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 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), as well as alphacoronaviruses. Spillover events, in which humans become infected with coronaviruses circulating in animal reservoirs, are common. Recently, three betacoronaviruses, including SARS-CoV-2, have crossed over from animals to humans, causing severe epidemics.

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

[0007] The name coronavirus comes from its crown-like appearance. Coronaviruses are a large group of viruses that have spike proteins on their surface that resemble the spikes of a crown. Antibodies elicited by natural infection or by the COVID-19 vaccines used for herd immunity in 2020–2023 primarily target the variable receptor-binding 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 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] Fusion peptide The fusion peptide is a domain located on the spike protein of coronaviruses that is involved in membrane fusion between the viral envelope and the host cell during infection.

[0018] Some parts of the S2 subunit are called cryptic epitopes, which are present on the surface but are buried and only accessible after a conformational change, making them hidden antigenic sites. Unlike the stem helix, which is always available for binding, antigenic sites such as the fusion peptide are considered cryptic, meaning they are hidden until the receptor (ACE2) engages. Recent omicron variants may protect cryptic epitopes by crowding the RBD in the down state, possibly stabilizing the S2 closed state.

[0019] 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).

[0020] 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. SARS-CoV-2 and betacoronaviruses, including alphacoronaviruses, 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.

[0021] 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]

[0022] 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]

[0023] 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.

[0024] 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.

[0025] 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.

[0026] 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.

[0027] The transmucosal treatment method of the present invention is effective at low dosages and combats weight loss.

[0028] Complementarity-Determining Regions (CDRs) Preferably, the CDR regions are according to Kabat et al. (1991), as described in Sequences of Proteins of Immunological Interest.

[0029] In a preferred embodiment, the present invention provides a heavy chain variable domain comprising a heavy chain CDR1 region comprising any one of SEQ ID NOs: 006 to 011, 013 to 018, 022 to 027, 033 to 038, 053 to 058, and 063 to 068; a heavy chain CDR2 region comprising any one of SEQ ID NOs: 082 to 087, 093 to 098, 115 to 120, 126 to 131, 170 to 175, and 191 to 196; and a heavy chain CDR3 region comprising any one of SEQ ID NOs: 202 to 207, 213 to 218, 266 to 271, 277 to 282, 299 to 304, and 310 to 315. The present invention provides antibodies comprising a light chain variable domain including a light chain CDR1 region comprising any one of SEQ ID NOs: 321 to 326, 353 to 358, 364 to 369, 386 to 391, 408 to 413, and 430 to 435, a light chain CDR2 region comprising any one of SEQ ID NOs: 439 to 444, 450 to 455, 461 to 466, 472 to 477, 489 to 494, and 523 to 528, and a light chain CDR3 region comprising any one of SEQ ID NOs: 534 to 539, 545 to 550, 556 to 561, 584 to 589, 613 to 618, and 634 to 639.

[0030] In a preferred embodiment, the present invention provides a heavy chain variable domain comprising a heavy chain CDR1 region comprising any one of SEQ ID NOs: 006 to 011, 013 to 018, 022 to 027, 033 to 038, 053 to 058, and 063 to 068; a heavy chain CDR2 region comprising any one of SEQ ID NOs: 082 to 087, 093 to 098, 115 to 120, 126 to 131, 170 to 175, and 191 to 196; and a heavy chain CDR3 region comprising any one of SEQ ID NOs: 202 to 207, 213 to 218, 266 to 271, 277 to 282, 299 to 304, and 310 to 315; Provided is an antibody comprising a light chain variable domain including a light chain CDR1 region comprising any one of SEQ ID NOs: 321 to 326, 353 to 358, 364 to 369, 386 to 391, 408 to 413, and 430 to 435; a light chain CDR2 region comprising any one of SEQ ID NOs: 439 to 444, 450 to 455, 461 to 466, 472 to 477, 489 to 494, and 523 to 528; and a light chain CDR3 region comprising any one of SEQ ID NOs: 534 to 539, 545 to 550, 556 to 561, 584 to 589, 613 to 618, and 634 to 639, wherein the antibody is administered to a mucosal epithelium.

[0031] In a preferred embodiment, the present invention provides a method for treating a coronavirus infection in an individual, the method comprising administering to an individual a heavy chain variable dopant comprising a heavy chain CDR1 region comprising any one of SEQ ID NOs: 006 to 011, 013 to 018, 022 to 027, 033 to 038, 053 to 058, or 063 to 068, a heavy chain CDR2 region comprising any one of SEQ ID NOs: 082 to 087, 093 to 098, 115 to 120, 126 to 131, 170 to 175, or 191 to 196, and a heavy chain CDR3 region comprising any one of SEQ ID NOs: 202 to 207, 213 to 218, 266 to 271, 277 to 282, 299 to 304, or 310 to 315. The method includes a step of administering to an individual in need thereof an antibody comprising a light chain variable domain including a main, a light chain CDR1 region comprising any one of SEQ ID NOs: 321 to 326, 353 to 358, 364 to 369, 386 to 391, 408 to 413, and 430 to 435, a light chain CDR2 region comprising any one of SEQ ID NOs: 439 to 444, 450 to 455, 461 to 466, 472 to 477, 489 to 494, and 523 to 528, and a light chain CDR3 region comprising any one of SEQ ID NOs: 534 to 539, 545 to 550, 556 to 561, 584 to 589, 613 to 618, and 634 to 639, wherein the antibody is administered to the mucosal epithelium.

[0032] In a preferred embodiment, the present invention provides a heavy chain variable domain comprising: a heavy chain CDR1 region comprising any one of SEQ ID NOs: 006 to 011, 013 to 018, 022 to 027, 033 to 038, 053 to 058, and 063 to 068; a heavy chain CDR2 region comprising any one of SEQ ID NOs: 082 to 087, 093 to 098, 115 to 120, 126 to 131, 170 to 175, and 191 to 196; and a heavy chain CDR3 region comprising any one of SEQ ID NOs: 202 to 207, 213 to 218, 266 to 271, 277 to 282, 299 to 304, and 310 to 315; 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: 321 to 326, 353 to 358, 364 to 369, 386 to 391, 408 to 413, and 430 to 435; a light chain CDR2 region comprising any one of SEQ ID NOs: 439 to 444, 450 to 455, 461 to 466, 472 to 477, 489 to 494, and 523 to 528; and a light chain CDR3 region comprising any one of SEQ ID NOs: 534 to 539, 545 to 550, 556 to 561, 584 to 589, 613 to 618, and 634 to 639.

[0033] In a preferred embodiment, the present invention provides an antibody for use in a method for treating a coronavirus infection in an individual, the antibody comprising a heavy chain CDR1 region comprising any one of SEQ ID NOs: 006 to 011, 013 to 018, 022 to 027, 033 to 038, 053 to 058, or 063 to 068; a heavy chain CDR2 region comprising any one of SEQ ID NOs: 082 to 087, 093 to 098, 115 to 120, 126 to 131, 170 to 175, or 191 to 196; and a heavy chain CDR2 region comprising any one of SEQ ID NOs: 202 to 207, 213 to 218, 266 to 271, 277 to 282, 299 to 304, or 310 to 315. and a light chain variable domain comprising a light chain CDR1 region comprising any one of SEQ ID NOs: 321 to 326, 353 to 358, 364 to 369, 386 to 391, 408 to 413, and 430 to 435, a light chain CDR2 region comprising any one of SEQ ID NOs: 439 to 444, 450 to 455, 461 to 466, 472 to 477, 489 to 494, and 523 to 528, and a light chain CDR3 region comprising any one of SEQ ID NOs: 534 to 539, 545 to 550, 556 to 561, 584 to 589, 613 to 618, and 634 to 639, and the antibody is administered to a mucosal epithelium.

[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 comprising SEQ ID NO: 013, a heavy chain CDR2 region comprising SEQ ID NO: 170, and a heavy chain CDR3 region comprising SEQ ID NO: 266; and a light chain variable domain comprising a light chain CDR1 region comprising SEQ ID NO: 386, a light chain CDR2 region comprising SEQ ID NO: 439, and a light chain CDR3 region comprising SEQ ID NO: 584.

[0035] 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: 013, a heavy chain CDR2 region consisting of SEQ ID NO: 170, and a heavy chain CDR3 region consisting of SEQ ID NO: 266, and a light chain variable domain comprising a light chain CDR1 region consisting of SEQ ID NO: 386, a light chain CDR2 region consisting of SEQ ID NO: 439, and a light chain CDR3 region consisting of SEQ ID NO: 584.

[0036] In a preferred embodiment, disclosed herein is an antibody for use in a method for the prophylactic treatment of a coronavirus infection in an individual, the antibody comprising a heavy chain CDR1 region comprising any one of SEQ ID NOs: 006-011, 013-018, 022-027, 033-038, 053-058, or 063-068; a heavy chain CDR2 region comprising any one of SEQ ID NOs: 082-087, 093-098, 115-120, 126-131, 170-175, or 191-196; and a heavy chain CDR2 region comprising any one of SEQ ID NOs: 202-207, 213-218, 266-271, 277-282, 299-304, or 310-315. 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: 321 to 326, 353 to 358, 364 to 369, 386 to 391, 408 to 413, and 430 to 435; a light chain CDR2 region including any one of SEQ ID NOs: 439 to 444, 450 to 455, 461 to 466, 472 to 477, 489 to 494, and 523 to 528; and a light chain CDR3 region including any one of SEQ ID NOs: 534 to 539, 545 to 550, 556 to 561, 584 to 589, 613 to 618, and 634 to 639, and 0.1 mg to 20 mg of the antibody is administered to the mucosa.

[0037] In a preferred embodiment, disclosed herein is an antibody for use in a method for the prophylactic treatment of a coronavirus infection in an individual, the antibody comprising a heavy chain CDR1 region comprising any one of SEQ ID NOs: 006-011, 013-018, 022-027, 033-038, 053-058, or 063-068; a heavy chain CDR2 region comprising any one of SEQ ID NOs: 082-087, 093-098, 115-120, 126-131, 170-175, or 191-196; and a heavy chain CDR2 region comprising any one of SEQ ID NOs: 202-207, 213-218, 266-271, 277-282, 299-304, or 310-315. 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: 321 to 326, 353 to 358, 364 to 369, 386 to 391, 408 to 413, and 430 to 435; a light chain CDR2 region including any one of SEQ ID NOs: 439 to 444, 450 to 455, 461 to 466, 472 to 477, 489 to 494, and 523 to 528; and a light chain CDR3 region including any one of SEQ ID NOs: 534 to 539, 545 to 550, 556 to 561, 584 to 589, 613 to 618, and 634 to 639, and 0.1 mg to 20 mg of the antibody is administered intranasally.

[0038] In a preferred embodiment, disclosed herein is an antibody for use in a method for the prophylactic treatment of coronavirus infection in an individual, the antibody comprising a heavy chain CDR1 region comprising any one of SEQ ID NOs: 006-011, 013-018, 022-027, 033-038, 053-058, 063-068, a heavy chain CDR2 region comprising any one of SEQ ID NOs: 082-087, 093-098, 115-120, 126-131, 170-175, 191-196, and a heavy chain CDR3 region comprising any one of SEQ ID NOs: 202-207, 213-218, 266-271, 277-282, 299-304, 310-315. The antibody comprises a light chain variable domain comprising a heavy chain variable domain including a CDR3 region; a light chain CDR1 region including any one of SEQ ID NOs: 321 to 326, 353 to 358, 364 to 369, 386 to 391, 408 to 413, and 430 to 435; a light chain CDR2 region including any one of SEQ ID NOs: 439 to 444, 450 to 455, 461 to 466, 472 to 477, 489 to 494, and 523 to 528; and a light chain CDR3 region including any one of SEQ ID NOs: 534 to 539, 545 to 550, 556 to 561, 584 to 589, 613 to 618, and 634 to 639, and 0.1 mg to 20 mg of the antibody is administered by oral inhalation.

[0039] In a preferred embodiment, disclosed herein is a composition comprising an antibody for use in a method for preventing or treating coronavirus infection, wherein the antibody comprises a heavy chain variable domain comprising: a heavy chain CDR1 region comprising any one of SEQ ID NOs: 006 to 011, 013 to 018, 022 to 027, 033 to 038, 053 to 058, or 063 to 068; a heavy chain CDR2 region comprising any one of SEQ ID NOs: 082 to 087, 093 to 098, 115 to 120, 126 to 131, 170 to 175, or 191 to 196; and a heavy chain CDR3 region comprising any one of SEQ ID NOs: 202 to 207, 213 to 218, 266 to 271, 277 to 282, 299 to 304, or 310 to 315. the antibody comprises a light chain variable domain comprising: a light chain CDR1 region comprising any one of SEQ ID NOs: 321 to 326, 353 to 358, 364 to 369, 386 to 391, 408 to 413, and 430 to 435; a light chain CDR2 region comprising any one of SEQ ID NOs: 439 to 444, 450 to 455, 461 to 466, 472 to 477, 489 to 494, and 523 to 528; and a light chain CDR3 region comprising any one of SEQ ID NOs: 534 to 539, 545 to 550, 556 to 561, 584 to 589, 613 to 618, and 634 to 639; 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.

[0040] Preferably, the antibody disclosed herein comprises a heavy chain variable domain comprising a heavy chain CDR1 region comprising any one of SEQ ID NOs: 006 to 011, 013 to 018, 022 to 027, 033 to 038, 053 to 058, and 063 to 068; a heavy chain CDR2 region comprising any one of SEQ ID NOs: 082 to 087, 093 to 098, 115 to 120, 126 to 131, 170 to 175, and 191 to 196; and a heavy chain CDR3 region comprising any one of SEQ ID NOs: 202 to 207, 213 to 218, 266 to 271, 277 to 282, 299 to 304, and 310 to 315.

[0041] Preferably, the antibody disclosed herein comprises a light chain variable domain comprising a light chain CDR1 region comprising any one of SEQ ID NOs: 321 to 326, 353 to 358, 364 to 369, 386 to 391, 408 to 413, and 430 to 435; a light chain CDR2 region comprising any one of SEQ ID NOs: 439 to 444, 450 to 455, 461 to 466, 472 to 477, 489 to 494, and 523 to 528; and a light chain CDR3 region comprising any one of SEQ ID NOs: 534 to 539, 545 to 550, 556 to 561, 584 to 589, 613 to 618, and 634 to 639.

[0042] Preferably, in the method for treatment of the present invention, the antibody comprises a heavy chain variable domain comprising a heavy chain CDR1 region comprising any one of SEQ ID NOs: 006 to 011, 013 to 018, 022 to 027, 033 to 038, 053 to 058, and 063 to 068; a heavy chain CDR2 region comprising any one of SEQ ID NOs: 082 to 087, 093 to 098, 115 to 120, 126 to 131, 170 to 175, and 191 to 196; and a heavy chain CDR3 region comprising any one of SEQ ID NOs: 202 to 207, 213 to 218, 266 to 271, 277 to 282, 299 to 304, and 310 to 315.

[0043] 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: 321 to 326, 353 to 358, 364 to 369, 386 to 391, 408 to 413, and 430 to 435; a light chain CDR2 region comprising any one of SEQ ID NOs: 439 to 444, 450 to 455, 461 to 466, 472 to 477, 489 to 494, and 523 to 528; and a light chain CDR3 region comprising any one of SEQ ID NOs: 534 to 539, 545 to 550, 556 to 561, 584 to 589, 613 to 618, and 634 to 639.

[0044] In an alternative preferred embodiment, the present invention provides a heavy chain variable fragment comprising a heavy chain CDR1 region comprising any one of SEQ ID NOs: 006 to 011, 014 to 018, 022 to 027, 033 to 038, 053 to 058, or 063 to 068; a heavy chain CDR2 region comprising any one of SEQ ID NOs: 082 to 087, 093 to 098, 115 to 120, 126 to 131, 171 to 175, or 191 to 196; and a heavy chain CDR3 region comprising any one of SEQ ID NOs: 202 to 207, 213 to 218, 267 to 271, 277 to 282, 299 to 304, or 310 to 315. and a light chain CDR3 region comprising any one of SEQ ID NOs: 534 to 539, 545 to 550, 556 to 561, 585 to 589, 613 to 618, and 634 to 639.

[0045] Embodiments with more than one CDR Preferably, the antibodies disclosed herein comprise a heavy chain variable domain comprising a heavy chain CDR2 region comprising any one of SEQ ID NOs: 082-087, 093-098, 115-120, 126-131, 170-175, and 191-196.

[0046] 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.

[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 intravenous administration.

[0048] 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.

[0049] In a preferred embodiment, the antibodies disclosed herein are administered intranasally.

[0050] 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.

[0051] In a preferred embodiment, the present disclosure provides a method of treating a coronavirus infection in an individual, the method comprising administering to an individual a heavy chain variable domain comprising a heavy chain CDR1 region comprising any one of SEQ ID NOs: 006-011, 013-018, 022-027, 033-038, 053-058, or 063-068, a heavy chain CDR2 region comprising any one of SEQ ID NOs: 082-087, 093-098, 115-120, 126-131, 170-175, or 191-196, and a heavy chain CDR3 region comprising any one of SEQ ID NOs: 202-207, 213-218, 266-271, 277-282, 299-304, or 310-315. and a light chain CDR1 region comprising any one of SEQ ID NOs: 321 to 326, 353 to 358, 364 to 369, 386 to 391, 408 to 413, and 430 to 435; a light chain CDR2 region comprising any one of SEQ ID NOs: 439 to 444, 450 to 455, 461 to 466, 472 to 477, 489 to 494, and 523 to 528; and a light chain CDR3 region comprising any one of SEQ ID NOs: 534 to 539, 545 to 550, 556 to 561, 584 to 589, 613 to 618, and 634 to 639, to an individual in need thereof, including transmucosal administration, preferably intranasal administration and / or oral inhalation.

[0052] In a preferred embodiment, the individual is infected with or at risk for coronavirus infection.

[0053] 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.

[0054] section Each of the following numbered sections represents a preferred embodiment of the present invention and is part of this specification. Verse 1: 1. A method for treating a coronavirus infection in an individual, comprising administering to an individual a heavy chain variable domain comprising: a heavy chain CDR1 region comprising any one of SEQ ID NOs: 006-011, 013-018, 022-027, 033-038, 053-058, or 063-068; a heavy chain CDR2 region comprising any one of SEQ ID NOs: 082-087, 093-098, 115-120, 126-131, 170-175, or 191-196; and a heavy chain CDR3 region comprising any one of SEQ ID NOs: 202-207, 213-218, 266-271, 277-282, 299-304, or 310-315; a light chain CDR1 region comprising any one of SEQ ID NOs: 1 to 326, 353 to 358, 364 to 369, 386 to 391, 408 to 413, and 430 to 435; a light chain CDR2 region comprising any one of SEQ ID NOs: 439 to 444, 450 to 455, 461 to 466, 472 to 477, 489 to 494, and 523 to 528; and a light chain CDR3 region comprising any one of SEQ ID NOs: 534 to 539, 545 to 550, 556 to 561, 584 to 589, 613 to 618, and 634 to 639 to an individual in need thereof, 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: 006 to 011, 013 to 018, 022 to 027, 033 to 038, 053 to 058, and 063 to 068, a heavy chain CDR2 region comprising any one of SEQ ID NOs: 082 to 087, 093 to 098, 115 to 120, 126 to 131, 170 to 175, and 191 to 196, and a heavy chain CDR3 region comprising any one of SEQ ID NOs: 202 to 207, 213 to 218, 266 to 271, 277 to 282, 299 to 304, and 310 to 315; 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: 439 to 444, 450 to 455, 461 to 466, 472 to 477, 489 to 494, and 523 to 528, and a light chain CDR3 region comprising any one of SEQ ID NOs: 534 to 539, 545 to 550, 556 to 561, 584 to 589, 613 to 618, and 634 to 639. Verse 3: 1. An antibody for use in a method for preventing or treating a coronavirus infection in an individual, the antibody comprising: a heavy chain CDR1 region comprising any one of SEQ ID NOs: 006 to 011, 013 to 018, 022 to 027, 033 to 038, 053 to 058, or 063 to 068; a heavy chain CDR2 region comprising any one of SEQ ID NOs: 082 to 087, 093 to 098, 115 to 120, 126 to 131, 170 to 175, or 191 to 196; and a heavy chain CDR3 region comprising any one of SEQ ID NOs: 202 to 207, 213 to 218, 266 to 271, 277 to 282, 299 to 304, or 310 to 315. 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: 321 to 326, 353 to 358, 364 to 369, 386 to 391, 408 to 413, and 430 to 435; a light chain CDR2 region comprising any one of SEQ ID NOs: 439 to 444, 450 to 455, 461 to 466, 472 to 477, 489 to 494, and 523 to 528; and a light chain CDR3 region comprising any one of SEQ ID NOs: 534 to 539, 545 to 550, 556 to 561, 584 to 589, 613 to 618, and 634 to 639. 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: 013, a heavy chain CDR2 region comprising SEQ ID NO: 170, and a heavy chain CDR3 region comprising SEQ ID NO: 266; a light chain variable domain comprising a light chain CDR1 region comprising SEQ ID NO: 386, a light chain CDR2 region comprising SEQ ID NO: 439, and a light chain CDR3 region comprising SEQ ID NO: 584. 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: 640 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: 641 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: 13. 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 antibody, wherein the antibody is defined in any one of claims 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.

[0055] 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.

[0056] 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.

[0057] 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.

[0058] 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.

[0059] 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.

[0060] 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.

[0061] 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.

[0062] 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.

[0063] 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.

[0064] 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.

[0065] 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.

[0066] Timing and Spacing Preferably, the antibody is administered at least once or at least twice per month.

[0067] Preferably, the antibody is administered at least once or at least twice per week.

[0068] 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.

[0069] 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.

[0070] 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.

[0071] 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.

[0072] Framework Area Preferably, the antibody comprises a heavy chain variable domain having the sequence of SEQ ID NO:640 and / or a light chain variable domain having the sequence of SEQ ID NO:641.

[0073] Preferably, the heavy chain variable domain of the antibody further comprises a heavy chain framework region FR1 of SEQ ID NO: 642, a heavy chain framework region FR2 of SEQ ID NO: 643, a heavy chain framework region FR3 of SEQ ID NO: 644, and / or a heavy chain framework region FR4 of SEQ ID NO: 645.

[0074] Preferably, the light chain variable domain further comprises a light chain framework region RF1 of SEQ ID NO: 646, a light chain framework region FR2 of SEQ ID NO: 647, a light chain framework region FR3 of SEQ ID NO: 648, and / or a light chain framework region FR4 of SEQ ID NO: 649.

[0075] Combination Compositions In a preferred embodiment, the inventors disclose a composition comprising a first binding fragment as disclosed herein and a second binding fragment, wherein the first fragment is selected from the group consisting of SEQ ID NOs: 006-011, 013-018, 022-027, 033-038, 053-058, 063-068, 082-087, 093-098, 115-120, 126-131, 170-175, 191-196, 202-207, 213-218, 266-268, 270-275, 280-282, 282-287, 291-296, 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, 330-331, 332-333, 334-335, 336-337, 338-339, 340-341, 342-343, 344-345, 346-347, 348-349, 350-351, 352-353, 354-355, 356-357, 358-359, 360-361, 362-363, 364-365, 366-367, 368-369, 71, 277 to 282, 299 to 304, 310 to 315, 321 to 326, 353 to 358, 364 to 369, 386 to 391, 408 to 413, 430 to 435, 439 to 444, 450 to 455, 461 to 466, 472 to 477, 489 to 494, 523 to 528, 534 to 539, 545 to 550, 556 to 561, 584 to 589, 613 to 618, and 634 to 639. More preferably, the first fragment comprises a sequence including one or more of SEQ ID NOs: 013, 170, 266, 386, 439, and 584. The second binding fragment comprises a sequence comprising any one of SEQ ID NOs: 650, 651, 652, 653, 654, or 655. Preferably, the fragment is selected from any one of the group comprising a full-length antibody, a Fab, a modified Fab, a Fab', a modified Fab', a F(ab')2, an Fv, a single domain antibody, a scFv, a scFv-Fc, a bivalent, trivalent, or tetravalent antibody, a Bis-scFv, a diabody, a triabody, a tetrabody, and epitope-binding fragments thereof.

[0076] In a preferred embodiment, the inventors disclose a composition comprising a first binding fragment as disclosed herein and a second binding fragment, wherein the first fragment is selected from the group consisting of SEQ ID NOs: 006-011, 013-018, 022-027, 033-038, 053-058, 063-068, 082-087, 093-098, 115-120, 126-131, 170-175, 191-196, 202-207, 213-218, 266-268, 270-275, 280-282, 282-287, 291-296, 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, 330-331, 332-333, 334-335, 336-337, 338-339, 340-341, 342-343, 344-345, 346-347, 348-349, 350-351, 352-353, 354-355, 356-357, 358-359, 360-361, 362-363, 364-365, 366-367, 368-369, 71, 277 to 282, 299 to 304, 310 to 315, 321 to 326, 353 to 358, 364 to 369, 386 to 391, 408 to 413, 430 to 435, 439 to 444, 450 to 455, 461 to 466, 472 to 477, 489 to 494, 523 to 528, 534 to 539, 545 to 550, 556 to 561, 584 to 589, 613 to 618, and 634 to 639. More preferably, the first fragment comprises one or more of SEQ ID NOs: 013, 170, 266, 386, 439, or 584. The second binding fragment comprises a sequence comprising any one of SEQ ID NOs: 656, 657, 658, 659, 660, or SEQ ID NO: 661. Preferably, the fragment is selected from any one of the group comprising a full-length antibody, a Fab, a modified Fab, a Fab', a modified Fab', a F(ab')2, an Fv, a single domain antibody, a scFv, a scFv-Fc, a bivalent, trivalent, or tetravalent antibody, a Bis-scFv, a diabody, a triabody, a tetrabody, and epitope-binding fragments thereof.

[0077] In a preferred embodiment, the inventors disclose a composition comprising a first binding fragment as disclosed herein and a second binding fragment, wherein the first fragment is selected from the group consisting of SEQ ID NOs: 006-011, 013-018, 022-027, 033-038, 053-058, 063-068, 082-087, 093-098, 115-120, 126-131, 170-175, 191-196, 202-207, 213-218, 266-268, 270-275, 280-282, 282-287, 291-296, 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, 330-331, 332-333, 334-335, 336-337, 338-339, 340-341, 342-343, 344-345, 346-347, 348-349, 350-351, 352-353, 354-355, 356-357, 358-359, 360-361, 362-363, 364-365, 366-367, 368-369, 71, 277 to 282, 299 to 304, 310 to 315, 321 to 326, 353 to 358, 364 to 369, 386 to 391, 408 to 413, 430 to 435, 439 to 444, 450 to 455, 461 to 466, 472 to 477, 489 to 494, 523 to 528, 534 to 539, 545 to 550, 556 to 561, 584 to 589, 613 to 618, and 634 to 639. More preferably, the first fragment comprises a sequence including one or more of SEQ ID NOs: 013, 170, 266, 386, 439, and 584. The second binding fragment comprises a sequence comprising any one of SEQ ID NOs: 662, 663, 664, 665, 666, or SEQ ID NO: 667. 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] In a preferred embodiment, the inventors disclose a composition comprising a first binding fragment as disclosed herein and a second binding fragment, wherein the first fragment is selected from the group consisting of SEQ ID NOs: 006-011, 013-018, 022-027, 033-038, 053-058, 063-068, 082-087, 093-098, 115-120, 126-131, 170-175, 191-196, 202-207, 213-218, 266-268, 270-275, 280-282, 282-287, 291-296, 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, 330-331, 332-333, 334-335, 336-337, 338-339, 340-341, 342-343, 344-345, 346-347, 348-349, 350-351, 352-353, 354-355, 356-357, 358-359, 360-361, 362-363, 364-365, 366-367, 368-369, 71, 277 to 282, 299 to 304, 310 to 315, 321 to 326, 353 to 358, 364 to 369, 386 to 391, 408 to 413, 430 to 435, 439 to 444, 450 to 455, 461 to 466, 472 to 477, 489 to 494, 523 to 528, 534 to 539, 545 to 550, 556 to 561, 584 to 589, 613 to 618, and 634 to 639. More preferably, the first fragment comprises a sequence including one or more of SEQ ID NOs: 013, 170, 266, 386, 439, and 584. The second binding fragment comprises a sequence comprising any one of SEQ ID NOs: 668, 669, 670, 671, 672, or 673. 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.

[0079] bispecific antibody In a preferred embodiment, the inventors disclose a bispecific antibody, which is capable of binding to the fusion peptide of a coronavirus. In a preferred embodiment, the inventors disclose a bispecific antibody, which is capable of binding to the fusion peptide of the spike protein of SARS-CoV-2.

[0080] 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.

[0081] 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 fusion peptide of SARS-CoV-2 and a second Fab capable of binding to the fusion peptide of SARS-CoV-2.

[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 the preferred SEQ ID NOs: 650, 651, 652, 653, 654-655, or the first Fab comprises a sequence comprising any one or more of the preferred SEQ ID NOs: 662, 663, 664, Alternatively, the first Fab comprises a sequence comprising one or more of preferred SEQ ID NOs: 668, 669, 670, 671, 672-673, and the second Fab comprises a sequence comprising one or more of preferred SEQ ID NOs: 013, 170, 266, 386, 439, or 584, or the second Fab comprises a sequence comprising one or more of preferred SEQ ID NOs: 656, 657, 658, 659, 660-661.

[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 fusion peptide 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: 013, 170, 266, 368, 439, or 584, and the second Fab comprises a sequence comprising any one or more of preferred SEQ ID NOs: 656, 657, 658, 659, 660-661. 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 fusion peptide of SARS-CoV-2 and a second Fab 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: 013, 170, 266, 368, 439, or 584, and the second Fab comprises a sequence comprising any one or more of preferred SEQ ID NOs: 650, 651, 652, 653, 654-655. 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, the inventors disclose an anti-SARS-CoV-2 bispecific antibody or antigen-binding fragment thereof having a first Fab capable of binding to the fusion peptide 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 the preferred SEQ ID NOs: 013, 170, 266, 368, 439, or 584, and the second Fab comprises a sequence comprising any one or more of the preferred SEQ ID NOs: 662, 663, 664, 665, 666-667. 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.

[0086] 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 fusion peptide 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 the preferred SEQ ID NOs: 013, 170, 266, 368, 439, or 584, and the second Fab comprises a sequence comprising any one or more of the preferred SEQ ID NOs: 668, 669, 670, 671, 672-673. 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.

[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: 013, 170 or 266, and the second Fab comprises any one or more of the sequences set forth in preferred SEQ ID NOs: 671, 672 or 673.

[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: 013, 170 or 266, and the second Fab comprises any one or more of the sequences set forth in preferred SEQ ID NOs: 653, 654 or 655.

[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: 013, 170 or 266, and the second Fab comprises any one or more of the sequences set forth in preferred SEQ ID NOs: 665, 666 or 667.

[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: 668, 669 or 670, and the second Fab comprises any one or more of the sequences set forth in preferred SEQ ID NOs: 386, 439 or 584.

[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: 650, 651 or 652, and the second Fab comprises any one or more of the sequences set forth in preferred SEQ ID NOs: 386, 439 or 584.

[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: 662, 663 or 664, and the second Fab comprises any one or more of the sequences set forth in preferred SEQ ID NOs: 386, 439 or 584.

[0093] 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: 013, 170 or 266, and the second Fab comprises any one or more of the sequences set forth in preferred SEQ ID NOs: 659, 660 or 661.

[0094] 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: 656, 657 or 658, and the second Fab comprises any one or more of the sequences set forth in preferred SEQ ID NOs: 386, 439 or 584.

[0095] The present invention also provides methods for the construction, expression and purification of bispecific antibodies capable of binding to the fusion peptide of SARS-CoV-2, and the use of bispecific antibodies in the field of medicine, in particular in the prevention and / or treatment of SARS-CoV-2 infection.

[0096] 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 a fusion peptide of a coronavirus, preferably SARS-CoV-2, and a second Fab capable of binding to a 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: 013, 170, 266, 368, 439, or 584, and the second Fab comprises a sequence comprising preferably any one or more of SEQ ID NOs: 656, 657, 658, 659, 660-661. Verse 27: 27. The bispecific antibody or antigen-binding fragment thereof according to clause 26, comprising a first Fab capable of binding to a fusion peptide of a coronavirus, preferably SARS-CoV-2, and a second Fab capable of binding to a 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: 013, 170, 266, 368, 439, and / or 584, and the second Fab comprises a sequence comprising preferably any one or more of SEQ ID NOs: 656, 657, 658, 659, 660, and / or 661. Verse 28: a first Fab that specifically binds to the fusion peptide 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: 013, the heavy chain CDR2 region of SEQ ID NO: 170, and the heavy chain CDR3 region of SEQ ID NO: 266, and a light chain variable region comprising, as CDRs, the light chain CDR1 region of SEQ ID NO: 368, the light chain CDR2 region of SEQ ID NO: 439, and the light chain CDR3 region of SEQ ID NO: 584; b.) the second Fab comprises a heavy chain variable region comprising, as CDRs, the heavy chain CDR1 region of SEQ ID NO: 656, the heavy chain CDR2 region of SEQ ID NO: 657, and the heavy chain CDR3 region of SEQ ID NO: 658, and a light chain variable region comprising, as CDRs, the light chain CDR1 region of SEQ ID NO: 659, the light chain CDR2 region of SEQ ID NO: 660, and the light chain CDR3 region of SEQ ID NO: 661; 28. The bispecific antibody or antigen-binding fragment thereof according to clause 26 or 27, Verse 29: a) a light chain comprising VL-CL domains and a heavy chain comprising VH-CH1-CH2-CH3 domains of an antibody or antigen-binding fragment thereof capable of binding to a fusion peptide 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 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 30: 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: 013, the heavy chain CDR2 region of SEQ ID NO: 170, and the heavy chain CDR3 region of SEQ ID NO: 266; 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: 368, the light chain CDR2 region of SEQ ID NO: 439, and the light chain CDR3 region of SEQ ID NO: 584; 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: 656, the heavy chain CDR2 region of SEQ ID NO: 657, and the heavy chain CDR3 region of SEQ ID NO: 658, 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: 659, the light chain CDR2 region of SEQ ID NO: 660, and the light chain CDR3 region of SEQ ID NO: 661. 29. A bispecific antibody or antigen-binding fragment thereof according to clause 29. Verse 31: a) a light chain comprising VL-CL domains and a heavy chain comprising VH-CH1-CH2-CH3 domains of an antibody capable of binding to a fusion peptide 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 fusion peptide of coronavirus, preferably SARS-CoV-2, comprises, as CDRs, the heavy chain CDR1 region of SEQ ID NO: 013, the heavy chain CDR2 region of SEQ ID NO: 170, and the heavy chain CDR3 region of SEQ ID NO: 266; 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: 368, the light chain CDR2 region of SEQ ID NO: 439, and the light chain CDR3 region of SEQ ID NO: 584; 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: 656, the heavy chain CDR2 region of SEQ ID NO: 657, and the heavy chain CDR3 region of SEQ ID NO: 658, 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: 659, the light chain CDR2 region of SEQ ID NO: 660, and the light chain CDR3 region of SEQ ID NO: 661, 32. A bispecific antibody or antigen-binding fragment thereof according to clause 31. Verse 33: a) a light chain comprising VL-CL domains and a heavy chain comprising VH-CH1-CH2-CH3 domains of an antibody capable of binding to a fusion peptide 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:640. 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: 684. 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:641. 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: 685. 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: 013, or a first nucleic acid sequence encoding an amino acid sequence that differs from SEQ ID NO: 013 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: 170, or a second nucleic acid sequence encoding an amino acid sequence that differs from SEQ ID NO: 170 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: 266, or a third nucleic acid sequence encoding an amino acid sequence that differs from SEQ ID NO: 266 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: 368, or a fourth nucleic acid sequence encoding an amino acid sequence that differs from SEQ ID NO: 368 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: 439, or an amino acid sequence that differs from SEQ ID NO: 439 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: 584, or an amino acid sequence that differs from SEQ ID NO: 584 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: 656, or an amino acid sequence that differs from SEQ ID NO: 656 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: 657, or an eighth nucleic acid sequence encoding an amino acid sequence that differs from SEQ ID NO: 657 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: 658, or an amino acid sequence that differs from SEQ ID NO: 658 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: 659, or an amino acid sequence that differs from SEQ ID NO: 659 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: 660, or an eleventh nucleic acid sequence encoding an amino acid sequence that differs from SEQ ID NO: 660 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: 661, or an amino acid sequence that differs from SEQ ID NO: 661 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 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.

[0097] 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, having a first Fab capable of binding to a fusion peptide of a coronavirus, preferably SARS-CoV-2, and a second Fab capable of binding to a stem helix of a coronavirus, preferably SARS-CoV-2, wherein the first Fab comprises a sequence comprising preferably any one or more of SEQ ID NOs: 013, 170, 266, 368, 439, or 584, and the second Fab comprises a sequence comprising preferably any one or more of SEQ ID NOs: 662, 663, 664, 665, 666 to 667. Verse 54: 54. The bispecific antibody or antigen-binding fragment thereof according to clause 53, comprising a first Fab capable of binding to a fusion peptide of a coronavirus, preferably SARS-CoV-2, and a second Fab capable of binding to a stem-helix 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: 013, 170, 266, 368, 439, and / or 584, and the second Fab comprises a sequence comprising preferably any one or more of SEQ ID NOs: 662, 663, 664, 665, 666, and / or 667. Verse 55: a first Fab that specifically binds to the fusion peptide of a coronavirus, preferably SARS-CoV-2, and a second Fab that specifically binds to the stem helix 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: 013, the heavy chain CDR2 region of SEQ ID NO: 170, and the heavy chain CDR3 region of SEQ ID NO: 266, and a light chain variable region comprising, as CDRs, the light chain CDR1 region of SEQ ID NO: 368, the light chain CDR2 region of SEQ ID NO: 439, and the light chain CDR3 region of SEQ ID NO: 584; b.) the second Fab comprises a heavy chain variable region comprising, as CDRs, the heavy chain CDR1 region of SEQ ID NO: 662, the heavy chain CDR2 region of SEQ ID NO: 663, and the heavy chain CDR3 region of SEQ ID NO: 664, and a light chain variable region comprising, as CDRs, the light chain CDR1 region of SEQ ID NO: 665, the light chain CDR2 region of SEQ ID NO: 666, and the light chain CDR3 region of SEQ ID NO: 667; 55. The bispecific antibody or antigen-binding fragment thereof according to clause 53 or 54, Verse 56: a) a light chain comprising VL-CL domains and a heavy chain comprising VH-CH1-CH2-CH3 domains of an antibody or antigen-binding fragment thereof capable of binding to a fusion peptide of a coronavirus, preferably SARS-CoV-2, and b) 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; 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 fusion peptide 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 fusion peptide of coronavirus, preferably SARS-CoV-2, comprises, as CDRs, the heavy chain CDR1 region of SEQ ID NO: 013, the heavy chain CDR2 region of SEQ ID NO: 170, and the heavy chain CDR3 region of SEQ ID NO: 266; 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: 368, the light chain CDR2 region of SEQ ID NO: 439, and the light chain CDR3 region of SEQ ID NO: 584; The VH domain of the antibody or antigen-binding fragment thereof capable of binding to the stem helix of coronavirus, preferably SARS-CoV-2, comprises, as CDRs, the heavy chain CDR1 region of SEQ ID NO: 662, the heavy chain CDR2 region of SEQ ID NO: 663, and the heavy chain CDR3 region of SEQ ID NO: 664, and the VL domain of the antibody or antigen-binding fragment thereof capable of binding to the stem helix of coronavirus, preferably SARS-CoV-2, comprises, as CDRs, the light chain CDR1 region of SEQ ID NO: 665, the light chain CDR2 region of SEQ ID NO: 666, and the light chain CDR3 region of SEQ ID NO: 667. 57. A bispecific antibody or antigen-binding fragment thereof according to clause 56. Verse 58: a) a light chain comprising VL-CL domains and a heavy chain comprising VH-CH1-CH2-CH3 domains of an antibody capable of binding to a fusion peptide of a coronavirus, preferably SARS-CoV-2, and b) 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 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 fusion peptide of coronavirus, preferably SARS-CoV-2, comprises, as CDRs, the heavy chain CDR1 region of SEQ ID NO: 013, the heavy chain CDR2 region of SEQ ID NO: 170, and the heavy chain CDR3 region of SEQ ID NO: 266; 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: 368, the light chain CDR2 region of SEQ ID NO: 439, and the light chain CDR3 region of SEQ ID NO: 584; The VH domain of the antibody capable of binding to the stem helix of coronavirus, preferably SARS-CoV-2, comprises as CDRs the heavy chain CDR1 region of SEQ ID NO: 662, the heavy chain CDR2 region of SEQ ID NO: 663, and the heavy chain CDR3 region of SEQ ID NO: 664, and the VL domain of the antibody capable of binding to the stem helix of coronavirus, preferably SARS-CoV-2, comprises as CDRs the light chain CDR1 region of SEQ ID NO: 665, the light chain CDR2 region of SEQ ID NO: 666, and the light chain CDR3 region of SEQ ID NO: 667. 59. A bispecific antibody or antigen-binding fragment thereof according to clause 58. Verse 60: a) a light chain comprising VL-CL domains and a heavy chain comprising VH-CH1-CH2-CH3 domains of an antibody capable of binding to a fusion peptide of a coronavirus, preferably SARS-CoV-2, and b) 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 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: 640. 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:688. 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:641. 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:689. Verse 65: 61. The bispecific antibody or antigen-binding fragment thereof according to any one of clauses 53 to 60, selected from any one of the group comprising a full length antibody, a Fab, a modified Fab, a Fab', a modified Fab', a F(ab')2, an Fv, a single domain antibody, a scFv, a scFv-Fc, a bivalent, trivalent or tetravalent antibody, a Bis-scFv, a diabody, a triabody, a tetrabody, and epitope-binding fragments thereof. Verse 66: 66. A method for the production of a bispecific antibody or antigen-binding fragment thereof according to any one of clauses 53 to 65, comprising: a) culturing a host cell comprising an expression vector comprising a polynucleotide encoding the bispecific antibody or antibody fragment according to any one of the preceding clauses under conditions that allow the production of said bispecific antibody or antigen-binding fragment thereof, and b) isolating the bispecific antibody or antigen-binding fragment thereof. A method comprising: Verse 67: 66. A nucleic acid molecule comprising a nucleic acid sequence encoding the bispecific antibody or antigen-binding fragment thereof, the heavy chain variable region and / or the light chain variable region of the bispecific antibody or antigen-binding fragment thereof according to any one of clauses 53 to 65, which is preferably an isolated nucleic acid molecule. Verse 68: 68. A bispecific antibody or antigen-binding fragment thereof according to any one of clauses 53 to 65 and clause 67, a nucleic acid molecule comprising a nucleic acid sequence encoding the heavy chain variable region and / or the light chain variable region of the bispecific antibody or antigen-binding fragment thereof, The nucleic acid sequence is (i) a first nucleic acid sequence encoding the amino acid sequence set forth in SEQ ID NO: 013, or a first nucleic acid sequence encoding an amino acid sequence that differs from SEQ ID NO: 013 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: 170, or a second nucleic acid sequence encoding an amino acid sequence that differs from SEQ ID NO: 170 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: 266, or a third nucleic acid sequence encoding an amino acid sequence that differs from SEQ ID NO: 266 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: 368, or a fourth nucleic acid sequence encoding an amino acid sequence that differs from SEQ ID NO: 368 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: 439, or an amino acid sequence that differs from SEQ ID NO: 439 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: 584, or an amino acid sequence that differs from SEQ ID NO: 584 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: 662, or an amino acid sequence that differs from SEQ ID NO: 662 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: 663, or an eighth nucleic acid sequence encoding an amino acid sequence that differs from SEQ ID NO: 663 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: 664, or an amino acid sequence that differs from SEQ ID NO: 664 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: 665, or an amino acid sequence that differs from SEQ ID NO: 665 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: 666, or an eleventh nucleic acid sequence encoding an amino acid sequence that differs from SEQ ID NO: 666 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: 667, or an amino acid sequence that differs from SEQ ID NO: 667 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]

[0098] [Figure 1] Survival after lethal challenge - Kaplan-Meier survival curves for the antibody according to the invention intraperitoneal prophylactic treatment group. Animals (n=10 per group) were treated at dose titration with an antibody having heavy chain CDR1 as SEQ ID NO: 013, heavy chain CDR2 as SEQ ID NO: 170, heavy chain CDR3 as SEQ ID NO: 266, light chain CDR1 as SEQ ID NO: 386, light chain CDR2 as SEQ ID NO: 439, and light chain CDR3 as SEQ ID NO: 584 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 2] Survival after lethal challenge - Kaplan-Meier survival curves for the antibody according to the invention intranasal prophylactic treatment group. Animals (n=10 per group) were treated at dose titration with an antibody having heavy chain CDR1 as SEQ ID NO: 013, heavy chain CDR2 as SEQ ID NO: 170, heavy chain CDR3 as SEQ ID NO: 266, light chain CDR1 as SEQ ID NO: 386, light chain CDR2 as SEQ ID NO: 439, and light chain CDR3 as SEQ ID NO: 584 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 3]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) were treated intranasally with a dose-adjusted antibody having heavy chain CDR1 as SEQ ID NO: 013, heavy chain CDR2 as SEQ ID NO: 170, heavy chain CDR3 as SEQ ID NO: 266, light chain CDR1 as SEQ ID NO: 386, light chain CDR2 as SEQ ID NO: 439, and light chain CDR3 as SEQ ID NO: 584. 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 4] 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 dose-adjusted antibody having heavy chain CDR1 as SEQ ID NO: 013, heavy chain CDR2 as SEQ ID NO: 170, heavy chain CDR3 as SEQ ID NO: 266, light chain CDR1 as SEQ ID NO: 386, light chain CDR2 as SEQ ID NO: 439, and light chain CDR3 as SEQ ID NO: 584. 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 5]Survival and Body Weight Change for Intranasal Administration—Antibodies According to the Invention Survival and body weight change for intranasal administration of two parental antibodies and a bispecific antibody according to the invention in a pre-exposure efficacy study in K18-hACE2 mice challenged with lethal SARS-CoV-2 delta. Animals (n=8 per group) were treated intranasally on day -1 with titrated doses of either parental mAb 1 (Figures 5A-5C), parental mAb 2 (Figures 5D-5F), or bispecific antibody (Figures 5G-5I). Animals were infected with 103.5 TCID50 of SARS-CoV-2 delta on day 0. A vehicle control group (n=10, PBS) was included. Figures 5A, 5D, and 5G show Kaplan-Meier survival curves for treatment groups. Lines are slightly shifted relative to the Y-axis for improved visual presentation. Figures 5B, 5E, and 5H show % body weight change relative to day 0. Error bars represent the mean and 95% confidence interval. If mice died or were euthanized during study follow-up, the last observed body weight was carried forward. Figures 5C, 5F, and 5I: Survival dose-response curves using Probit regression. A vehicle control group was included in the modeling at a dose of 0 mg / kg. Point-by-point 95% reference confidence intervals for the predicted doses are shown. Asterisks indicate significant differences compared to the vehicle control group. [Figure 6A] Linear epitope mapping for the spike fusion peptide domains of alpha- and beta-coronaviruses. Linear epitope mapping for the spike fusion peptide 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 contain multiple smaller portions of the epitope. Epitope footprint and paratope hotspots for antibodies against the fusion peptide domain of SARS-CoV-2. CDRs of the heavy and light chains are shown. [Figure 6B]Linear epitope mapping for the spike fusion peptide domains of alpha- and beta-coronaviruses. Linear epitope mapping for the spike fusion peptide 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 contain multiple smaller portions of the epitope. Epitope footprint and paratope hotspots for antibodies against the fusion peptide domain of SARS-CoV-2. CDRs of the heavy and light chains are shown. DETAILED DESCRIPTION OF THE INVENTION

[0099] 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.

[0100] 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.

[0101] Prevention of infection is preferably achieved by administration of the antibodies disclosed herein prior to exposure to coronavirus, i.e., pre-exposure prophylaxis.

[0102] 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.

[0103] An infected individual may be asymptomatic, or alternatively, an infected individual may be symptomatic.

[0104] Preferably, post-exposure prophylaxis involves administration of the antibodies disclosed herein after exposure to the coronavirus to prevent symptomatic disease.

[0105] Preferably, post-exposure prophylaxis involves administration of the antibodies disclosed herein after exposure to the coronavirus to prevent severe disease, particularly hospitalization.

[0106] 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.

[0107] 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.

[0108] coronavirus As used herein, the term "coronavirus" includes reference to positive-sense, single-stranded RNA viruses belonging to the Coronaviridae family.

[0109] Preferably, the antibodies disclosed herein are capable of specifically binding to the fusion peptide of a coronavirus, in particular SARS-CoV-2.

[0110] Preferably, the antibodies disclosed herein are capable of specifically binding to the fusion peptide of a SARS-CoV-2 variant of concern.

[0111] Preferably, the antibodies disclosed herein are capable of neutralizing coronaviruses, particularly SARS-CoV-2.

[0112] 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.

[0113] 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."

[0114] 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."

[0115] 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.

[0116] 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 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.

[0117] 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).

[0118] Preferably, the individual is a mammal, more preferably a human.

[0119] 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.

[0120] 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.

[0121] 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.

[0122] 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 divided into subclasses or isotypes. For example, the IgG class is further divided 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.

[0123] 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 fusion peptides of spike proteins. The antibodies disclosed herein bind to conserved epitopes in the fusion peptide 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.

[0124] The antibodies disclosed herein can be used in isolated or non-isolated form.

[0125] Preferably, the compositions of the present invention comprise a single anti-coronavirus antibody disclosed herein.

[0126] Preferably, the antibodies of the invention disclosed herein are capable of cross-neutralizing coronaviruses, particularly SARS-CoV-2.

[0127] 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.

[0128] 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.

[0129] Preferably, the coronaviruses described herein include SARS-CoV-2 and variants.

[0130] 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.

[0131] Preferably, the antibodies disclosed herein are human antibodies.

[0132] 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.

[0133] 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.

[0134] 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.

[0135] 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.

[0136] Preferably, the antibodies of the invention disclosed herein are capable of cross-neutralizing coronaviruses, particularly SARS-CoV-2.

[0137] 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.

[0138] Preferably, the antibody is an IgG antibody, preferably an IgG1 antibody.

[0139] 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.

[0140] Preferably, the antibody is an IgA antibody.

[0141] 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.

[0142] Preferably, the antibody is an IgM antibody.

[0143] 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.

[0144] For the avoidance of doubt, the term "anti-coronavirus antibody" can be used interchangeably with "coronavirus antibody."

[0145] 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.

[0146] 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.

[0147] 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.

[0148] 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.

[0149] 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.

[0150] 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.

[0151] Preferably, the antibodies of the invention are administered transmucosally.

[0152] 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 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.

[0153] Preferably, the antibodies of the invention are administered intranasally.

[0154] 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.

[0155] 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.

[0156] 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.

[0157] Preferably, the antibodies of the invention are administered by oral inhalation.

[0158] 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).

[0159] 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.

[0160] 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.

[0161] 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.

[0162] Preferably, the antibodies of the invention are administered by oral pharyngeal administration.

[0163] 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.

[0164] 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.

[0165] 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.

[0166] 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.

[0167] 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).

[0168] 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.

[0169] 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.

[0170] 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.

[0171] 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.

[0172] 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.

[0173] In a preferred embodiment, the binding interaction between the binding molecule, preferably an antibody, and the fusion peptide is mediated exclusively by the variable sequences of the light and heavy chains.

[0174] The antibodies disclosed herein can specifically bind to coronavirus in attenuated or inactivated form or in live and / or infectious form.

[0175] The antibodies disclosed herein can also specifically bind to one or more fragments of a coronavirus.

[0176] The antibodies disclosed herein preferably comprise heavy chain CDR1 sequences comprising any one of the following SEQ ID NOs: 006 to 011, 013 to 018, 022 to 027, 033 to 038, 053 to 058, and 063 to 068. A CDR comprising the amino acid sequence of SEQ ID NO: 013 is particularly preferred.

[0177] The antibodies disclosed herein preferably comprise a heavy chain CDR2 sequence comprising any one of the following SEQ ID NOs: 082 to 087, 093 to 098, 115 to 120, 126 to 131, 170 to 175, and 191 to 196. A CDR comprising the amino acid sequence of SEQ ID NO: 170 is particularly preferred.

[0178] The antibodies disclosed herein preferably comprise a heavy chain CDR3 sequence comprising any one of the following SEQ ID NOs: 202 to 207, 213 to 218, 266 to 271, 277 to 282, 299 to 304, and 310 to 315. A CDR comprising the amino acid sequence of SEQ ID NO: 266 is particularly preferred.

[0179] The antibodies disclosed herein preferably comprise a light chain CDR1 sequence comprising any one of the following SEQ ID NOs: 321 to 326, 353 to 358, 364 to 369, 386 to 391, 408 to 413, and 430 to 435. A CDR comprising the amino acid sequence of SEQ ID NO: 386 is particularly preferred.

[0180] The antibodies disclosed herein preferably comprise a light chain CDR2 sequence comprising any one of the following SEQ ID NOs: 439 to 444, 450 to 455, 461 to 466, 472 to 477, 489 to 494, and 523 to 528. A CDR comprising the amino acid sequence of SEQ ID NO: 439 is particularly preferred.

[0181] The antibodies disclosed herein preferably comprise a light chain CDR3 sequence comprising any one of the following SEQ ID NOs: 534 to 539, 545 to 550, 556 to 561, 584 to 589, 613 to 618, and 634 to 639. A CDR comprising the amino acid sequence of SEQ ID NO: 584 is particularly preferred.

[0182] The antibodies disclosed herein comprise a heavy chain variable domain further comprising at least the heavy chain framework region FR1 of SEQ ID NO:642.

[0183] 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:643.

[0184] 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:644.

[0185] The antibodies disclosed herein comprise a heavy chain variable domain that further comprises at least the heavy chain framework region FR4 of SEQ ID NO:645.

[0186] In a preferred embodiment, the heavy chain variable domain of the antibody comprises a heavy chain framework region FR1 of SEQ ID NO: 642, a heavy chain framework region FR2 of SEQ ID NO: 643, a heavy chain framework region FR3 of SEQ ID NO: 644, and / or a heavy chain framework region FR4 of SEQ ID NO: 645, preferably all of the heavy chain framework regions FR1 to FR4.

[0187] 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:646.

[0188] 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:647.

[0189] 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:648.

[0190] 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:649.

[0191] In a preferred embodiment, the light chain variable domain of the antibody comprises a heavy chain framework region FR1 of SEQ ID NO: 646, a heavy chain framework region FR2 of SEQ ID NO: 647, a heavy chain framework region FR3 of SEQ ID NO: 648, and / or a light chain framework region FR4 of SEQ ID NO: 649, preferably all of the light chain framework regions FR1 to FR4.

[0192] Most preferably, the antibodies disclosed herein comprise a heavy chain variable domain having the sequence of SEQ ID NO:640 and / or a light chain variable domain having the sequence of SEQ ID NO:641.

[0193] 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: 640. 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: 641. 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).

[0194] Preferably, the antibodies disclosed herein comprise a heavy chain variable domain having the sequence of SEQ ID NO: 640 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.

[0195] Preferably, the antibodies disclosed herein comprise a light chain variable domain having the sequence of SEQ ID NO: 641 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, said amino acid insertions, deletions or substitutions for the heavy and / or light chain variable domains are not within the CDRs.

[0197] 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).

[0198] Heavy chain CDR1 region (SEQ ID NO: 001 to SEQ ID NO: 076) Sequence number 001 DTFS Sequence number 002 DTFSD Sequence number 003 DTFSDY Sequence number 004 DTFSDYR Sequence number 005 DTFSDYRI Sequence number 006 DTFSDYRIH Sequence number 007 DTFSDYRIHW Sequence number 008 DTFSDYRIHWV Sequence number 009 DTFSDYRIHWVR Sequence number 010 DTFSDYRIHWVRQ Sequence number 011 DTFSDYRIHWVRQA Sequence number 012 DYRI SEQ ID NO: 013 DYRIH Sequence number 014 DYRIHW Sequence number 015 DYRIHWV Sequence number 016 DYRIHWVR Sequence number 017 DYRIHWVRQ Sequence number 018 DYRIHWVRQA SEQ ID NO: 019 FSDY Sequence number 020 FSDYR SEQ ID NO: 021 FSDYRI SEQ ID NO: 022 FSDYRIH Sequence number 023 FSDYRIHW SEQ ID NO: 024 FSDYRIHWV Sequence number 025 FSDYRIHWVR SEQ ID NO: 026 FSDYRIHWVRQ Sequence number 027 FSDYRIHWVRQA Sequence number 028 GDTFS SEQ ID NO: 029 GDTFSD Sequence number 030 GDTFSDY Sequence number 031 GDTFSDYR SEQ ID NO: 032 GDTFSDYRI SEQ ID NO: 033 GDTFSDYRIH Sequence number 034 GDTFSDYRIHW Sequence number 035 GDTFSDYRIHWV Sequence number 036 GDTFSDYRIHWVR Sequence number 037 GDTFSDYRIHWVRQ Sequence number 038 GDTFSDYRIHWVRQA Sequence number 039 HWVR Sequence number 040 HWVRQ Sequence number 041 HWVRQA SEQ ID NO. 042 IHWV SEQ ID NO. 043 IHWVR SEQ ID NO. 044 IHWVRQ Sequence number 045 IHWVRQA SEQ ID NO. 046 RIHW SEQ ID NO. 047 RIHWV SEQ ID NO. 048 RIHWVR SEQ ID NO. 049 RIHWVRQ Sequence number 050 RIHWVRQA Sequence number 051 SDYR SEQ ID NO: 052 SDYRI SEQ ID NO: 053 SDYRIH Sequence number 054 SDYRIHW SEQ ID NO. 055 SDYRIHWV SEQ ID NO. 056 SDYRIHWVR SEQ ID NO: 057 SDYRIHWVRQ Sequence number 058 SDYRIHWVRQA SEQ ID NO. 059 TFSD Sequence number 060 TFSDY Sequence number 061 TFSDYR SEQ ID NO: 062 TFSDYRI SEQ ID NO: 063 TFSDYRIH Sequence number 064 TFSDYRIHW Sequence number 065 TFSDYRIHWV Sequence number 066 TFSDYRIHWVR Sequence number 067 TFSDYRIHWVRQ Sequence number 068 TFSDYRIHWVRQA SEQ ID NO. 069 WVRQ Sequence number 070 WVRQA SEQ ID NO. 071 YRIH SEQ ID NO. 072 YRIHW SEQ ID NO. 073 YRIHWV SEQ ID NO. 074 YRIHWVR SEQ ID NO. 075 YRIHWVRQ Sequence number 076 YRIHWVRQA Heavy chain CDR2 region (SEQ ID NO: 077 to SEQ ID NO: 196) Sequence number 077 EWMGRMNPKSGDTNFA Sequence number 078 EWMGRMNPKSGDTNFAQ Sequence number 079 EWMGRMNPKSGDTNFAQK Sequence number 080 EWMGRMNPKSGDTNFAQKF Sequence number 081 EWMGRMNPKSGDTNFAQKFQ Sequence number 082 EWMGRMNPKSGDTNFAQKFQG Sequence number 083 EWMGRMNPKSGDTNFAQKFQGR Sequence number 084 EWMGRMNPKSGDTNFAQKFQGRV Sequence number 085 EWMGRMNPKSGDTNFAQKFQGRVT Sequence number 086 EWMGRMNPKSGDTNFAQKFQGRVTM Sequence number 087 EWMGRMNPKSGDTNFAQKFQGRVTMT Sequence number 088 GRMNPKSGDTNFA Sequence number 089 GRMNPKSGDTNFAQ Sequence number 090 GRMNPKSGDTNFAQK Sequence number 091 GRMNPKSGDTNFAQKF Sequence number 092 GRMNPKSGDTNFAQKFQ Sequence number 093 GRMNPKSGDTNFAQKFQG Sequence number 094 GRMNPKSGDTNFAQKFQGR Sequence number 095 GRMNPKSGDTNFAQKFQGRV Sequence number 096 GRMNPKSGDTNFAQKFQGRVT Sequence number 097 GRMNPKSGDTNFAQKFQGRVTM Sequence number 098 GRMNPKSGDTNFAQKFQGRVTMT Sequence number 099 KSGDTNFA Sequence number 100 KSGDTNFAQ Sequence number 101 KSGDTNFAQK Sequence number 102 KSGDTNFAQKF Sequence number 103 KSGDTNFAQKFQ Sequence number 104 KSGDTNFAQKFQG Sequence number 105 KSGDTNFAQKFQGR Sequence number 106 KSGDTNFAQKFQGRV Sequence number 107 KSGDTNFAQKFQGRVT Sequence number 108 KSGDTNFAQKFQGRVTM Sequence number 109 KSGDTNFAQKFQGRVTMT SEQ ID NO: 110 LEWMGRMNPKSGDTNFA Sequence number 111 LEWMGRMNPKSGDTNFAQ SEQ ID NO: 112 LEWMGRMNPKSGDTNFAQK SEQ ID NO: 113 LEWMGRMNPKSGDTNFAQKF SEQ ID NO: 114 LEWMGRMNPKSGDTNFAQKFQ SEQ ID NO: 115 LEWMGRMNPKSGDTNFAQKFQG SEQ ID NO: 116 LEWMGRMNPKSGDTNFAQKFQGR SEQ ID NO: 117 LEWMGRMNPKSGDTNFAQKFQGRV SEQ ID NO: 118 LEWMGRMNPKSGDTNFAQKFQGRVT SEQ ID NO: 119 LEWMGRMNPKSGDTNFAQKFQGRVTM SEQ ID NO: 120 LEWMGRMNPKSGDTNFAQKFQGRVTMT SEQ ID NO: 121 MGRMNPKSGDTNFA Sequence number 122 MGRMNPKSGDTNFAQ SEQ ID NO: 123 MGRMNPKSGDTNFAQK SEQ ID NO: 124 MGRMNPKSGDTNFAQKF Sequence number 125 MGRMNPKSGDTNFAQKFQ SEQ ID NO: 126 MGRMNPKSGDTNFAQKFQG SEQ ID NO: 127 MGRMNPKSGDTNFAQKFQGR SEQ ID NO: 128 MGRMNPKSGDTNFAQKFQGRV SEQ ID NO: 129 MGRMNPKSGDTNFAQKFQGRVT Sequence number 130 MGRMNPKSGDTNFAQKFQGRVTM SEQ ID NO: 131 MGRMNPKSGDTNFAQKFQGRVTMT SEQ ID NO: 132 MNPKSGDTNFA Sequence number 133 MNPKSGDTNFAQ SEQ ID NO: 134 MNPKSGDTNFAQK SEQ ID NO: 135 MNPKSGDTNFAQKF SEQ ID NO: 136 MNPKSGDTNFAQKFQ SEQ ID NO: 137 MNPKSGDTNFAQKFQG SEQ ID NO: 138 MNPKSGDTNFAQKFQGR SEQ ID NO: 139 MNPKSGDTNFAQKFQGRV SEQ ID NO: 140 MNPKSGDTNFAQKFQGRVT SEQ ID NO: 141 MNPKSGDTNFAQKFQGRVTM SEQ ID NO: 142 MNPKSGDTNFAQKFQGRVTMT SEQ ID NO: 143 NPKSGDTNFA Sequence number 144 NPKSGDTNFAQ SEQ ID NO: 145 NPKSGDTNFAQK Sequence number 146 NPKSGDTNFAQKF Sequence number 147 NPKSGDTNFAQKFQ Sequence number 148 NPKSGDTNFAQKFQG Sequence number 149 NPKSGDTNFAQKFQGR Sequence number 150 NPKSGDTNFAQKFQGRV Sequence number 151 NPKSGDTNFAQKFQGRVT Sequence number 152 NPKSGDTNFAQKFQGRVTM SEQ ID NO: 153 NPKSGDTNFAQKFQGRVTMT SEQ ID NO: 154 PKSGDTNFA Sequence number 155 PKSGDTNFAQ SEQ ID NO: 156 PKSGDTNFAQK Sequence number 157 PKSGDTNFAQKF Sequence number 158 PKSGDTNFAQKFQ Sequence number 159 PKSGDTNFAQKFQG Sequence number 160 PKSGDTNFAQKFQGR SEQ ID NO: 161 PKSGDTNFAQKFQGRV SEQ ID NO: 162 PKSGDTNFAQKFQGRVT SEQ ID NO: 163 PKSGDTNFAQKFQGRVTM SEQ ID NO: 164 PKSGDTNFAQKFQGRVTMT SEQ ID NO: 165 RMNPKSGDTNFA Sequence number 166 RMNPKSGDTNFAQ Sequence number 167 RMNPKSGDTNFAQK Sequence number 168 RMNPKSGDTNFAQKF Sequence number 169 RMNPKSGDTNFAQKFQ Sequence number 170 RMNPKSGDTNFAQKFQG Sequence number 171 RMNPKSGDTNFAQKFQGR Sequence number 172 RMNPKSGDTNFAQKFQGRV Sequence number 173 RMNPKSGDTNFAQKFQGRVT Sequence number 174 RMNPKSGDTNFAQKFQGRVTM SEQ ID NO: 175 RMNPKSGDTNFAQKFQGRVTMT Sequence number 176 SGDTNFAQ Sequence number 177 SGDTNFAQK Sequence number 178 SGDTNFAQKF Sequence number 179 SGDTNFAQKFQ Sequence number 180 SGDTNFAQKFQG Sequence number 181 SGDTNFAQKFQGR Sequence number 182 SGDTNFAQKFQGRV Sequence number 183 SGDTNFAQKFQGRVT Sequence number 184 SGDTNFAQKFQGRVTM Sequence number 185 SGDTNFAQKFQGRVTMT SEQ ID NO: 186 WMGRMNPKSGDTNFA Sequence number 187 WMGRMNPKSGDTNFAQ Sequence number 188 WMGRMNPKSGDTNFAQK Sequence number 189 WMGRMNPKSGDTNFAQKF Sequence number 190 WMGRMNPKSGDTNFAQKFQ Sequence number 191 WMGRMNPKSGDTNFAQKFQG Sequence number 192 WMGRMNPKSGDTNFAQKFQGR Sequence number 193 WMGRMNPKSGDTNFAQKFQGRV Sequence number 194 WMGRMNPKSGDTNFAQKFQGRVT Sequence number 195 WMGRMNPKSGDTNFAQKFQGRVTM SEQ ID NO: 196 WMGRMNPKSGDTNFAQKFQGRVTMT Heavy chain CDR3 region (SEQ ID NO: 197 to SEQ ID NO: 315) SEQ ID NO: 197 ASLLIVGGFDPL SEQ ID NO: 198 ASLLIVGGFDPLD SEQ ID NO: 199 ASLLIVGGFDPLDD SEQ ID NO: 200 ASLLIVGGFDPLDDF Sequence number 201 ASLLIVGGFDPLDDFE SEQ ID NO: 202 ASLLIVGGFDPLDDFEV SEQ ID NO: 203 ASLLIVGGFDPLDDFEVW SEQ ID NO: 204 ASLLIVGGFDPLDDFEVWG SEQ ID NO: 205 ASLLIVGGFDPLDDFEVWGQ SEQ ID NO: 206 ASLLIVGGFDPLDDFEVWGQG SEQ ID NO: 207 ASLLIVGGFDPLDDFEVWGQGT SEQ ID NO: 208 CASLLIVGGFDPL SEQ ID NO: 209 CASLLIVGGFDPLD SEQ ID NO: 210 CASLLIVGGFDPLDD SEQ ID NO: 211 CASLLIVGGFDPLDDF SEQ ID NO: 212 CASLLIVGGFDPLDDFE SEQ ID NO: 213 CASLLIVGGFDPLDDFEV SEQ ID NO: 214 CASLLIVGGFDPLDDFEVW SEQ ID NO: 215 CASLLIVGGFDPLDDFEVWG SEQ ID NO: 216 CASLLIVGGFDPLDDFEVWGQ SEQ ID NO: 217 CASLLIVGGFDPLDDFEVWGQG SEQ ID NO: 218 CASLLIVGGFDPLDDFEVWGQGT SEQ ID NO: 219 GFDPLDD SEQ ID NO: 220 GFDPLDDF SEQ ID NO: 221 GFDPLDDFE SEQ ID NO: 222 GFDPLDDFEV SEQ ID NO: 223 GFDPLDDFEVW SEQ ID NO: 224 GFDPLDDFEVWG SEQ ID NO: 225 GFDPLDDFEVWGQ SEQ ID NO: 226 GFDPLDDFEVWGQG SEQ ID NO: 227 GFDPLDDFEVWGQGT SEQ ID NO: 228 GGFDPL SEQ ID NO: 229 GGFDPLD SEQ ID NO: 230 GGFDPLDD SEQ ID NO: 231 GGFDPLDDF SEQ ID NO: 232 GGFDPLDDFE SEQ ID NO: 233 GGFDPLDDFEV SEQ ID NO: 234 GGFDPLDDFEVW SEQ ID NO: 235 GGFDPLDDFEVWG SEQ ID NO: 236 GGFDPLDDFEVWGQ SEQ ID NO: 237 GGFDPLDDFEVWGQG SEQ ID NO: 238 GGFDPLDDFEVWGQGT SEQ ID NO: 239 IVGGFDPL SEQ ID NO: 240 IVGGFDPLD SEQ ID NO: 241 IVGGFDPLDD SEQ ID NO: 242 IVGGFDPLDDF SEQ ID NO: 243 IVGGFDPLDDFE SEQ ID NO: 244 IVGGFDPLDDFEV SEQ ID NO: 245 IVGGFDPLDDFEVW SEQ ID NO: 246 IVGGFDPLDDFEVWG SEQ ID NO: 247 IVGGFDPLDDFEVWGQ SEQ ID NO: 248 IVGGFDPLDDFEVWGQG SEQ ID NO: 249 IVGGFDPLDDFEVWGQGT Sequence number 250 LIVGGFDPL SEQ ID NO: 251 LIVGGFDPLD SEQ ID NO: 252 LIVGGFDPLDD SEQ ID NO: 253 LIVGGFDPLDDF SEQ ID NO: 254 LIVGGFDPLDDFE SEQ ID NO: 255 LIVGGFDPLDDFEV SEQ ID NO: 256 LIVGGFDPLDDFEVW SEQ ID NO: 257 LIVGGFDPLDDFEVWG SEQ ID NO: 258 LIVGGFDPLDDFEVWGQ SEQ ID NO: 259 LIVGGFDPLDDFEVWGQG SEQ ID NO: 260 LIVGGFDPLDDFEVWGQGT SEQ ID NO: 261 LLIVGGFDPL SEQ ID NO: 262 LLIVGGFDPLD SEQ ID NO: 263 LLIVGGFDPLDD SEQ ID NO: 264 LLIVGGFDPLDDF SEQ ID NO: 265 LLIVGGFDPLDDFE SEQ ID NO: 266 LLIVGGFDPLDDFEV SEQ ID NO: 267 LLIVGGFDPLDDFEVW SEQ ID NO: 268 LLIVGGFDPLDDFEVWG SEQ ID NO: 269 LLIVGGFDPLDDFEVWGQ SEQ ID NO: 270 LLIVGGFDPLDDFEVWGQG SEQ ID NO: 271 LLIVGGFDPLDDFEVWGQGT SEQ ID NO: 272 SLLIVGGFDPL SEQ ID NO: 273 SLLIVGGFDPLD SEQ ID NO: 274 SLLIVGGFDPLDD SEQ ID NO: 275 SLLIVGGFDPLDDF SEQ ID NO: 276 SLLIVGGFDPLDDFE SEQ ID NO: 277 SLLIVGGFDPLDDFEV SEQ ID NO: 278 SLLIVGGFDPLDDFEVW SEQ ID NO: 279 SLLIVGGFDPLDDFEVWG SEQ ID NO: 280 SLLIVGGFDPLDDFEVWGQ SEQ ID NO: 281 SLLIVGGFDPLDDFEVWGQG SEQ ID NO: 282 SLLIVGGFDPLDDFEVWGQGT SEQ ID NO: 283 VGGFDPL SEQ ID NO: 284 VGGFDPLD SEQ ID NO: 285 VGGFDPLDD SEQ ID NO: 286 VGGFDPLDDF Sequence number 287 VGGFDPLDDFE SEQ ID NO: 288 VGGFDPLDDFEV SEQ ID NO: 289 VGGFDPLDDFEVW SEQ ID NO: 290 VGGFDPLDDFEVWG SEQ ID NO: 291 VGGFDPLDDFEVWGQ SEQ ID NO: 292 VGGFDPLDDFEVWGQG SEQ ID NO: 293 VGGFDPLDDFEVWGQGT SEQ ID NO: 294 YCASLLIVGGFDPL SEQ ID NO: 295 YCASLLIVGGFDPLD SEQ ID NO: 296 YCASLLIVGGFDPLDD SEQ ID NO: 297 YCASLLIVGGFDPLDDF Sequence number 298 YCASLLIVGGFDPLDDFE SEQ ID NO: 299 YCASLLIVGGFDPLDDFEV SEQ ID NO: 300 YCASLLIVGGFDPLDDFEVW SEQ ID NO: 301 YCASLLIVGGFDPLDDFEVWG SEQ ID NO: 302 YCASLLIVGGFDPLDDFEVWGQ SEQ ID NO: 303 YCASLLIVGGFDPLDDFEVWGQG SEQ ID NO: 304 YCASLLIVGGFDPLDDFEVWGQGT SEQ ID NO: 305 YYCASLLIVGGFDPL SEQ ID NO: 306 YYCASLLIVGGFDPLD SEQ ID NO: 307 YYCASLLIVGGFDPLDD SEQ ID NO: 308 YYCASLLIVGGFDPLDDF Sequence number 309 YYCASLLIVGGFDPLDDFE SEQ ID NO: 310 YYCASLLIVGGFDPLDDFEV SEQ ID NO: 311 YYCASLLIVGGFDPLDDFEVW SEQ ID NO: 312 YYCASLLIVGGFDPLDDFEVWG SEQ ID NO: 313 YYCASLLIVGGFDPLDDFEVWGQ SEQ ID NO: 314 YYCASLLIVGGFDPLDDFEVWGQG SEQ ID NO: 315 YYCASLLIVGGFDPLDDFEVWGQGT Light chain CDR1 region (SEQ ID NO: 316 to SEQ ID NO: 435) SEQ ID NO:316 CSGTSSDVGG SEQ ID NO: 317 CSGTSSDVGGY SEQ ID NO: 318 CSGTSSDVGGYN SEQ ID NO: 319 CSGTSSDVGGYNF SEQ ID NO: 320 CSGTSSDVGGYNFV SEQ ID NO: 321 CSGTSSDVGGYNFVS Sequence number 322 CSGTSSDVGGYNFVSW SEQ ID NO: 323 CSGTSSDVGGYNFVSWY SEQ ID NO: 324 CSGTSSDVGGYNFVSWYQ Sequence number 325 CSGTSSDVGGYNFVSWYQH SEQ ID NO: 326 CSGTSSDVGGYNFVSWYQHH SEQ ID NO: 327 DVGGY SEQ ID NO: 328 DVGGYN SEQ ID NO: 329 DVGGYNF SEQ ID NO: 330 DVGGYNFV SEQ ID NO: 331 DVGGYNFVS Sequence number 332 DVGGYNFVSW SEQ ID NO: 333 DVGGYNFVSWY SEQ ID NO: 334 DVGGYNFVSWYQ Sequence number 335 DVGGYNFVSWYQH Sequence number 336 DVGGYNFVSWYQHH SEQ ID NO: 337 GTSSDVGG SEQ ID NO: 338 GTSSDVGGY Sequence number 339 GTSSDVGGYN SEQ ID NO: 340 GTSSDVGGYNF SEQ ID NO: 341 GTSSDVGGYNFV SEQ ID NO: 342 GTSSDVGGYNFVS Sequence number 343 GTSSDVGGYNFVSW SEQ ID NO: 344 GTSSDVGGYNFVSWY Sequence number 345 GTSSDVGGYNFVSWYQ Sequence number 346 GTSSDVGGYNFVSWYQH Sequence number 347 GTSSDVGGYNFVSWYQHH SEQ ID NO:348 ISCSGTSSDVGG SEQ ID NO: 349 ISCSGTSSDVGGY SEQ ID NO: 350 ISCSGTSSDVGGYN SEQ ID NO: 351 ISCSGTSSDVGGYNF SEQ ID NO: 352 ISCSGTSSDVGGYNFV SEQ ID NO: 353 ISCSGTSSDVGGYNFVS Sequence number 354 ISCSGTSSDVGGYNFVSW SEQ ID NO: 355 ISCSGTSSDVGGYNFVSWY SEQ ID NO: 356 ISCSGTSSDVGGYNFVSWYQ Sequence number 357 ISCSGTSSDVGGYNFVSWYQH Sequence number 358 ISCSGTSSDVGGYNFVSWYQHH SEQ ID NO:359 SCSGTSSDVGG SEQ ID NO: 360 SCSGTSSDVGGY SEQ ID NO: 361 SCSGTSSDVGGYN SEQ ID NO: 362 SCSGTSSDVGGYNF SEQ ID NO: 363 SCSGTSSDVGGYNFV SEQ ID NO: 364 SCSGTSSDVGGYNFVS Sequence number 365 SCSGTSSDVGGYNFVSW SEQ ID NO: 366 SCSGTSSDVGGYNFVSWY SEQ ID NO: 367 SCSGTSSDVGGYNFVSWYQ Sequence number 368 SCSGTSSDVGGYNFVSWYQH Sequence number 369 SCSGTSSDVGGYNFVSWYQHH SEQ ID NO:370 SDVGG SEQ ID NO: 371 SDVGGY SEQ ID NO:372 SDVGGYN SEQ ID NO: 373 SDVGGYNF SEQ ID NO:374 SDVGGYNFV SEQ ID NO: 375 SDVGGYNFVS Sequence number 376 SDVGGYNFVSW SEQ ID NO:377 SDVGGYNFVSWY SEQ ID NO:378 SDVGGYNFVSWYQ Sequence number 379 SDVGGYNFVSWYQH Sequence number 380 SDVGGYNFVSWYQHH SEQ ID NO: 381 SGTSSDVGG SEQ ID NO: 382 SGTSSDVGGY SEQ ID NO: 383 SGTSSDVGGYN SEQ ID NO: 384 SGTSSDVGGYNF Sequence number 385 SGTSSDVGGYNFV Sequence number 386 SGTSSDVGGYNFVS Sequence number 387 SGTSSDVGGYNFVSW SEQ ID NO: 388 SGTSSDVGGYNFVSWY Sequence number 389 SGTSSDVGGYNFVSWYQ Sequence number 390 SGTSSDVGGYNFVSWYQH Sequence number 391 SGTSSDVGGYNFVSWYQHH SEQ ID NO: 392 SSDVGG Sequence number 393 SSDVGGY Sequence number 394 SSDVGGYN Sequence number 395 SSDVGGYNF Sequence number 396 SSDVGGYNFV Sequence number 397 SSDVGGYNFVS Sequence number 398 SSDVGGYNFVSW Sequence number 399 SSDVGGYNFVSWY Sequence number 400 SSDVGGYNFVSWYQ Sequence number 401 SSDVGGYNFVSWYQH Sequence number 402 SSDVGGYNFVSWYQHH SEQ ID NO: 403 TISCSGTSSDVGG SEQ ID NO: 404 TISCSGTSSDVGGY Sequence number 405 TISCSGTSSDVGGYN SEQ ID NO: 406 TISCSGTSSDVGGYNF Sequence number 407 TISCSGTSSDVGGYNFV Sequence number 408 TISCSGTSSDVGGYNFVS Sequence number 409 TISCSGTSSDVGGYNFVSW SEQ ID NO: 410 TISCSGTSSDVGGYNFVSWY SEQ ID NO: 411 TISCSGTSSDVGGYNFVSWYQ Sequence number 412 TISCSGTSSDVGGYNFVSWYQH SEQ ID NO: 413 TISCSGTSSDVGGYNFVSWYQHH SEQ ID NO:414 TSSDVGG SEQ ID NO: 415 TSSDVGGY SEQ ID NO: 416 TSSDVGGYN SEQ ID NO: 417 TSSDVGGYNF SEQ ID NO: 418 TSSDVGGYNFV Sequence number 419 TSSDVGGYNFVS Sequence number 420 TSSDVGGYNFVSW SEQ ID NO:421 TSSDVGGYNFVSWY SEQ ID NO:422 TSSDVGGYNFVSWYQ Sequence number 423 TSSDVGGYNFVSWYQH Sequence number 424 TSSDVGGYNFVSWYQHH SEQ ID NO: 425 VTISCSGTSSDVGG SEQ ID NO: 426 VTISCSGTSSDVGGY SEQ ID NO: 427 VTISCSGTSSDVGGYN SEQ ID NO: 428 VTISCSGTSSDVGGYNF SEQ ID NO: 429 VTISCSGTSSDVGGYNFV SEQ ID NO: 430 VTISCSGTSSDVGGYNFVS Sequence number 431 VTISCSGTSSDVGGYNFVSW SEQ ID NO: 432 VTISCSGTSSDVGGYNFVSWY SEQ ID NO: 433 VTISCSGTSSDVGGYNFVSWYQ Sequence number 434 VTISCSGTSSDVGGYNFVSWYQH Sequence number 435 VTISCSGTSSDVGGYNFVSWYQHH Light chain CDR2 region (SEQ ID NO: 436 to SEQ ID NO: 528) SEQ ID NO: 436 EVTK SEQ ID NO: 437 EVTKR SEQ ID NO: 438 EVTKRP SEQ ID NO: 439 EVTKRPS SEQ ID NO: 440 EVTKRPSG SEQ ID NO: 441 EVTKRPSGV SEQ ID NO: 442 EVTKRPSGVP SEQ ID NO: 443 EVTKRPSGVPD SEQ ID NO: 444 EVTKRPSGVPDR SEQ ID NO: 445 ILIYEV SEQ ID NO: 446 ILIYEVT SEQ ID NO: 447 ILIYEVTK SEQ ID NO: 448 ILIYEVTKR SEQ ID NO: 449 ILIYEVTKRP Sequence number 450 ILIYEVTKRPS SEQ ID NO: 451 ILIYEVTKRPSG SEQ ID NO: 452 ILIYEVTKRPSGV SEQ ID NO: 453 ILIYEVTKRPSGVP SEQ ID NO: 454 ILIYEVTKRPSGVPD SEQ ID NO: 455 ILIYEVTKRPSGVPDR SEQ ID NO: 456 IYEV SEQ ID NO: 457 IYEVT SEQ ID NO: 458 IYEVTK SEQ ID NO: 459 IYEVTKR Sequence number 460 IYEVTKRP SEQ ID NO: 461 IYEVTKRPS SEQ ID NO: 462 IYEVTKRPSG Sequence number 463 IYEVTKRPSGV SEQ ID NO: 464 IYEVTKRPSGVP SEQ ID NO: 465 IYEVTKRPSGVPD SEQ ID NO: 466 IYEVTKRPSGVPDR SEQ ID NO. 467 KILIYEV SEQ ID NO: 468 KILIYEVT SEQ ID NO: 469 KILIYEVTK SEQ ID NO: 470 KILIYEVTKR SEQ ID NO: 471 KILIYEVTKRP SEQ ID NO: 472 KILIYEVTKRPS SEQ ID NO: 473 KILIYEVTKRPSG SEQ ID NO: 474 KILIYEVTKRPSGV SEQ ID NO: 475 KILIYEVTKRPSGVP SEQ ID NO: 476 KILIYEVTKRPSGVPD SEQ ID NO: 477 KILIYEVTKRPSGVPDR SEQ ID NO: 478 KRPS SEQ ID NO: 479 KRPSG Sequence number 480 KRPSGV SEQ ID NO: 481 KRPSGVP SEQ ID NO:482 KRPSGVPD SEQ ID NO:483 KRPSGVPDR SEQ ID NO: 484 LIYEV Sequence number 485 LIYEVT Sequence number 486 LIYEVTK Sequence number 487 LIYEVTKR Sequence number 488 LIYEVTKRP Sequence number 489 LIYEVTKRPS SEQ ID NO: 490 LIYEVTKRPSG Sequence number 491 LIYEVTKRPSGV SEQ ID NO: 492 LIYEVTKRPSGVP SEQ ID NO: 493 LIYEVTKRPSGVPD SEQ ID NO: 494 LIYEVTKRPSGVPDR SEQ ID NO: 495 PSGV SEQ ID NO: 496 PSGVP SEQ ID NO: 497 PSGVPD SEQ ID NO:498 PSGVPDR SEQ ID NO: 499 RPSG Sequence number 500 RPSGV SEQ ID NO:501 RPSGVP SEQ ID NO:502 RPSGVPD SEQ ID NO:503 RPSGVPDR SEQ ID NO: 504 TKRP Sequence number 505 TKRPS SEQ ID NO:506 TKRPSG Sequence number 507 TKRPSGV SEQ ID NO:508 TKRPSGVP SEQ ID NO:509 TKRPSGVPD SEQ ID NO: 510 TKRPSGVPDR Sequence number 511 VTKR SEQ ID NO: 512 VTKRP SEQ ID NO: 513 VTKRPS SEQ ID NO:514 VTKRPSG SEQ ID NO. 515 VTKRPSGV SEQ ID NO: 516 VTKRPSGVP SEQ ID NO:517 VTKRPSGVPD SEQ ID NO:518 VTKRPSGVPDR SEQ ID NO:519 YEVT SEQ ID NO:520 YEVTK SEQ ID NO: 521 YEVTKR SEQ ID NO:522 YEVTKRP SEQ ID NO:523 YEVTKRPS SEQ ID NO:524 YEVTKRPSG SEQ ID NO: 525 YEVTKRPSGV SEQ ID NO:526 YEVTKRPSGVP SEQ ID NO:527 YEVTKRPSGVPD SEQ ID NO:528 YEVTKRPSGVPDR Light chain CDR3 region (SEQ ID NOs: 529 to 639) SEQ ID NO:529 ADYYCSSYGG SEQ ID NO: 530 ADYYCSSYGGT SEQ ID NO:531 ADYYCSSYGGTN SEQ ID NO: 532 ADYYCSSYGGTNN SEQ ID NO:533 ADYYCSSYGGTNNL SEQ ID NO:534 ADYYCSSYGGTNNLL Sequence number 535 ADYYCSSYGGTNNLLF SEQ ID NO:536 ADYYCSSYGGTNNLLFG SEQ ID NO:537 ADYYCSSYGGTNNLLFGG SEQ ID NO:538 ADYYCSSYGGTNNLLFGGG SEQ ID NO: 539 ADYYCSSYGGTNNLLFGGGT SEQ ID NO:540 CSSYGG SEQ ID NO:541 CSSYGGT SEQ ID NO:542 CSSYGGTN SEQ ID NO:543 CSSYGGTNN SEQ ID NO:544 CSSYGGTNNL SEQ ID NO: 545 CSSYGGTNNLL Sequence number 546 CSSYGGTNNLLF SEQ ID NO:547 CSSYGGTNNLLFG SEQ ID NO:548 CSSYGGTNNLLFGG SEQ ID NO: 549 CSSYGGTNNLLFGGG Sequence number 550 CSSYGGTNNLLFGGGT SEQ ID NO:551 DYYCSSYGG SEQ ID NO: 552 DYYCSSYGGT Sequence number 553 DYYCSSYGGTN Sequence number 554 DYYCSSYGGTNN Sequence number 555 DYYCSSYGGTNNL SEQ ID NO: 556 DYYCSSYGGTNNLL Sequence number 557 DYYCSSYGGTNNLLF Sequence number 558 DYYCSSYGGTNNLLFG Sequence number 559 DYYCSSYGGTNNLLFGG Sequence number 560 DYYCSSYGGTNNLLFGGG SEQ ID NO: 561 DYYCSSYGGTNNLLFGGGT SEQ ID NO: 562 GGTN SEQ ID NO: 563 GGTNN SEQ ID NO:564 GGTNNL SEQ ID NO: 565 GGTNNLL SEQ ID NO:566 GGTNNLLF SEQ ID NO:567 GGTNNLLFG SEQ ID NO:568 GGTNNLLFGG SEQ ID NO: 569 GGTNNLLFGGG SEQ ID NO: 570 GGTNNLLFGGGT SEQ ID NO:571 GTNN Sequence number 572 GTNNL SEQ ID NO:573 GTNNLL Sequence number 574 GTNNLLF Sequence number 575 GTNNLLFG SEQ ID NO:576 GTNNLLFGG Sequence number 577 GTNNLLFGGG SEQ ID NO:578 GTNNLLFGGGT SEQ ID NO:579 SSYGG SEQ ID NO:580 SSYGGT SEQ ID NO:581 SSYGGTN SEQ ID NO:582 SSYGGTNN SEQ ID NO:583 SSYGGTNNL SEQ ID NO:584 SSYGGTNNLL Sequence number 585 SSYGGTNNLLF SEQ ID NO:586 SSYGGTNNLLFG SEQ ID NO:587 SSYGGTNNLLFGG SEQ ID NO:588 SSYGGTNNLLFGGG SEQ ID NO:589 SSYGGTNNLLFGGGT SEQ ID NO:590 SYGG SEQ ID NO:591 SYGGT SEQ ID NO:592 SYGGTN SEQ ID NO:593 SYGGTNN SEQ ID NO:594 SYGGTNNL SEQ ID NO:595 SYGGTNNLL SEQ ID NO:596 SYGGTNNLLF SEQ ID NO:597 SYGGTNNLLFG SEQ ID NO:598 SYGGTNNLLFGG Sequence number 599 SYGGTNNLLFGGG Sequence number 600 SYGGTNNLLFGGGT Sequence number 601 TNNL SEQ ID NO: 602 TNNLL Sequence number 603 TNNLLF Sequence number 604 TNNLLFG Sequence number 605 TNNLLFGG Sequence number 606 TNNLLFGGG Sequence number 607 TNNLLFGGGT Sequence number 608 YCSSYGG SEQ ID NO: 609 YCSSYGGT Sequence number 610 YCSSYGGTN SEQ ID NO: 611 YCSSYGGTNN SEQ ID NO: 612 YCSSYGGTNNL SEQ ID NO: 613 YCSSYGGTNNLL Sequence number 614 YCSSYGGTNNLLF SEQ ID NO: 615 YCSSYGGTNNLLFG SEQ ID NO: 616 YCSSYGGTNNLLFGG SEQ ID NO: 617 YCSSYGGTNNLLFGGG SEQ ID NO: 618 YCSSYGGTNNLLFGGGT SEQ ID NO: 619 YGGT Sequence number 620 YGGTN SEQ ID NO: 621 YGGTNN SEQ ID NO: 622 YGGTNNL SEQ ID NO: 623 YGGTNNLL Sequence number 624 YGGTNNLLF SEQ ID NO: 625 YGGTNNLLFG SEQ ID NO: 626 YGGTNNLLFGG SEQ ID NO: 627 YGGTNNLLFGGG SEQ ID NO: 628 YGGTNNLLFGGGT SEQ ID NO: 629 YYCSSYGG SEQ ID NO: 630 YYCSSYGGT Sequence number 631 YYCSSYGGTN Sequence number 632 YYCSSYGGTNN Sequence number 633 YYCSSYGGTNNL SEQ ID NO: 634 YYCSSYGGTNNLL Sequence number 635 YYCSSYGGTNNLLF Sequence number 636 YYCSSYGGTNNLLFG SEQ ID NO: 637 YYCSSYGGTNNLLFGG SEQ ID NO: 638 YYCSSYGGTNNLLFGGG Sequence number 639 YYCSSYGGTNNLLFGGGT Heavy Chain Variable Domain (SEQ ID NO: 640) QMQLMQSGAEVKKPGASVTVSCKASGDTFSDYRIHWVRQAPGQGLEWMGRMNPKSGDTNFAQKFQGRVTMTRDMSINTAYMTLSGLTFDDTALYYCASLLIVGGDPLDDFEVWGQGTMVTISS Light Chain Variable Domain (SEQ ID NO: 641) QSALTQPPSASGSPGQSVTISCSGTSSDVGGYNFVSWYQHHPGKAPKILIYEVTKRPSGVPDRFSGSKSGNTASLTVSGLQAEDEADYYCSSYGGTNNLLFGGGTKLTVL Heavy chain FR1 region (SEQ ID NO: 642) Sequence number 642 QMQLMQSGAEVKKPGASVTVSCKASGDTFS Heavy chain FR2 region (SEQ ID NO: 643) Sequence number 643 WVRQAPGQGLEWMG Heavy chain FR3 region (SEQ ID NO: 644) SEQ ID NO: 644 RVTMTRDMSINTAYMTLSGLTFDDTALYYCAS Heavy chain FR4 region (SEQ ID NO: 645) SEQ ID NO: 645 WGQGTMVTISS Light chain FR1 region (SEQ ID NO: 646) SEQ ID NO: 646 QSALTQPPSASGSPGQSVTISC Light chain FR2 region (SEQ ID NO: 647) SEQ ID NO: 647 WYQHHPGKAPKILIY Light chain FR3 region (SEQ ID NO: 648) SEQ ID NO: 648 GVPDRFSGSKSGNTASLTVSGLQAEDEADYYC Light chain FR4 region (SEQ ID NO: 649) SEQ ID NO: 649 FGGGTKLTVL Heavy chain CDR1 region (SEQ ID NO: 650) SEQ ID NO: 650 YFYLH Heavy chain CDR2 region (SEQ ID NO: 651) Sequence number 651 IINPRGDGTRYAQKFQG Heavy chain CDR3 region (SEQ ID NO: 652) SEQ ID NO: 652 GADHGAFDI Light chain CDR1 region (SEQ ID NO: 653) SEQ ID NO: 653 RASQSVRRNYFA Light chain CDR2 region (SEQ ID NO: 654) SEQ ID NO: 654 DASTRAT Light chain CDR3 region (SEQ ID NO: 655) SEQ ID NO: 655 QQYDSSPPMYI Heavy chain CDR1 region (SEQ ID NO: 656) Sequence number 656 GYAMH Heavy chain CDR2 region (SEQ ID NO: 657) Sequence number 657 VISRDARNKYYADSVKG Heavy chain CDR3 region (SEQ ID NO: 658) SEQ ID NO: 658 LIIPGITEPGSPDALDI Light chain CDR1 region (SEQ ID NO: 659) Sequence number 659 RASQDISKWLA Light chain CDR2 region (SEQ ID NO: 660) SEQ ID NO: 660 AASSLQS Light chain CDR3 region (SEQ ID NO: 661) Sequence number 661 QQASSFPWSIT Heavy chain CDR1 region (SEQ ID NO: 662) SEQ ID NO: 662 SHYMH Heavy chain CDR2 region (SEQ ID NO: 663) Sequence number 663 IINPSGSGTAYGQKFQG Heavy chain CDR3 region (SEQ ID NO: 664) SEQ ID NO: 664 GSGGLFAY Light chain CDR1 region (SEQ ID NO: 665) SEQ ID NO: 665 RASQIVRSNYLA Light chain CDR2 region (SEQ ID NO: 666) SEQ ID NO: 666 GASSRAT Light chain CDR3 region (SEQ ID NO: 667) Sequence number 667 LQYDSSPPTYI Heavy chain CDR1 region (SEQ ID NO: 668) SEQ ID NO: 668 SYYMH Heavy chain CDR2 region (SEQ ID NO: 669) Sequence number 669 LITPSGDDTYYAQRFQG Heavy chain CDR3 region (SEQ ID NO: 670) SEQ ID NO: 670 MSRAGGFDV Light chain CDR1 region (SEQ ID NO: 671) SEQ ID NO: 671 RASQSITGRYLA Light chain CDR2 region (SEQ ID NO: 672) SEQ ID NO: 672 GESSRVT Light chain CDR3 region (SEQ ID NO: 673) Sequence number 673 QHFASSPPTYT Heavy chain (SEQ ID NO: 674) SEQ ID NO: 674 QEQLVQSGAEVKKPGASVKVSCKSSGFTFSYFYLHWVRQAPGQGLEWMGIINPRGDGTRYAQKFQGRVTMTRDASTGTLYMELRSLRSEDTAVYYCARGADHGAFDIWGQGTM VTVSSASVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECD KTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEK TISKAKGQPREPQVCTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK Light chain (SEQ ID NO: 675) SEQ ID NO: 675 EIVLTQSPGTLSLSPGERATLSCRASQSVRRNYFAWYQQKRGQAPRLLIYDASTRATGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQYDSSPPMYIFGQGTKL EIKSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSC Arm heavy chain (SEQ ID NO: 676) SEQ ID NO: 676 QMQLMQSGAEVKKPGASVTVSCKASGDTFSDYRIHWVRQAPGQGLEWMGRMNPKSGDTNFAQKFQGRVTMTRDMSINTAYMTLSGLTFDDTALYYCASLLIVGGFDPLDDFEV WGQGTMVTISSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSC DKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIE KTISKAKGQPREPQVYTLPPCRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK Arm light chain (SEQ ID NO: 677) SEQ ID NO: 677 QSALTQPPSASGSPGQSVTISCSGTSSDVGGYNFVSWYQHHPGKAPKILIYEVTKRPSGVPDRFSGSKSGNTASLTVSGLQAEDEADYYCSSYGGTNNLLFGGGTKLT VLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS Heavy chain arm (SEQ ID NO: 678) SEQ ID NO: 678 QEQLVQSGAEVKKPGASVKVSCKSSGFTFSYFYLHWVRQAPGQGLEWMGIINPRGDGTRYAQKFQGRVTMTRDASTGTLYMELRSLRSEDTAVYYCARGADHGAFDIWGQGT MVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKT HTCPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKT ISKAKGQPREPQVYTLPPCRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK Light chain arm (SEQ ID NO: 679) SEQ ID NO: 679 EIVLTQSPGTLSLSPGERATLSCRASQSVRRNYFAWYQQKRGQAPRLLIYDASTRATGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQYDSSPPMYIFGQGTKLE IKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC Arm heavy chain (SEQ ID NO: 680) SEQ ID NO: 680 QMQLMQSGAEVKKPGASVTVSCKASGDTFSDYRIHWVRQAPGQGLEWMGRMNPKSGDTNFAQKFQGRVTMTRDMSINTAYMTLSGLTFDDTALYYCASLLIVGGFDPLDDFEVW GQGTMVTISGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTEC SDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALAPIE KTISKAKGQPREPQVCTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK Arm light chain (SEQ ID NO: 681) SEQ ID NO: 681 QSALTQPPSASGSPGQSVTISCSGTSSDVGGYNFVSWYQHHPGKAPKILIYEVTKRPSGVPDRFSGSKSGNTASLTVSGLQAEDEADYYCSSYGGTNNLLFGGGTKL TVLSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSC Arm heavy chain (SEQ ID NO: 682) SEQ ID NO: 682 QSALTQPPSASGSPGQSVTISCSGTSSDVGGYNFVSWYQHHPGKAPKILIYEVTKRPSGVPDRFSGSKSGNTASLTVSGLQAEDEADYYCSSYGGTNNLLFGGGTKLTVL GQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECSDKTHT CPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTI SKAKGQPREPQVCTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK Light chain (SEQ ID NO: 683) SEQ ID NO: 683 QMQLMQSGAEVKKPGASVTVSCKASGDTFSDYRIHWVRQAPGQGLEWMGRMNPKSGDTNFAQKFQGRVTMTRDMSINTAYMTLSGLTFDDTALYYCASLLIVGGFDPLDDFEV WGQGTMVTISSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSC Heavy chain variable domain arm (SEQ ID NO: 684) SEQ ID NO: 684 QVQLVESGGGVVQPGRSLRLSCAASGLTFSGYAMHWVRQAPGKGLEWVAVISRDAR NKYYADSVKGRFTISRDNSKKTVYLEMNSLRVEDTAVYYCAILIIPGITEPGSPDALDI WGQGTMVSVSS Light Chain Variable Domain (SEQ ID NO: 685) SEQ ID NO: 685 DIQMTQSPSSMSASVGDRVTITCRASQDISKWLAWYQQRPGKAPKLLIYAASSLQSGV PSRFSGSGSGTDFTLTISSLQPEDFATYYCQQASSFPWSITFGQGTRLEIR Heavy chain variable domain arm (SEQ ID NO: 686) SEQ ID NO: 686 QEQLVQSGAEVKKPGASVKVSCKSSGFTFSYFYLHWVRQAPGQGLEWMGIINPRGD GTRYAQKFQGRVTMTRDASTGTLYMELRSLRSEDTAVYYCARGADHGAFDIWGQGT MVTVSS Light Chain Variable Domain (SEQ ID NO: 687) SEQ ID NO: 687 EIVLTQSPGTLSLSPGERATLSCRASQSVRRNYFAWYQQKRGQAPRLLIYDASTRATG IPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQYDSSPPMYIFGQGTKLEIK Heavy chain variable domain arm (SEQ ID NO: 688) SEQ ID NO: 688 QVQLVQSGAEVKKPAASVKVSCKASGDTFSSHYMHWVRQAPGQGPEWMGIINPSGS GTAYGQKFQGRLTMTRDTSTSTVYMELSSLTSDDTAVYYCGGGSGGLFAYWGQGTL VTVSS Light Chain Variable Domain (SEQ ID NO: 689) SEQ ID NO: 689 EIVLTQSPGTLSLSPGERATLSCRASQIVRSNYLAWYQQKPGQAPRLLIYGASSRATGT PDRFSGGGSGTDFTLTINRLEPEDFAVYYCLQYDSSPPTYIFGQGTKLEIK [Example]

[0199] Example 1 Antibodies according to the invention (intraperitoneal) 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: 013, heavy chain CDR2 as SEQ ID NO: 170, heavy chain CDR3 as SEQ ID NO: 266, light chain CDR1 as SEQ ID NO: 386, light chain CDR2 as SEQ ID NO: 439, and light chain CDR3 as SEQ ID NO: 584 after systemic intraperitoneal administration in a SARS-CoV-2 Delta K18 hACE2 Tg mouse model.

[0200] On day -1 of the study, animals (n=10 per group) were treated intraperitoneally with a range of test monoclonal antibodies (10-0.2 mg / kg) or PBS control as vehicle. On day 0 of the study, animals were treated with 10 3.5 TCID 50 were loaded with the SARS-CoV-2 delta variant.

[0201] Materials and Methods The monoclonal antibody according to the present invention was buffered with phosphate buffered saline (PBS) and diluted with PBS to a final concentration for administration (10-0.2 mg / kg in 200 μL), thereby preparing a monoclonal antibody dilution.

[0202] Monoclonal antibody dilutions were made at various concentrations, such that administration of 200 μL relative to the average body weight per cage for the dosing group on day -5 resulted in monoclonal antibody doses ranging from 10 mg / kg, 5 mg / kg, 1.7 mg / kg, 0.5 mg / kg to 0.2 mg / kg.

[0203] Antibodies were stored at -85°C ± 10°C, diluents were stored at 4°C until use, and the temperature of the storage units was monitored.

[0204] 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.

[0205] 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 16–21 g on day −5. Sixty mice, 8–9 weeks old on the day of test material administration, were used; all were female. Ten animals were assigned to six treatment groups based on their weight on day −5 to create groups with similar average body weights. 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).

[0206] 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 IP at doses of 0.5 mg / kg or higher. A total of 60 mice, 8-9 weeks old at the time of administration of the test material, were assigned to six experimental groups according to Table 1 (see below). Mice were allowed a period of more than 3 days for acclimatization.

[0207] [Table 1]

[0208] Female SARS-CoV-2 DeltaK18 hACE2 mice were treated with antibody at doses ranging from 10 mg / kg to 0.2 mg / kg, based on the average body weight per cage, via the intraperitoneal route. On day 0, all mice received a lethal dose (10 3.5 TCID 50) and monitored for weight loss and mortality until the study was terminated on day 14.

[0209] Antibody administration Upon arrival, the test antibodies were stored at −80° C. Appropriate doses according to the treatment schedule (Table 1) were prescribed assuming an average body weight per cage.

[0210] 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.

[0211] Virus administration The viral material was stored at -80°C and kept at -10°C 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.

[0212] Laboratory analysis The actual dose of virus administered was verified by back titrating the inoculum and titrating replicate samples on Vero cells.

[0213] Final Inspection Mice were euthanized by cervical dislocation at the end of the study on day 14. No complete necropsy was performed.

[0214] Data analysis and statistical methods Survival fraction, survival time, and body weight change (area under the curve) at day 14 were compared with the control group using Fisher's exact test, log-rank, and Welch's t-test, respectively. All groups were compared with the vehicle (PBS solution) control group. P values ​​were adjusted according to Bonferroni (for two comparisons with vehicle), following a stepwise approach (starting with the highest antibody dose and conditionally testing lower doses if the previous step was statistically significant).

[0215] Statistical analysis was performed using R, and statistical significance was set at α = 0.05.

[0216] survival rate Prophylactic treatment with test antibody at 0.5 mg / kg or higher provided a statistically significant increase in survival compared to vehicle (see Figure 1). Animals treated with 0.5 mg / kg had a 60% survival rate, with 1.7 and 5 mg / kg test antibody having a 100% survival rate, and 10 mg / kg having a 90% survival rate. Meanwhile, the control group showed 0% survival at day 7, with a median survival of 6 days.

[0217] Prophylactic treatment with 0.2 mg / kg or greater of the test antibody resulted in a significant improvement in survival.

[0218] 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.

[0219] conclusion In this lethal SARS-CoV-2 DeltaK18 hACE2 Tg mouse model, prophylactic intraperitoneal administration of test antibody at 0.5 mg / kg or higher provided significantly improved survival and reduced weight loss compared to vehicle, while all animals in the control group died.

[0220] Example 2 Antibodies according to the invention (intranasal) The aim of this study was to evaluate the pre-exposure efficacy of a monoclonal antibody according to the invention after systemic intranasal administration in a SARS-CoV-2 delta K18 hACE2 Tg mouse model.

[0221] On day -1 of the study, animals (n=10 per group) were treated with a range of intranasal doses of test monoclonal antibodies (10-0.2 mg / kg) or PBS control. On day 0 of the study, animals were treated with 10 3.5 TCID 50 were loaded with the SARS-CoV-2 delta variant.

[0222] Materials and Methods The monoclonal antibodies according to the invention, buffered in PBS, were diluted in PBS to the final concentration for administration (10-0.2 mg / kg in 50 μL).

[0223] Monoclonal antibody dilutions were made at various concentrations so that administration of 50 μL relative to the average body weight per cage in the dosing group resulted in antibody doses ranging from 10 mg / kg, 5 mg / kg, 1.7 mg / kg, 0.5 mg / kg to 0.2 mg / kg.

[0224] Antibodies were stored at -85°C ± 10°C, diluents were stored at 4°C until use, and the temperature of the storage units was monitored.

[0225] 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.

[0226] animal The SARS-CoV-2 Delta K18 hACE2 Tg mouse model was used, weighing approximately 16–22 g on the day of test material administration. Sixty mice, 8–9 weeks old on the day of test material administration, were used; all were female. Ten animals were assigned to six treatment groups based on their body weight on day -5 to create groups with similar body weights. 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, 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.

[0227] 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, 8-9 weeks old at the time of dosing with the test material, were transferred to the animal facility and assigned to six experimental groups according to Table 2 (see below). Mice were given treatment for a period of 3 days.

[0228] [Table 2]

[0229] Female SARS-CoV-2 DeltaK18 hACE2 mice were treated with antibody at doses of 10 mg / kg to 0.2 mg / kg per cage, based on the average group weight, via the intranasal route. On day 0, all mice received a lethal dose (10 3.5 TCID 50) and monitored for survival and weight loss until the study was terminated on day 14.

[0230] Antibody administration Upon arrival, the test antibodies were stored at −80° C. Appropriate doses according to the treatment schedule (Table 2) were prescribed based on the average weight per cage.

[0231] 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.

[0232] Virus administration The viral material was stored at -80°C and thawed prior to 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.

[0233] Laboratory analysis The administered virus dose was verified by back titrating the inoculum and titrating replicate samples on Vero cells.

[0234] Final Inspection Mice were euthanized by cervical dislocation at the end of the study on day 14. No complete necropsy was performed.

[0235] Data analysis and statistical methods Survival fraction, survival time, and body weight change (area under the curve) at day 14 were compared with corresponding control groups using Fisher's exact test, log-rank, and Welch's t-test, respectively. All groups were compared to the vehicle (PBS solution) control group. P values ​​were adjusted according to Bonferroni (for two comparisons with vehicle), following a stepwise approach (starting with the highest antibody dose and conditionally testing lower doses if the previous step was statistically significant).

[0236] Statistical analysis was performed using R, and statistical significance was set at α = 0.05.

[0237] survival rate Prophylactic treatment with test antibody at 0.5 mg / kg or higher provided a statistically significant increase in survival compared to vehicle controls (see Figure 2), while the control group showed 0% survival at day 7, with a median survival of 6 days.

[0238] Prophylactic treatment with test antibody at doses above 0.2 mg / kg resulted in a significant improvement in survival.

[0239] 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 test antibody at greater than 0.2 g / kg resulted in a significant reduction in weight loss compared to the control group.

[0240] conclusion In this lethal SARS-CoV-2 DeltaK18 hACE2 Tg mouse model, prophylactic intranasal administration of test antibodies at 0.5 mg / kg or higher significantly improved survival and reduced weight loss compared to vehicle. In contrast, all animals in the control group had died by day 7.

[0241] Example 3 Antibodies according to the present invention The objective of this study is to evaluate the pre- and post-exposure efficacy of a monoclonal antibody according to the invention having a heavy chain CDR1 as SEQ ID NO: 013, a heavy chain CDR2 as SEQ ID NO: 170, a heavy chain CDR3 as SEQ ID NO: 266, a light chain CDR1 as SEQ ID NO: 386, a light chain CDR2 as SEQ ID NO: 439, and a light chain CDR3 as SEQ ID NO: 584 after intranasal administration in an NHP model in which each subject has been pre-screened for the absence of serum antibodies against the SARS-CoV-2 virus.

[0242] NHP subjects are treated with an intranasal dose of a composition comprising the antibody described above. The fixed, or nominal, dose of antibody administered to each NHP subject is 0.01 mg to 10 mg (e.g., 0.01 to 2.0 mg ("low dose") or 2.0 to 10 mg ("high dose") or a control (e.g., vehicle only) on day -1 of the study.

[0243] Materials and Methods Monoclonal antibodies according to the invention are provided in sodium acetate buffer (e.g., 20 mM sodium acetate, 75 mM sodium chloride, 5% sucrose, pH 5.5) and diluted to a final concentration for administration ranging from a nominal dose of 0.01 mg to 2.0 mg or 2.0 to 10 mg, administered in 50 μL to 500 μL per nostril.

[0244] NHP subjects assigned to the control group (sodium acetate buffer) will receive vehicle (e.g., 20 mM sodium acetate, 75 mM sodium chloride, 5% sucrose, pH 5.5), administered as 50 µL–500 µL per nostril.

[0245] All NHPs will receive an intranasal challenge with SARS-CoV-2 on day 0.

[0246] Samples will be collected during the study, and sample types will include nasal sampling (e.g., nasal swabs, washes, or scrapes), serum and / or bronchoalveolar lavage (BAL) samples. Sampling periods will encompass the study period within the time frame of Day -1 to end of study (follow-up period of 7 to 21 days).

[0247] Samples are analyzed for viral quantification (RT-PCR and / or TCID 50 The antibodies are analyzed for antibody titer and neutralization titer by ELISA, HPLC, or similar methods, and in vitro neutralization assays (e.g., pseudovirion neutralization).

[0248] animal NHP subjects are used at a weight of 3-12 kg (e.g., 5 kg per animal) at the start of the study. Four to 12 animals are assigned to each treatment group.

[0249] Study design The dose levels of monoclonal antibodies applied in this example are based on maximum doses and lower doses based on allometric calculations derived from mouse studies.

[0250] [Table 3]

[0251] NHP subjects will receive intranasal administration of a composition containing an antibody at a dose (nominal dose) of 0.01 mg to 2.0 mg or 2.0 mg to 10 mg. On day 0, all NHP subjects will be challenged with a dose of SARS-CoV-2 and will be observed for clinical symptoms (e.g., weight loss and / or temperature) and virological measures until the end of the study (e.g., 7 to 21 days).

[0252] Antibody administration Test antibody or vehicle alone is administered (eg, by pipette or spray) to each nostril using a volume of 50 μL to 500 μL per nostril according to the treatment schedule (Table 3).

[0253] Virus administration The viral material is thawed prior to administration. Once thawed, the material is diluted and each animal receives 50 μL to 500 μL of viral solution per nostril. NHP subjects are administered 10 2 ~10 8 TCID 50 of SARS-CoV-2 (e.g., 10 5 TCID 50 They are infected with SARS-CoV-2.

[0254] Data analysis and statistical methods Clinical symptoms (eg, weight loss and temperature) and virological measures are used to compare treated and control groups.

[0255] Results - Clinical symptoms Prophylactic treatment with low and high dose antibody administration provides a reduction in clinical symptom measures compared to control groups.

[0256] Results – Virological Measurements Prophylactic treatment with low and high doses of antibody administration provides a reduction in virological measurements compared to the control group, with all control NHPs showing a viral load in samples following SARS-CoV-2 challenge.

[0257] conclusion In this SARS-CoV-2 NHP model, prophylactic intranasal administration of 2.0-10 mg (high dose) and 0.01-2.0 mg (low dose) of a test antibody according to the invention provides a reduction in virological measures and / or a reduction in clinical symptoms compared to control NHPs.

[0258] Example 4 Compositions comprising antibodies according to the present invention The objective of this study is to evaluate the pre- and post-exposure efficacy of a composition comprising a first monoclonal antibody according to the invention ("first antibody") having a heavy chain CDR1 as SEQ ID NO: 013, a heavy chain CDR2 as SEQ ID NO: 170, a heavy chain CDR3 as SEQ ID NO: 266, a light chain CDR1 as SEQ ID NO: 386, a light chain CDR2 as SEQ ID NO: 439, and a light chain CDR3 as SEQ ID NO: 584, and a second monoclonal antibody according to the invention ("second antibody") having a heavy chain CDR1 as SEQ ID NO: 656, a heavy chain CDR2 as SEQ ID NO: 657, a heavy chain CDR3 as SEQ ID NO: 658, a light chain CDR1 as SEQ ID NO: 659, a light chain CDR2 as SEQ ID NO: 660, and a light chain CDR3 as SEQ ID NO: 661, following intranasal administration in an NHP model in which each subject has been pre-screened for the absence of serum antibodies to the SARS-CoV-2 virus.

[0259] NHP subjects are treated with an intranasal dose of a composition containing the above-described antibodies (either a single antibody or a combination of both antibodies). The fixed, or nominal, dose of antibody administered to each NHP subject is 0.003 mg to 5 mg or a control (e.g., vehicle only) on day -1 of the study.

[0260] Materials and Methods Monoclonal antibodies according to the invention are provided in sodium acetate buffer (e.g., 20 mM sodium acetate, 75 mM sodium chloride, 5% sucrose, pH 5.5) and diluted to a final concentration for administration ranging from a nominal dose of 0.003 mg to 5 mg, administered in 50 μL to 500 μL per nostril.

[0261] NHP subjects assigned to the control group (sodium acetate buffer) will receive vehicle (e.g., 20 mM sodium acetate, 75 mM sodium chloride, 5% sucrose, pH 5.5), administered as 50 µL–500 µL per nostril.

[0262] All NHPs will receive an intranasal challenge with SARS-CoV-2 on day 0.

[0263] Samples will be collected during the study, and sample types will include nasal sampling (e.g., nasal swabs, washes, or scrapes), serum and / or bronchoalveolar lavage (BAL) samples. Sampling periods will encompass the duration of the study within the time frame of Day -1 to the end of the study (e.g., a 7- to 21-day follow-up period).

[0264] Samples are analyzed for viral quantification (RT-PCR and / or TCID 50 The antibody titer and neutralization titer are analyzed by ELISA, HPLC, or similar methods.

[0265] animal NHP subjects are used at a weight of 3-12 kg (e.g., 5 kg per animal) at the start of the study. Four to 12 animals are assigned to each treatment group.

[0266] Study design The dose levels of the monoclonal antibodies and combinations applied in this example are based on maximum dosages and on allometric calculations derived from mouse studies.

[0267] [Table 4]

[0268] NHP subjects will receive intranasal administration of a composition containing either a single antibody at a nominal dose of 0.003-5 mg (treatment group 1 or 2). Another NHP subject will receive intranasal administration of a composition containing the first and second antibodies at a nominal dose of 0.003-5 mg (treatment group 3). On day 0, all NHP subjects will be challenged with a dose of SARS-CoV-2 and will be observed for clinical symptoms (e.g., weight loss and / or temperature) and virological measures until the end of the study (e.g., 7-21 days).

[0269] Antibody administration According to the treatment schedule (Table 4), antibody compositions (treatment groups 1-3) or vehicle alone are administered (e.g., by pipette or spray) to each nostril using volumes of 50 μL to 500 μL per nostril.

[0270] Virus administration The viral material is thawed prior to administration. Once thawed, the material is diluted and each animal receives 50 μL to 500 μL of viral solution per nostril. NHP subjects are administered 10 2 ~10 8 TCID 50 of SARS-CoV-2 (e.g., 10 5 TCID 50 They are infected with SARS-CoV-2.

[0271] Data analysis and statistical methods Clinical symptoms (e.g., weight loss and temperature) and virological measures are used to compare treatment groups (e.g., treatment groups 1-3) with the control group and / or between treatment groups (e.g., treatment group 1 versus treatment group 3).

[0272] Results - Clinical symptoms Prophylactic treatment with treatments (shown in Table 4) provides a reduction in clinical symptom measures compared to the control group or between treatment groups.

[0273] Results – Virological Measurements Prophylactic treatment with treatments (shown in Table 4) provides a reduction in virological measures compared to the control group or between treatment groups.

[0274] conclusion In this SARS-CoV-2 NHP model, prophylactic intranasal administration of a treatment according to the invention (shown in Table 4) provides a reduction in clinical symptoms and / or a reduction in virological measures compared to control NHPs and in comparisons between treatments.

[0275] Example 5 Bispecific antibodies according to the present invention The aim of this intranasal mouse study was to evaluate the pre- and post-exposure efficacy compared to a parent antibody of a bispecific monoclonal antibody according to the invention 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 SEQ ID NOs: 013, 170, 266, 386, 439, or 584, and the second Fab comprises a sequence comprising any one or more of SEQ ID NOs: 656, 657, 658, 659, 660-661.

[0276] 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.

[0277] 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.

[0278] 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.

[0279] All mice will receive an intranasal challenge with SARS-CoV-2 delta on day 0.

[0280] 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.

[0281] 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.

[0282] [Table 5]

[0283] Mice are treated with compositions containing either the parental or bispecific antibodies at doses ranging from 0.001 to 15 mg / kg via the intranasal route. On day 0, all mice are challenged with a dose of SARS-CoV-2 and monitored for survival and body weight until the end of the study (e.g., 10-15 days).

[0284] 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.

[0285] Virus administration The viral material was thawed prior to administration. Once thawed, the material was diluted and each animal received 25 μL to 50 μL of virus per nostril. Mice were 10 2 ~10 8 TCID 50 of SARS-CoV-2 (e.g., 10 3.5 TCID 50 They are infected with SARS-CoV-2.

[0286] Laboratory analysis The inoculum is returned to the laboratory and replicate samples are titrated on Vero cells to verify the administered virus dose.

[0287] 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).

[0288] Data analysis and statistical methods Survival fraction, survival time, and weight change (area under the curve) are compared with corresponding control groups using Fisher's exact test, log-rank, and Welch's t-test, respectively. All groups are compared to the vehicle control group. P values ​​are adjusted according to Bonferroni (for triplicate comparisons with vehicle), followed by a stepwise approach within the antibody (starting with the highest dose and conditionally testing lower doses if the previous step is statistically significant).

[0289] Combination Index A survival dose-response curve was fitted for each of the treatments (parental mAb 1, parental mAb 2, and bispecific) and the ED 50 The efficacy of the bispecific antibody is then compared to the parental antibodies by calculating a combination index.

[0290] 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.

[0291] 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.

[0292] Prophylactic treatment with the parental or bispecific antibodies (shown in Table 5) provides a statistically significant reduction in weight loss compared to the control group.

[0293] 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.

[0294] Example 5a Bispecific antibodies according to the present invention The aim of this intranasal mouse study was to evaluate the pre- and post-exposure efficacy of a bispecific monoclonal antibody according to the invention, 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, compared to the parent antibodies. The first parent antibody ("Parent mAb 1") has the following CDRs represented by SEQ ID NOs: 013, 170, 266, 386, 439, and 584. The second parent antibody ("Parent mAb 2") has the following CDRs represented by SEQ ID NOs: 650, 651, 652, 653, 654, and 655. The bispecific antibody according to the invention has a first Fab comprising a sequence comprising any one or more of SEQ ID NOs: 013, 170, 266, 386, 439, and 584, and a second Fab comprising a sequence comprising any one or more of SEQ ID NOs: 650, 651, 652, 653, 654, and 655.

[0295] Animals (n=8 per group) were treated on day −1 of the study with intranasal doses including 0.007 mg / kg to 1.7 mg / kg (parental mAb 1 or parental mAb 2), or 0.002 mg / kg to 10 mg / kg (bispecific antibody), as described above, or vehicle control.

[0296] Materials and Methods The monoclonal parent antibodies and bispecific antibodies according to the invention were provided in sodium acetate buffer (20 mM sodium acetate, 75 mM sodium chloride, 5% sucrose, pH 5.5), and the bispecific antibodies according to the invention were provided in phosphate buffered saline (PBS), and all antibodies were diluted to a final concentration for administration ranging from 0.007 mg / kg to 1.7 mg / kg (parent mAb 1 or parent mAb 2), or 0.002 mg / kg to 10 mg / kg (bispecific antibody), and administered in 25 μL per nostril.

[0297] Mice assigned to the control group received vehicle (phosphate buffered saline, PBS), administered at 25 μL per nostril.

[0298] 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.

[0299] 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 13.6-22.6 g on day 0. A total of 144 mice, all female, were used, aged 7-11 weeks. Eight animals were assigned to each of the 17 treatment groups, creating groups with similar average body weights. Eight mice were assigned to the vehicle control group. 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 a red plastic tunnel. 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. Each animal was weighed daily beginning on the day of infection (day 0).

[0300] Study design The dose levels of the parental antibodies and their combinations applied in this example were based on previous experience with these antibodies, which demonstrated intranasal protection IN at doses of 0.5 mg / kg or higher (parental mAb 1) and 0.2 mg / kg or higher (parental mAb 2). The efficacy of the two parental bispecific antibodies was expected to be comparable, and the bispecific antibodies were included at doses one dose step up and down from the parental antibody dose range. A total of 144 mice, aged 7 to 11 weeks, were assigned to 18 experimental groups according to Table 6 (see below). Mice were allowed more than 3 days for acclimatization.

[0301] [Table 6]

[0302] Mice were treated with compositions containing either the parental or bispecific antibodies at doses ranging from 0.002 to 10 mg / kg via the intranasal route. On day 0, all mice were challenged with a dose of SARS-CoV-2 and monitored for survival and body weight until the end of the study (day 14).

[0303] Antibody administration Upon arrival, the test antibodies were stored at −80° C.±10° C. Appropriate doses according to the treatment schedule (Table 6) were prescribed based on the average weight per cage.

[0304] 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.

[0305] Virus administration The virus material was stored at -80°C ± 10°C and thawed before administration. 3.5TCID 50 The material was diluted in cold PBS equivalent to 50 μL / mL. Animals were anesthetized with isoflurane (4% v / v in 2 L / min O) as needed. Each animal received approximately 10 3.5 TCID 50 Approximately 50 μL (25 μL in each nostril) of virus, equivalent to 100 μL of nasal inoculation, was given by intranasal inoculation.

[0306] Final Inspection Mice were euthanized by cervical dislocation at the end of the study on day 14. No complete necropsy was performed.

[0307] Data analysis and statistical methods Survival fraction, survival time, and body weight change (using area under the curve) at day 14 were compared with the vehicle control group using Fisher's exact test, log-rank, and Welch's t-test, respectively. P values ​​were adjusted according to Bonferroni for the four comparisons to vehicle control, followed by a stepwise approach starting with the highest dose within an arm and conditionally testing lower doses if the previous dose was statistically significant.

[0308] Probit regression was used to estimate survival dose-response curves without assuming parallel dose-response curves for treatments using the survival fraction at day 14. The model was used to estimate the median effective dose and to determine the ratio between treatments.

[0309] Statistical analysis was performed using R, and statistical significance was set at α = 0.05.

[0310] survival rate The vehicle control group showed 0% survival and a median survival time of 7 days. Prophylactic intranasal treatment with 0.11 mg / kg or more of either parental antibody intranasally provided a statistically significant increase in survival compared to the control group (see Figures 5A and 5D). Prophylactic treatment with 0.16 mg / kg or more of the bispecific antibody provided a statistically significant increase in survival compared to the control group (Figure 5G).

[0311] Prophylactic intranasal treatment with 0.03 mg / kg or more of either the parental antibody or 0.039 mg / kg or more of the bispecific antibody intranasally provides a statistically significant increase in survival time compared to the control group.

[0312] Survival dose-response curves Probit regression was used to estimate survival dose-response curves without assuming parallel dose-response curves between treatments. A vehicle control group was included in the model at a dose of 0 mg / kg. Probit regression models were used to estimate the antibody dose corresponding to 50% survival (ED 50 ) was estimated (see Table 7 and Figures 5C, 5F, and 5I). The intranasal median effective dose was lowest for the bispecific antibody at 0.013 mg / kg. Both parental antibodies were within a 4-fold ratio, with a significantly higher ED compared to the bispecific antibody. 50 No significant differences in were determined.

[0313] [Table 7]

[0314] 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.

[0315] Prophylactic intranasal treatment with either the parental antibody at ≥0.03 mg / kg or the bispecific antibody at ≥0.039 mg / kg resulted in a significant reduction in weight loss compared to the control group (Figures 5B, 5E, and 5H).

[0316] conclusion In this SARS-CoV-2 delta mouse model, prophylactic intranasal administration of bispecific antibodies according to the invention (shown in Table 6) provides a significant increase in survival rate at doses of 0.16 mg / kg and above and a reduction in weight loss at doses of 0.039 mg / kg and above compared to the control group. Furthermore, the median effective dose of the bispecific antibody was not significantly different from that of the parent antibody (Table 7).

[0317] Example 6 Compositions Comprising Antibody Combinations According to the Invention The purpose of this intranasal mouse study is to evaluate the pre- and post-exposure efficacy of a composition comprising a first antibody of the invention capable of binding to the stem helix of SARS-CoV-2 and a second antibody capable of binding to the fusion peptide of SARS-CoV-2. The first antibody ("first mAb") has a heavy chain CDR1 as SEQ ID NO: 013, a heavy chain CDR2 as SEQ ID NO: 170, a heavy chain CDR3 as SEQ ID NO: 266, a light chain CDR1 as SEQ ID NO: 386, a light chain CDR2 as SEQ ID NO: 439, and a light chain CDR3 as SEQ ID NO: 584. The second antibody ("second mAb") has a heavy chain CDR1 as SEQ ID NO: 662, a heavy chain CDR2 as SEQ ID NO: 663, a heavy chain CDR3 as SEQ ID NO: 664, a light chain CDR1 as SEQ ID NO: 6665, a light chain CDR2 as SEQ ID NO: 666, and a light chain CDR3 as SEQ ID NO: 667.

[0318] Animals are treated on day -1 of the study with either an intranasal dose (0.001 mg / kg to 15 mg / kg) containing the first mAb or the second mAb, or a composition containing the first and second antibodies described above, or a vehicle control.

[0319] Materials and Methods The first and second mAbs are provided in sodium acetate buffer (e.g., 20 mM sodium acetate, 75 mM sodium chloride, 5% sucrose, pH 5.5) and diluted to a final concentration for administration ranging from 0.001 mg / kg to 15 mg / kg, administered as 25 μL to 50 μL per nostril.

[0320] 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.

[0321] All mice will receive an intranasal challenge with SARS-CoV-2 delta on day 0.

[0322] 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.

[0323] Study design The dose levels for the first or second mAb, or compositions comprising the first and second antibodies described above, in this example are based on maximum dosages and on previous results of efficacy studies in mice using the parent antibody alone, with a 3- or 4-fold dilution curve applied for each parent antibody or bispecific antibody, starting at a maximum dose of 15 mg / kg or less.

[0324] [Table 8]

[0325] Mice are treated via intranasal administration with a composition containing either the first or second antibody or a composition containing the combination at a dose ranging from 0.001 to 15 mg / kg. On day 0, all mice are challenged with a dose of SARS-CoV-2 and monitored for survival and body weight until the end of the study (e.g., 10-15 days).

[0326] Antibody administration According to the treatment schedule (Table 8), a composition containing the first or second antibody or combination, or vehicle alone, is administered (e.g., by pipette or spray) to each nostril using a volume of 25 μL to 50 μL per nostril.

[0327] Virus administration The viral material was thawed prior to administration. Once thawed, the material was diluted and each animal received 25 μL to 50 μL of viral solution per nostril. Mice were 10 2 ~10 8 TCID 50 of SARS-CoV-2 (e.g., 10 3.5 TCID 50 They are infected with SARS-CoV-2.

[0328] Laboratory analysis The inoculum is returned to the laboratory and replicate samples are titrated on Vero cells to verify the administered virus dose.

[0329] 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).

[0330] Data analysis and statistical methods Survival fraction, survival time, and weight change (area under the curve) are compared with corresponding control groups using Fisher's exact test, log-rank, and Welch's t-test, respectively. All groups are compared to the vehicle control group. P values ​​are adjusted according to Bonferroni (for triplicate comparisons with vehicle), followed by a stepwise approach within the antibody (starting with the highest dose and conditionally testing lower doses if the previous step is statistically significant).

[0331] Combination Index A survival dose-response curve was fitted for each of the treatments (compositions containing the first or second antibody or combination) and the ED 50The effectiveness of the combination is then compared to the first and second antibodies by calculating a combination index.

[0332] survival rate Prophylactic treatment with a composition comprising the first or second antibody or combination (shown in Table 8) compared to the control group provides statistically significant protection against mortality and significant improvement in survival compared to the control group.

[0333] 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.

[0334] Prophylactic treatment with a composition containing the combination (shown in Table 8) provides a statistically significant reduction in weight loss compared to the control group.

[0335] conclusion In this SARS-CoV-2 delta mouse model, prophylactic intranasal administration of a composition comprising a combination according to the invention (shown in Table 8) provides a significant improvement in survival and reduced weight loss compared to the control group. The combination index results suggest at least comparable, additive, or possibly synergistic effects for the combination compared to the first or second antibody administered alone.

[0336] Example 7 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.

[0337] 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.

[0338] [Table 9]

[0339] ELISA binding assays (Table 10) show that antibodies according to the invention were able to effectively bind to all spike antigens tested from alpha- and beta-coronaviruses, including SARS-CoV-2 delta, Omicron XBB.1.5, and hCoV NL63. The anti-S1 antibody tested bound only to SARS-CoV-2 delta and showed no breadth among the alpha-coronavirus NL63, as expected from the literature. An isotype control antibody showed no binding to any coronavirus antigens.

[0340] Example 8 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 from SARS-CoV-2, SARS-CoV-1, MERS, hCoV NL63, hCoV HKU, hCoV OC43, and hCoV 229E (V-Plex COVID-19 Coronavirus Panel 3 (IgG) Kit cat# K15399U-2) (Table 11), as well as a set of Omicron variants (V-Plex SARS-CoV-2 Panel 34 (IgG) Kit cat# K15690U-2) (Table 12). 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). The 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 (which is the range between the lower and upper limits of quantitation (LLOQ and ULOQ)) was reported.

[0341] [Table 10]

[0342] MSD binding arrays (see Table 11) demonstrated that the antibodies effectively bound spike antigens from the betacoronaviruses SARS-CoV-2, SARS-CoV-1, MERS, HKU1, and OC43. Alphacoronaviruses, including 229E and NL63, also bound. The antibodies did not bind to the SARS-CoV-2 receptor-binding domain (RBD) on the S1 portion of the spike used in this study. In contrast, the anti-S1 antibodies tested here bound well to the RBD domain and wild-type SARS-CoV-2, as expected from the literature, and did not show breadth among other coronaviruses. This suggests that the antibodies in accordance with the present invention exhibit breadth of binding across the spikes of all beta- and alphacoronaviruses tested, while the anti-RBD antibodies bind only to the SARS-CoV-2 spike and RBD domain. The isotype control showed no binding to any coronavirus antigens.

[0343] [Table 11]

[0344] MSD binding arrays demonstrated that antibodies according to the present invention were able to bind to all omicron variants tested in this assay at concentrations up to 10 ng / mL (Table 12), thus retaining activity among the omicron variants tested, as confirmed by XBB.1.5 in an ELISA binding assay (Table 10). While the anti-S1 control antibody bound to the SARS-CoV-2 spike at lower concentrations than antibodies according to the present invention, early omicron variants bound only at extrapolated high concentrations, indicating low affinity for these variants, and later omicron variants such as BQ.1 and XBB.1 did not bind. This indicates that antibodies according to the present invention can bind to recent omicron variants, while anti-RBD antibodies do not bind to these variants. A negative isotype control antibody showed no binding to coronavirus antigens.

[0345] Example 9 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.

[0346] [Table 12]

[0347] Overall, the antibodies according to the invention are able to neutralize the tested viruses SARS-CoV-1, SARS-CoV-2 and MERS at various concentrations (see Table 13), with IC for SARS-CoV-1. 50 This confirms that this antibody has neutralizing activity across these betacoronaviruses, as demonstrated by binding assessment by MSD (Table 11). The SARS-CoV-2 positive serum control showed a high MN 50 A titer is required.

[0348] Example 10 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 BA4, Omicron BQ.1.1, Omicron XBB.1, hCoV NL63, and hCoV 229E.

[0349] [Table 13]

[0350] Overall, the antibodies according to the invention were able to neutralize all alpha- and betaviruses tested at various concentrations (see Table 14), with an overall trend of higher IC for omicron variants.50 This demonstrates that the antibodies according to the invention exhibit breadth of neutralization across alpha- and beta-coronaviruses, as expected based on the binding data.

[0351] Example 11 Epitope mapping of the antibody according to the present invention The objective of this study was to precisely determine the amino acid sequences in spike proteins across various alpha- and beta-coronaviruses to which antibodies of the present invention bind. 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) 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.

[0352] The antibodies according to the invention bound to a peptide array of alpha- and beta-coronaviruses in specific epitopes of 10-20 amino acids (see Figure 6). This method allowed for a high degree of confidence in the definition of the epitopes, although there was some variability in the outer regions of the epitopes for each strain, as indicated by asterisks.

[0353] Example 13 Affinity binding assessment of bispecific antibodies by MSD The goal of this study was to evaluate the relative binding affinity to 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 that uses electrochemiluminescent labels conjugated to the detection antibodies.

[0354] The first parent antibody ("Parent mAb 1") has the following CDRs set forth in SEQ ID NOs: 013, 170, 266, 386, 439, and 584. The second parent antibody ("Parent mAb 2") has the following CDRs set forth in SEQ ID NOs: 650, 651, 652, 653, 654, and 655. A bispecific antibody according to the invention has a first Fab comprising a sequence comprising any one or more of SEQ ID NOs: 013, 170, 266, 386, 439, and 584, and a second Fab comprising a sequence comprising any one or more of SEQ ID NOs: 650, 651, 652, 653, 654, and 655.

[0355] Three versions of the bispecific antibody were produced and all were tested with the parental mAb 1 and parental mAb 2 combination.

[0356] In this assay, up to 10 trimeric viral antigens are individually printed onto spots in each well of the plate, allowing for a multiplexed assay readout. The antibody of the present invention binds to the viral antigen through Fab-mediated recognition, and then an anti-human IgG sulfotag detection antibody is added, recognizing the human IgG Fab. Addition of a read buffer containing substrate and passage of current through the plate electrodes initiates an electrochemical (current), chemiluminescent (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 20), as well as a set of Omicron variants (V-Plex SARS-CoV-2 Panel 34 (IgG) Kit cat# K15690U-2) (Table 21). 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.

[0357] [Table 14]

[0358] MSD binding arrays (see Table 20) demonstrated that the bispecific antibodies could effectively bind to all spike antigens from alpha- and beta-coronaviruses. The alphacoronavirus NL63 did not bind to the parental mAb 2 anti-stem helix antibody, but bound only to the anti-fusion peptide parental mAb 1 and the bispecific antibody, albeit at slightly higher concentrations than the parental mAb 1. The alphacoronavirus 229E bound to all antibodies tested, but only to the parental mAb 1 at lower concentrations. The SARS-CoV-2 spike RBD antigen bound only to the parental mAb 2 (stem helix mAb) and the bispecific antibody at higher concentrations. The control anti-S1 mAb bound only to the SARS-CoV-2 spike and RBD at lower concentrations. The data suggest that the bispecific antibodies retain the binding capabilities of their respective parental antibodies, as demonstrated here by the retention of alphacoronavirus binding activity using the Fab arm of parental mAb 1.

[0359] [Table 15]

[0360] MSD binding arrays (see Table 21) showed that the three bispecific antibodies could effectively bind to all spike antigens from SARS-CoV-2 omicron variants at various concentrations. Only parental mAb 2 retained efficient binding to late omicron variants (<1000 pg / ml), and parental mAb 1 required high antibody concentrations, while the bispecific antibodies retained binding at lower concentrations, suggesting that the bispecific antibodies could utilize the Fab arm of parental Fab 2.

[0361] Example 14 Neutralization of live virus by bispecific antibodies The objective of this study was to evaluate the ability of bispecific antibodies to neutralize live coronaviruses and pseudovirions. Bispecific antibodies were tested for functional activity in live virus microneutralization assays against MERS, SARS-CoV-1, SARS-CoV-2 Wuhan, Delta, and Omicron BA.4 / 5.

[0362] The first parent antibody ("Parent mAb 1") has the following CDRs set forth in SEQ ID NOs: 013, 170, 266, 386, 439, and 584. The second parent antibody ("Parent mAb 2") has the following CDRs set forth in SEQ ID NOs: 650, 651, 652, 653, 654, and 655. A bispecific antibody according to the invention comprises a first Fab comprising a sequence comprising any one or more of SEQ ID NOs: 013, 170, 266, 386, 439, and 584, and a second Fab comprising a sequence comprising any one or more of SEQ ID NOs: 650, 651, 652, 653, 654, and 655.

[0363] The bispecific antibody BISPECIFIC 1 comprises the first Fab and second Fab described above.

[0364] Briefly, serial dilutions of antibodies were pre-incubated with the corresponding viruses and then added to the respective cell lines (Vero: MERS and SARS-CoV-1, all others 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 50 ) reported. The bispecific antibodies were tested for neutralizing activity in the range of 0.02 to 500 μg / ml compared to individual control antibodies.

[0365] [Table 16]

[0366] Overall, the bispecific antibodies were able to neutralize the tested viruses SARS-CoV-1, SARS-CoV-2 Wuhan, Delta and Omicron BA.4 / 5, and MERS at various concentrations (see Table 22). The anti-stem helix parental mAb 2 neutralized SARS-CoV-2 Wuhan and MERS with a lower IC compared to the anti-fusion peptide parental mAb 1. 50 This trend was also observed for bispecific antibodies, indicating that the antibodies can utilize the anti-stem helix Fab arm. For SARS-CoV-2 delta, Omicron BA4.5, and SARS-CoV-1, the parental and bispecific antibodies had similar IC 50 Overall, this demonstrates that the bispecific antibody retains the affinity derived from both parental Fab arms and retains neutralizing activity across the betacoronaviruses tested.

[0367] Example 15 Neutralization of pseudovirions by bispecific antibodies Pseudovirion neutralization assays against pseudotyped virus particles were performed.

[0368] The first parent antibody ("Parent mAb 1") has the following CDRs set forth in SEQ ID NOs: 013, 170, 266, 386, 439, and 584. The second parent antibody ("Parent mAb 2") has the following CDRs set forth in SEQ ID NOs: 650, 651, 652, 653, 654, and 655. A bispecific antibody according to the invention has a first Fab comprising a sequence comprising any one or more of SEQ ID NOs: 013, 170, 266, 386, 439, and 584, and a second Fab comprising a sequence comprising any one or more of SEQ ID NOs: 650, 651, 652, 653, 654, and 655.

[0369] The bispecific antibody BISPECIFIC 1 comprises a first Fab and a second Fab as described above.

[0370] The advantage over live virus assays is that pseudotyped viruses can be used in laboratories with low biosafety levels, allowing for more rapid production and testing of novel variants of concern. 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. After incubation, cells were washed and lysed, and luciferase activity in the cell lysates was measured using the Nano-Glo Luciferase Assay System and 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 against SARS-CoV-2 Wuhan, Delta, Omicron BA4 / 5, BQ.1, and XBB.1, hCoV NL63, and hCoV 229E in the range of 0.01 to 250 μg / ml, compared to individual control antibodies.

[0371] [Table 17]

[0372] Pseudovirion neutralization assays (see Table 23) show that parental antibody 2 according to the invention is able to neutralize Omicron variants, especially late-arising variants such as XBB.1.1, at lower concentrations than the anti-fusogenic peptide parental antibody mAb 1, with the exception of SARS-CoV-2 Wuhan, which is neutralized by the bispecific antibody at the lowest concentration compared to the parental antibody, and the bispecific antibody has an IC intermediate between the respective parental mAbs. 50The overall trend of bispecific antibodies retaining the neutralizing activity of their parental antibodies was confirmed by live virus assays (Table 22), demonstrating that the bispecific antibodies retain the neutralizing affinity derived from each Fab arm of the parental antibodies.

Claims

1. An anti-coronavirus, preferably anti-SARS-CoV-2 bispecific antibody or antigen-binding fragment thereof, having a first Fab capable of binding to a fusion peptide of a coronavirus, preferably SARS-CoV-2, and a second Fab capable of binding to a stem helix of a coronavirus, preferably SARS-CoV-2, wherein the first Fab comprises a sequence comprising preferably any one or more of SEQ ID NOs: 013, 170, 266, 368, 439, or 584, and the second Fab comprises a sequence comprising preferably any one or more of SEQ ID NOs: 662, 663, 664, 665, 666-667.

2. 2. The bispecific antibody or antigen-binding fragment thereof of claim 1, comprising a first Fab capable of binding to a fusion peptide of a coronavirus, preferably SARS-CoV-2, and a second Fab capable of binding to the stem helix of a coronavirus, preferably SARS-CoV-2, wherein the first Fab comprises a sequence comprising preferably any one or more of SEQ ID NOs: 013, 170, 266, 368, 439, and / or 584, and the second Fab comprises a sequence comprising preferably any one or more of SEQ ID NOs: 662, 663, 664, 665, 666, and / or 667.

3. a first Fab that specifically binds to the fusion peptide of a coronavirus, preferably SARS-CoV-2, and a second Fab that specifically binds to the stem helix 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: 013, the heavy chain CDR2 region of SEQ ID NO: 170, and the heavy chain CDR3 region of SEQ ID NO: 266, and a light chain variable region comprising, as CDRs, the light chain CDR1 region of SEQ ID NO: 368, the light chain CDR2 region of SEQ ID NO: 439, and the light chain CDR3 region of SEQ ID NO: 584; b.) the second Fab comprises a heavy chain variable region comprising, as CDRs, the heavy chain CDR1 region of SEQ ID NO: 662, the heavy chain CDR2 region of SEQ ID NO: 663, and the heavy chain CDR3 region of SEQ ID NO: 664, and a light chain variable region comprising, as CDRs, the light chain CDR1 region of SEQ ID NO: 665, the light chain CDR2 region of SEQ ID NO: 666, and the light chain CDR3 region of SEQ ID NO: 667; 3. The bispecific antibody or antigen-binding fragment thereof according to claim 1 or 2,

4. a) a light chain comprising VL-CL domains and a heavy chain comprising VH-CH1-CH2-CH3 domains of an antibody or antigen-binding fragment thereof capable of binding to a fusion peptide of a coronavirus, preferably SARS-CoV-2, and b) 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; wherein the constant domains CL and CH1 from an antibody or antigen-binding fragment thereof capable of binding to the fusion peptide of a coronavirus, preferably SARS-CoV-2, are replaced by each other.

4. The bispecific antibody or antigen-binding fragment thereof according to claim 1 .

5. 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: 013, the heavy chain CDR2 region of SEQ ID NO: 170, and the heavy chain CDR3 region of SEQ ID NO: 266; 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: 368, the light chain CDR2 region of SEQ ID NO: 439, and the light chain CDR3 region of SEQ ID NO: 584; The VH domain of the antibody or antigen-binding fragment thereof capable of binding to the stem helix of coronavirus, preferably SARS-CoV-2, comprises, as CDRs, the heavy chain CDR1 region of SEQ ID NO: 662, the heavy chain CDR2 region of SEQ ID NO: 663, and the heavy chain CDR3 region of SEQ ID NO: 664, and the VL domain of the antibody or antigen-binding fragment thereof capable of binding to the stem helix of coronavirus, preferably SARS-CoV-2, comprises, as CDRs, the light chain CDR1 region of SEQ ID NO: 665, the light chain CDR2 region of SEQ ID NO: 666, and the light chain CDR3 region of SEQ ID NO:

667. The bispecific antibody or antigen-binding fragment thereof according to claim 4.

6. a) a light chain comprising VL-CL domains and a heavy chain comprising VH-CH1-CH2-CH3 domains of an antibody capable of binding to a fusion peptide of a coronavirus, preferably SARS-CoV-2, and b) 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 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; 4. The bispecific antibody or antigen-binding fragment thereof according to claim 1 .

7. 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: 013, the heavy chain CDR2 region of SEQ ID NO: 170, and the heavy chain CDR3 region of SEQ ID NO: 266; 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: 368, the light chain CDR2 region of SEQ ID NO: 439, and the light chain CDR3 region of SEQ ID NO: 584; The VH domain of the antibody capable of binding to the stem helix of coronavirus, preferably SARS-CoV-2, comprises as CDRs the heavy chain CDR1 region of SEQ ID NO: 662, the heavy chain CDR2 region of SEQ ID NO: 663, and the heavy chain CDR3 region of SEQ ID NO: 664, and the VL domain of the antibody capable of binding to the stem helix of coronavirus, preferably SARS-CoV-2, comprises as CDRs the light chain CDR1 region of SEQ ID NO: 665, the light chain CDR2 region of SEQ ID NO: 666, and the light chain CDR3 region of SEQ ID NO:

667. The bispecific antibody or antigen-binding fragment thereof according to claim 6.

8. a) a light chain comprising VL-CL domains and a heavy chain comprising VH-CH1-CH2-CH3 domains of an antibody capable of binding to a fusion peptide of a coronavirus, preferably SARS-CoV-2, and b) 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 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; 4. The bispecific antibody or antigen-binding fragment thereof according to claim 1 .

9. 9. The bispecific antibody or antigen-binding fragment thereof according to claim 1 , 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:

640.

10. 9. The bispecific antibody or antigen-binding fragment thereof according to claim 1 , 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:

688.

11. 9. The bispecific antibody or antigen-binding fragment thereof according to claim 1 , 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:

641.

12. 9. The bispecific antibody or antigen-binding fragment thereof of claim 1 , 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:

689.

13. 9. The bispecific antibody or antigen-binding fragment thereof according to any one of claims 1 to 8, wherein the bispecific antibody or antigen-binding fragment thereof is selected from the group consisting of 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.

14. 14. A method for the production of a bispecific antibody or antigen-binding fragment thereof according to any one of claims 1 to 13, comprising: a) culturing a host cell comprising an expression vector comprising a polynucleotide encoding the bispecific antibody or antibody fragment of any one of claims 1 to 13 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:

15. 14. A nucleic acid molecule, preferably an isolated nucleic acid molecule, comprising a nucleic acid sequence encoding the bispecific antibody or antigen-binding fragment thereof, or 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 claims 1 to 13.

16. a nucleic acid sequence encoding the bispecific antibody or antigen-binding fragment thereof, or 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 claims 1 to 65 and claim 15; The nucleic acid sequence is (i) a first nucleic acid sequence encoding the amino acid sequence set forth in SEQ ID NO: 013, or a first nucleic acid sequence encoding an amino acid sequence that differs from SEQ ID NO: 013 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: 170, or a second nucleic acid sequence encoding an amino acid sequence that differs from SEQ ID NO: 170 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: 266, or a third nucleic acid sequence encoding an amino acid sequence that differs from SEQ ID NO: 266 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: 368, or a fourth nucleic acid sequence encoding an amino acid sequence that differs from SEQ ID NO: 368 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: 439, or an amino acid sequence that differs from SEQ ID NO: 439 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: 584, or an amino acid sequence that differs from SEQ ID NO: 584 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: 662, or an amino acid sequence that differs from SEQ ID NO: 662 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: 663, or an eighth nucleic acid sequence encoding an amino acid sequence that differs from SEQ ID NO: 663 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: 664, or an amino acid sequence that differs from SEQ ID NO: 664 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: 665, or an amino acid sequence that differs from SEQ ID NO: 665 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: 666, or an eleventh nucleic acid sequence encoding an amino acid sequence that differs from SEQ ID NO: 666 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: 667, or an amino acid sequence that differs from SEQ ID NO: 667 by at most 1, 2, or 3 amino acids. A nucleic acid molecule comprising at least one of the following:

17. 14. The bispecific antibody or antigen-binding fragment thereof of any one of claims 1 to 13 for use in a method for treating a coronavirus infection in a subject.

18. A composition comprising a bispecific antibody and / or antigen-binding fragment thereof according to any one of claims 1 to 13.

19. 19. The composition of claim 18, further comprising a second medicament for simultaneous, separate, or sequential administration.

20. 20. The composition of claim 19, wherein the second medicament comprises a second antibody or an antigen-binding fragment thereof.

21. 20. The composition of claim 19, wherein the second medicament comprises a bispecific antibody or a bispecific antigen-binding fragment thereof.

22. 20. The composition of claim 19, further comprising a pharmaceutically acceptable excipient or carrier.

23. 19. The composition of claim 18, comprising an effective amount of a bispecific antibody or antigen-binding fragment thereof, a nucleic acid molecule, or a vector, and optionally a pharmaceutically acceptable excipient or carrier, 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.

24. 19. The composition of claim 18 for use as a medicine.

25. 19. An inhalation device comprising the composition of claim 18.

26. 14. The bispecific antibody or antigen-binding fragment thereof according to any one of claims 1 to 13, which is bivalent.

27. 14. The bispecific antibody or antigen-binding fragment thereof according to any one of claims 1 to 13, which is 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.

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