 454 to 459, 465 to 470, 476 to 481, and 508 to 513; and a light chain CDR3 region comprising any one of SEQ ID NOs: 529 to 534, 540 to 545, 573 to 578, 601 to 606, 612 to 617, and 623 to 628, 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 antibody comprising a heavy chain CDR1 region comprising any one of SEQ ID NOs: 004 to 009, 015 to 020, 029 to 034, 039 to 044, 047 to 052, or 065 to 070, a heavy chain CDR2 region comprising any one of SEQ ID NOs: 082 to 087, 082 to 087, 103 to 108, 125 to 130, 136 to 141, 147 to 152, or 191 to 196, and a heavy chain CDR3 region comprising any one of SEQ ID NOs: 210 to 215, 221 to 226, 245 to 250, 256 to 261, 286 to 291, or 297 to 302. the antibody comprising a light chain variable domain; a light chain CDR1 region comprising any one of SEQ ID NOs: 319 to 324, 330 to 335, 350 to 355, 372 to 377, 383 to 388, and 415 to 420; a light chain CDR2 region comprising any one of SEQ ID NOs: 432 to 437, 443 to 448, 454 to 459, 465 to 470, 476 to 481, and 508 to 513; and a light chain CDR3 region comprising any one of SEQ ID NOs: 529 to 534, 540 to 545, 573 to 578, 601 to 606, 612 to 617, and 623 to 628 to an individual in need thereof, 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: 004 to 009, 015 to 020, 029 to 034, 039 to 044, 047 to 052, and 065 to 070; a heavy chain CDR2 region comprising any one of SEQ ID NOs: 082 to 087, 103 to 108, 125 to 130, 136 to 141, 147 to 152, and 191 to 196; and a heavy chain CDR3 region comprising any one of SEQ ID NOs: 210 to 215, 221 to 226, 245 to 250, 256 to 261, 286 to 291, and 297 to 302; 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: 319 to 324, 330 to 335, 350 to 355, 372 to 377, 383 to 388, and 415 to 420; a light chain CDR2 region comprising any one of SEQ ID NOs: 432 to 437, 443 to 448, 454 to 459, 465 to 470, 476 to 481, and 508 to 513; and a light chain CDR3 region comprising any one of SEQ ID NOs: 529 to 534, 540 to 545, 573 to 578, 601 to 606, 612 to 617, and 623 to 628.

[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: 004 to 009, 015 to 020, 029 to 034, 039 to 044, 047 to 052, or 065 to 070; a heavy chain CDR2 region comprising any one of SEQ ID NOs: 082 to 087, 103 to 108, 125 to 130, 136 to 141, 147 to 152, or 191 to 196; and a heavy chain CDR2 region comprising any one of SEQ ID NOs: 210 to 215, 221 to 226, 245 to 250, 256 to 261, 286 to 291, or 297 to 302. a heavy chain variable domain comprising a heavy chain CDR3 region comprising any one of SEQ ID NOs: 319 to 324, 330 to 335, 350 to 355, 372 to 377, 383 to 388, and 415 to 420; a light chain CDR2 region comprising any one of SEQ ID NOs: 432 to 437, 443 to 448, 454 to 459, 465 to 470, 476 to 481, and 508 to 513; and a light chain variable domain comprising a light chain CDR3 region comprising any one of SEQ ID NOs: 529 to 534, 540 to 545, 573 to 578, 601 to 606, 612 to 617, and 623 to 628, 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: 029, a heavy chain CDR2 region comprising SEQ ID NO: 136, and a heavy chain CDR3 region comprising SEQ ID NO: 256, and a light chain variable domain comprising a light chain CDR1 region comprising SEQ ID NO: 372, a light chain CDR2 region comprising SEQ ID NO: 432, and a light chain CDR3 region comprising SEQ ID NO: 573.

[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: 029, a heavy chain CDR2 region consisting of SEQ ID NO: 136, and a heavy chain CDR3 region consisting of SEQ ID NO: 256, and a light chain variable domain comprising a light chain CDR1 region consisting of SEQ ID NO: 372, a light chain CDR2 region consisting of SEQ ID NO: 432, and a light chain CDR3 region consisting of SEQ ID NO: 573.

[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: 004-009, 015-020, 029-034, 039-044, 047-052, 065-070, a heavy chain CDR2 region comprising any one of SEQ ID NOs: 082-087, 103-108, 125-130, 136-141, 147-152, 191-196, and a heavy chain CDR2 region comprising any one of SEQ ID NOs: 210-215, 221-226, 245-250, 256-261, 286-291, 297-302. 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: 319 to 324, 330 to 335, 350 to 355, 372 to 377, 383 to 388, and 415 to 420; a light chain CDR2 region including any one of SEQ ID NOs: 432 to 437, 443 to 448, 454 to 459, 465 to 470, 476 to 481, and 508 to 513; and a light chain CDR3 region including any one of SEQ ID NOs: 529 to 534, 540 to 545, 573 to 578, 601 to 606, 612 to 617, and 623 to 628, 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: 004-009, 015-020, 029-034, 039-044, 047-052, 065-070, a heavy chain CDR2 region comprising any one of SEQ ID NOs: 082-087, 103-108, 125-130, 136-141, 147-152, 191-196, and a heavy chain CDR2 region comprising any one of SEQ ID NOs: 210-215, 221-226, 245-250, 256-261, 286-291, 297-302. 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: 319 to 324, 330 to 335, 350 to 355, 372 to 377, 383 to 388, and 415 to 420; a light chain CDR2 region including any one of SEQ ID NOs: 432 to 437, 443 to 448, 454 to 459, 465 to 470, 476 to 481, and 508 to 513; and a light chain CDR3 region including any one of SEQ ID NOs: 529 to 534, 540 to 545, 573 to 578, 601 to 606, 612 to 617, and 623 to 628, and is administered intranasally in an amount of 0.1 mg to 20 mg.

[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: 004-009, 015-020, 029-034, 039-044, 047-052, 065-070, a heavy chain CDR2 region comprising any one of SEQ ID NOs: 082-087, 103-108, 125-130, 136-141, 147-152, 191-196, and a heavy chain CDR3 region comprising any one of SEQ ID NOs: 210-215, 221-226, 245-250, 256-261, 286-291, 297-302. 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: 319 to 324, 330 to 335, 350 to 355, 372 to 377, 383 to 388, and 415 to 420; a light chain CDR2 region including any one of SEQ ID NOs: 432 to 437, 443 to 448, 454 to 459, 465 to 470, 476 to 481, and 508 to 513; and a light chain CDR3 region including any one of SEQ ID NOs: 529 to 534, 540 to 545, 573 to 578, 601 to 606, 612 to 617, and 623 to 628, 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: 004-009, 015-020, 029-034, 039-044, 047-052, and 065-070; a heavy chain CDR2 region comprising any one of SEQ ID NOs: 082-087, 103-108, 125-130, 136-141, 147-152, and 191-196; and a heavy chain CDR3 region comprising any one of SEQ ID NOs: 210-215, 221-226, 245-250, 256-261, 286-291, and 297-302. the antibody comprises a light chain variable domain comprising: a light chain CDR1 region comprising any one of SEQ ID NOs: 319 to 324, 330 to 335, 350 to 355, 372 to 377, 383 to 388, and 415 to 420; a light chain CDR2 region comprising any one of SEQ ID NOs: 432 to 437, 443 to 448, 454 to 459, 465 to 470, 476 to 481, and 508 to 513; and a light chain CDR3 region comprising any one of SEQ ID NOs: 529 to 534, 540 to 545, 573 to 578, 601 to 606, 612 to 617, and 623 to 628; the route of administration 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: 004 to 009, 015 to 020, 029 to 034, 039 to 044, 047 to 052, and 065 to 070; a heavy chain CDR2 region comprising any one of SEQ ID NOs: 082 to 087, 103 to 108, 125 to 130, 136 to 141, 147 to 152, and 191 to 196; and a heavy chain CDR3 region comprising any one of SEQ ID NOs: 210 to 215, 221 to 226, 245 to 250, 256 to 261, 286 to 291, and 297 to 302.

[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: 319 to 324, 330 to 335, 350 to 355, 372 to 377, 383 to 388, and 415 to 420; a light chain CDR2 region comprising any one of SEQ ID NOs: 432 to 437, 443 to 448, 454 to 459, 465 to 470, 476 to 481, and 508 to 513; and a light chain CDR3 region comprising any one of SEQ ID NOs: 529 to 534, 540 to 545, 573 to 578, 601 to 606, 612 to 617, and 623 to 628.

[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: 004 to 009, 015 to 020, 029 to 034, 039 to 044, 047 to 052, and 065 to 070; a heavy chain CDR2 region comprising any one of SEQ ID NOs: 082 to 087, 103 to 108, 125 to 130, 136 to 141, 147 to 152, and 191 to 196; and a heavy chain CDR3 region comprising any one of SEQ ID NOs: 210 to 215, 221 to 226, 245 to 250, 256 to 261, 286 to 291, and 297 to 302.

[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: 319 to 324, 330 to 335, 350 to 355, 372 to 377, 383 to 388, and 415 to 420; a light chain CDR2 region comprising any one of SEQ ID NOs: 432 to 437, 443 to 448, 454 to 459, 465 to 470, 476 to 481, and 508 to 513; and a light chain CDR3 region comprising any one of SEQ ID NOs: 529 to 534, 540 to 545, 573 to 578, 601 to 606, 612 to 617, and 623 to 628.

[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: 004 to 009, 015 to 020, 030 to 034, 039 to 044, 047 to 052, and 050 to 054, a heavy chain CDR2 region comprising any one of SEQ ID NOs: 082 to 087, 103 to 108, 125 to 130, 137 to 141, 147 to 152, and 132 to 136, and a heavy chain CDR3 region comprising any one of SEQ ID NOs: 210 to 215, 221 to 226, 245 to 250, 257 to 261, 286 to 291, and 252 to 256. and a light chain CDR3 region comprising any one of SEQ ID NOs: 529 to 534, 540 to 545, 574 to 578, 601 to 606, 612 to 617, and 569 to 573.

[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, 103-108, 125-130, 136-141, 147-152, 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: 004-009, 015-020, 029-034, 039-044, 047-052, or 065-070; a heavy chain CDR2 region comprising any one of SEQ ID NOs: 082-087, 103-108, 125-130, 136-141, 147-152, or 191-196; and a heavy chain CDR3 region comprising any one of SEQ ID NOs: 210-215, 221-226, 245-250, 256-261, 286-291, or 297-302. and a light chain CDR3 region comprising any one of SEQ ID NOs: 529 to 534, 540 to 545, 573 to 578, 601 to 606, 612 to 617, and 623 to 628.

[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: 004-009, 015-020, 029-034, 039-044, 047-052, or 065-070; a heavy chain CDR2 region comprising any one of SEQ ID NOs: 082-087, 103-108, 125-130, 136-141, 147-152, or 191-196; and a heavy chain CDR3 region comprising any one of SEQ ID NOs: 210-215, 221-226, 245-250, 256-261, 286-291, or 297-302; and a light chain CDR3 region comprising one of SEQ ID NOs: 529 to 534, 540 to 545, 573 to 578, 601 to 606, 612 to 617, or 623 to 628, wherein the antibody is administered mucosally. Verse 2: a heavy chain variable domain comprising a heavy chain CDR1 region comprising any one of SEQ ID NOs: 004 to 009, 015 to 020, 029 to 034, 039 to 044, 047 to 052, and 065 to 070, a heavy chain CDR2 region comprising any one of SEQ ID NOs: 082 to 087, 103 to 108, 125 to 130, 136 to 141, 147 to 152, and 191 to 196, and a heavy chain CDR3 region comprising any one of SEQ ID NOs: 210 to 215, 221 to 226, 245 to 250, 256 to 261, 286 to 291, and 297 to 302; 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: 324, 330 to 335, 350 to 355, 372 to 377, 383 to 388, and 415 to 420; a light chain CDR2 region comprising any one of SEQ ID NOs: 432 to 437, 443 to 448, 454 to 459, 465 to 470, 476 to 481, and 508 to 513; and a light chain CDR3 region comprising any one of SEQ ID NOs: 529 to 534, 540 to 545, 573 to 578, 601 to 606, 612 to 617, and 623 to 628. Verse 3: 1. An antibody for use in a method for preventing or treating a coronavirus infection in an individual, comprising: a heavy chain CDR1 region comprising any one of SEQ ID NOs: 004 to 009, 015 to 020, 029 to 034, 039 to 044, 047 to 052, or 065 to 070; a heavy chain CDR2 region comprising any one of SEQ ID NOs: 082 to 087, 103 to 108, 125 to 130, 136 to 141, 147 to 152, or 191 to 196; and a heavy chain CDR3 region comprising any one of SEQ ID NOs: 210 to 215, 221 to 226, 245 to 250, 256 to 261, 286 to 291, or 297 to 302. 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: 319 to 324, 330 to 335, 350 to 355, 372 to 377, 383 to 388, and 415 to 420; a light chain CDR2 region comprising any one of SEQ ID NOs: 432 to 437, 443 to 448, 454 to 459, 465 to 470, 476 to 481, and 508 to 513; and a light chain CDR3 region comprising any one of SEQ ID NOs: 529 to 534, 540 to 545, 573 to 578, 601 to 606, 612 to 617, and 623 to 628. 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: 029, a heavy chain CDR2 region comprising SEQ ID NO: 136, and a heavy chain CDR3 region comprising SEQ ID NO: 256; a light chain variable domain comprising a light chain CDR1 region comprising SEQ ID NO: 372, a light chain CDR2 region comprising SEQ ID NO: 432, and a light chain CDR3 region comprising SEQ ID NO: 573. 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 disclosed herein and a second binding fragment, wherein the first fragment is selected from the group consisting of SEQ ID NOs: 004-009, 015-020, 029-034, 039-044, 047-052, 065-070, 082-087, 103-108, 125-130, 136-141, 147-152, 191-196, 210-215, 221-226, 245-248, 250-252, 260-262, 270-274, 280-282, 290-292, 300-304, 310-312, 320-322, 330-332, 340-342, 350-352, 360-362, 370-372, 380-382, 390-404, 410-412, 420-422, 430-432, 440-442, 450-462, 460-472, 470-482, 480-492, 500-512, 510-512, 520-522, 530-542, 540-542, 550-552, 560-562, 570-572, 580-582, 590-604, 600-604, 610-612, 620-622, 630-632, 6 More preferably, the first fragment comprises a sequence comprising any one of SEQ ID NOs: 029, 136, 256, 372, 432, 573, 574, 575, 580, 581, 590, 601, 612, 617, 623, 628, 630, 635, 640, 645, 650, 651, 652, 653, 654, 655, 660, 661, 662, 663, 664, 665, 670, 671, 672, 673, 674, 675, 676, 677, 678, 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, 733, 734, 735, 736, 737, 738, 739, 740, 745, 746, 747, 748, 749, 7 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 disclosed herein and a second binding fragment, wherein the first fragment is selected from the group consisting of SEQ ID NOs: 004-009, 015-020, 029-034, 039-044, 047-052, 065-070, 082-087, 103-108, 125-130, 136-141, 147-152, 191-196, 210-215, 221-226, 245-248, 250-252, 260-262, 270-274, 280-282, 290-292, 300-304, 310-312, 320-322, 330-332, 340-342, 350-352, 360-362, 370-372, 380-382, 390-404, 410-412, 420-422, 430-432, 440-442, 450-462, 460-472, 470-482, 480-492, 500-512, 510-512, 520-522, 530-542, 540-542, 550-552, 560-562, 570-572, 580-582, 590-604, 600-604, 610-612, 620-622, 630-632, 6 More preferably, the first fragment comprises a sequence comprising any one of SEQ ID NOs: 029, 136, 256, 372, 432, 573, 574, 575, 580, 581, 590, 601, 612, 617, 623, 628, 630, 635, 640, 645, 650, 651, 652, 653, 654, 655, 660, 661, 662, 663, 664, 665, 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, 733, 734, 735, 736, 737, 738, 739, 740, 745, 746, 747, 748, 7 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 disclosed herein and a second binding fragment, wherein the first fragment is selected from the group consisting of SEQ ID NOs: 004-009, 015-020, 029-034, 039-044, 047-052, 065-070, 082-087, 103-108, 125-130, 136-141, 147-152, 191-196, 210-215, 221-226, 245-248, 250-252, 260-262, 270-274, 280-282, 290-292, 300-304, 310-312, 320-322, 330-332, 340-342, 350-352, 360-362, 370-372, 380-382, 390-404, 410-412, 420-422, 430-432, 440-442, 450-462, 460-472, 470-482, 480-492, 500-512, 510-512, 520-522, 530-542, 540-542, 550-552, 560-562, 570-572, 580-582, 590-604, 600-604, 610-612, 620-622, 630-632, 6 More preferably, the first fragment comprises a sequence comprising any one of SEQ ID NOs: 029, 136, 256, 372, 432, 573, 574, 575, 580, 581, 590, 601, 612, 617, 623, 628, 630, 635, 640, 645, 650, 651, 652, 653, 654, 655, 660, 661, 662, 663, 664, 665, 670, 671, 672, 673, 674, 675, 676, 677, 678, 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, 733, 734, 735, 736, 737, 738, 739, 740, 745, 746, 747, 748, 749, 7 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 disclosed herein and a second binding fragment, wherein the first fragment is selected from the group consisting of SEQ ID NOs: 004-009, 015-020, 029-034, 039-044, 047-052, 065-070, 082-087, 103-108, 125-130, 136-141, 147-152, 191-196, 210-215, 221-226, 245-248, 250-252, 260-262, 270-274, 280-282, 290-292, 300-304, 310-312, 320-322, 330-332, 340-342, 350-352, 360-362, 370-372, 380-382, 390-404, 410-412, 420-422, 430-432, 440-442, 450-462, 460-472, 470-482, 480-492, 500-512, 510-512, 520-522, 530-542, 540-542, 550-552, 560-562, 570-572, 580-582, 590-604, 600-604, 610-612, 620-622, 630-632, 6 More preferably, the first fragment comprises a sequence comprising any one of SEQ ID NOs: 029, 136, 256, 372, 432, and 573. 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: 029, 136, 256, 372, 432, or 573, 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, 691, 692, 693, 694, 695, 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 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: 029, 136, 256, 372, 432, or 573, 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: 029, 136, 256, 372, 432, or 573, 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: 029, 136, 256, 372, 432, or 573, 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: 029, 136, 256, 372, 432, or 573, 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: 029, 136 or 256, 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: 029, 136 or 256, 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: 029, 136 or 256, 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: 029, 136 or 256, 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: 372, 432 or 573.

[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: 372, 432 or 573.

[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: 372, 432 or 573.

[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: 372, 432 or 573.

[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, 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: 029, 136, 256, 372, 432, or 573, 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: 029, 136, 256, 372, 432, and / or 573, 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: 029, the heavy chain CDR2 region of SEQ ID NO: 136, and the heavy chain CDR3 region of SEQ ID NO: 256, and a light chain variable region comprising, as CDRs, the light chain CDR1 region of SEQ ID NO: 372, the light chain CDR2 region of SEQ ID NO: 432, and the light chain CDR3 region of SEQ ID NO: 573; 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: 029, a heavy chain CDR2 region of SEQ ID NO: 136, and a heavy chain CDR3 region of SEQ ID NO: 256; 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: 372, a light chain CDR2 region of SEQ ID NO: 432, and a light chain CDR3 region of SEQ ID NO: 573; 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: 029, a heavy chain CDR2 region of SEQ ID NO: 136, and a heavy chain CDR3 region of SEQ ID NO: 256; 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: 372, a light chain CDR2 region of SEQ ID NO: 432, and a light chain CDR3 region of SEQ ID NO: 573; 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: 029, or a first nucleic acid sequence encoding an amino acid sequence that differs from SEQ ID NO: 029 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: 136, or a second nucleic acid sequence encoding an amino acid sequence that differs from SEQ ID NO: 136 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: 256, or a third nucleic acid sequence encoding an amino acid sequence that differs from SEQ ID NO: 256 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: 372, or a fourth nucleic acid sequence encoding an amino acid sequence that differs from SEQ ID NO: 372 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: 432, or a fifth nucleic acid sequence encoding an amino acid sequence that differs from SEQ ID NO: 432 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: 573, or an amino acid sequence that differs from SEQ ID NO: 573 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 that differs 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: 029, 136, 256, 372, 432, or 573, 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: 029, 136, 256, 372, 432, and / or 573, 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: 029, the heavy chain CDR2 region of SEQ ID NO: 136, and the heavy chain CDR3 region of SEQ ID NO: 256, and a light chain variable region comprising, as CDRs, the light chain CDR1 region of SEQ ID NO: 372, the light chain CDR2 region of SEQ ID NO: 432, and the light chain CDR3 region of SEQ ID NO: 573; 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: 029, a heavy chain CDR2 region of SEQ ID NO: 136, and a heavy chain CDR3 region of SEQ ID NO: 256; 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: 372, a light chain CDR2 region of SEQ ID NO: 432, and a light chain CDR3 region of SEQ ID NO: 573; 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: 029, a heavy chain CDR2 region of SEQ ID NO: 136, and a heavy chain CDR3 region of SEQ ID NO: 256; 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: 372, a light chain CDR2 region of SEQ ID NO: 432, and a light chain CDR3 region of SEQ ID NO: 573; 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, wherein the nucleic acid molecule 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: 029, or a first nucleic acid sequence encoding an amino acid sequence that differs from SEQ ID NO: 029 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: 136, or a second nucleic acid sequence encoding an amino acid sequence that differs from SEQ ID NO: 136 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: 256, or a third nucleic acid sequence encoding an amino acid sequence that differs from SEQ ID NO: 256 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: 372, or a fourth nucleic acid sequence encoding an amino acid sequence that differs from SEQ ID NO: 372 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: 432, or a fifth nucleic acid sequence encoding an amino acid sequence that differs from SEQ ID NO: 432 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: 573, or an amino acid sequence that differs from SEQ ID NO: 573 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 inoculation - 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 line has been 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-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. Lines are slightly shifted on the y-axis to improve visual presentation. [Figure 3] Survival rate after lethal challenge - Kaplan-Meier survival curves for intraperitoneal prophylactic treatment groups with antibodies according to the invention. Animals (n=10 per group, except for n=9 at 0.5 mg / kg) were treated at dose-adjusted intervals with an antibody having heavy chain CDR1 as SEQ ID NO: 029, heavy chain CDR2 as SEQ ID NO: 136, heavy chain CDR3 as SEQ ID NO: 256, light chain CDR1 as SEQ ID NO: 372, light chain CDR2 as SEQ ID NO: 432, and light chain CDR3 as SEQ ID NO: 573 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 offset on the X- and Y-axes to improve visual presentation. [Figure 4] Survival rate after lethal challenge - Kaplan-Meier survival curves for the intranasal prophylactic treatment group with an antibody according to the invention. Animals (n=10 per group) were treated at dose titration with an antibody having heavy chain CDR1 as SEQ ID NO: 029, heavy chain CDR2 as SEQ ID NO: 136, heavy chain CDR3 as SEQ ID NO: 256, light chain CDR1 as SEQ ID NO: 372, light chain CDR2 as SEQ ID NO: 432, and light chain CDR3 as SEQ ID NO: 573 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 offset on the Y-axis to improve visual representation. [Figure 5]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 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 6] 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 7] Body Weight Change - Antibody According to the Invention Body weight change (%) relative to day 0 for the intraperitoneal prophylactic treatment group. On day -1, animals (n=10 per group, except for n=9 at 0.5 mg / kg) were treated intranasally with titrated antibody having heavy chain CDR1 as SEQ ID NO: 029, heavy chain CDR2 as SEQ ID NO: 136, heavy chain CDR3 as SEQ ID NO: 256, light chain CDR1 as SEQ ID NO: 372, light chain CDR2 as SEQ ID NO: 432, and light chain CDR3 as SEQ ID NO: 573. On day 0, animals were infected with 103.5 TCID50 of SARS-CoV-2 delta. A vehicle control group (PBS) was included. Bars represent 95% CI of the mean. If mice died or were euthanized during study follow-up, the last observed body weight was carried forward. [Figure 8]Body Weight Change - Antibody According to the Invention Body weight change (%) relative to day 0 for the intranasal prophylactic treatment group. On day -1, animals (n=10 per group) were treated intranasally with a titrated antibody having heavy chain CDR1 as SEQ ID NO: 029, heavy chain CDR2 as SEQ ID NO: 136, heavy chain CDR3 as SEQ ID NO: 256, light chain CDR1 as SEQ ID NO: 372, light chain CDR2 as SEQ ID NO: 432, and light chain CDR3 as SEQ ID NO: 573. On day 0, animals were infected with 103.5 TCID50 of SARS-CoV-2 delta. 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 9] Linear epitope mapping to the spike helix domains of alpha- and beta-coronaviruses. Amino acids highlighted in gray were identified as epitopes for antibody binding to the spike protein. Target sequences for the seven strains tested are aligned by Clustal. * may include multiple smaller portions of the epitope. Epitope footprint (B) and paratope hotspots (A) for antibodies against the SH domain of SARS-CoV-2. CDR regions for the heavy and light chains are shown. 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: 004-009, 015-020, 029-034, 039-044, 047-052, 065-070, 082-087, 103-108, 125-130, 136-141, 147-152, 191-196, 210-215, 221-226, 245-250, 251-252, 261-262, 262-263, 263-264, 264-265, 265-266, 266-267, 267-268, 270-272, 271-273, 272-274, 275-276, 277-278, 278-279, 280-282, 281-283, 282-284, 283-285, 284-286, 285-287, 286-288, 287-289, 291-296, 300-302, 304-306, 305-307, 306-308, 307-309, 310-311, 312-313, 314-315, 316-317, 318-319, 320-321, 322-323, 324-325, 326-327, 328 and a CDR sequence comprising any one of 6 to 261, 286 to 291, 297 to 302, 319 to 324, 330 to 335, 350 to 355, 372 to 377, 383 to 388, 415 to 420, 432 to 437, 443 to 448, 454 to 459, 465 to 470, 476 to 481, 508 to 513, 529 to 534, 540 to 545, 573 to 578, 601 to 606, 612 to 617, and 623 to 628. More preferably, the first antibody comprises CDR sequences comprising any one or more of SEQ ID NOs: 029, 136, 256, 372, 432, or 573, 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: 004-009, 015-020, 029-034, 039-044, 047-052, 065-070, 082-087, 103-108, 125-130, 136-141, 147-152, 191-196, 210-215, 221-226, 245-250, 251-252, 261-262, 262-264, 263-265, 264-266, 265-267, 266-268, 267-269, 270-271, 272-273, 274-275, 276-277, 278-279, 280-282, 284-285, 286-287, 288-289, 290-291, 300-302, 304-306, 308-309, 310-311, 312-313, 314-315, 316-317, 318-319, 320-321, 322-323, 324-325, 326-327, 328-329, 330-335, 332-336, 334-337, 336-338, 340-341, 342-343, 344 and 623 to 628. More preferably, the first antibody comprises a CDR sequence comprising one or more of SEQ ID NOs: 029, 136, 256, 372, 432, and 573. More preferably, the first antibody comprises CDR sequences comprising any one or more of SEQ ID NOs: 055, 137, 257, 363, 480, or 574, 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: 004-009, 015-020, 029-034, 039-044, 047-052, 065-070, 082-087, 103-108, 125-130, 136-141, 147-152, 191-196, 210-215, 221-226, 245-250, 251-252, 261-262, 262-263, 263-264, 264-265, 265-266, 266-267, 267-268, 270-272, 271-273, 272-274, 275-276, 277-278, 278-279, 280-282, 281-283, 282-284, 283-285, 284-286, 285-287, 286-288, 287-289, 291-296, 300-302, 304-306, 305-307, 306-308, 307-309, 310-311, 312-313, 314-315, 316-317, 318-319, 320-321, 322-323, 324-325, 326-327, 328 and a CDR sequence comprising any one of 6 to 261, 286 to 291, 297 to 302, 319 to 324, 330 to 335, 350 to 355, 372 to 377, 383 to 388, 415 to 420, 432 to 437, 443 to 448, 454 to 459, 465 to 470, 476 to 481, 508 to 513, 529 to 534, 540 to 545, 573 to 578, 601 to 606, 612 to 617, and 623 to 628. More preferably, the first antibody comprises CDR sequences comprising any one or more of SEQ ID NOs: 029, 136, 256, 372, 432, or 573, 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: 004-009, 015-020, 029-034, 039-044, 047-052, 065-070, 082-087, 103-108, 125-130, 136-141, 147-152, 191-196, 210-215, 221-226, 245-250, 251-252, 261-262, 262-263, 263-264, 264-265, 265-266, 266-267, 267-268, 270-272, 271-273, 272-274, 275-276, 277-278, 278-279, 280-282, 281-283, 282-284, 283-285, 284-286, 285-287, 286-288, 287-289, 291-296, 300-302, 304-306, 305-307, 306-308, 307-309, 310-311, 312-313, 314-315, 316-317, 318-319, 320-321, 322-323, 324-325, 326-327, 328 and a CDR sequence comprising any one of 6 to 261, 286 to 291, 297 to 302, 319 to 324, 330 to 335, 350 to 355, 372 to 377, 383 to 388, 415 to 420, 432 to 437, 443 to 448, 454 to 459, 465 to 470, 476 to 481, 508 to 513, 529 to 534, 540 to 545, 573 to 578, 601 to 606, 612 to 617, and 623 to 628. More preferably, the first antibody comprises CDR sequences comprising any one or more of SEQ ID NOs: 029, 136, 256, 372, 432, or 573, 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 heavy chain CDR1 sequences comprising any one of the following SEQ ID NOs: 004 to 009, 015 to 020, 029 to 034, 039 to 044, 047 to 052, and 065 to 070. A CDR comprising the amino acid sequence of SEQ ID NO: 029 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, 103 to 108, 125 to 130, 136 to 141, 147 to 152, and 191 to 196. A CDR comprising the amino acid sequence of SEQ ID NO: 136 is particularly preferred.

[0179] The antibodies disclosed herein preferably comprise heavy chain CDR3 sequences comprising any one of the following SEQ ID NOs: 210 to 215, 221 to 226, 245 to 250, 256 to 261, 286 to 291, and 297 to 302. A CDR comprising the amino acid sequence of SEQ ID NO: 256 is particularly preferred.

[0180] The antibodies disclosed herein preferably comprise a light chain CDR1 sequence comprising any one of the following SEQ ID NOs: 319 to 324, 330 to 335, 350 to 355, 372 to 377, 383 to 388, and 415 to 420. A CDR comprising the amino acid sequence of SEQ ID NO: 372 is particularly preferred.

[0181] The antibodies disclosed herein preferably comprise a light chain CDR2 sequence comprising any one of the following SEQ ID NOs: 432 to 437, 443 to 448, 454 to 459, 465 to 470, 476 to 481, and 508 to 513. A CDR comprising the amino acid sequence of SEQ ID NO: 432 is particularly preferred.

[0182] The antibodies disclosed herein preferably comprise a light chain CDR3 sequence comprising any one of the following SEQ ID NOs: 529 to 534, 540 to 545, 573 to 578, 601 to 606, 612 to 617, and 623 to 628. A CDR comprising the amino acid sequence of SEQ ID NO: 573 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 FTSY Sequence number 002 FTSYY Sequence number 003 FTSYYM Sequence number 004 FTSYYMH Sequence number 005 FTSYYMHW Sequence number 006 FTSYYMHWV Sequence number 007 FTSYYMHWVR Sequence number 008 FTSYYMHWVRQ Sequence number 009 FTSYYMHWVRQA Sequence number 010 GYTFT SEQ ID NO: 011 GYTFTS SEQ ID NO: 012 GYTFTSY SEQ ID NO: 013 GYTFTSYY SEQ ID NO: 014 GYTFTSYYM SEQ ID NO: 015 GYTFTSYYMH Sequence number 016 GYTFTSYYMHW SEQ ID NO: 017 GYTFTSYYMHWV SEQ ID NO: 018 GYTFTSYYMHWVR SEQ ID NO: 019 GYTFTSYYMHWVRQ Sequence number 020 GYTFTSYYMHWVRQA Sequence number 021 HWVR Sequence number 022 HWVRQ Sequence number 023 HWVRQA SEQ ID NO. 024 MHWV Sequence number 025 MHWVR SEQ ID NO. 026 MHWVRQ Sequence number 027 MHWVRQA SEQ ID NO: 028 SYYM SEQ ID NO. 029 SYYMH Sequence number 030 SYYMHW SEQ ID NO: 031 SYYMHWV Sequence number 032 SYYMHWVR SEQ ID NO: 033 SYYMHWVRQ Sequence number 034 SYYMHWVRQA SEQ ID NO: 035 TFTS Sequence number 036 TFTSY Sequence number 037 TFTSYY Sequence number 038 TFTSYYM Sequence number 039 TFTSYYMH Sequence number 040 TFTSYYMHW Sequence number 041 TFTSYYMHWV Sequence number 042 TFTSYYMHWVR Sequence number 043 TFTSYYMHWVRQ Sequence number 044 TFTSYYMHWVRQA SEQ ID NO: 045 TSYY SEQ ID NO: 046 TSYYM SEQ ID NO: 047 TSYYMH SEQ ID NO: 048 TSYYMHW SEQ ID NO: 049 TSYYMHWV Sequence number 050 TSYYMHWVR SEQ ID NO: 051 TSYYMHWVRQ Sequence number 052 TSYYMHWVRQA SEQ ID NO. 053 WVRQ Sequence number 054 WVRQA Sequence number 055 YMHW Sequence number 056 YMHWV Sequence number 057 YMHWVR Sequence number 058 YMHWVRQ Sequence number 059 YMHWVRQA Sequence number 060 YTFT SEQ ID NO: 061 YTFTS SEQ ID NO: 062 YTFTSY SEQ ID NO: 063 YTFTSYY SEQ ID NO: 064 YTFTSYYM Sequence number 065 YTFTSYYMH Sequence number 066 YTFTSYYMHW Sequence number 067 YTFTSYYMHWV Sequence number 068 YTFTSYYMHWVR Sequence number 069 YTFTSYYMHWVRQ Sequence number 070 YTFTSYYMHWVRQA SEQ ID NO: 071 YYMH Sequence number 072 YYMHW SEQ ID NO: 073 YYMHWV Sequence number 074 YYMHWVR SEQ ID NO: 075 YYMHWVRQ Sequence number 076 YYMHWVRQA Heavy chain CDR2 region (SEQ ID NO: 077 to SEQ ID NO: 196) SEQ ID NO: 077 EWMGLITPSGDDTYYA SEQ ID NO: 078 EWMGLITPSGDDTYYAQ Sequence number 079 EWMGLITPSGDDTYYAQR Sequence number 080 EWMGLITPSGDDTYYAQRF Sequence number 081 EWMGLITPSGDDTYYAQRFQ Sequence number 082 EWMGLITPSGDDTYYAQRFQG Sequence number 083 EWMGLITPSGDDTYYAQRFQGR Sequence number 084 EWMGLITPSGDDTYYAQRFQGRV Sequence number 085 EWMGLITPSGDDTYYAQRFQGRVT Sequence number 086 EWMGLITPSGDDTYYAQRFQGRVTM Sequence number 087 EWMGLITPSGDDTYYAQRFQGRVTMT Sequence number 088 GDDTYYAQ Sequence number 089 GDDTYYAQR Sequence number 090 GDDTYYAQRF Sequence number 091 GDDTYYAQRFQ Sequence number 092 GDDTYYAQRFQG Sequence number 093 GDDTYYAQRFQGR Sequence number 094 GDDTYYAQRFQGRV Sequence number 095 GDDTYYAQRFQGRVT Sequence number 096 GDDTYYAQRFQGRVTM Sequence number 097 GDDTYYAQRFQGRVTMT SEQ ID NO: 098 GLITPSGDDTYYA Sequence number 099 GLITPSGDDTYYAQ Sequence number 100 GLITPSGDDTYYAQR SEQ ID NO: 101 GLITPSGDDTYYAQRF SEQ ID NO: 102 GLITPSGDDTYYAQRFQ SEQ ID NO: 103 GLITPSGDDTYYAQRFQG SEQ ID NO: 104 GLITPSGDDTYYAQRFQGR SEQ ID NO: 105 GLITPSGDDTYYAQRFQGRV SEQ ID NO: 106 GLITPSGDDTYYAQRFQGRVT SEQ ID NO: 107 GLITPSGDDTYYAQRFQGRVTM SEQ ID NO: 108 GLITPSGDDTYYAQRFQGRVTMT Sequence number 109 ITPSGDDTYYA SEQ ID NO: 110 ITPSGDDTYYAQ Sequence number 111 ITPSGDDTYYAQR SEQ ID NO: 112 ITPSGDDTYYAQRF SEQ ID NO: 113 ITPSGDDTYYAQRFQ SEQ ID NO: 114 ITPSGDDTYYAQRFQG SEQ ID NO: 115 ITPSGDDTYYAQRFQGR SEQ ID NO: 116 ITPSGDDTYYAQRFQGRV SEQ ID NO: 117 ITPSGDDTYYAQRFQGRVT Sequence number 118 ITPSGDDTYYAQRFQGRVTM SEQ ID NO: 119 ITPSGDDTYYAQRFQGRVTMT SEQ ID NO: 120 LEWMGLITPSGDDTYYA SEQ ID NO: 121 LEWMGLITPSGDDTYYAQ SEQ ID NO: 122 LEWMGLITPSGDDTYYAQR SEQ ID NO: 123 LEWMGLITPSGDDTYYAQRF SEQ ID NO: 124 LEWMGLITPSGDDTYYAQRFQ SEQ ID NO: 125 LEWMGLITPSGDDTYYAQRFQG SEQ ID NO: 126 LEWMGLITPSGDDTYYAQRFQGR SEQ ID NO: 127 LEWMGLITPSGDDTYYAQRFQGRV SEQ ID NO: 128 LEWMGLITPSGDDTYYAQRFQGRVT SEQ ID NO: 129 LEWMGLITPSGDDTYYAQRFQGRVTM SEQ ID NO: 130 LEWMGLITPSGDDTYYAQRFQGRVTMT SEQ ID NO: 131 LITPSGDDTYYA SEQ ID NO: 132 LITPSGDDTYYAQ Sequence number 133 LITPSGDDTYYAQR SEQ ID NO: 134 LITPSGDDTYYAQRF Sequence number 135 LITPSGDDTYYAQRFQ SEQ ID NO: 136 LITPSGDDTYYAQRFQG SEQ ID NO: 137 LITPSGDDTYYAQRFQGR SEQ ID NO: 138 LITPSGDDTYYAQRFQGRV Sequence number 139 LITPSGDDTYYAQRFQGRVT SEQ ID NO: 140 LITPSGDDTYYAQRFQGRVTM SEQ ID NO: 141 LITPSGDDTYYAQRFQGRVTMT SEQ ID NO: 142 MGLITPSGDDTYYA SEQ ID NO: 143 MGLITPSGDDTYYAQ SEQ ID NO: 144 MGLITPSGDDTYYAQR SEQ ID NO: 145 MGLITPSGDDTYYAQRF SEQ ID NO: 146 MGLITPSGDDTYYAQRFQ SEQ ID NO: 147 MGLITPSGDDTYYAQRFQG SEQ ID NO: 148 MGLITPSGDDTYYAQRFQGR SEQ ID NO: 149 MGLITPSGDDTYYAQRFQGRV SEQ ID NO: 150 MGLITPSGDDTYYAQRFQGRVT SEQ ID NO: 151 MGLITPSGDDTYYAQRFQGRVTM SEQ ID NO: 152 MGLITPSGDDTYYAQRFQGRVTMT SEQ ID NO: 153 PSGDDTYYA SEQ ID NO: 154 PSGDDTYYAQ Sequence number 155 PSGDDTYYAQR SEQ ID NO: 156 PSGDDTYYAQRF SEQ ID NO: 157 PSGDDTYYAQRFQ SEQ ID NO: 158 PSGDDTYYAQRFQG SEQ ID NO: 159 PSGDDTYYAQRFQGR SEQ ID NO: 160 PSGDDTYYAQRFQGRV SEQ ID NO: 161 PSGDDTYYAQRFQGRVT SEQ ID NO: 162 PSGDDTYYAQRFQGRVTM SEQ ID NO: 163 PSGDDTYYAQRFQGRVTMT Sequence number 164 SGDDTYYA Sequence number 165 SGDDTYYAQ Sequence number 166 SGDDTYYAQR Sequence number 167 SGDDTYYAQRF Sequence number 168 SGDDTYYAQRFQ Sequence number 169 SGDDTYYAQRFQG Sequence number 170 SGDDTYYAQRFQGR SEQ ID NO: 171 SGDDTYYAQRFQGRV Sequence number 172 SGDDTYYAQRFQGRVT Sequence number 173 SGDDTYYAQRFQGRVTM SEQ ID NO: 174 SGDDTYYAQRFQGRVTMT Sequence number 175 TPSGDDTYYA SEQ ID NO: 176 TPSGDDTYYAQ Sequence number 177 TPSGDDTYYAQR SEQ ID NO: 178 TPSGDDTYYAQRF Sequence number 179 TPSGDDTYYAQRFQ Sequence number 180 TPSGDDTYYAQRFQG Sequence number 181 TPSGDDTYYAQRFQGR SEQ ID NO: 182 TPSGDDTYYAQRFQGRV Sequence number 183 TPSGDDTYYAQRFQGRVT Sequence number 184 TPSGDDTYYAQRFQGRVTM Sequence number 185 TPSGDDTYYAQRFQGRVTMT SEQ ID NO: 186 WMGLITPSGDDTYYA Sequence number 187 WMGLITPSGDDTYYAQ Sequence number 188 WMGLITPSGDDTYYAQR Sequence number 189 WMGLITPSGDDTYYAQRF Sequence number 190 WMGLITPSGDDTYYAQRFQ Sequence number 191 WMGLITPSGDDTYYAQRFQG Sequence number 192 WMGLITPSGDDTYYAQRFQGR SEQ ID NO: 193 WMGLITPSGDDTYYAQRFQGRV Sequence number 194 WMGLITPSGDDTYYAQRFQGRVT Sequence number 195 WMGLITPSGDDTYYAQRFQGRVTM SEQ ID NO: 196 WMGLITPSGDDTYYAQRFQGRVTMT Heavy chain CDR3 region (SEQ ID NO: 197 to SEQ ID NO: 302) SEQ ID NO: 197 AGGF SEQ ID NO: 198 AGGFD SEQ ID NO: 199 AGGFDV Sequence number 200 AGGFDVW Sequence number 201 AGGFDVWG SEQ ID NO: 202 AGGFDVWGQ SEQ ID NO: 203 AGGFDVWGQG SEQ ID NO: 204 AGGFDVWGQGT Sequence number 205 AKMSRA Sequence number 206 AKMSRAG Sequence number 207 AKMSRAGG SEQ ID NO: 208 AKMSRAGGF Sequence number 209 AKMSRAGGFD SEQ ID NO: 210 AKMSRAGGFDV Sequence number 211 AKMSRAGGFDVW Sequence number 212 AKMSRAGGFDVWG SEQ ID NO: 213 AKMSRAGGFDVWGQ Sequence number 214 AKMSRAGGFDVWGQG Sequence number 215 AKMSRAGGFDVWGQGT SEQ ID NO. 216 CAKMSRA SEQ ID NO: 217 CAKMSRAG SEQ ID NO: 218 CAKMSRAGG SEQ ID NO: 219 CAKMSRAGGF Sequence number 220 CAKMSRAGGFD SEQ ID NO: 221 CAKMSRAGGFDV Sequence number 222 CAKMSRAGGFDVW Sequence number 223 CAKMSRAGGFDVWG SEQ ID NO: 224 CAKMSRAGGFDVWGQ Sequence number 225 CAKMSRAGGFDVWGQG Sequence number 226 CAKMSRAGGFDVWGQGT SEQ ID NO: 227 GFDV SEQ ID NO: 228 GFDVW Sequence number 229 GFDVWG SEQ ID NO: 230 GFDVWGQ SEQ ID NO: 231 GFDVWGQG SEQ ID NO: 232 GFDVWGQGT SEQ ID NO: 233 GGFD SEQ ID NO: 234 GGFDV SEQ ID NO: 235 GGFDVW SEQ ID NO: 236 GGFDVWG SEQ ID NO: 237 GGFDVWGQ SEQ ID NO: 238 GGFDVWGQG SEQ ID NO: 239 GGFDVWGQGT Sequence number 240 KMSRA Sequence number 241 KMSRAG SEQ ID NO: 242 KMSRAGG SEQ ID NO: 243 KMSRAGGF SEQ ID NO: 244 KMSRAGGFD SEQ ID NO: 245 KMSRAGGFDV Sequence number 246 KMSRAGGFDVW Sequence number 247 KMSRAGGFDVWG SEQ ID NO: 248 KMSRAGGFDVWGQ Sequence number 249 KMSRAGGFDVWGQG Sequence number 250 KMSRAGGFDVWGQGT Sequence number 251 MSRA SEQ ID NO: 252 MSRAG SEQ ID NO: 253 MSRAGG SEQ ID NO: 254 MSRAGGF SEQ ID NO: 255 MSRAGGFD SEQ ID NO: 256 MSRAGGFDV Sequence number 257 MSRAGGFDVW Sequence number 258 MSRAGGFDVWG SEQ ID NO: 259 MSRAGGFDVWGQ Sequence number 260 MSRAGGFDVWGQG SEQ ID NO: 261 MSRAGGFDVWGQGT SEQ ID NO: 262 RAGG SEQ ID NO: 263 RAGGF SEQ ID NO: 264 RAGGFD SEQ ID NO: 265 RAGGFDV Sequence number 266 RAGGFDVW Sequence number 267 RAGGFDVWG SEQ ID NO: 268 RAGGFDVWGQ SEQ ID NO: 269 RAGGFDVWGQG SEQ ID NO: 270 RAGGFDVWGQGT SEQ ID NO: 271 SRAG SEQ ID NO: 272 SRAGG SEQ ID NO: 273 SRAGGF SEQ ID NO: 274 SRAGGFD SEQ ID NO: 275 SRAGGFDV SEQ ID NO: 276 SRAGGFDVW Sequence number 277 SRAGGFDVWG SEQ ID NO: 278 SRAGGFDVWGQ SEQ ID NO: 279 SRAGGFDVWGQG SEQ ID NO: 280 SRAGGFDVWGQGT Sequence number 281 YCAKMSRA Sequence number 282 YCAKMSRAG SEQ ID NO: 283 YCAKMSRAGG SEQ ID NO: 284 YCAKMSRAGGF Sequence number 285 YCAKMSRAGGFD Sequence number 286 YCAKMSRAGGFDV Sequence number 287 YCAKMSRAGGFDVW Sequence number 288 YCAKMSRAGGFDVWG Sequence number 289 YCAKMSRAGGFDVWGQ Sequence number 290 YCAKMSRAGGFDVWGQG Sequence number 291 YCAKMSRAGGFDVWGQGT Sequence number 292 YYCAKMSRA Sequence number 293 YYCAKMSRAG SEQ ID NO: 294 YYCAKMSRAGG SEQ ID NO: 295 YYCAKMSRAGGF Sequence number 296 YYCAKMSRAGGFD Sequence number 297 YYCAKMSRAGGFDV Sequence number 298 YYCAKMSRAGGFDVW Sequence number 299 YYCAKMSRAGGFDVWG Sequence number 300 YYCAKMSRAGGFDVWGQ Sequence number 301 YYCAKMSRAGGFDVWGQG Sequence number 302 YYCAKMSRAGGFDVWGQGT Light chain CDR1 region (SEQ ID NO: 303 to SEQ ID NO: 420) Sequence number 303 ASQSIT Sequence number 304 ASQSITG Sequence number 305 ASQSITGR SEQ ID NO: 306 ASQSITGRY SEQ ID NO: 307 ASQSITGRYL SEQ ID NO: 308 ASQSITGRYLA Sequence number 309 ASQSITGRYLAW SEQ ID NO: 310 ASQSITGRYLAWY SEQ ID NO: 311 ASQSITGRYLAWYQ SEQ ID NO: 312 ASQSITGRYLAWYQQ SEQ ID NO: 313 ASQSITGRYLAWYQQK SEQ ID NO: 314 ATLSCRASQSIT SEQ ID NO: 315 ATLSCRASQSITG SEQ ID NO: 316 ATLSCRASQSITGR SEQ ID NO: 317 ATLSCRASQSITGRY SEQ ID NO: 318 ATLSCRASQSITGRYL SEQ ID NO: 319 ATLSCRASQSITGRYLA SEQ ID NO: 320 ATLSCRASQSITGRYLAW SEQ ID NO: 321 ATLSCRASQSITGRYLAWY SEQ ID NO: 322 ATLSCRASQSITGRYLAWYQ SEQ ID NO: 323 ATLSCRASQSITGRYLAWYQQ SEQ ID NO: 324 ATLSCRASQSITGRYLAWYQQK SEQ ID NO: 325 CRASQSIT SEQ ID NO: 326 CRASQSITG SEQ ID NO: 327 CRASQSITGR SEQ ID NO: 328 CRASQSITGRY SEQ ID NO: 329 CRASQSITGRYL SEQ ID NO: 330 CRASQSITGRYLA SEQ ID NO: 331 CRASQSITGRYLAW SEQ ID NO: 332 CRASQSITGRYLAWY SEQ ID NO: 333 CRASQSITGRYLAWYQ SEQ ID NO: 334 CRASQSITGRYLAWYQQ SEQ ID NO: 335 CRASQSITGRYLAWYQQK SEQ ID NO: 336 ITGR SEQ ID NO: 337 ITGRY SEQ ID NO: 338 ITGRYL SEQ ID NO: 339 ITGRYLA SEQ ID NO: 340 ITGRYLAW SEQ ID NO: 341 ITGRYLAWY SEQ ID NO: 342 ITGRYLAWYQ SEQ ID NO: 343 ITGRYLAWYQQ SEQ ID NO: 344 ITGRYLAWYQQK SEQ ID NO: 345 LSCRASQSIT SEQ ID NO: 346 LSCRASQSITG SEQ ID NO: 347 LSCRASQSITGR SEQ ID NO: 348 LSCRASQSITGRY SEQ ID NO: 349 LSCRASQSITGRYL SEQ ID NO: 350 LSCRASQSITGRYLA SEQ ID NO: 351 LSCRASQSITGRYLAW SEQ ID NO: 352 LSCRASQSITGRYLAWY SEQ ID NO: 353 LSCRASQSITGRYLAWYQ SEQ ID NO: 354 LSCRASQSITGRYLAWYQQ SEQ ID NO: 355 LSCRASQSITGRYLAWYQQK Sequence number 356 QSIT SEQ ID NO:357 QSITG SEQ ID NO: 358 QSITGR SEQ ID NO:359 QSITGRY Sequence number 360 QSITGRYL SEQ ID NO: 361 QSITGRYLA SEQ ID NO: 362 QSITGRYLAW SEQ ID NO: 363 QSITGRYLAWY SEQ ID NO: 364 QSITGRYLAWYQ Sequence number 365 QSITGRYLAWYQQ Sequence number 366 QSITGRYLAWYQQK SEQ ID NO: 367 RASQSIT SEQ ID NO: 368 RASQSITG SEQ ID NO: 369 RASQSITGR SEQ ID NO: 370 RASQSITGRY SEQ ID NO: 371 RASQSITGRYL SEQ ID NO: 372 RASQSITGRYLA SEQ ID NO: 373 RASQSITGRYLAW SEQ ID NO: 374 RASQSITGRYLAWY SEQ ID NO: 375 RASQSITGRYLAWYQ SEQ ID NO: 376 RASQSITGRYLAWYQQ Sequence number 377 RASQSITGRYLAWYQQK SEQ ID NO: 378 SCRASQSIT SEQ ID NO: 379 SCRASQSITG SEQ ID NO: 380 SCRASQSITGR SEQ ID NO: 381 SCRASQSITGRY SEQ ID NO: 382 SCRASQSITGRYL SEQ ID NO: 383 SCRASQSITGRYLA SEQ ID NO: 384 SCRASQSITGRYLAW SEQ ID NO: 385 SCRASQSITGRYLAWY SEQ ID NO: 386 SCRASQSITGRYLAWYQ SEQ ID NO: 387 SCRASQSITGRYLAWYQQ SEQ ID NO: 388 SCRASQSITGRYLAWYQQK Sequence number 389 SITG SEQ ID NO. 390 SITGR SEQ ID NO: 391 SITGRY SEQ ID NO: 392 SITGRYL SEQ ID NO: 393 SITGRYLA SEQ ID NO. 394 SITGRYLAW SEQ ID NO: 395 SITGRYLAWY SEQ ID NO: 396 SITGRYLAWYQ Sequence number 397 SITGRYLAWYQQ Sequence number 398 SITGRYLAWYQQK Sequence number 399 SQSIT Sequence number 400 SQSITG Sequence number 401 SQSITGR SEQ ID NO: 402 SQSITGRY SEQ ID NO: 403 SQSITGRYL SEQ ID NO: 404 SQSITGRYLA Sequence number 405 SQSITGRYLAW SEQ ID NO: 406 SQSITGRYLAWY SEQ ID NO: 407 SQSITGRYLAWYQ SEQ ID NO: 408 SQSITGRYLAWYQQ Sequence number 409 SQSITGRYLAWYQQK SEQ ID NO: 410 TLSCRASQSIT SEQ ID NO: 411 TLSCRASQSITG SEQ ID NO: 412 TLSCRASQSITGR SEQ ID NO: 413 TLSCRASQSITGRY SEQ ID NO: 414 TLSCRASQSITGRYL SEQ ID NO: 415 TLSCRASQSITGRYLA SEQ ID NO: 416 TLSCRASQSITGRYLAW SEQ ID NO: 417 TLSCRASQSITGRYLAWY SEQ ID NO: 418 TLSCRASQSITGRYLAWYQ SEQ ID NO: 419 TLSCRASQSITGRYLAWYQQ SEQ ID NO: 420 TLSCRASQSITGRYLAWYQQK Light chain CDR2 region (SEQ ID NO: 421 to SEQ ID NO: 513) SEQ ID NO: 421 ESSR SEQ ID NO: 422 ESSRV SEQ ID NO: 423 ESSRVT SEQ ID NO: 424 ESSRVTG SEQ ID NO: 425 ESSRVTGI SEQ ID NO: 426 ESSRVTGIP SEQ ID NO: 427 ESSRVTGIPD SEQ ID NO: 428 ESSRVTGIPDR SEQ ID NO: 429 GESS SEQ ID NO: 430 GESSR SEQ ID NO: 431 GESSRV SEQ ID NO: 432 GESSRVT SEQ ID NO: 433 GESSRVTG SEQ ID NO: 434 GESSRVTGI SEQ ID NO: 435 GESSRVTGIP SEQ ID NO: 436 GESSRVTGIPD SEQ ID NO: 437 GESSRVTGIPDR SEQ ID NO: 438 LLMYGE SEQ ID NO: 439 LLMYGES SEQ ID NO: 440 LLMYGESS SEQ ID NO: 441 LLMYGESSR SEQ ID NO: 442 LLMYGESSRV SEQ ID NO: 443 LLMYGESSRVT SEQ ID NO: 444 LLMYGESSRVTG SEQ ID NO: 445 LLMYGESSRVTGI SEQ ID NO: 446 LLMYGESSRVTGIP SEQ ID NO: 447 LLMYGESSRVTGIPD SEQ ID NO: 448 LLMYGESSRVTGIPDR SEQ ID NO: 449 LMYGE SEQ ID NO: 450 LMYGES SEQ ID NO: 451 LMYGESS SEQ ID NO: 452 LMYGESSR SEQ ID NO: 453 LMYGESSRV SEQ ID NO: 454 LMYGESSRVT SEQ ID NO: 455 LMYGESSRVTG SEQ ID NO: 456 LMYGESSRVTGI SEQ ID NO: 457 LMYGESSRVTGIP SEQ ID NO: 458 LMYGESSRVTGIPD SEQ ID NO: 459 LMYGESSRVTGIPDR SEQ ID NO: 460 MYGE SEQ ID NO: 461 MYGES SEQ ID NO: 462 MYGESS SEQ ID NO: 463 MYGESSR SEQ ID NO: 464 MYGESSRV SEQ ID NO: 465 MYGESSRVT SEQ ID NO: 466 MYGESSRVTG SEQ ID NO: 467 MYGESSRVTGI SEQ ID NO: 468 MYGESSRVTGIP SEQ ID NO: 469 MYGESSRVTGIPD SEQ ID NO: 470 MYGESSRVTGIPDR SEQ ID NO: 471 RLLMYGE SEQ ID NO: 472 RLLMYGES SEQ ID NO: 473 RLLMYGESS SEQ ID NO: 474 RLLMYGESSR SEQ ID NO: 475 RLLMYGESSRV SEQ ID NO: 476 RLLMYGESSRVT SEQ ID NO: 477 RLLMYGESSRVTG SEQ ID NO: 478 RLLMYGESSRVTGI SEQ ID NO: 479 RLLMYGESSRVTGIP SEQ ID NO: 480 RLLMYGESSRVTGIPD SEQ ID NO: 481 RLLMYGESSRVTGIPDR SEQ ID NO: 482 RVTG SEQ ID NO: 483 RVTGI SEQ ID NO: 484 RVTGIP SEQ ID NO: 485 RVTGIPD SEQ ID NO: 486 RVTGIPDR Sequence number 487 SRVT SEQ ID NO: 488 SRVTG SEQ ID NO: 489 SRVTGI SEQ ID NO: 490 SRVTGIP SEQ ID NO:491 SRVTGIPD SEQ ID NO: 492 SRVTGIPDR SEQ ID NO: 493 SSRV SEQ ID NO:494 SSRVT SEQ ID NO: 495 SSRVTG SEQ ID NO:496 SSRVTGI SEQ ID NO:497 SSRVTGIP SEQ ID NO:498 SSRVTGIPD SEQ ID NO: 499 SSRVTGIPDR Sequence number 500 VTGI Sequence number 501 VTGIP SEQ ID NO:502 VTGIPD SEQ ID NO:503 VTGIPDR SEQ ID NO: 504 YGES Sequence number 505 YGESS Sequence number 506 YGESSR SEQ ID NO:507 YGESSRV Sequence number 508 YGESSRVT Sequence number 509 YGESSRVTG SEQ ID NO: 510 YGESSRVTGI SEQ ID NO:511 YGESSRVTGIP SEQ ID NO: 512 YGESSRVTGIPD SEQ ID NO: 513 YGESSRVTGIPDR Light chain CDR3 region (SEQ ID NO: 514 to SEQ ID NO: 628) Sequence number 514 ASSP SEQ ID NO:515 ASSPP SEQ ID NO:516 ASSPPT SEQ ID NO:517 ASSPPTY SEQ ID NO:518 ASSPPTYT SEQ ID NO:519 ASSPPTYTF SEQ ID NO:520 ASSPPTYTFG SEQ ID NO:521 ASSPPTYTFGQ SEQ ID NO:522 ASSPPTYTFGQG SEQ ID NO: 523 ASSPPTYTFGQGT SEQ ID NO:524 AVYYCQHFASS SEQ ID NO: 525 AVYYCQHFASSP SEQ ID NO:526 AVYYCQHFASSPP SEQ ID NO:527 AVYYCQHFASSPPT SEQ ID NO:528 AVYYCQHFASSPPTY SEQ ID NO:529 AVYYCQHFASSPPTYT SEQ ID NO: 530 AVYYCQHFASSPPTYTF SEQ ID NO:531 AVYYCQHFASSPPTYTFG SEQ ID NO:532 AVYYCQHFASSPPTYTFGQ SEQ ID NO: 533 AVYYCQHFASSPPTYTFGQG SEQ ID NO: 534 AVYYCQHFASSPPTYTFGQGT Sequence number 535 CQHFASS SEQ ID NO:536 CQHFASSP SEQ ID NO:537 CQHFASSPP SEQ ID NO:538 CQHFASSPPT SEQ ID NO:539 CQHFASSPPTY SEQ ID NO:540 CQHFASSPPTYT SEQ ID NO:541 CQHFASSPPTYTF SEQ ID NO:542 CQHFASSPPTYTFG SEQ ID NO:543 CQHFASSPPTYTFGQ SEQ ID NO:544 CQHFASSPPTYTFGQG SEQ ID NO: 545 CQHFASSPPTYTFGQGT SEQ ID NO:546 FASS SEQ ID NO:547 FASSP SEQ ID NO:548 FASSPP SEQ ID NO:549 FASSPPT SEQ ID NO:550 FASSPPTY SEQ ID NO:551 FASSPPTYT SEQ ID NO:552 FASSPPTYTF SEQ ID NO:553 FASSPPTYTFG SEQ ID NO:554 FASSPPTYTFGQ SEQ ID NO: 555 FASSPPTYTFGQG SEQ ID NO: 556 FASSPPTYTFGQGT SEQ ID NO. 557 HFASS SEQ ID NO: 558 HFASSP SEQ ID NO:559 HFASSPP SEQ ID NO:560 HFASSPPT SEQ ID NO:561 HFASSPPTY SEQ ID NO:562 HFASSPPTYT SEQ ID NO:563 HFASSPPTYTF SEQ ID NO:564 HFASSPPTYTFG SEQ ID NO:565 HFASSPPTYTFGQ SEQ ID NO:566 HFASSPPTYTFGQG SEQ ID NO:567 HFASSPPTYTFGQGT Sequence number 568 QHFASS Sequence number 569 QHFASSP SEQ ID NO:570 QHFASSPP SEQ ID NO:571 QHFASSPPT SEQ ID NO:572 QHFASSPPTY Sequence number 573 QHFASSPPTYT Sequence number 574 QHFASSPPTYTF Sequence number 575 QHFASSPPTYTFG Sequence number 576 QHFASSPPTYTFGQ Sequence number 577 QHFASSPPTYTFGQG Sequence number 578 QHFASSPPTYTFGQGT SEQ ID NO:579 SPPT SEQ ID NO:580 SPPTY SEQ ID NO:581 SPPTYT SEQ ID NO:582 SPPTYTF SEQ ID NO:583 SPPTYTFG SEQ ID NO:584 SPPTYTFGQ SEQ ID NO:585 SPPTYTFGQG SEQ ID NO:586 SPPTYTFGQGT SEQ ID NO:587 SSPP SEQ ID NO:588 SSPPT SEQ ID NO:589 SSPPTY SEQ ID NO:590 SSPPTYT SEQ ID NO:591 SSPPTYTF SEQ ID NO:592 SSPPTYTFG SEQ ID NO:593 SSPPTYTFGQ SEQ ID NO:594 SSPPTYTFGQG SEQ ID NO: 595 SSPPTYTFGQGT Sequence number 596 VYYCQHFASS Sequence number 597 VYYCQHFASSP Sequence number 598 VYYCQHFASSPP Sequence number 599 VYYCQHFASSPPT Sequence number 600 VYYCQHFASSPPTY Sequence number 601 VYYCQHFASSPPTYT Sequence number 602 VYYCQHFASSPPTYTF Sequence number 603 VYYCQHFASSPPTYTFG Sequence number 604 VYYCQHFASSPPTYTFGQ Sequence number 605 VYYCQHFASSPPTYTFGQG Sequence number 606 VYYCQHFASSPPTYTFGQGT Sequence number 607 YCQHFASS Sequence number 608 YCQHFASSP Sequence number 609 YCQHFASSPP SEQ ID NO: 610 YCQHFASSPPT SEQ ID NO: 611 YCQHFASSPPTY SEQ ID NO: 612 YCQHFASSPPTYT SEQ ID NO: 613 YCQHFASSPPTYTF SEQ ID NO: 614 YCQHFASSPPTYTFG SEQ ID NO: 615 YCQHFASSPPTYTFGQ SEQ ID NO: 616 YCQHFASSPPTYTFGQG SEQ ID NO: 617 YCQHFASSPPTYTFGQGT SEQ ID NO: 618 YYCQHFASS SEQ ID NO: 619 YYCQHFASSP SEQ ID NO: 620 YYCQHFASSPP SEQ ID NO: 621 YYCQHFASSPPT SEQ ID NO: 622 YYCQHFASSPPTY SEQ ID NO: 623 YYCQHFASSPPTYT SEQ ID NO: 624 YYCQHFASSPPTYTF SEQ ID NO: 625 YYCQHFASSPPTYTFG SEQ ID NO: 626 YYCQHFASSPPTYTFGQ SEQ ID NO: 627 YYCQHFASSPPTYTFGQG SEQ ID NO: 628 YYCQHFASSPPTYTFGQGT Heavy Chain Variable Domain (SEQ ID NO: 629) QVQLVQSGAEVKKPGASVKVSCQTSGYTFTSYYMHWVRQAPGQGLEWMGLITPSGDDTYYAQRFQGRVTMTRDTSTSPTYMELSSLTSEDTAVYYCAKMSRAGGFDVWGQGTLVTVSS Light Chain Variable Domain (SEQ ID NO: 630) EVVLTQSPGTLSLSPGERATLSCRASQSITGRYLAWYQQKPGQAPRLLMYGESSRVTGIPDRFSGGGSGTDFTLTISRLEPEDFAVYYCQHFASSPPTYTFGQGTKLEIR Heavy chain FR1 region (SEQ ID NO: 631) Sequence number 631 QVQLVQSGAEVKKPGASVKVSCQTSGYTFT Heavy chain FR2 region (SEQ ID NO: 632) Sequence number 632 WVRQAPGQGLEWMG Heavy chain FR3 region (SEQ ID NO: 633) SEQ ID NO: 633 RVTMTRDTSTSPTYMELSSLTSEDTAVYYCAK Heavy chain FR4 region (SEQ ID NO: 634) Sequence number 634 WGQGTLVTVSS Light chain FR1 region (SEQ ID NO: 635) SEQ ID NO: 635 EVVLTQSPGTLSLSPGERATLSC Light chain FR2 region (SEQ ID NO: 636) SEQ ID NO: 636 WYQQKPGQAPRLLMY Light chain FR3 region (SEQ ID NO: 637) SEQ ID NO: 637 GIPDRFSGGGSGTDFTLTISRLEPEDFAVYYC Light chain FR4 region (SEQ ID NO: 638) SEQ ID NO: 638 FGQGTKLEIR 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 SHYMH Heavy chain CDR2 region (SEQ ID NO: 652) Sequence number 652 IINPSGSGTAYGQKFQG Heavy chain CDR3 region (SEQ ID NO: 653) SEQ ID NO: 653 GSGGLFAY Light chain CDR1 region (SEQ ID NO: 654) SEQ ID NO: 654 RASQIVRSNYLA Light chain CDR2 region (SEQ ID NO: 655) Sequence number 655 GASSRAT Light chain CDR3 region (SEQ ID NO: 656) Sequence number 656 LQYDSSPPTYI Heavy chain CDR1 region (SEQ ID NO: 657) SEQ ID NO: 657 YFYLH Heavy chain CDR2 region (SEQ ID NO: 658) Sequence number 658 IINPRGDGTRYAQKFQG Heavy chain CDR3 region (SEQ ID NO: 659) SEQ ID NO: 659 GADHGAFDI Light chain CDR1 region (SEQ ID NO: 660) SEQ ID NO: 660 RASQSVRRNYFA Light chain CDR2 region (SEQ ID NO: 661) SEQ ID NO: 661 DASTRAT Light chain CDR3 region (SEQ ID NO: 662) SEQ ID NO: 662 QQYDSSPPMYI 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 inoculated 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, such that administration of 200 μL relative to the average body weight per cage in the dosing group 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.

[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] JPEG2026505051000002.jpg55170JPEG2026505051000003.jpg55170

[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 ) SARS-CoV-2 delta variant and monitored for weight loss and mortality until the end of the study 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 rates, survival times, and body weight changes (area under the curve) at day 11 were compared with the control group using Fisher's exact test, log-rank test, 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 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 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 0.5 mg / kg or higher of the test antibody resulted in a significant reduction in 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 inoculated 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) SARS-CoV-2 delta variant and monitored for weight loss and mortality until the end of the study 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 mL 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 rates, survival times, and body weight changes (area under the curve) at day 11 were compared with the control group using Fisher's exact test, log-rank test, 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. In contrast, the control group demonstrated 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: 029, heavy chain CDR2 as SEQ ID NO: 136, heavy chain CDR3 as SEQ ID NO: 256, light chain CDR1 as SEQ ID NO: 372, light chain CDR2 as SEQ ID NO: 432, and light chain CDR3 as SEQ ID NO: 573 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, except for n=9 in the 0.5 mg / kg 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 inoculated 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 so that administration of 200 μL to a 20 g mouse 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 model 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 16.7-22.4 g on the day of virus inoculation. Fifty-nine mice, 7-11 weeks of age, all female, were used. Ten animals were assigned to five treatment groups, with nine animals assigned to the 0.5 mg / kg group. All mice were housed in individually ventilated cages (IVCs) with corncob bedding, tissue or shredded paper as nesting material, wooden chew blocks, easily accessible food pellets and 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).

[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 59 mice, 7-11 weeks old, were assigned to six experimental groups according to Table 3 (see below). Mice were allowed more than 3 days for acclimatization.

[0254] [Table 3]

[0255] Female SARS-CoV-2 delta K18 hACE2 mice were treated with antibody at doses of 10 mg / kg to 0.2 mg / kg, based on a mouse body weight of 20 g, via intraperitoneal administration. On day 0, all mice received a lethal dose (10 3.5 TCID 50 ) SARS-CoV-2 delta variant and monitored for weight loss and mortality until the end of the study on day 12.

[0256] Antibody administration Upon arrival, the test antibodies were stored at −80° C.±10° C. Appropriate doses according to the treatment schedule (Table 3) were formulated assuming a body weight of 20 g per mouse.

[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 12. 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 12 were compared with the vehicle (PBS solution) control group using Fisher's exact test, log-rank test, and Welch's t-test, respectively. P values ​​were adjusted according to Bonferroni (for two-way comparisons with vehicle) and followed 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 test antibody at 5 mg / kg or higher provided a statistically significant increase in survival compared to vehicle (see Figure 3). Animals treated with test antibody at 5 mg / kg and 10 mg / kg had a 90% survival rate, while the control group had a 0% survival rate at day 7. The median survival time in the control group was 6.5 days.

[0264] Prophylactic treatment with test antibody at 1.7 mg / kg or higher resulted in a significant improvement in survival.

[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 1.7 mg / kg or higher of 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 test antibody at ≥5 mg / kg significantly improved survival and reduced body weight loss by ≥1.7 mg / kg compared to vehicle, while all animals in the control group died.

[0267] Example 4 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: 029, heavy chain CDR2 as SEQ ID NO: 136, heavy chain CDR3 as SEQ ID NO: 256, light chain CDR1 as SEQ ID NO: 372, light chain CDR2 as SEQ ID NO: 432, and light chain CDR3 as SEQ ID NO: 573 after intranasal 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 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 inoculated with the SARS-CoV-2 delta variant.

[0269] 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).

[0270] Monoclonal dilutions were made at various concentrations so that administration of 50 μL to a 20 g mouse 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 mouse model, weighing approximately 17.2–21.9 g on the day of test material administration. Sixty mice, 7–11 weeks old, all female, were used. Ten animals were assigned to six treatment groups based on day -5 weight, creating groups with similar mean body weight 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).

[0274] 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 mice, 7-11 weeks old, were transported to the animal facility and assigned to six experimental groups according to Table 4 (see below). Mice were allowed more than 3 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 a mouse body weight of 20 g, via the intranasal route. On day 0, all mice received a lethal dose (10 3.5 TCID 50 ) SARS-CoV-2 delta variant and monitored for mortality and weight loss until the study was terminated on day 12.

[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 assuming a mouse weight of 20 g.

[0278] 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.

[0279] 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.

[0280] Laboratory analysis The administered virus dose 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 12. 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 12 were compared with the vehicle (PBS solution) control group using Fisher's exact test, log-rank test, and Welch's t-test, respectively. P values ​​were adjusted according to Bonferroni (for two-way comparisons with vehicle) and followed 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] Survival rate – intranasal Prophylactic treatment with 5 mg / kg or higher of test antibody provided a statistically significant increase in survival compared to vehicle (see Figure 4). Animals treated with 5 mg / kg and 10 mg / kg of test antibody had an 80% survival rate, while the control group had a 0% survival rate at day 7. The median survival time in the control group was 6.5 days.

[0285] Prophylactic treatment with 0.2 mg / kg or greater of the test antibody resulted in a significant improvement in survival.

[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 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 mg / kg or higher of the test antibody resulted in a significant reduction in body weight loss compared to the control group.

[0287] conclusion In this lethal SARS-CoV-2 DeltaK18 hACE2 Tg mouse model, prophylactic intranasal administration of test antibody at ≥5 mg / kg significantly improved survival and reduced body weight loss by ≥0.2 mg / kg compared to vehicle. In contrast, all animals in the control group died by day 7.

[0288] Example 5 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: 029, 136, 256, 372, 432, and 573. The second parent antibody ("Parent mAb 2") has the following CDRs represented by SEQ ID NOs: 645, 646, 647, 648, 649-650. Bispecific antibodies according to the invention have a first Fab comprising a sequence comprising any one or more of SEQ ID NOs: 029, 136, 256, 372, 432, or 573, and a second Fab comprising a sequence comprising any one or more of SEQ ID NOs: 645, 646, 647, 648, 649-650.

[0289] 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.

[0290] 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.

[0291] 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.

[0292] All mice will receive an intranasal inoculation of SARS-CoV-2 delta on day 0.

[0293] 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.

[0294] 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.

[0295] [Table 5]

[0296] 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 inoculated 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).

[0297] Antibody administration According to the treatment schedule (Table 5), 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.

[0298] 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., 103.5 TCID 50 They are infected with SARS-CoV-2.

[0299] Laboratory analysis The inoculum is returned to the laboratory and replicate samples are titrated on Vero cells to verify the administered virus dose.

[0300] 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).

[0301] 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 test, 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).

[0302] 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 parent antibody.

[0303] survival rate Prophylactic treatment with parental mAb 1, parental mAb 2, or the bispecific antibody (shown in Table 5) provided statistically significant protection against mortality compared to the control group and a significant improvement in survival compared to the control group.

[0304] 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.

[0305] 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.

[0306] conclusion In this SARS-CoV-2 delta mouse model, prophylactic intranasal administration of bispecific antibodies according to the invention (shown in Table 5) 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.

[0307] Example 6 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.

[0308] 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.

[0309] [Table 6]

[0310] ELISA binding assays (Table 6) showed that the antibodies according to the invention were able to effectively bind to all spike antigens tested from betacoronaviruses, including SARS-CoV-2 delta and omicron XBB.1.5. The alphacoronavirus NL63 spike antigen did not bind to the antibodies. The anti-S1 antibody tested bound only to SARS-CoV-2 delta, not to the omicron variants, and showed no binding among the alphacoronavirus NL63, as expected from the literature. The isotype control antibody showed no binding to any coronavirus antigens.

[0311] Example 7 Affinity binding evaluation of the antibody according to the present invention by 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 of 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 7), as well as a set of Omicron variants (V-Plex SARS-CoV-2 Panel 34 (IgG) Kit cat# K15690U-2) (Table 8). 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.

[0312] [Table 7]

[0313] MSD binding arrays (see Table 7) demonstrated that the antibodies of the present invention can effectively bind to all spike antigens from betacoronaviruses, including SARS-CoV-2, MERS, HKU1, and OC43. Alphacoronaviruses, including 229E and NL63, and the SARS-CoV-2 receptor-binding domain (RBD) on the S1 portion of the spike did not bind to the antibodies. The anti-S1 antibodies tested here bound extremely well to the RBD domain and wild-type SARS-CoV-2, as expected from the literature, and did not show binding breadth among the other alphacoronaviruses tested. This suggests that the antibodies of the present invention exhibit efficient binding across the spikes of all betacoronaviruses tested, while the anti-RBD antibodies bind only to the spike and RBD domain of SARS-CoV-2. An isotype control antibody did not show any binding to coronavirus antigens.

[0314] [Table 8]

[0315] MSD binding arrays demonstrated that the antibodies of the present invention effectively bound all omicron variants tested in this assay, even at concentrations lower than SARS-CoV-2 Wuhan (Table 8), thus retaining activity among omicron variants. The BA.1 and BN.1 variants of SARS-CoV-2 bound particularly at low concentrations. The anti-S1 control antibody bound to the SARS-CoV-2 spike at lower concentrations than the antibodies of the present invention, but bound early omicron variants only at very high concentrations and did not bind later omicron variants such as BQ.1 and XBB.1. This indicates that the antibodies of the present invention can bind to recent omicron variants, while the anti-RBD antibody does not bind to these variants. The isotype control antibody showed no binding to coronavirus antigens.

[0316] Example 8 Live virus neutralization of antibodies according to the invention 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, serial dilutions of antibodies were pre-incubated with the corresponding viruses 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 enzyme-tagged detection antibodies were 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.

[0317] [Table 9]

[0318] Overall, the antibodies according to the invention are able to neutralize the tested viruses SARS-CoV-1, SARS-CoV-2 and MERS-CoV at various concentrations (see Table 9), with IC for MERS-CoV. 50 Convalescent sera showed high MN concentrations, confirming that the antibodies had neutralizing activity across these betacoronaviruses as demonstrated by binding assays. 50 It was only able to neutralize SARS-CoV-2 Wuhan at a titer.

[0319] Example 9 Neutralization of pseudovirions by antibodies of the present invention 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 BA.4, hCoV NL63, and hCoV 229E.

[0320] [Table 10]

[0321] 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 10), with an overall trend of higher IC for omicron variants and delta.50 As expected based on the lack of binding in the MSD binding assay, antibodies according to the invention did not neutralize the alphacoronaviruses hCoV NL63 and 229E.

[0322] Example 10 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.

[0323] The antibodies bound to peptide arrays of alpha- and beta-coronaviruses in specific epitopes of 10-20 amino acids (see Figure 9). 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: 004-009, 015-020, 029-034, 039-044, 047-052, 065-070; a heavy chain CDR2 region comprising any one of SEQ ID NOs: 082-087, 103-108, 125-130, 136-141, 147-152, 191-196; and a heavy chain CDR3 region comprising any one of SEQ ID NOs: 210-215, 221-226, 245-250, 256-261, 286-291, 297-302; 4, 330-335, 350-355, 372-377, 383-388, 415-420; a light chain CDR2 region comprising any one of SEQ ID NOs: 432-437, 443-448, 454-459, 465-470, 476-481, 508-513; and a light chain CDR3 region comprising any one of SEQ ID NOs: 529-534, 540-545, 573-578, 601-606, 612-617, 623-628 to an individual in need thereof, 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: 004 to 009, 015 to 020, 029 to 034, 039 to 044, 047 to 052, and 065 to 070, a heavy chain CDR2 region comprising any one of SEQ ID NOs: 082 to 087, 103 to 108, 125 to 130, 136 to 141, 147 to 152, and 191 to 196, and a heavy chain CDR3 region comprising any one of SEQ ID NOs: 210 to 215, 221 to 226, 245 to 250, 256 to 261, 286 to 291, and 297 to 302; and SEQ ID NO:

319. 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: 324 to 335, 350 to 355, 372 to 377, 383 to 388, and 415 to 420; a light chain CDR2 region comprising any one of SEQ ID NOs: 432 to 437, 443 to 448, 454 to 459, 465 to 470, 476 to 481, and 508 to 513; and a light chain CDR3 region comprising any one of SEQ ID NOs: 529 to 534, 540 to 545, 573 to 578, 601 to 606, 612 to 617, and 623 to 628.

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: 004-009, 015-020, 029-034, 039-044, 047-052, 065-070, a heavy chain CDR2 region comprising any one of SEQ ID NOs: 082-087, 103-108, 125-130, 136-141, 147-152, 191-196, and a heavy chain CDR3 region comprising any one of SEQ ID NOs: 210-215, 221-226, 245-250, 256-261, 286-291, 297-302. a light chain CDR1 region comprising any one of SEQ ID NOs: 319-324, 330-335, 350-355, 372-377, 383-388, and 415-420; a light chain CDR2 region comprising any one of SEQ ID NOs: 432-437, 443-448, 454-459, 465-470, 476-481, and 508-513; and a light chain CDR3 region comprising any one of SEQ ID NOs: 529-534, 540-545, 573-578, 601-606, 612-617, and 623-628, 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: 029, a heavy chain CDR2 region comprising SEQ ID NO: 136, and a heavy chain CDR3 region comprising SEQ ID NO: 256, and a light chain variable domain comprising a light chain CDR1 region comprising SEQ ID NO: 372, a light chain CDR2 region comprising SEQ ID NO: 432, and a light chain CDR3 region comprising SEQ ID NO:

573.

5. 5. The method, composition or antibody according to any one of claims 1 to 4, wherein the 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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