Methods of treating or preventing sars-cov-2 infection and covid-19 with anti-sars-cov-2 spike glycoprotein antibodies

Abstract

The present invention provides methods for preventing and treating SARS-CoV-2 infection, COVID-19, or symptoms thereof. The methods of the present invention feature administration of one or more antigen-binding molecules (e.g., antibodies) that bind to a surface protein (e.g., spike protein) of SARS-CoV-2.

Description

[0001] Cross-reference of related applications

[0002] This patent application claims the benefit of the following U.S. Provisional Patent Application Nos., filed June 3, 2020: 63 / 034,348; 63 / 036,956; filed June 9, 2020: 63 / 038,274; filed June 12, 2020: 63 / 043,336; filed August 3, 2020: 63 / 060,592; filed August 7, 2020: 63 / 062,961; filed August 14, 2020: 63 / 065,799; and filed September 29, 2020: 63 / 060,592. 63 / 084,881, 63 / 085,066 submitted on September 29, 2020, 63 / 089,399 submitted on October 8, 2020, 63 / 090,690 submitted on October 12, 2020, 63 / 094,133 submitted on October 20, 2020, 63 / 105,779 submitted on October 26, 2020, 63 / 106,696 submitted on October 28, 2020, 63 / 112,140 submitted on November 10, 2020, 63 / 116,773 submitted on November 20, 2020, and November 20, 2020 Submissions dated March 30, 2020: 63 / 119,593; December 1, 2020: 63 / 120,065; December 14, 2020: 63 / 124,980; December 29, 2020: 63 / 131,627; January 25, 2021: 63 / 141,423; January 26, 2021: 63 / 141,952; January 27, 2021: 63 / 142,471; February 2, 2021: 63 / 144,789; February 18, 2021: 63 / 150,978; March 2021: 63 / 119,593; December 29, 2020: 63 / 131,627; January 25, 2021: 63 / 141,423; January 26, 2021: 63 / 141,952; January 27, 2021: 63 / 142,471; February 2, 2021: 63 / 144,789; February 18, 2021: 63 / 150,978; March 2021: 63 / 150,978; December 25, 2021: 63 / 141,423; January 26, 2021: 63 / 141,952; January 27, 2021: 63 / 142,471; February 2, 2021: 63 / 144,789; February 18, 2021: 63 / 150,978; December 25, 2021: The entire contents of each of these U.S. provisional patent applications are incorporated herein by reference for all purposes. (The applications are: 63 / 162,504 filed on March 17, 2021; 63 / 162,996 filed on March 18, 2021; 63 / 164,488 filed on March 22, 2021; 63 / 165,654 filed on March 24, 2021; 63 / 166,187 filed on March 25, 2021; 63 / 173,468 filed on April 11, 2021; 63 / 185,301 filed on May 6, 2021; and 63 / 186,029 filed on May 7, 2021.)

[0003] Federal government-funded research or development

[0004] This invention was carried out with government support under Agreement HHSO100201700020C granted by the U.S. Department of Health and Human Services. The government holds certain rights to this invention.

[0005] Reference to the sequence list

[0006] This patent application is incorporated by reference to the sequence list created on June 2, 2021, containing 72,328 bytes, and filed in a computer-readable form as document 10807WO01-Sequence. Technical Field

[0007] This invention belongs to the medical field and relates to methods and pharmaceutical compositions for treating SARS-CoV-2 infection and COVID-19 by administering antigen-binding molecules that bind to the surface proteins of SARS-CoV-2 (e.g., anti-SARS-CoV-2 spike glycoprotein antibodies and their antigen-binding fragments, or combinations of such antibodies or antigen-binding fragments). Background Technology

[0008] Coronaviruses are a family of enveloped, single-stranded RNA viruses. In recent decades, two highly pathogenic coronavirus strains have been identified in humans: Severe Acute Respiratory Syndrome Coronavirus (SARS-CoV) and Middle East Respiratory Syndrome Coronavirus (MERS-CoV). These viruses have been found to cause severe, and sometimes fatal, respiratory illness.

[0009] Coronaviruses possess an RNA genome encased in a nucleocapsid (N) protein surrounded by an outer envelope. This envelope consists of membrane (M) and envelope (E) proteins (which are involved in viral assembly) and a spike (S) protein (which mediates entry into host cells). The S protein forms a large trimeric spike, providing the coronavirus's signature crown-like appearance. The S protein trimer binds to a host receptor and, following initiation by cellular proteases, mediates host-virus membrane fusion. The S protein appears to be central to SARS-CoV-2 viral infection. The SARS-CoV-2 S protein binds with high affinity to the host receptor angiotensin-converting enzyme 2 (ACE2), and ACE2 has been shown to function as a host cell entry receptor in cellular analyses and animal models.

[0010] Given the potential key role of the S protein in the pathogenesis of SARS-CoV-2, significant efforts are underway to develop antibodies and vaccines that target this protein. Summary of the Invention

[0011] This disclosure provides methods for improving one or more clinical parameters of COVID-19. In some embodiments, the method includes administering a therapeutic composition to a subject in need, wherein the therapeutic composition comprises at least one antigen-binding molecule that binds to the surface protein of SARS-CoV-2. In some embodiments, the subject is a person suffering from laboratory-confirmed SARS-CoV-2 and one or more COVID-19 symptoms. In some cases, the one or more COVID-19 symptoms include fever, cough, or shortness of breath.

[0012] In some implementations, subjects were selected from groups consisting of: (a) COVID-19 patients requiring low-flow oxygen supplementation; (b) COVID-19 patients requiring high-intensity oxygen therapy but not mechanical ventilation; and (c) COVID-19 patients requiring mechanical ventilation. In some cases, subjects were hospitalized due to one or more COVID-19 symptoms. In some cases, subjects were outpatients (i.e., treated on an outpatient basis).

[0013] This disclosure also provides methods for preventing SARS-CoV-2 infection or COVID-19 in subjects. In some cases, the method includes administering a prophylactic composition to the subject, wherein the prophylactic composition comprises at least one antigen-binding molecule that binds to a surface protein of SARS-CoV-2 (e.g., the SARS-CoV-2 spike protein).

[0014] In some implementations, the subjects are individuals who are at high risk of SARS-CoV-2 infection but are not infected. In other implementations, the subjects at high risk of SARS-CoV-2 infection are healthcare workers, first responders, or family members of individuals who have tested positive for SARS-CoV-2 infection.

[0015] In some embodiments, the therapeutic or prophylactic composition comprises a first antigen-binding molecule and a second antigen-binding molecule, the first antigen-binding molecule binding to a first epitope on the surface protein of SARS-CoV-2 and the second antigen-binding molecule binding to a second epitope on the surface protein of SARS-CoV-2, wherein the first epitope and the second epitope do not overlap structurally.

[0016] In some embodiments, the therapeutic or prophylactic composition further comprises a third antigen-binding molecule that binds to a third epitope on the surface protein of SARS-CoV-2, wherein the third epitope does not structurally overlap with the first and second epitopes.

[0017] In some embodiments, the therapeutic or prophylactic composition comprises a first antigen-binding molecule and a second antigen-binding molecule, the first antigen-binding molecule binding to a first epitope on the surface protein of SARS-CoV-2, and the second antigen-binding molecule binding to a second epitope on the surface protein of SARS-CoV-2, wherein the first antigen-binding molecule and the second antigen-binding molecule are capable of simultaneously binding to the surface protein of SARS-CoV-2. In some embodiments, the therapeutic or prophylactic composition further comprises a third antigen-binding molecule binding to a third epitope on the surface protein of SARS-CoV-2, wherein the first antigen-binding molecule, the second antigen-binding molecule, and the third antigen-binding molecule are capable of simultaneously binding to the surface protein of SARS-CoV-2. In some embodiments, a) the first antigen-binding molecule comprises three heavy chain complementarity-determining regions (CDRs) (HCDR1, HCDR2, and HCDR3) contained within a heavy chain variable region (HCVR) containing the amino acid sequence shown in SEQ ID NO:2, and three light chain complementarity-determining regions (CDRs) (LCDR1, LCDR2, and LCDR3) contained within a light chain variable region (LCVR) containing the amino acid sequence shown in SEQ ID NO:10; b) the second antigen-binding molecule comprises three heavy chain complementarity-determining regions (CDRs) (HCDR1, HCDR2, and HCDR3) contained within a heavy chain variable region (HCVR) containing the amino acid sequence shown in SEQ ID NO:22, and three light chain complementarity-determining regions (CDRs) (LCDR1, LCDR2, and LCDR3) contained within a light chain variable region (LCVR) containing the amino acid sequence shown in SEQ ID NO:22, and three light chain complementarity-determining regions ... The amino acid sequence shown in NO:30 is contained within the light chain variable region (LCVR); and c) the third antigen-binding molecule contains three heavy chain complementarity-determining regions (CDRs) (HCDR1, HCDR2, and HCDR3) contained within the heavy chain variable region (HCVR) containing the amino acid sequence shown in SEQ ID NO:73, and three light chain complementarity-determining regions (CDRs) (LCDR1, LCDR2, and LCDR3) contained within the light chain variable region (LCVR) containing the amino acid sequence shown in SEQ ID NO:81.

[0018] In any of the various embodiments, the surface protein of SARS-CoV-2 is a spike (S) protein containing a receptor-binding domain comprising an amino acid sequence having at least 80% identity with SEQ ID NO:59.

[0019] In some embodiments, the antigen-binding molecule is an anti-SARS-CoV-2 spike glycoprotein antibody or an antigen-binding fragment thereof, which contains six complementarity-determining regions HCDR1-HCDR2-HCDR3-LCDR1-LCDR2-LCDR3, which are contained within heavy chain variable region (HCVR) and light chain variable region (LCVR) amino acid sequence pairs comprising amino acid sequences selected from the group consisting of SEQ ID NO: 2 / 10, 22 / 30, 42 / 50 and 73 / 81. In some embodiments, the anti-SARS-CoV-2 spike glycoprotein antibody or antigen-binding fragment comprises six complementarity-determining regions HCDR1-HCDR2-HCDR3-LCDR1-LCDR2-LCDR3, each containing an amino acid sequence selected from the group consisting of: SEQ ID NO:4-6-8-12-14-16, 24-26-28-32-34-36, 44-46-48-52-34-54, and 75-77-79-83-85-87. In some embodiments, the anti-SARS-CoV-2 spike glycoprotein antibody or antigen-binding fragment comprises an HCVR / LCVR amino acid sequence pair, which contains an amino acid sequence selected from the group consisting of: SEQ ID NO:2 / 10, 22 / 30, 42 / 50, and 73 / 81. In some embodiments, the anti-SARS-CoV-2 spike glycoprotein antibody comprises a human IgG heavy chain constant region. In some cases, the anti-SARS-CoV-2 spike glycoprotein antibody contains the heavy chain constant region of the IgG1 or IgG4 isotype. In some cases, the anti-SARS-CoV-2 spike glycoprotein antibody contains heavy chain and light chain amino acid sequence pairs selected from the group consisting of: SEQ ID NO: 18 / 20, 38 / 40, 56 / 58, and 89 / 91.

[0020] In some embodiments, the antigen-binding molecule is an anti-SARS-CoV-2 spike glycoprotein antibody having the same binding and / or blocking properties as a reference antibody, the reference antibody comprising an HCVR / LCVR amino acid sequence pair comprising amino acid sequences selected from the group consisting of: SEQ ID NO: 2 / 10, 22 / 30, 42 / 50, and 73 / 81. In some embodiments, the antigen-binding molecule is an anti-SARS-CoV-2 spike glycoprotein antibody having the same binding and / or blocking properties as a reference antibody, the reference antibody comprising a heavy chain and a light chain amino acid sequence pair comprising amino acid sequences selected from the group consisting of: SEQ ID NO: 18 / 20, 38 / 40, 56 / 58, and 89 / 91.

[0021] In some embodiments, the first antigen-binding molecule is a first anti-SARS-CoV-2 spike glycoprotein antibody or its antigen-binding fragment thereof, which contains six complementarity-determining regions HCDR1-HCDR2-HCDR3-LCDR1-LCDR2-LCDR3, which are contained within the heavy chain variable region (HCVR) and light chain variable region (LCVR) amino acid sequence pairs containing the amino acid sequence of SEQ ID NO:2 / 10, and the second antigen-binding molecule is a second anti-SARS-CoV-2 spike glycoprotein antibody or its antigen-binding fragment thereof, which contains six complementarity-determining regions HCDR1-HCDR2-HCDR3-LCDR1-LCDR2-LCDR3, which are contained within the heavy chain variable region (HCVR) and light chain variable region (LCVR) amino acid sequence pairs containing the amino acid sequence of SEQ ID NO:22 / 30. In some cases, the first anti-SARS-CoV-2 spike glycoprotein antibody or antigen-binding fragment contains six complementarity-determining regions HCDR1-HCDR2-HCDR3-LCDR1-LCDR2-LCDR3, each containing the amino acid sequence SEQ ID NO:4-6-8-12-14-16, and the second anti-SARS-CoV-2 spike glycoprotein antibody or antigen-binding fragment contains six complementarity-determining regions HCDR1-HCDR2-HCDR3-LCDR1-LCDR2-LCDR3, each containing the amino acid sequence SEQ ID NO:24-26-28-32-34-36. In some embodiments, the first anti-SARS-CoV-2 spike glycoprotein antibody or antigen-binding fragment comprises an HCVR / LCVR amino acid sequence pair containing the amino acid sequence of SEQ ID NO:2 / 10, and the second anti-SARS-CoV-2 spike glycoprotein antibody or antigen-binding fragment comprises an HCVR / LCVR amino acid sequence pair containing the amino acid sequence of SEQ ID NO:22 / 30. In some embodiments, the first and second anti-SARS-CoV-2 spike glycoprotein antibodies comprise a human IgG heavy chain constant region. In some embodiments, the first and second anti-SARS-CoV-2 spike glycoprotein antibodies comprise a heavy chain constant region of an IgG1 or IgG4 isotype.In some cases, the first anti-SARS-CoV-2 spike glycoprotein antibody comprises a heavy chain and a light chain, the heavy chain comprising the amino acid sequence of SEQ ID NO:18 and the light chain comprising the amino acid sequence of SEQ ID NO:20, and the second anti-SARS-CoV-2 spike glycoprotein antibody comprises a heavy chain and a light chain, the heavy chain comprising the amino acid sequence of SEQ ID NO:38 and the light chain comprising the amino acid sequence of SEQ ID NO:40.

[0022] In some embodiments, the first antigen-binding molecule is a first anti-SARS-CoV-2 spike glycoprotein antibody or an antigen-binding fragment thereof, having the same binding and / or blocking properties as a reference antibody, the reference antibody comprising an HCVR / LCVR amino acid sequence pair comprising the amino acid sequence of SEQ ID NO:2 / 10, and the second antigen-binding molecule is a second anti-SARS-CoV-2 spike glycoprotein antibody or an antigen-binding fragment thereof, having the same binding and / or blocking properties as the reference antibody, the reference antibody comprising an HCVR / LCVR amino acid sequence pair comprising the amino acid sequence of SEQ ID NO:22 / 30. In some embodiments, the first antigen-binding molecule is a first anti-SARS-CoV-2 spike glycoprotein antibody or an antigen-binding fragment thereof, having the same binding and / or blocking properties as a reference antibody, the reference antibody comprising a heavy chain and a light chain pair comprising the amino acid sequence of SEQ ID NO:18 / 20, and the second antigen-binding molecule is a first anti-SARS-CoV-2 spike glycoprotein antibody or an antigen-binding fragment thereof, having the same binding and / or blocking properties as a reference antibody, the reference antibody comprising a heavy chain and a light chain pair comprising the amino acid sequence of SEQ ID NO:38 / 40.

[0023] In some embodiments, the antigen-binding molecule is an anti-SARS-CoV-2 spike glycoprotein antibody or its antigen-binding fragment, comprising six complementarity-determining regions HCDR1-HCDR2-HCDR3-LCDR1-LCDR2-LCDR3, which are contained within a heavy chain variable region (HCVR) containing the amino acid sequence of SEQ ID NO:42 and a light chain variable region (LCVR) containing the amino acid sequence of SEQ ID NO:50. In some cases, the anti-SARS-CoV-2 spike glycoprotein antibody or antigen-binding fragment comprises six complementarity-determining regions HCDR1-HCDR2-HCDR3-LCDR1-LCDR2-LCDR3, which are each contained within the amino acid sequence of SEQ ID NO:44-46-48-52-34-54. In some cases, the anti-SARS-CoV-2 spike glycoprotein antibody or antigen-binding fragment comprises HCVR and LCVR, where the HCVR contains the amino acid sequence of SEQ ID NO:42 and the LCVR contains the amino acid sequence of SEQ ID NO:50. In some embodiments, the anti-SARS-CoV-2 spike glycoprotein antibody comprises a human IgG heavy chain constant region. In some cases, the anti-SARS-CoV-2 spike glycoprotein antibody comprises a heavy chain constant region of an IgG1 or IgG4 isotype. In some cases, the anti-SARS-CoV-2 spike glycoprotein antibody comprises a heavy chain and a light chain, wherein the heavy chain comprises the amino acid sequence of SEQ ID NO:56 and the light chain comprises the amino acid sequence of SEQ ID NO:58.

[0024] In some embodiments, the antigen-binding molecule is an anti-SARS-CoV-2 spike glycoprotein antibody having the same binding and / or blocking properties as a reference antibody, the reference antibody comprising an HCVR / LCVR amino acid sequence pair comprising the amino acid sequences of SEQ ID NO:42 / 50. In some embodiments, the antigen-binding molecule is an anti-SARS-CoV-2 spike glycoprotein antibody having the same binding and / or blocking properties as a reference antibody, the reference antibody comprising a heavy chain and a light chain pair comprising the amino acid sequences of SEQ ID NO:56 / 58.

[0025] In any of the various embodiments of the methods discussed above or herein, the therapeutic or prophylactic composition comprises 1 mg to 10 g of an antigen-binding molecule. In some cases, the therapeutic or prophylactic composition comprises about 1.2 g of mAb10933 and about 1.2 g of mAb10987. In some cases, the therapeutic or prophylactic composition comprises about 1.2 g of mAb10985. In some cases, the therapeutic or prophylactic composition comprises about 4.0 g of mAb10933 and about 4.0 g of mAb10987. In some cases, the therapeutic or prophylactic composition comprises about 150 mg of mAb10933 and about 150 mg of mAb10987. In some cases, the therapeutic or prophylactic composition comprises about 150 mg of mAb10985. In some cases, the therapeutic or prophylactic composition comprises about 300 mg of mAb10933 and about 300 mg of mAb10987. In some cases, the therapeutic or preventative composition contains about 300 mg of mAb10985. In some cases, the therapeutic or preventative composition contains about 600 mg of mAb10933 and about 600 mg of mAb10987. In some cases, the therapeutic or preventative composition contains about 600 mg of mAb10985. In some cases, the therapeutic or preventative composition contains 150 mg to 1200 mg of mAb10933 and 150 mg to 1200 mg of mAb10987. In some cases, the therapeutic or preventative composition also contains 150 mg to 1200 mg of mAb10985. In some cases, the therapeutic or preventative composition contains about 1.2 g of mAb10989.

[0026] In some embodiments, the therapeutic or prophylactic composition is administered to the subject via intravenous infusion or subcutaneous injection. In some embodiments, this disclosure provides a method for treating a subject infected with SARS-CoV-2, comprising administering 1.2 g of mAb10987 and 1.2 g of mAb10933 via intravenous infusion. In some embodiments, this disclosure provides a method for treating a subject suffering from COVID-19, comprising administering 1.2 g of mAb10987 and 1.2 g of mAb10933 via intravenous infusion. In some embodiments, this disclosure provides a method for treating a subject infected with SARS-CoV-2, comprising administering 600 mg of mAb10987 and 600 mg of mAb10933 via intravenous infusion. In some embodiments, this disclosure provides a method for treating a subject suffering from COVID-19, comprising administering 600 mg of mAb10987 and 600 mg of mAb10933 via intravenous infusion. In some embodiments, this disclosure provides a method for treating a subject infected with SARS-CoV-2, the method comprising administering 4 g of mAb10987 and 4 g of mAb10933 via intravenous infusion. In some embodiments, this disclosure provides a method for treating a subject suffering from COVID-19, the method comprising administering 4 g of mAb10987 and 4 g of mAb10933 via intravenous infusion. In some embodiments, this disclosure provides a method for treating a subject infected with SARS-CoV-2, the method comprising administering 300 mg of mAb10987 and 300 mg of mAb10933 via intravenous infusion. In some embodiments, this disclosure provides a method for treating a subject suffering from COVID-19, the method comprising administering 300 mg of mAb10987 and 300 mg of mAb10933 via intravenous infusion. In some embodiments, this disclosure provides a method for treating a subject infected with SARS-CoV-2, the method comprising administering 150 mg of mAb10987 and 150 mg of mAb10933 via intravenous infusion. In some embodiments, this disclosure provides a method for treating a subject suffering from COVID-19, the method comprising administering 150 mg of mAb10987 and 150 mg of mAb10933 via intravenous infusion. In some embodiments, this disclosure provides a method for treating a subject infected with SARS-CoV-2, the method comprising administering 600 mg of mAb10987 and 600 mg of mAb10933 via subcutaneous injection.In some embodiments, this disclosure provides a method for treating a subject with COVID-19, comprising administering 600 mg of mAb10987 and 600 mg of mAb10933 via subcutaneous injection. In some embodiments, this disclosure provides a method for treating a subject infected with SARS-CoV-2, comprising administering 300 mg of mAb10987 and 300 mg of mAb10933 via subcutaneous injection. In some embodiments, this disclosure provides a method for treating a subject with COVID-19, comprising administering 300 mg of mAb10987 and 300 mg of mAb10933 via subcutaneous injection. In the above embodiments, mAb10987 and mAb10933 may be administered simultaneously, for example, by combining the antibodies in an IV bag prior to a single infusion, or by combining the antibodies into a syringe prior to a single injection. Alternatively, the two antibodies may be administered as two separate subcutaneous injections. In the above embodiments, the subject may be at high risk of clinical complications.

[0027] In any of the various implementation schemes, the subject exhibits one or more efficacy parameters after administration of the therapeutic composition, the one or more efficacy parameters being selected from the group consisting of: (a) a reduction in SARS-CoV-2 virus shedding from baseline; (b) an improvement of at least 1 point in clinical status using a 7-point ordinal scale; (c) a reduction or elimination of the need for supplemental oxygen; (d) a reduction or elimination of the need for mechanical ventilation; (e) prevention of COVID-19-related mortality; (f) prevention of all-cause mortality; and (g) a change in the serum concentration of one or more disease-related biomarkers. In some cases, the 7-point ordinal scale is: [1] death; [2] hospitalization requiring invasive mechanical ventilation or extracorporeal membrane oxygenation; [3] hospitalization requiring non-invasive ventilation or a high-flow oxygen device; [4] hospitalization requiring supplemental oxygen; [5] hospitalization not requiring supplemental oxygen—requiring ongoing medical care (COVID-19 related or otherwise); [6] hospitalization not requiring supplemental oxygen—no longer requiring ongoing medical care; and [7] not hospitalized. In some cases, one or more efficacy parameters are measured 21 days after administration of the first dose of the therapeutic composition. In some cases, the reduction in SARS-CoV-2 virus shedding from baseline is determined by real-time quantitative PCR (RT-qPCR) of nasopharyngeal swab samples, nasal cavity samples, or saliva samples. In some cases, changes in serum concentrations of one or more disease-associated biomarkers are observed, such as changes in C-reactive protein, lactate dehydrogenase, D-dimer, or ferritin.

[0028] In any of the various implementation schemes, following administration of the therapeutic composition, the subject exhibited fewer than 5 COVID-19 related medical care visits, telemedicine visits, hospital admissions, and / or intensive care unit (ICU) admissions. In some cases, the subject exhibited fewer than 5 COVID-19 related medical care visits, telemedicine visits, hospital admissions, and / or ICU admissions within 29 days of administration of the first dose of the therapeutic composition. In some cases, the subject exhibited fewer than 4, 3, 2, or 1 COVID-19 related medical care visits, telemedicine visits, hospital admissions, and / or ICU admissions.

[0029] In some implementations, subjects tested negative for SARS-CoV-2 within 2 days to 3 weeks after the first administration of the therapeutic composition. In some cases, a negative SARS-CoV-2 test was determined by RT-qPCR in nasopharyngeal swab samples, nasal cavity samples, or saliva samples.

[0030] In some embodiments, these methods further include administering an additional therapeutic agent to the subject. In some cases, the additional therapeutic agent is an antiviral compound. In some embodiments, the antiviral compound is remdesivir. In some cases, the additional therapeutic agent is an IL-6 or IL-6R blocker. In some embodiments, the additional therapeutic agent is tocilizumab or thalidomide. In some cases, the additional therapeutic agent is a steroidal compound. In some embodiments, the additional therapeutic agent is administered prior to the therapeutic composition. In some embodiments, the additional therapeutic agent is administered after or simultaneously with the therapeutic composition. In any of the various embodiments of the methods discussed above or herein, the subject may be seronegative for SARS-CoV-2 infection.

[0031] In one aspect, this disclosure provides a method for improving one or more clinical parameters of SARS-CoV-2 infection, the method comprising administering a therapeutic composition to a subject suffering from SARS-CoV-2 infection, wherein the therapeutic composition comprises a first anti-SARS-CoV-2 spike glycoprotein antibody or an antigen-binding fragment thereof and a second anti-SARS-CoV-2 spike glycoprotein antibody or an antigen-binding fragment thereof, the first anti-SARS-CoV-2 spike glycoprotein antibody or an antigen-binding fragment thereof comprising three heavy chain complementarity-determining regions (HCDRs) and three light chain complementarity-determining regions (LCDRs), the three HCDRs and the three LCDRs being contained within a heavy chain variable region (HCVR) and a light chain variable region (LCVR) amino acid sequence pair, the HCVR and LCVR amino acid sequence pair comprising the amino acid sequence of SEQ ID NO:2 / 10, the second anti-SARS-CoV-2 spike glycoprotein antibody or an antigen-binding fragment thereof comprising three HCDRs and three LCDRs, the three HCDRs and the three LCDRs being contained within an HCVR and LCVR amino acid sequence pair, the HCVR and LCVR amino acid sequence pair comprising the amino acid sequence of SEQ ID NO:2 / 10. The amino acid sequence NO:22 / 30, wherein the therapeutic composition, when administered to a comparable group of seronegative subjects, more rapidly relieved at least one symptom of SARS-CoV-2 infection when administered to a group of seronegative subjects compared to a group of subjects administered a placebo.

[0032] In one aspect, this disclosure provides a method for improving one or more clinical parameters of SARS-CoV-2 infection, wherein the method comprises administering a therapeutic composition to a subject suffering from SARS-CoV-2 infection, wherein the therapeutic composition comprises a first anti-SARS-CoV-2 spike glycoprotein antibody or an antigen-binding fragment thereof and a second anti-SARS-CoV-2 spike glycoprotein antibody or an antigen-binding fragment thereof, the first anti-SARS-CoV-2 spike glycoprotein antibody or an antigen-binding fragment thereof comprising three heavy chain complementarity-determining regions (HCDRs) and three light chain complementarity-determining regions (LCDRs), the three HCDRs and the three LCDRs being contained within a heavy chain variable region (HCVR) and a light chain variable region (LCVR) amino acid sequence pair, the HCVR and LCVR amino acid sequence pair comprising the amino acid sequence of SEQ ID NO:2 / 10, the second anti-SARS-CoV-2 spike glycoprotein antibody or an antigen-binding fragment thereof comprising three HCDRs and three LCDRs, the three HCDRs and the three LCDRs being contained within an HCVR and LCVR amino acid sequence pair, the HCVR and LCVR amino acid sequence pair comprising the amino acid sequence of SEQ ID NO:2 / 10. The amino acid sequence NO:22 / 30, wherein the therapeutic composition, when administered to a seronegative subject population, more rapidly relieved at least one symptom of SARS-CoV-2 infection compared to a comparable seropositive subject population.

[0033] In one aspect, this disclosure provides a method for improving one or more clinical parameters of SARS-CoV-2 infection, the method comprising administering a therapeutic composition to a subject suffering from SARS-CoV-2 infection, wherein the therapeutic composition comprises a first anti-SARS-CoV-2 spike glycoprotein antibody or an antigen-binding fragment thereof and a second anti-SARS-CoV-2 spike glycoprotein antibody or an antigen-binding fragment thereof, the first anti-SARS-CoV-2 spike glycoprotein antibody or an antigen-binding fragment thereof comprising three heavy chain complementarity-determining regions (HCDRs) and three light chain complementarity-determining regions (LCDRs), the three HCDRs and the three LCDRs being contained within a heavy chain variable region (HCVR) and a light chain variable region (LCVR) amino acid sequence pair, the HCVR and LCVR amino acid sequence pair comprising the amino acid sequence of SEQ ID NO:2 / 10, the second anti-SARS-CoV-2 spike glycoprotein antibody or an antigen-binding fragment thereof comprising three HCDRs and three LCDRs, the three HCDRs and the three LCDRs being contained within an HCVR and LCVR amino acid sequence pair, the HCVR and LCVR amino acid sequence pair comprising the amino acid sequence of SEQ ID NO:2 / 10. The amino acid sequence of NO:22 / 30, wherein the therapeutic composition reduced the viral load in the subject population 7 days after administration (day 7) compared to the day of administration (day 0).

[0034] In some implementations, compared with a comparable group of subjects treated with placebo, the time-weighted mean change in nasopharyngeal (NP) viral load from baseline to day 7 was at least 0.86 log10 copies / mL lower in patients treated with 0.6 g primary anti-SARS-CoV-2 spike glycoprotein antibody and 0.6 g secondary anti-SARS-CoV-2 spike glycoprotein antibody (p < 0.0001).

[0035] In some implementations, compared with a comparable group of subjects treated with placebo, the nasopharyngeal (NP) viral load in the seronegative subject group treated with 1.2g primary anti-SARS-CoV-2 spike glycoprotein antibody and 1.2g secondary anti-SARS-CoV-2 spike glycoprotein antibody was reduced by at least 1.04 log10 copies / mL from baseline to day 7 (p<0.0001).

[0036] In some implementations, compared with a comparable population of subjects treated with placebo, patients treated with 0.6g of primary anti-SARS-CoV-2 spike glycoprotein antibody and 0.6g of secondary anti-SARS-CoV-2 spike glycoprotein antibody showed a mean decrease of at least 0.71 log10 copies / mL in nasopharyngeal (NP) viral load from baseline to day 7 (p<0.0001).

[0037] In some implementations, compared with a comparable population of subjects treated with placebo, patients treated with 1.2g of primary anti-SARS-CoV-2 spike glycoprotein antibody and 1.2g of secondary anti-SARS-CoV-2 spike glycoprotein antibody showed a mean decrease of 0.86 log10 copies / mL in nasopharyngeal (NP) viral load from baseline to day 7 (p<0.0001).

[0038] In one aspect, this disclosure provides a method for improving one or more clinical parameters of SARS-CoV-2 infection, the method comprising administering a therapeutic composition to a subject suffering from SARS-CoV-2 infection, wherein the therapeutic composition comprises a first anti-SARS-CoV-2 spike glycoprotein antibody or an antigen-binding fragment thereof and a second anti-SARS-CoV-2 spike glycoprotein antibody or an antigen-binding fragment thereof, the first anti-SARS-CoV-2 spike glycoprotein antibody or an antigen-binding fragment thereof comprising three heavy chain complementarity-determining regions (HCDRs) and three light chain complementarity-determining regions (LCDRs), the three HCDRs and the three LCDRs being contained within a heavy chain variable region (HCVR) and a light chain variable region (LCVR) amino acid sequence pair, the HCVR and LCVR amino acid sequence pair comprising the amino acid sequence of SEQ ID NO:2 / 10, the second anti-SARS-CoV-2 spike glycoprotein antibody or an antigen-binding fragment thereof comprising three HCDRs and three LCDRs, the three HCDRs and the three LCDRs being contained within an HCVR and LCVR amino acid sequence pair, the HCVR and LCVR amino acid sequence pair comprising the amino acid sequence of SEQ ID NO:2 / 10. The amino acid sequence of NO:22 / 30, wherein the therapeutic composition reduces viral load in the subject population.

[0039] In some embodiments of the methods discussed above or herein, administration of the therapeutic composition comprises administering 0.6 g of a primary anti-SARS-CoV-2 spike glycoprotein antibody and 0.6 g of a secondary anti-SARS-CoV-2 spike glycoprotein antibody, and wherein the administration produces a mean viral load reduction of at least 3.00 log10 copies / mL on day 7 after administration, compared to the baseline viral load measured on day 0 prior to administration. In some cases, the reduction is at least 3.50 log10 copies / mL. In some cases, the reduction is at least 3.90 log10 copies / mL.

[0040] In some embodiments, administration of the therapeutic composition comprises administering 1.2 g of a primary anti-SARS-CoV-2 spike glycoprotein antibody and 1.2 g of a secondary anti-SARS-CoV-2 spike glycoprotein antibody, wherein the administration produces a mean viral load reduction of at least 3.50 log10 copies / mL on day 7 after administration, compared to the baseline viral load measured on day 0 prior to administration. In some cases, the reduction is at least 3.75 log10 copies / mL. In some cases, the reduction is at least 4.09 log10 copies / mL.

[0041] In one aspect, this disclosure provides a method for improving one or more clinical parameters of SARS-CoV-2 infection, the method comprising administering a therapeutic composition to a subject suffering from SARS-CoV-2 infection, wherein the therapeutic composition comprises a first anti-SARS-CoV-2 spike glycoprotein antibody or an antigen-binding fragment thereof and a second anti-SARS-CoV-2 spike glycoprotein antibody or an antigen-binding fragment thereof, the first anti-SARS-CoV-2 spike glycoprotein antibody or an antigen-binding fragment thereof comprising three heavy chain complementarity-determining regions (HCDRs) and three light chain complementarity-determining regions (LCDRs), the three HCDRs and the three LCDRs being contained within a heavy chain variable region (HCVR) and a light chain variable region (LCVR) amino acid sequence pair, the HCVR and LCVR amino acid sequence pair comprising the amino acid sequence of SEQ ID NO:2 / 10, the second anti-SARS-CoV-2 spike glycoprotein antibody or an antigen-binding fragment thereof comprising three HCDRs and three LCDRs, the three HCDRs and the three LCDRs being contained within an HCVR and LCVR amino acid sequence pair, the HCVR and LCVR amino acid sequence pair comprising the amino acid sequence of SEQ ID NO:2 / 10. The amino acid sequence of NO:22 / 30, wherein, compared with a comparable group of subjects treated with placebo, the therapeutic composition reduced the median time to symptom relief (defined as symptoms becoming milder or disappearing) by 4 days in a group of subjects treated with either 0.6g of primary anti-SARS-CoV-2 spike glycoprotein antibody and 0.6g of secondary anti-SARS-CoV-2 spike glycoprotein antibody or 1.2g of primary anti-SARS-CoV-2 spike glycoprotein antibody and 1.2g of secondary anti-SARS-CoV-2 spike glycoprotein antibody. In some embodiments, subjects and / or subject groups include unhospitalized subjects with COVID-19.

[0042] Any of the various methods discussed above or herein may be reformulated as (i) methods for treating and / or preventing SARS-CoV-2 infection and / or COVID-19, and / or for treating, preventing, and reducing the severity or progression of SARS-CoV-2 infection and / or COVID-19 or their symptoms, or (ii) the use of antigen-binding molecules or antibodies (and antigen-binding fragments) in the preparation of medicaments for treating and / or preventing SARS-CoV-2 infection and / or COVID-19, and / or for treating, preventing, and reducing the severity or progression of SARS-CoV-2 infection and / or COVID-19 or their symptoms. Specifically, this disclosure includes the use of antigen-binding molecules that bind to the surface protein of SARS-CoV-2 (including the anti-SARS-CoV-2 spike glycoprotein antibodies or their antigen-binding fragments discussed herein) for the prevention and treatment of SARS-CoV-2 infection and COVID-19 and / or for treating, preventing, and reducing the severity or progression of SARS-CoV-2 infection and / or COVID-19 or their symptoms. This disclosure also includes the use of antigen-binding molecules that bind to the surface protein of SARS-CoV-2 (including the anti-SARS-CoV-2 spike glycoprotein antibodies or antigen-binding fragments thereof discussed herein) in the preparation of medicaments for the prevention and treatment of SARS-CoV-2 infection and COVID-19 and / or for the treatment, prevention, and reduction of the severity or progression of SARS-CoV-2 infection and / or COVID-19. When methods are discussed herein with reference to combinations of two anti-SARS-CoV-2 spike protein antibodies, such combinations include the use of a first such antibody or antigen-binding fragment thereof in the preparation of a medicament used in combination with a second such antibody or antigen-binding fragment thereof (or a third or fourth such antibody or antigen-binding fragment), and the use of a second such antibody or antigen-binding fragment thereof (or a third or fourth such antibody or antigen-binding fragment thereof) in the preparation of a medicament used in combination with a first such antibody.

[0043] In various embodiments, any feature or component of the embodiments discussed above or herein may be combined, and such combinations are covered within the scope of this disclosure. Any particular value discussed above or herein may be combined with another related value discussed above or herein to enumerate a range of values ​​having an upper and lower end indicating a range, and such ranges are covered within the scope of this disclosure.

[0044] Other implementation methods will become apparent upon reading the detailed implementation details that follow. Attached Figure Description

[0045] Figure 1A and Figure 1BThis section provides an overview of the study design for evaluating the prophylactic efficacy of anti-SARS-CoV-2 spike glycoprotein antibodies in a rhesus monkey model of SARS-CoV-2 infection. Figure 1A The effects of anti-SARS-CoV-2 spike glycoprotein antibody prophylaxis on viral genomic RNA (gRNA) and subgenomic RNA (sgRNA) in nasopharyngeal swabs and bronchoalveolar lavage (BAL) fluid. Figure 1B ).

[0046] Figure 2A , Figure 2B , Figure 2C and Figure 2D This section provides an overview of the study design for evaluating the prophylactic and therapeutic efficacy of anti-SARS-CoV-2 spike glycoprotein antibodies in a rhesus monkey model of SARS-CoV-2 infection. Figure 2A The effect of anti-SARS-CoV-2 spike glycoprotein antibody on the prevention of viral gRNA and sgRNA in nasopharyngeal and oral swabs. Figure 2B The effect of anti-SARS-CoV-2 spike glycoprotein antibody therapy on viral gRNA and sgRNA in nasopharyngeal and oral swabs. Figure 2C ) and representative images of lung tissue pathology in treated and placebo animals ( Figure 2D ).

[0047] Figure 3A and Figure 3B Showing from Figures 2A to 2D The results of RNA sequence analysis of the viral RNA studied are shown. Figure 3A The frequency of all amino acid changes identified in the spike protein across all viral sequences is shown; each point represents the frequency of the corresponding amino acid change in a specific viral sample, and the samples are grouped based on treatment regimen: isotype control (placebo), therapeutic antibody administered before (prophylactic) or after viral challenge (therapeutic). Figure 3B Detailed genomic information on all amino acid variations identified within the spike protein sequence across all samples is presented; for each sample, the frequency of all mutations has been calculated, and these frequencies are shown as a percentage of the viral population with amino acid variations in the input virus, or as a range of frequency percentages (minimum % to maximum %) in the viral populations isolated from the placebo, prophylactic, and therapeutic groups.

[0048] Figure 4A , Figure 4B and Figure 4C This section provides an overview of the study design for evaluating the therapeutic and prophylactic efficacy of anti-SARS-CoV-2 spike glycoprotein antibodies in a Syrian golden hamster model of SARS-CoV-2 infection. Figure 4AThe effect of anti-SARS-CoV-2 spike glycoprotein antibody treatment or prevention on weight loss. Figure 4B The study also investigated the effects of anti-SARS-CoV-2 spike glycoprotein antibody therapy on the levels of gRNA and sgRNA in the lungs.

[0049] Figure 5 This is a schematic overview of the research design discussed in Example 2.

[0050] Figure 6 A CONSORT diagram illustrating the screening, randomization, and treatment of subjects in the study discussed in Example 2 is shown.

[0051] Figure 7 The relationship between baseline serum antibody status and baseline viral load in the placebo group of the study discussed in Example 2 is shown.

[0052] Figure 8 The changes in viral load over time in the placebo group in the study discussed in Example 2 are shown, broken down by baseline serum antibody status.

[0053] Figure 9 The proportion of patients in the placebo group who had ≥1 COVID-19 related medical care visit (MAV) by day 29 in the study discussed in Example 2 is shown.

[0054] Figure 10A and Figure 10B This shows the daily time-weighted average (TWA) change in viral load from baseline during treatment with REGEN-COV in the study discussed in Example 2. 10 (Copies / mL) (Forest Plot).

[0055] Figure 11 This shows the daily TWA (log) change in viral load from baseline during treatment with REGEN-COV in the study discussed in Example 2. 10 (Copies / mL) (Plot). The figure shows the change in mean viral load from baseline to day 7 at each visit in the overall population (modified full analysis set, excluding patients who were negative for SARS-CoV-2 by qualitative reverse transcription polymerase chain reaction at baseline) and in groups defined by baseline antibody status and baseline viral load (in log10 copies / mL). Figure C The bars in the table represent standard error. The limit of detection (dashed line) is 714 copies / mL (2.85 log10 copies / mL). IV, intravenous (intravenous); SE, standard error.

[0056] Figure 12The duration of RT-qPCR remaining negative by baseline viral load category is shown in the study discussed in Example 2.

[0057] Figure 13 The proportion of patients with high viral load at each visit in the study discussed in Example 2 is shown.

[0058] Figure 14A , Figure 14B and Figure 14C The proportion of patients with COVID-19-related MAV in the study discussed in Example 2 is shown.

[0059] Figure 15A , Figure 15B and Figure 15C The viral load of patients with and without ≥1 COVID-19-related MAV by day 29 in the study discussed in Example 2 is shown.

[0060] Figure 16 The study showed that seronegative patients (n=217) had significantly higher viral loads than those who had developed their own SARS-CoV-2 antibodies (seropositive) at the time of randomization.

[0061] Figure 17 The study showed that among hospitalized COVID-19 patients receiving low-flow supplemental oxygen, seropositive patients had a lower cumulative incidence of death or mechanical ventilation compared to seronegative patients.

[0062] Figure 18 Clinical outcomes in cohort 1 are shown categorized by serological status and viral load. Patients with negative serological status or high viral load at baseline had worse clinical outcomes.

[0063] Figure 19 A seamless phase 1 / 2 / 3 study design was demonstrated in hospitalized patients with COVID-19.

[0064] Figure 20 This shows the results for the full analysis set (FAS), the modified full analysis set (mFAS), and viral load >10. 6 Or viral load >10 7 The number of patients who are serum negative, serum positive, or have borderline results or missing data (“Other”).

[0065] Figure 21 The mean viral load is shown for seronegative patients (circles), seronegative patients (squares), and other patients (borderline results or missing data; triangles). TWA, time-weighted mean; CI, confidence interval.

[0066] Figure 22The mean viral load is shown for patients treated intravenously with placebo (circle), 1.2g mAb10933 + 1.2g mAb10987 (total 2.4g; square), or 4g mAb10933 + 4g mAb10987 (total 8g; triangle).

[0067] Figure 23 The graph shows the change in mean viral load from baseline in patients treated intravenously with placebo (circles), 1.2g mAb10933 + 1.2g mAb10987 (total 2.4g; squares), or 4g mAb10933 + 4g mAb10987 (total 8g; triangles). Patients are categorized by viral load as follows: >10 4 copies / mL, >10 5 copies / mL, >10 6 copies / mL and >10 7 Copy / mL.

[0068] Figure 24 The graph shows the change in mean viral load over time in patients treated intravenously with placebo (circles), 1.2g mAb10933 + 1.2g mAb10987 (total 2.4g; squares), or 4g mAb10933 + 4g mAb10987 (total 8g; triangles). Patients are categorized by baseline viral load as follows: >10 4 copies / mL, >10 5 copies / mL, >10 6 copies / mL and >10 7 Copy / mL.

[0069] Figure 25 The graph shows the change in mean viral load over time in patients treated intravenously with placebo (circles), 1.2g mAb10933 + 1.2g mAb10987 (total 2.4g; squares), or 4g mAb10933 + 4g mAb10987 (total 8g; triangles). Patients are categorized by serological status and the following clinical trials: 2066 (inpatient study; Example 1) and 2067 (non-asylum / outpatient study; Example 2).

[0070] Figure 26The graph shows the change in mean viral load from baseline in patients treated intravenously with placebo (circles), 1.2g mAb10933 + 1.2g mAb10987 (total 2.4g; squares), or 4g mAb10933 + 4g mAb10987 (total 8g; triangles). Patients are categorized by serological status and the following clinical trials: 2066 (inpatient study; Example 1) and 2067 (non-asylum / outpatient study; Example 2).

[0071] Figure 27 The graph shows the change in mean viral load over time in patients treated intravenously with placebo (circles), 1.2g mAb10933 + 1.2g mAb10987 (total 2.4g; squares), or 4g mAb10933 + 4g mAb10987 (total 8g; triangles). Patients are categorized by baseline viral load and the following clinical trials: 2066 (inpatient study; Example 1) and 2067 (outpatient / outpatient study; Example 2).

[0072] Figure 28 The graph shows the change in mean viral load from baseline in patients treated intravenously with placebo (circles), 1.2g mAb10933 + 1.2g mAb10987 (total 2.4g; squares), or 4g mAb10933 + 4g mAb10987 (total 8g; triangles). Patients are categorized by baseline viral load and the following clinical trials: 2066 (inpatient study; Example 1) and 2067 (outpatient / outpatient study; Example 2).

[0073] Figure 29 The neutralizing percentages of mAb10933 (REGN10933) alone, mAb10987 (REGN10987) alone, and mAb10933+mAb10987 (REGN10933+REGN10987) combination against pseudotyped VSV expressing the B.1.1.7 SARS-CoV-2 variant (also known as the "UK variant") are shown. Both antibodies, alone or in combination, can neutralize the virus.

[0074] Figure 30 The results showed that in placebo and mAb10933+mAb10987 (also collectively referred to as REGEN-COV) TM Weekly viral load in individual symptomatic subjects (solid symbols) and asymptomatic subjects (hollow symbols) in both treatment groups. REGEN-COV TM Infections in the control group lasted no more than one week, while approximately 40% of infections in the placebo group lasted three to four weeks, as assessed by measuring viral load.

[0075] Figure 31 A schematic diagram of a phase 3 study of non-hospitalized patients treated with REGEN-COV or placebo is shown.

[0076] Figure 32 The revisions to the Phase 3 cohort recruitment are shown, modifying the trial to include 2400 mg and 1200 mg doses.

[0077] Figure 33 The clinical efficacy of REGEN-COV was demonstrated, comparing the treatment effects of placebo, 2400 mg, and 1200 mg intravenous treatment groups. Treatment significantly reduced...

[0078] COVID-19-related hospitalizations or all-cause mortality and duration of symptoms, and similar therapeutic effects at both dose levels (the point estimate for the 2400 mg dose has a high confidence level due to the large event size).

[0079] Figure 34 Balanced baseline demographics are shown for the Phase 3 cohort 1mFAS (patients ≥18 years old who were SARS-CoV-2 PCR positive at baseline and had ≥1 serious COVID-19 risk factor) of outpatients treated with 8000 mg REGEN-COV, 2400 mg REGEN-COV, 1200 mg REGEN-COV, or placebo (PBO). Phase 3 patients had higher baseline viral load and higher baseline seronegativity than high-risk patients in Phase 1 / 2.

[0080] Figure 35 Kaplan-Meier curves are shown for the time to COVID-19-related hospitalization or all-cause mortality up to day 29 following administration of 1200 mg mAb10933 + 1200 mg mAb10987 (intravenous) in subjects with ≥1 severe COVID-19 risk factor. Compared to placebo, the risk of COVID-19 hospitalization or all-cause mortality was reduced by 71% across the entire modified full analysis set (mFAS).

[0081] Figure 36 Kaplan-Meier curves are shown for the time to COVID-19-related hospitalization or all-cause mortality up to day 29 following administration of 600 mg mAb10933 + 600 mg mAb10987 (intravenous) in subjects with ≥1 severe COVID-19 risk factor. Compared to placebo, the risk of COVID-19 hospitalization or all-cause mortality was reduced by 71% across the entire modified full analysis set (mFAS).

[0082] Figure 37This study shows the number of COVID-19-related hospitalizations or all-cause deaths up to day 29 following administration of either 1200 mg mAb10933 + 1200 mg mAb10987 (IV) or 600 mg mAb10933 + 600 mg mAb10987 (IV) in subjects with ≥1 severe COVID-19 risk factor. Results were consistent between the two treatment groups.

[0083] Figure 38 Kaplan-Meier curves for the time to symptom resolution of COVID-19 are shown in subjects with ≥1 severe COVID-19 risk factor. HR: hazard ratio. The median time to symptom resolution was 14 days in the placebo group and 10 days in each of the 1.2g and 2.4g treatment groups.

[0084] Figure 39 The time to symptom resolution is shown for outpatients with ≥1 severe COVID-19 risk factor. Improvement in symptom resolution was consistent between the modified full analysis set (mFAS) and the group with high viral load or seronegative status at baseline.

[0085] Figure 40 Serious adverse events (SAEs) and adverse events of particular concern (SAEIs) are presented in outpatients in Phase 3 Cohort 1 treated intravenously with 1200 mg REGEN-COV, 2400 mg REGEN-COV, 8000 mg REGEN-COV, or placebo (PBO). Safety was acceptable in all treatment groups, and no serious safety issues were identified. Specifically, SAEs and AESIs occurred more frequently in the placebo group compared to any REGEN-COV treatment group; no safety imbalances were observed between the different REGEN-COV dose groups; no safety signals were observed in safety laboratories (chemistry, hematology); more patients in the placebo group experienced treatment-term adverse events (TEAEs) with fatal consequences compared to any REGEN-COV treatment group; and very few patients in the REGEN-COV dose groups experienced infusion-related reactions (IRRs) and hypersensitivity-related AESIs.

[0086] Figure 41Serious adverse events (SAEs) were observed in outpatients of Phase 3 Cohort 1 treated intravenously with 1200 mg REGEN-COV, 2400 mg REGEN-COV, 8000 mg REGEN-COV, or placebo (PBO). More than one patient in each treatment group experienced an SAE. The placebo group had a higher frequency of SAEs compared to any REGEN-COV dose group, with more frequently reported events consistent with COVID-19 and related complications, and the lower frequency of events in the REGEN-COV dose group was consistent with treatment benefit.

[0087] Figure 42 Adverse events of particular concern (AESIs) (e.g., infusion-related reactions or hypersensitivity reactions) in outpatients of Phase 3 cohort 1 treated intravenously with 1200 mg REGEN-COV, 2400 mg REGEN-COV, 8000 mg REGEN-COV, or placebo (PBO) are shown. >1 patient experienced an AESI in any treatment group. The incidence of infusion-related reactions or hypersensitivity reactions was low across all dose groups.

[0088] Figure 43 The changes in viral load from baseline (log10 copies / mL) on day 7 after treatment with mAb10933 and mAb10987 are shown in outpatients with one or more severe COVID-19 risk factors.

[0089] Figure 44 This shows the change in viral load (log10 copies / mL) from baseline on day 7 after treatment with mAb10933 and mAb10987 in outpatients with one or more severe COVID-19 risk factors. N: Number of subjects; SD: Standard deviation; D7: Day 7; mFAS: Modified full analysis set; PA6: Protocol revision 6, which modified the clinical trial protocol to remove the 8g dose and introduce a 1.2g dose.

[0090] Figure 45 Demographic and baseline characteristics of seronegative IV patients (seronegative mFAS) are shown. These groups are well balanced.

[0091] Figure 46 Serum negative

[0092] Demographic and baseline characteristics of patients receiving subcutaneous injections (serum-negative mFAS). These groups are well-balanced.

[0093] Figure 47The least-squares mean of viral load over time from baseline is shown in patients treated with mAb10933+mAb10987 via intravenous (IV) or subcutaneous (SC).

[0094] Figure 48 The changes in viral load from baseline are shown in phase 1 / 2 and phase 3 trials in 2067 seronegative patients (Example 2; outpatients) and in 20145 patients (Example 7; dose range discovery). The changes in viral load from baseline were comparable between studies.

[0095] Figure 49 Safety data from the clinical trial described in Example 7 are shown. All treatment groups were well tolerated, and no safety signals were identified.

[0096] Figure 50 Treatment-period adverse events observed in patients receiving subcutaneous treatment during the clinical trial described in Example 7 are shown. All doses were well tolerated, with few treatment-period adverse events. Of the observed events, these were not serious.

[0097] Figure 51 A comparison is shown between the clinical trial described in Example 2 (“Analysis Set 2067”) and the clinical trial described in Example 7 (“Analysis Set 20145”). Data for Analysis Set 2067 are provided before and after the correction of the dose groups from 2400 mg and 8000 mg to 1200 mg and 2400 mg (original Ph3 and corrected Ph3, respectively). Analysis of the change in viral load from baseline to day 7 (log10 copies / mL) shows that the reduction in viral load was similar before and after the correction and across all doses.

[0098] Figure 52 The mean viral load is shown for patients with and without hospitalization / death outcomes. Viral loads for patients treated with placebo (PBO), 2.4g REGEN-COV, or 8.0g REGEN-COV are shown before PA6. Viral loads for patients treated with PBO, 1.2g REGEN-COV, or 2.4g REGEN-COV are shown at or after PA6.

[0099] Figure 53 The spaghetti plot shows viral load in individual patients with and without hospitalization / death outcomes (placebo, 1.2g REGEN-COV, and 2.4g REGEN-COV).

[0100] Figure 54The spaghetti plot shows viral loads in individual patients with and without hospitalization / death outcomes (placebo, 2.4 g REGEN-COV, and 8.0 g REGEN-COV).

[0101] Figure 55 Box plots of viral load at baseline and day 7 post-treatment are shown for individual patients with and without hospitalization / death outcomes (placebo, 1.2 g REGEN-COV, and 2.4 g REGEN-COV).

[0102] Figure 56 Box plots of viral load at baseline and day 7 post-treatment are shown for individual patients with and without hospitalization / death outcomes (placebo, 2.4 g REGEN-COV, and 8.0 g REGEN-COV).

[0103] Figure 57 An overview of the clinical trial described in Example 7 is shown.

[0104] Figure 58 The changes in viral load on day 7 after treatment are shown in all patients, seronegative patients, and seronegative patients (divided into those with and without COVID-19-related events). Patients who received placebo and experienced events had higher baseline viral loads and slower viral clearance, while seronegative patients who experienced events had similarly high baseline viral loads and delayed clearance, suggesting they may have had an ineffective antibody response.

[0105] Figure 59 The hierarchy of hypothesis testing in the phase 3 prevention trial described in Example 4 is shown, along with the treatment efficacy at each endpoint. REGEN-COV significantly prevented infection, improved disease progression, and reduced viral load.

[0106] Figure 60 The symptomatic infection endpoint in the Phase 3 prevention trial described in Example 4 is shown. REGEN-COV significantly reduced symptomatic COVID-19 in all three definitions.

[0107] Figure 61 The cumulative incidence of symptomatic infection in the Phase 3 prevention trial described in Example 4 is shown. REGEN-COV prevented the onset of symptomatic infection starting 1 day after administration.

[0108] Figure 62 The weekly onset of symptomatic infection is shown in the Phase 3 prevention trial described in Example 4. REGEN-COV reduced the overall risk of symptomatic infection by 81%, by 72% in the first week, and by 93% from week 2 to week 4.

[0109] Figure 63 This demonstrates that in the Phase 3 prevention trial described in Example 4, REGEN-COV significantly reduced the number of weeks of symptomatic infection.

[0110] Figure 64 This demonstrates that in the Phase 3 prevention trial described in Example 4, REGEN-COV significantly reduced the number of weeks of symptomatic infection.

[0111] Figure 65 This demonstrates that in the Phase 3 prevention trial described in Example 4, REGEN-COV significantly reduced the total number of weeks of infection.

[0112] Figure 66 This illustrates the hierarchy of hypothesis testing in the Phase 3 first-line therapy trial described in Example 4. REGEN-COV significantly prevented...

[0113] Asymptomatic infection progresses to disease and reduces viral load. Treatment is more effective after the first three days.

[0114] Figure 67 The mean viral load over time in asymptomatic patients in the Phase 3 first-line therapy trial described in Example 4 (2069) is shown compared to symptomatic patients in the modified Phase 3 trial of Example 2 (2067). Early treatment with REGEN-COV provided a greater reduction in viral load over time.

[0115] Figure 68 The mean concentrations of mAb10933 and mAb10987 in the serum of the sentinel and safety cohorts over time were shown in the Phase 3 first-line therapy trial described in Example 4, following a single 1200 mg dose.

[0116] Figure 69 This demonstrates that REGEN-COV has an acceptable and well-tolerated safety profile in the Phase 3 trial of Example 4, with no severe or serious safety issues.

[0117] Figure 70 It was shown that in the phase 3 trial of Example 4, adverse events during the treatment period occurred in ≥2% of any treatment group.

[0118] Figure 71 As shown, serious adverse events are rare, and no COVID-related serious adverse events were observed in patients treated with REGEN-COV.

[0119] Figure 72This study shows the cumulative incidence of symptomatic infection, calculated on a study day basis, in a clinical trial evaluating the ability of REGEN-COV to prevent COVID-19 symptoms. Subcutaneous administration of REGEN-COV reduced the risk of symptomatic SARS-CoV-2 infection by 81.4%.

[0120] Figure 73 This study presents the cumulative incidence of symptomatic infection, calculated on a study day basis, in a clinical trial assessing the ability of REGEN-COV to prevent COVID-19 symptoms. During the efficacy evaluation period, treatment with 1200 mg REGEN-COV subcutaneously (SC) resulted in a 31.5% relative risk reduction in the progression from asymptomatic to symptomatic infection (29 / 100 [29.0%], compared to 44 / 104 [42.3%] in the placebo group; p = 0.0380), with a more significant effect observed after 3 days or longer of REGEN-COV administration (76.4% relative risk reduction).

[0121] Figure 74 This illustrates the change in mean viral load over time in PCR-positive and serologically negative patients at baseline during the clinical trial described in Example 7.

[0122] Figure 75 The mean concentration of total REGEN-COV in serum following a single intravenous (IV) and subcutaneous (SC) injection is shown in the clinical trial described in Example 7, recruited from out-of-bed PCR-positive patients.

[0123] Figure 76 The number of patients assigned to different treatment groups is shown in a clinical trial designed to study REGEN-COV treatment in non-hospitalized patients.

[0124] Figure 77A , Figure 77B and Figure 77C : Figure 77A The clinical efficacy of REGEN-COV at an IV dose of 1200 mg was demonstrated in relation to hospitalization or all-cause mortality. Treatment significantly reduced hospitalization or all-cause mortality. Figure 77B The clinical efficacy of REGEN-COV at an IV dose of 2400 mg was demonstrated in relation to hospitalization or all-cause mortality. Treatment significantly reduced hospitalization or all-cause mortality. Figure 77C The clinical efficacy of REGEN-COV at 1200 mg IV dose and 2400 mg IV dose was demonstrated in terms of symptom resolution time. Treatment reduced the median number of days to symptom resolution by 4 days.

[0125] Figure 78The demographic and baseline medical characteristics of patients assigned to different treatment groups are shown in a clinical trial designed to study REGEN-COV treatment in non-hospitalized patients.

[0126] Figure 79 The effects of different treatment groups on each of the endpoints of a phase 3 clinical trial in non-hospitalized adult patients with COVID-19 are shown.

[0127] Figure 80 An overview of serious adverse events and adverse events of particular concern is provided in patients treated intravenously with 1200 mg REGEN-COV, 2400 mg REGEN-COV, 8000 mg REGEN-COV or placebo.

[0128] Figure 81 A schematic overview of a study design evaluating treatment with REGEN-COV in adult non-hospitalized patients with COVID-19 is shown.

[0129] Figure 82 The changes in viral load over time in the placebo group are shown, broken down by baseline serum antibody status.

[0130] Figure 83A , Figure 83B and Figure 83C : Figure 83A A forest plot showing COVID-19-related hospitalizations or all-cause mortality up to day 29 in non-hospitalized adults with one or more severe COVID-19 risk factors is presented. Figure 83B These data are broken down according to the risk factors defined in the plan, and Figure 83C These data are broken down by combination of other risk factors.

[0131] Figure 84A and Figure 84B : Figure 84A This shows the proportion of patients who received a single intravenous dose of 1200 mg REGEN-COV and were hospitalized for COVID-19 or died from all causes between day 4 and day 29. Figure 84B This shows the proportion of patients who received a single intravenous dose of 2400 mg REGEN-COV and were hospitalized for COVID-19 or died from all causes between day 4 and day 29.

[0132] Figure 85 The time to symptom resolution was shown for outpatients with one or more serious COVID-19 risk factors who were treated intravenously with 1200 mg REGEN-COV or 2400 mg REGEN-COV.

[0133] Figure 86A, Figure 86B and Figure 86C : Figure 86A This study illustrates the change in viral load over time in outpatients with one or more severe COVID-19 risk factors treated intravenously with 1200 mg REGEN-COV or 2400 mg REGEN-COV. Both doses significantly reduced viral load compared to placebo. Figure 86B The changes in viral load in these patients over time are shown, broken down by baseline serum antibody status (seronegative and seropositive). Figure 86C The changes in viral load over time for these patients are shown, categorized by baseline viral load (>10). 4 copies / mL, >10 5 copies / mL, >10 6 copies / mL and >10 7 (Copies / mL) subdivision.

[0134] Figure 87 The changes in viral load from baseline (log10 copies / mL) on day 7 in outpatients with one or more serious COVID-19 risk factors treated intravenously with 1200 mg REGEN-COV or 2400 mg REGEN-COV are shown.

[0135] Figure 88A , Figure 88B and Figure 88C : Figure 88A The study shows the change in viral load over time in outpatients with one or more severe COVID-19 risk factors treated intravenously with 2400 mg REGEN-COV or 8000 mg REGEN-COV. Both doses significantly reduced viral load compared to placebo. Figure 88B The changes in viral load in these patients over time are shown, broken down by baseline serum antibody status (seronegative and seropositive). Figure 88C The changes in viral load over time for these patients are shown, categorized by baseline viral load (>10). 4 copies / mL, >10 5 copies / mL, >10 6 copies / mL and >10 7 (Copies / mL) subdivision.

[0136] Figure 89 The order of testing for the primary hierarchical analysis is shown, indicating the order of evaluation for the primary and secondary endpoints.

[0137] Figure 90 The protocol-defined risk factors for severe COVID-19 are shown in the REGEN-COV clinical trial evaluating non-hospitalized patients.

[0138] Figure 91 Demographic and baseline medical characteristics of patients receiving 8000 mg REGEN-COV or placebo are shown.

[0139] Figure 92 The proportion of patients in the placebo group who had at least one COVID-19-related hospitalization or all-cause death, categorized by baseline viral load, is shown.

[0140] Figure 93 The viral load in the placebo group is shown (with hospitalization / death, without hospitalization / death, and breakdown by baseline serum antibody status).

[0141] Figure 94 The percentage of patients with one or more COVID-19-related hospitalizations and / or all-cause deaths is shown.

[0142] Figure 95 The percentage of patients who had one or more medical care visits or died from all causes after treatment with REGEN-COV is shown.

[0143] Figure 96 Results of hospitalized patients during the clinical trial process were presented in outpatients with one or more severe COVID-19 risk factors.

[0144] Figure 97 This shows the proportion of patients treated with 1200 mg REGEN-COV, 2400 mg REGEN-COV, or placebo who had one or more COVID-19-related hospitalizations, emergency room visits, or all-cause deaths.

[0145] Figure 98 This shows the proportion of patients who had one or more COVID-19-related hospitalizations and / or all-cause deaths among those treated with 8000 mg REGEN-COV or placebo.

[0146] Figure 99 This shows the proportion of patients treated with 8000 mg REGEN-COV or placebo who had one or more COVID-19-related medical care visits or all-cause death.

[0147] Figure 100 Treatment-period adverse events leading to death were shown in patients treated with 1200 mg REGEN-COV, 2400 mg REGEN-COV, 8000 mg REGEN-COV, or placebo.

[0148] Figure 101An overview of serious adverse events and adverse events of particular concern during the treatment period is provided in patients treated with 1200 mg REGEN-COV, 2400 mg REGEN-COV, 8000 mg REGEN-COV, or placebo.

[0149] Figure 102 Adverse events of particular concern are shown in patients receiving 1200 mg REGEN-COV, 2400 mg REGEN-COV, 8000 mg REGEN-COV, or placebo who require medical care in a healthcare facility.

[0150] Figure 103 The mean pharmacokinetic parameters of serum mAb10933 and mAb10987 are shown in patients treated with 1200 mg REGEN-COV, 2400 mg REGEN-COV, 8000 mg REGEN-COV or placebo. Detailed Implementation

[0151] Before describing the invention, it should be understood that the invention is not limited to the specific methods and experimental conditions described, as such methods and conditions can vary. It should also be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting, as the scope of the invention is limited only by the appended claims.

[0152] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. As used herein, when referring to a specific listed numerical value, the term “about” means that the value may differ from the listed value by no more than 1%. For example, as used herein, the expression “about 100” includes 99 and 101 and all values ​​in between (e.g., 99.1, 99.2, 99.3, 99.4, etc.).

[0153] Although any methods and materials similar to or equivalent to those described herein may be used in the practice or testing of this invention, preferred methods and materials are now described. All patents, applications, and non-patent publications referenced in this specification are incorporated herein by reference in their entirety.

[0154] Methods for preventing and treating SARS-CoV-2 infection and COVID-19

[0155] This invention provides methods for preventing and treating SARS-CoV-2 infection and COVID-19 in subjects in need of treatment by administering one or more antigen-binding molecules that bind to the surface protein of SARS-CoV-2, including anti-SARS-CoV-2 spike glycoprotein antibodies or antigen-binding fragments thereof discussed herein. In some cases, the subject is a hospitalized COVID-19 patient. In some cases, the subject is an outpatient (i.e., not bedridden) who has tested positive for SARS-CoV-2 infection. In some cases, the subject is a patient with laboratory-confirmed SARS-CoV-2 and one or more COVID-19 symptoms (such as fever, cough, or shortness of breath). In some cases, the subject is (a) a COVID-19 patient requiring low-flow oxygen supplementation; (b) a COVID-19 patient requiring high-intensity oxygen therapy but not mechanical ventilation; or (c) a COVID-19 patient requiring mechanical ventilation. In some cases, the subject is a non-hospitalized person with COVID-19 symptoms. In some cases, the subjects are uninfected individuals, such as uninfected individuals in high-risk exposure groups (e.g., healthcare workers or first responders) or uninfected individuals who have been in close contact with a subject infected with SARS-CoV-2 (e.g., roommates or family members infected with COVID-19). In some cases, the subjects are at high risk of COVID-19 complications or are more likely to be infected with SARS-CoV-2, such as the elderly, immunocompromised individuals, and those who frequently respond poorly to vaccines. In some embodiments, the present invention provides methods for treating, preventing, and reducing the severity or progression of SARS-CoV-2 infection and / or COVID-19.

[0156] This invention also includes the use of antigen-binding molecules that bind to the surface protein of SARS-CoV-2 (including the anti-SARS-CoV-2 spike glycoprotein antibodies or antigen-binding fragments thereof discussed herein) for the prevention and treatment of SARS-CoV-2 infection and COVID-19, and / or for the treatment, prevention, and reduction of the severity or progression of SARS-CoV-2 infection and / or COVID-19, or their symptoms. This invention also includes the use of antigen-binding molecules that bind to the surface protein of SARS-CoV-2 (including the anti-SARS-CoV-2 spike glycoprotein antibodies or antigen-binding fragments thereof discussed herein) in the preparation of medicaments for the prevention and treatment of SARS-CoV-2 infection and COVID-19, and / or for the treatment, prevention, and reduction of the severity or progression of SARS-CoV-2 infection and / or COVID-19. When methods are discussed herein with reference to combinations of two anti-SARS-CoV-2 spike protein antibodies, such combinations include the use of a first such antibody or antigen-binding fragment thereof in the preparation of a medicament used in combination with a second such antibody or antigen-binding fragment thereof, and the use of a second such antibody or antigen-binding fragment thereof in the preparation of a medicament used in combination with a first such antibody.

[0157] As used herein, a therapeutic or preventative agent for a disorder or condition (e.g., an anti-SARS-CoV-2 spike glycoprotein antibody) means a compound that, in a statistical sample, reduces the incidence of a disorder or condition in a treated sample relative to an untreated control sample, or delays the onset of a disorder or condition relative to an untreated control sample. As used herein, the term "treatment" includes improvement or elimination of an identified condition. In either case, prevention or treatment can be identified in the diagnosis provided by a physician or other healthcare provider and the anticipated outcome of administering the therapeutic or preventative agent.

[0158] Typically, treatment or prevention of a disease or condition as described in this disclosure is achieved by administering one or more anti-SARS-CoV-2 spike glycoprotein antibodies or antigen-binding fragments thereof in an effective amount. An effective amount of reagent refers to the amount that effectively achieves the desired therapeutic or preventative outcome at the required dose and time period. The therapeutically effective amount of reagent in this disclosure may vary depending on factors such as an individual's disease state, age, sex, and weight, as well as the reagent's ability to elicit the desired response in the individual. A preventatively effective amount refers to the amount that effectively achieves the desired preventative outcome at the required dose and time period.

[0159] In some implementations, anti-SARS-CoV-2 spike glycoprotein antibodies or antigen-binding fragments thereof can be used to treat, prevent, or reduce SARS-CoV-2 infection or the progression of COVID-19. In some cases, anti-SARS-CoV-2 spike glycoprotein antibodies or antigen-binding fragments thereof block the interaction with the spike protein receptor-binding domain (RBD) of angiotensin-converting enzyme 2 (ACE2), thereby reducing the infectivity of host cells. Blocking viral entry leads to a reduction in SARS-CoV-2 RNA replication and corresponding viral shedding in affected tissues. Therefore, in some implementations, anti-SARS-CoV-2 spike glycoprotein antibodies or antigen-binding fragments thereof will reduce viral shedding in the upper respiratory tract. In some implementations, viral shedding is measured from samples collected from the patient's upper respiratory tract 7 to 29 days after the start of administration (e.g., days 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, or 29 after the start of administration). In some cases, the reduction in SARS-CoV-2 viral shedding from baseline is determined by RT-qPCR in nasopharyngeal swab samples, nasal cavity samples, or saliva samples.

[0160] In some cases, anti-SARS-CoV-2 spike glycoprotein antibodies or their antigen-binding fragments have improved the clinical status of patients (e.g., patients diagnosed with SARS-CoV-2 infection or COVID-19). In some implementations, the improvement in clinical status is based on a 7-point ordinal scale (assessing clinical status from death[1] to non-hospitalization[7]) used to assess changes in clinical status. Utilizing an ordinal scale that ranks multiple clinical outcomes of interest (e.g., death, mechanical ventilation, etc.) according to their clinical importance is an appropriate method for assessing the efficacy of trials in severely and / or critically ill patients with COVID-19. In some cases, administration of anti-SARS-CoV-2 spike glycoprotein antibodies or their antigen-binding fragments has improved the clinical status of patients by at least 1 or 2 points. In some cases, administration of anti-SARS-CoV-2 spike glycoprotein antibodies or their antigen-binding fragments has resulted in a reduction in mortality and / or oxygen therapy use, and / or an increase in the number of days without ventilation in such patients. As discussed above, the following ordinal scales can be used to assess improvements in clinical status:

[0161] [1] Death

[0162] [2] Hospitalization, requiring invasive mechanical ventilation or ECMO

[0163] [3] Hospitalization, requiring non-invasive ventilation or high-flow oxygen device

[0164] [4] Hospitalization, requiring supplemental oxygen supply

[0165] [5] Hospitalization, no supplemental oxygen required—requires continuous medical care

[0166] (COVID-19 related or other)

[0167] [6] Hospitalized, no need for supplemental oxygen supply—no longer requiring continuous medical care.

[0168] [7] No hospitalization.

[0169] In some cases, following administration of the anti-SARS-CoV-2 spike glycoprotein antibody or its antigen-binding fragment, subjects exhibited fewer than 5 COVID-19-related medical care visits, telemedicine visits, hospital admissions, and / or intensive care unit (ICU) admissions. In some cases, following administration of the first dose of the anti-SARS-CoV-2 spike glycoprotein antibody or its antigen-binding fragment, subjects exhibited fewer than 5 (e.g., fewer than 5, fewer than 4, fewer than 3, fewer than 2, or fewer than 1) COVID-19-related medical care visits, telemedicine visits, hospital admissions, and / or ICU admissions within a 7- to 42-day period (e.g., 21 to 42 days). In some implementations, subjects exhibited fewer than 5 COVID-19-related visits within a 29-day period following the first dose. In some cases, subjects exhibited fewer than 4 COVID-19-related medical care visits, telemedicine visits, hospital admissions, and / or ICU admissions. In some cases, participants experienced fewer than 3 COVID-19 related medical care visits, telemedicine visits, hospital admissions, and / or intensive care unit (ICU) admissions. In some cases, participants experienced fewer than 2 COVID-19 related medical care visits, telemedicine visits, hospital admissions, and / or ICU admissions. In some cases, participants experienced no more than 1 COVID-19 related medical care visit, telemedicine visit, hospital admission, and / or ICU admission.

[0170] In some cases, after administration of an anti-SARS-CoV-2 spike glycoprotein antibody or its antigen-binding fragment, subjects tested negative for SARS-CoV-2 within 2 days to 3 weeks after the first administration of the therapeutic composition. In some cases, SARS-CoV-2 testing was negative by RT-qPCR in nasopharyngeal swab samples, nasal cavity samples, or saliva samples.

[0171] In any of the various implementation schemes discussed above or herein (e.g., prophylactic or therapeutic administration of a combination of mAb10933 and mAb10987), the administration of one or more anti-SARS-CoV-2 spike glycoprotein antibodies may result in any or more of the following:

[0172] (a) Time-weighted average decrease in viral shedding from baseline (log 10 (Copies / mL), as measured by RT-qPCR in nasopharyngeal (NP) swabs;

[0173] (b) Time-weighted average decrease in viral shedding from baseline (log 10 (Copies / mL), as measured by RT-qPCR in nasal swabs;

[0174] (c) Time-weighted average decrease in viral shedding from baseline (log 10 (copy / mL), such as when measured by RT-qPCR in saliva samples;

[0175] (d) Using a 7-point ordinal scale, the clinical condition has improved by at least 1 point relative to baseline;

[0176] (e) The number of COVID-19 related medical care visits was reduced compared to the control group (COVID-19 related medical care visits are defined as hospital, emergency room (ER) visits, emergency care visits, physician office visits or telemedicine visits where the primary cause of the visits is COVID-19);

[0177] (f) Compared to the control, the time for RT-qPCR to be negative in NP swabs and not subsequently become positive by RT-qPCR was reduced;

[0178] (g) Compared with the control, the incidence of hospitalization or the length of hospital stay was reduced;

[0179] (h) Compared to the control group, the incidence of ICU admission or the number of days spent in the ICU was reduced;

[0180] (i) The incidence or duration of mechanical ventilation was reduced compared to the control.

[0181] (j) The duration of COVID-19 symptoms was reduced compared to the control group;

[0182] (k) Compared to the control, the time it took for all tested samples to be negative for RT-qPCR and for any subsequent tested samples (nasopharyngeal swabs, nasal swabs, saliva) to not become positive for RT-qPCR was reduced;

[0183] (l) The incidence of subsequent signs or symptoms of SARS-CoV-2 infection (narrow or broad sense) decreased;

[0184] (m) Time-weighted average daily viral load reduction (e.g., a reduction of 0.4 or more log10 copies / mL by day 7, a reduction of 0.5 or more log10 copies / mL by day 7, or a reduction of 0.6 or more log10 copies / mL by day 7, or a reduction of 0.5 or more log10 copies / mL by day 11, or a reduction of 0.6 or more log10 copies / mL by day 11, or a reduction of 0.7 or more log10 copies / mL by day 11); and

[0185] (n) Time-weighted average viral load decrease from baseline (log10 copies / mL).

[0186] In some implementations, administration of anti-SARS-CoV-2 spike glycoprotein antibodies is more effective in subjects without an effective amount of anti-SARS-CoV-2 antibodies in their blood (“seronegative” subjects) than in subjects with an effective amount of anti-SARS-CoV-2 antibodies in their blood (“seropositive” subjects). In the embodiments provided herein, serological status (i.e., seronegative, seropositive, or indeterminate) is determined by assessing the presence of serum anti-SARS-CoV-2 antibodies: anti-spike[S1]IgA (Euroimmun IgA assay), anti-spike[S1]IgG (Euroimmun IgG assay), and anti-nucleocapsid IgG (Abbot IgG assay). Study participants are analyzed in groups seronegative (if all available tests are negative), seropositive (if any of these tests are positive), or seroindeterminate (missing or indeterminate results). If the antibody level in a sample is below the limit of quantitation for detection, the test is classified as negative. As described herein, the methods described herein can have different effects in seronegative subjects compared to a comparable seronegative subject population (e.g., greater reduction in viral load, faster time to symptom relief, and fewer post-administration healthcare visits).

[0187] Antigen-binding molecules and antibodies against SARS-CoV-2 spike glycoprotein

[0188] The methods and uses of the present invention utilize antigen-binding molecules that bind to the surface proteins of SARS-CoV-2. In some embodiments, the antigen-binding molecule is an anti-SARS-CoV-2 spike glycoprotein antibody or an antigen-binding fragment thereof.

[0189] The variable region, CDR, and amino acid and nucleotide sequences of the heavy and light chains of exemplary antibodies that bind to the SARS-CoV-2 spike protein are shown in Tables 1 and 2 below. Other amino acid and nucleotide sequences of the variable region, CDR, and heavy and light chains of exemplary antibodies and antigen-binding fragments that bind to the SARS-CoV-2 spike protein and can be used in the methods described herein are available in U.S. Patent No. 10,787,501, the entire contents of which are incorporated herein by reference.

[0190] Table 1: Amino Acid Sequence Identifiers

[0191]

[0192]

[0193] Table 2: Nucleic Acid Sequence Identifiers

[0194]

[0195] In various embodiments, the anti-SARS-CoV-2 spike glycoprotein antibody or antigen-binding fragment used for the methods or uses discussed herein is an antibody or antigen-binding fragment comprising six CDRs (HCDR1-HCDR2-HCDR3-LCDR1-LCDR2-LCDR3) of any one or more of the antibodies listed in Table 1. In some cases, the anti-SARS-CoV-2 spike glycoprotein antibody or antigen-binding fragment comprises a CDR belonging to a heavy chain variable region (HCVR) and light chain variable region pair comprising an amino acid sequence selected from the group consisting of: SEQ ID NO: 2 / 10, 22 / 30, 42 / 50, and 73 / 81. Methods and techniques for identifying CDRs within HCVR and LCVR amino acid sequences are well known in the art and can be used to identify CDRs within specific HCVR and / or LCVR amino acid sequences disclosed herein. Exemplary specifications that can be used to identify the boundaries of CDRs include, for example, the Kabat definition, the Chothia definition, and the AbM definition. Generally, the Kabat definition is based on sequence variability, the Chothia definition is based on the location of structural loop regions, and the AbM definition is a compromise between the Kabat and Chothia approaches. See, for example, Kabat, “Sequences of Proteins of Immunological Interest,” National Institutes of Health, Bethesda, Maryland, 1991; Al-Lazikani et al., J Mol Biol, Vol. 273: 927–948, 1997; and Martin et al., PNAS (USA), Vol. 86: 9268–9272, 1989. Public databases can also be used to identify CDR sequences within antibodies.

[0196] In some embodiments, the anti-SARS-CoV-2 spike glycoprotein antibody or antigen-binding fragment comprises HCDR1-HCDR2-HCDR3-LCDR1-LCDR2-LCDR3 domains, which respectively comprise amino acid sequences selected from the group consisting of: SEQ ID NO: 4-6-8-12-14-16, 24-26-28-32-34-36, 44-46-48-52-34-54 and 75-77-79-83-85-87.

[0197] In some embodiments, the anti-SARS-CoV-2 spike glycoprotein antibody or antigen-binding fragment comprises an HCVR / LCVR amino acid sequence pair, which comprises an amino acid sequence selected from the group consisting of: SEQ ID NO: 2 / 10, 22 / 30, 42 / 50, and 73 / 81.

[0198] In some embodiments, the anti-SARS-CoV-2 spike glycoprotein antibody comprises a heavy chain (HC) and a light chain (LC) pair, the HC and LC pair comprising an amino acid sequence selected from the group consisting of: SEQ ID NO: 18 / 20, 38 / 40, 56 / 58 and 89 / 91.

[0199] In some embodiments, the anti-SARS-CoV-2 spike glycoprotein antibody binds to an epitope within the SARS-CoV-2 spike protein receptor-binding domain (RBD) (amino acids 1-1273 of NCBI accession number (MN908947.3), SEQ ID NO:59). In some cases, the antibody (e.g., mAb10989) binds residues 467-513 of the RBD (DISTEIYQAGSTPCNGVEGFNCYFPLQSYGFQPTNGVGYQPYRVVVL) (SEQ ID NO:60). In some cases, the antibody (e.g., mAb10987) binds residues 432-452 of the RBD (CVIAWNSNNLDSKVGGNYNYL) (SEQ ID NO:61). In some cases, antibodies (e.g., mAb10933) bind to residues 467-510 of the RBD (DISTEIYQAGSTPCNGVEGFNCYFPLQSYGFQPTNGVGYQPYRV) (SEQ ID NO:62).

[0200] In some embodiments, the anti-SARS-CoV-2 spike glycoprotein antibody is mAb10933. In some embodiments, the anti-SARS-CoV-2 spike glycoprotein antibody is mAb10987. In some embodiments, the anti-SARS-CoV-2 spike glycoprotein antibody is mAb10989. In various embodiments, the anti-SARS-CoV-2 spike glycoprotein antibody is an antibody comprising the CDR, HCVR, and LCVR or heavy and light chains (e.g., the amino acid sequences shown in Table 1) of mAb10933. In various embodiments, the anti-SARS-CoV-2 spike glycoprotein antibody is an antibody comprising the CDR, HCVR, and LCVR or heavy and light chains (e.g., the amino acid sequences shown in Table 1) of mAb10987. In various embodiments, the anti-SARS-CoV-2 spike glycoprotein antibody is an antibody comprising the CDR, HCVR, and LCVR or heavy and light chains (e.g., the amino acid sequences shown in Table 1) of mAb10989. The antibodies provided in this article are interchangeably identified as “mAb” followed by a number or “REGN” followed by a number. For example, mAb10933 and REGN10933 refer to the same antibody (the amino acid sequence provided in Table 1 and the nucleic acid sequence provided in Table 2). Similarly, mAb10987 and REGN10987 are equivalent, mAb10989 and REGN10989 are equivalent, and mAb10985 and REGN10985 are equivalent. Additionally, mAb10933 can be referred to as cassicimarab, and mAb10987 can be referred to as edevimarab. The combination of cassicimarab and edevimarab is referred to as REGEN-COV.

[0201] In some embodiments, the methods and uses discussed herein include compositions comprising a first antigen-binding molecule (e.g., an antibody) and a second antigen-binding molecule (e.g., an antibody), the first antigen-binding molecule binding to a first epitope on the surface protein of SARS-CoV-2, and the second antigen-binding molecule binding to a second epitope on the surface protein of SARS-CoV-2, wherein the first and second epitopes do not overlap structurally. In some embodiments, the methods and uses discussed herein include combinations of two or more anti-SARS-CoV-2 spike glycoprotein antibodies or antigen-binding fragments thereof. In some cases, the two antibodies or antigen-binding fragments used in combination bind to non-overlapping epitopes of the RBD. In some embodiments, the combination includes mAb10987 and mAb10933. In some embodiments, the combination includes mAb10987 and mAb10989. In some embodiments, the combination includes mAb10933 and mAb10987 and mAb10985. In various embodiments, the combination includes a first anti-SARS-CoV-2 spike glycoprotein antibody, a second anti-SARS-CoV-2 spike glycoprotein antibody, and optionally a third anti-SARS-CoV-2 spike glycoprotein antibody. The first anti-SARS-CoV-2 spike glycoprotein antibody is an antibody containing the CDR, HCVR, and LCVR or heavy and light chains (e.g., the amino acid sequences shown in Table 1) of mAb10933. The second anti-SARS-CoV-2 spike glycoprotein antibody is an antibody containing the CDR, HCVR, and LCVR or heavy and light chains (e.g., the amino acid sequences shown in Table 1) of mAb10987. The third anti-SARS-CoV-2 spike glycoprotein antibody is an antibody containing the CDR, HCVR, and LCVR or heavy and light chains (e.g., the amino acid sequences shown in Table 1) of mAb10985. In various embodiments, the combination includes a first anti-SARS-CoV-2 spike glycoprotein antibody and a second anti-SARS-CoV-2 spike glycoprotein antibody, wherein the first anti-SARS-CoV-2 spike glycoprotein antibody is an antibody containing the CDR, HCVR, and LCVR or heavy and light chains (e.g., the amino acid sequences shown in Table 1) of mAb10989, and the second anti-SARS-CoV-2 spike glycoprotein antibody is an antibody containing the CDR, HCVR, and LCVR or heavy and light chains (e.g., the amino acid sequences shown in Table 1) of mAb10987.In some implementations, combinations of antigen-binding molecules (e.g., antibodies, such as mAb10987 and mAb10933, mAb10987 and mAb10989, or mAb10987 and mAb10933 and mAb10985) can reduce the frequency of escape mutants (e.g., SARS-CoV-2 viruses with one or more mutations in the S protein to reduce the efficacy of treatment, for example, by reducing antibody binding to the S protein). In the presence of mAb10933 (cassicimarab) or mAb10987 (edevimab), escape variants were identified after two passages in cell cultures of recombinant VSV encoding the SARS-CoV-2 spike protein; however, no escape variants were identified after two passages in the presence of both cassicimarab and edevimab. This antibody combination was also effective against variant SARS-CoV-2 viruses. For example, the ability of the combination of mAb10933 and mAb10987 to neutralize pseudotyped VSV, which expresses a SARS-CoV-2 variant called B.1.1.7, also known as the "UK variant," is being evaluated. This variant is rapidly increasing and may have different effects than wild-type SARS-CoV-2, including more severe symptoms and potential resistance to vaccines and / or therapeutics. It is classified in part by the following mutations in the spike protein: HV 69-70 deletion, Y144 deletion, N501Y, A570D, P681H, T716I, S982A, and D1118H. The combination of cassicimarab and edevimab has been shown to effectively neutralize the virus. Figure 29 In fact, cascisizumab and edevimab, alone and together, retain neutralizing activity against pseudoviruses expressing all spike protein substitutions found in the B.1.1.7 lineage (UK source) and against pseudoviruses expressing only N501Y found in B.1.1.7 and other circulating lineages. cascisizumab and edevimab together retain neutralizing activity against pseudoviruses expressing all spike protein substitutions or single substitutions of K417N, E484K, or N501Y found in the B.1.1351 lineage (South African source) and against K417T+E484K substitution found in the P.1 lineage (Brazil source), although, as mentioned above, cascisizumab alone, rather than edevimab, reduces activity against pseudoviruses expressing K417N or E484K. E484k substitution is also found in the B.1.526 lineage (New York source).

[0202] Table 3A: Cacireximab and Idvimab, alone and together, preserved their effectiveness against the B.1.427 / B.1.429 lineage. The neutralizing activity of L452R substitution found in (California source) .

[0203]

[0204]

[0205] Table 3B: Neutralization data of pseudoviruses replaced by SARS-CoV-2 variants using both cassicimarab and edevimab

[0206]

[0207] a Detection of pseudoviruses expressing the entire variant spike protein. The following variations of the wild-type spike protein were found in the variants: del69-70, del145, N501Y, A570D, D614G, P681H, T716I, S982A, and D1118H.

[0208] b Detection of pseudoviruses expressing the entire variant spike protein. The following variations of the wild-type spike protein were found in the variants: D80Y, D215Y, del241-243, K417N, E484K, N501Y, D614G, and A701V.

[0209] c No change: Susceptibility decreased by <2 times.

[0210] d Not all isolates in the New York lineage have the E484K substitution (as of February 2021).

[0211] Certain variants showed reduced susceptibility to casciretumab alone, including those with spike protein amino acid substitutions of K417E (182-fold), K417N (7-fold), K417R (61-fold), Y453F (>438-fold), L455F (80-fold), E484K (25-fold), F486V (>438-fold), and Q493K (>438-fold). Variants showing reduced susceptibility to edevimab alone included those with substitutions of K444N (>755-fold), K444Q (>548-fold), K444T (>1,033-fold), and V445A (548-fold). Casciretumab together showed reduced susceptibility to variants with K444T (6-fold) and V445A (5-fold) substitutions. In a neutralization assay using pseudotyped VSVs identified in circulating SARS-CoV-2, variants showing reduced susceptibility to cassicimarumab alone included those with Q409E (4-fold), G476S (5-fold), and S494P (5-fold) substitutions, while variants showing reduced susceptibility to edevimab alone included those with N439K (463-fold) substitution. Other substitutions detected in the pseudovirus assay that also showed reduced activity against cassicimarumab alone included E484Q (9-fold) and Q493E (446-fold). Both cassicimarumab and edevimab retained activity against all detected variants. In some embodiments, this disclosure provides a method for treating SARS-CoV-2 infection, the method comprising administering mAb10933 and mAb10987, wherein the SARS-CoV-2 is a variant of SARS-CoV-2, which, for example, contains HV 69-70 deletion, Y144 deletion, Q409E, K417E, K417N, K417R, N439K, Y453F, L455F, G476S, E484K, E484Q, F486V, Q493K, Q493E, S494P, N501Y, A570D, P681H, T716I, S982A, or D1118H, or any combination thereof. In the clinical trial of Example 2, interim data showed that only one variant (G446V) was present in subjects with an allele fraction ≥15%, detected in 3 out of 66 subjects with nucleotide sequencing data, with each subject detected at a single time point (2 subjects in the placebo group and the 2400 mg cassicimarab and edevimab group were detected at baseline, and 1 subject in the 8000 mg cassicimarab and edevimab group was detected on day 25). In the VSV pseudoparticle neutralization assay, the susceptibility of the G446V variant to edevimab was reduced to 1 / 135 compared to wild-type, but susceptibility to cassicimarab alone and to cassicimarab and edevimab together was retained.

[0212] In some embodiments, the methods and uses discussed herein include compositions comprising a first antigen-binding molecule (e.g., an antibody) and a second antigen-binding molecule (e.g., an antibody), the first antigen-binding molecule binding to a first epitope on the surface protein of SARS-CoV-2 and the second antigen-binding molecule binding to a second epitope on the surface protein of SARS-CoV-2, wherein the first antigen-binding molecule and the second antigen-binding molecule are capable of simultaneously binding to the surface protein of SARS-CoV-2.

[0213] In some embodiments, one, two, three, four or more antibodies or antigen-binding fragments thereof may be administered in combination (e.g., simultaneously or sequentially). Exemplary combinations include mAb10933 and mAb10987, mAb10989 and mAb10987, mAb10933 and mAb10989, and mAb10933 and mAb10987 and mAb10985.

[0214] As used herein, “antibody binding to SARS-CoV-2 spike protein” or “anti-SARS-CoV-2 spike glycoprotein antibody” or “anti-SARS-CoV-2 spike protein antibody” includes an antibody that binds to a soluble fragment of the SARS-CoV-2 spike protein and its antigen-binding fragment, and may also bind to an epitope within the receptor-binding domain (RBD) of the spike protein. Other antibodies that may be used alone or in combination with each other or with one or more of the antibodies disclosed herein for use in the context of the methods disclosed herein include, for example, LY-CoV555 (Eli Lilly); 47D11 (Wang et al., Nature Communications, article 2251); B38, H4, B5 and / or H2 (Wu et al., 10.1126 / science.abc 2241, 2020); STI-1499 (Sorrento Therapeutics); VIR-7831 and VIR-7832 (VirBiotherapeutics).

[0215] As used herein, the term "antibody" means any antigen-binding molecule or molecular complex that includes at least one complementarity-determining region (CDR) that specifically binds to or interacts with a particular antigen (e.g., the SARS-CoV-2 spike protein). The term "antibody" also refers to an immunoglobulin molecule comprising four polypeptide chains, two heavy (H) chains and two light (L) chains interconnected by disulfide bonds, and their polymers (e.g., IgM). Each heavy chain includes a heavy chain variable region (abbreviated herein as HCVR or V). H The heavy-chain constant region comprises three structural domains C. H 1. C H 2 and CH 3. Each light chain includes a light chain variable region (abbreviated as LCVR or V in this document). L The light chain constant region includes a structural domain (C0) and a light chain constant region. L 1) V can be H District and V L The region is further subdivided into highly variable areas called Complementary Determinant Regions (CDRs), interspersed with more conservative areas called Framing Regions (FRs). Each V H and V L It consists of three CDRs and four FRs, arranged in the following order from the amino terminus to the carboxyl terminus: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. In different embodiments of the invention, the FRs of the anti-SARS-CoV-2 spike protein antibody (or its antigen-binding portion) may be identical to the human germline sequence, or may be natural or artificially modified. The common amino acid sequence can be defined based on the side-by-side analysis of two or more CDRs.

[0216] As used herein, the term "antibody" also includes the antigen-binding fragment of the complete antibody molecule. As used herein, the terms "antigen-binding portion" of an antibody, "antigen-binding fragment," etc., encompass any naturally occurring, enzymatically obtainable, synthetic, or genetically engineered polypeptide or glycoprotein that specifically binds to an antigen to form a complex. The antigen-binding fragment of an antibody can be derived from the complete antibody molecule, for example, using any suitable standard technique, such as proteolytic digestion or recombinant genetic engineering techniques involving the manipulation and expression of DNA encoding variable and optionally constant domains of the antibody. Such DNA is known and / or readily available from, for example, commercial sources, DNA libraries (containing, for example, phage-antibody libraries), or can be synthesized. DNA can be sequenced and manipulated by chemical methods or by molecular biological techniques, for example, arranging one or more variable and / or constant domains into suitable conformations, or introducing codons, generating cysteine ​​residues, modifying, adding, or deleting amino acids, etc.

[0217] Non-limiting examples of antigen-binding fragments include: (i) Fab fragments; (ii) F(ab')2 fragments; (iii) Fd fragments; (iv) Fv fragments; (v) single-chain Fv (scFv) molecules; (vi) dAb fragments; and (vii) minimal recognition units composed of amino acid residues mimicking the hypervariable region of an antibody (e.g., a separated complementarity-determining region (CDR), such as a CDR3 peptide) or a restricted FR3-CDR3-FR4 peptide. Other engineered molecules, such as domain-specific antibodies, single-domain antibodies, domain-deficient antibodies, chimeric antibodies, CDR-grafted antibodies, biantibodies, triantibodies, tetraantibodies, microantibodies, nanobodies (e.g., monovalent nanobodies, bivalent nanobodies, etc.), small modular immunopharmaceuticals (SMIPs), and shark variable IgNAR domains, are also encompassed within the term "antigen-binding fragments" as used herein.

[0218] Antigen-binding fragments of antibodies typically include at least one variable domain. Variable domains can have any size or amino acid composition and will generally include at least one CDR adjacent to or within one or more frame sequences. L V associated with the structural domain H In the antigen-binding fragment of the domain, V H Domain and V L Domains can be positioned relative to each other in any suitable arrangement. For example, a variable region can be a dimer and contain V. H -V H V H -V L or V L -V dimer. Alternatively, the antigen-binding fragment of the antibody may contain monomeric V. H or V L Structural domain.

[0219] In some embodiments, the antigen-binding fragment of the antibody may contain at least one variable domain covalently linked to at least one constant domain. Non-limiting exemplary configurations of variable and constant domains that can be found in the antigen-binding fragment of the antibody of the present invention include: (i) V H -C H 1; (ii)V H -C H 2; (iii)V H -C H 3; (iv)V H -C H 1-C H 2; (v)V H -C H 1-C H 2-C H 3;(vi)VH -C H 2-C H 3;(vii)V H -C L (viii)V L -C H 1; (ix)V L -C H 2; (x)V L -C H 3;(xi)V L -C H 1-C H 2;(xii)V L -C H 1-C H 2-C H 3; (xiii)V L -C H 2-C H 3; and (xiv)V L -C L In any configuration of the variable and constant domains (including any of the exemplary configurations listed above), the variable and constant domains may be directly connected to each other or may be connected via full or partial hinge or connector regions. The hinge region may consist of at least two (e.g., 5, 10, 15, 20, 40, 60, or more) amino acids, which result in flexible or semi-flexible connections between adjacent variable and / or constant domains in a single polypeptide molecule. Furthermore, the antigen-binding fragment of the antibody of the present invention may comprise a homodimer or heterodimer (or other multimer) of any of the variable and constant domain configurations listed above, which are non-covalently associated with each other and / or with one or more monomers V. H or V L Domain covalent association (e.g., via one or more disulfide bonds).

[0220] Like intact antibody molecules, antigen-binding fragments can be monospecific or multispecific (e.g., bispecific). Multispecific antigen-binding fragments of antibodies typically comprise at least two distinct variable domains, each capable of specifically binding to a single antigen or different epitopes on the same antigen. Any multispecific antibody form (including the exemplary bispecific antibody forms disclosed herein) can be adapted to the antigen-binding fragment context of the antibodies of this invention using conventional techniques available in the art.

[0221] In some embodiments of the invention, the anti-SARS-CoV-2 spike protein antibody of the present invention is a human antibody. As used herein, the term "human antibody" is intended to include antibodies having variable and constant regions derived from human germline immunoglobulin sequences. The human antibodies of the present invention may include amino acid residues not encoded by human germline immunoglobulin sequences (e.g., mutations introduced by random or site-specific mutagenesis in vitro or by somatic mutations in vivo), such as in CDRs, particularly in CDR3. However, as used herein, the term "human antibody" is not intended to include antibodies in which a germline CDR sequence derived from another mammalian species (e.g., mouse) has been grafted onto a human scaffold region sequence.

[0222] In some embodiments, the antibodies of the present invention may be recombinant human antibodies. As used herein, the term "recombinant human antibody" is intended to encompass all human antibodies prepared, expressed, generated, or isolated in a recombinant manner, such as antibodies expressed using a recombinant expression vector transfected into host cells (further described below), antibodies isolated from a library of recombinant combined human antibodies (further described below), antibodies isolated from animals (e.g., mice) that are transgenic with respect to human immunoglobulin genes (see, for example, Taylor et al., Nucl Acids Res, Vol. 20: pp. 6287-6295, 1992), or antibodies prepared, expressed, generated, or isolated by any other method involving splicing a human immunoglobulin gene sequence into another DNA sequence. Such recombinant human antibodies have variable and constant regions derived from human germline immunoglobulin sequences. However, in some embodiments, such recombinant human antibodies undergo in vitro mutagenesis (or, when using animals with transgenic human Ig sequences, in vivo somatic cell mutagenesis), and therefore the V of the recombinant antibody is... H District and V L The amino acid sequence of the region is as follows: Although it is derived from human lineage V H Sequence and V L The sequence is related to it, but it may not be naturally present in human antibody germline libraries.

[0223] Human antibodies can exist in two forms associated with hinge heterogeneity. In one form, the immunoglobulin molecule comprises a stable four-chain construct of approximately 150-160 kDa, where the dimers are held together by interchain heavy chain disulfide bonds. In the second form, the dimers are not linked by interchain disulfide bonds and form a molecule of approximately 75-80 kDa, consisting of covalently coupled light and heavy chains (half-antibodies). These forms are extremely difficult to separate even after affinity purification.

[0224] The frequency of the second form occurring in various intact IgG isotypes is based on, but not limited to, structural differences associated with the isotype of the antibody's hinge region. A single amino acid substitution in the hinge region of the human IgG4 hinge can significantly reduce the occurrence of the second form (Angal et al., Molecular Immunology, Vol. 30: p. 105, 1993) to levels typically observed with the human IgG1 hinge. This invention covers the hinge, C... H Zone 2 or C H Region 3 contains one or more mutated antibodies, which may be, for example, desired in production to increase the yield of the desired antibody form.

[0225] The antibodies of this invention can be isolated antibodies. As used herein, "isolated antibody" means an antibody that has been identified and isolated and / or recovered from at least one component of its natural environment. For example, an antibody that has been isolated or removed from at least one component of an organism, or from tissues or cells where the antibody is naturally present or produced, is an "isolated antibody" for the purposes of this invention. Isolated antibodies also include in situ antibodies within recombinant cells. Isolated antibodies are antibodies that have undergone at least one purification or isolation step. According to some embodiments, isolated antibodies may be substantially free of other cellular material and / or chemicals.

[0226] This invention includes antibodies that neutralize and / or block the SARS-CoV-2 spike protein. As used herein, “neutralizing” or “blocking” antibodies are intended to refer to antibodies that bind to the SARS-CoV-2 spike protein: (i) inhibit the activity of the SARS-CoV-2 spike protein to any detectable extent, for example, inhibiting the ability of SARS-CoV-S to bind to receptors such as ACE2, to be cleaved by proteases such as TMPRSS2, or to mediate viral entry into host cells or viral replication in host cells.

[0227] Compared to the corresponding germline sequences of derived antibodies, the anti-SARS-CoV-2 spike protein antibodies disclosed herein may include one or more amino acid substitutions, insertions, and / or deletions in the frame and / or CDR regions of the heavy and light chain variable domains. Such mutations can be readily identified by comparing the amino acid sequences disclosed herein with germline sequences available from, for example, public antibody sequence databases. This invention comprises antibodies derived from any amino acid sequence disclosed herein and their antigen-binding fragments, wherein one or more amino acids in one or more frame and / or CDR regions are mutated to one or more corresponding residues of the germline sequence of the derived antibody, or mutated to one or more corresponding residues of another human germline sequence, or mutated to a conserved amino acid substitution of one or more corresponding germline residues (such sequence changes are collectively referred to herein as "germination mutations"). Starting from the heavy and light chain variable region sequences disclosed herein, those skilled in the art can readily generate numerous antibody and antigen-binding fragments comprising one or more individual germline mutations or combinations thereof. In some embodiments, V H Domain and / or V L All frame and / or CDR residues within the domain are mutated back to residues found in the original germline sequence of the derived antibody. In other embodiments, only certain residues are mutated back to the original germline sequence, for example, mutated residues found only in the first 8 amino acids of FR1 or the last 8 amino acids of FR4, or mutated residues found only in CDR1, CDR2, or CDR3. In other embodiments, one or more frame and / or CDR residues are mutated to one or more corresponding residues of a different germline sequence (i.e., a germline sequence different from the germline sequence of the original derived antibody). Furthermore, the antibody of the present invention may contain any combination of two or more germline mutations within the frame and / or CDR region, for example, wherein certain individual residues are mutated to corresponding residues of a specific germline sequence, while certain other residues different from the original germline sequence may be maintained or mutated to corresponding residues of a different germline sequence. Once obtained, antibody-antigen binding fragments containing one or more germline mutations can be readily detected for one or more desired properties, such as improved binding specificity, increased binding affinity, improved or enhanced antagonistic or agonistic biological properties (as applicable), reduced immunogenicity, etc. Antibody-antigen binding fragments obtained in this general manner are covered within the scope of this invention.

[0228] The present invention also includes antibodies against the SARS-CoV-2 spike protein, comprising variants of any one of the disclosed HCVR, LCVR, and / or CDR amino acid sequences having one or more conserved substitutions. For example, the present invention includes antibodies against the SARS-CoV-2 spike protein having HCVR, LCVR, and / or CDR amino acid sequences having, for example, 10 or fewer, 8 or fewer, 6 or fewer, 4 or fewer, etc., conserved amino acid substitutions relative to any one of the disclosed HCVR, LCVR, and / or CDR amino acid sequences.

[0229] The term "epitope" refers to an antigenic determinant that interacts with a specific antigen-binding site in the variable region of an antibody molecule, known as a complementary site. A single antigen can have more than one epitope. Therefore, different antibodies can bind to different regions on the antigen and can have different biological effects. Epitopes can be conformational or linear. Conformational epitopes are generated by spatially juxtaposed amino acids from different segments of a linear polypeptide chain. Linear epitopes are generated by adjacent amino acid residues in a polypeptide chain. In some cases, epitopes can contain portions of sugars, phosphoryl groups, or sulfonyl groups on the antigen.

[0230] When referring to nucleic acids or fragments thereof, the terms "substantially identical" or "substantially the same" mean that, when optimally aligned with another nucleic acid (or its complementary strand) by appropriate nucleotide insertions or deletions, the nucleotide sequence identity is at least about 95% of the nucleotide bases, and more preferably, at least about 96%, 97%, 98%, or 99%, as measured by any well-known sequence identity algorithm such as FASTA, BLAST, or Gap, as discussed below. In some cases, nucleic acid molecules having substantial identity with a reference nucleic acid molecule may encode polypeptides having the same or substantially similar amino acid sequences as the polypeptides encoded by the reference nucleic acid molecule.

[0231] When applied to peptides, the terms "substantially similar" or "substantially alike" mean that, when optimally aligned using procedures such as GAP or BESTFIT with default gap weights, two peptide sequences share at least 95% sequence identity, and more preferably, at least 98% or 99% sequence identity. Preferably, the dissimilar residue positions differ due to conserved amino acid substitutions. A "conserved amino acid substitution" is an amino acid substitution in which one amino acid residue is replaced by another amino acid residue with a side chain (R group) having similar chemical properties (e.g., charge or hydrophobicity). Overall, conserved amino acid substitutions do not substantially alter the functional properties of the protein. In cases where two or more amino acid sequences differ from each other due to conserved substitutions, the percentage of sequence identity or degree of similarity can be adjusted upwards to correct for the conservatism of the substitution. Methods for making such adjustments are well known to those skilled in the art. See, for example, Pearson, WR, Methods Mol Biol, Vol. 24: pp. 307-331, 1994, incorporated herein by reference. Examples of groups of amino acids with side chains having similar chemical properties include (1) aliphatic side chains: glycine, alanine, valine, leucine, and isoleucine; (2) aliphatic-hydroxy side chains: serine and threonine; (3) amide-containing side chains: asparagine and glutamine; (4) aromatic side chains: phenylalanine, tyrosine, and tryptophan; (5) basic side chains: lysine, arginine, and histidine; (6) acidic side chains: aspartic acid and glutamic acid; and (7) sulfur-containing side chains are cysteine ​​and methionine. Preferred conserved amino acid substituents are: valine-leucine-isoleucine, phenylalanine-tyrosine, lysine-arginine, alanine-valine, glutamic acid-aspartic acid, and asparagine-glutamine. Alternatively, a conservative substitution is any variation with a positive value in the PAM250 log-likelihood matrix published in Gonnet et al., Science, Vol. 256: pp. 1443-1445, 1992, which is incorporated herein by reference. A “moderately conservative” substitution is any variation with a non-negative value in the PAM250 log-likelihood matrix.

[0232] Sequence analysis software is commonly used to measure the sequence similarity, also known as sequence identity, of peptides. Protein analysis software uses similarity metrics assigned to various substitutions, deletions, and other modifications (including conserved amino acid substitutions) to match similar sequences. For example, GCG software contains programs such as Gap and Bestfit, which can be used with default parameters to determine sequence homology or sequence identity between closely related peptides, such as homologous peptides from different biological species or between wild-type proteins and their mutant counterparts. See, for example, GCG version 6.1. Peptide sequences can also be compared using FASTA, a program in GCG version 6.1, with default or recommended parameters. FASTA (e.g., FASTA2 and FASTA3) provides alignment of the best overlapping region between the query sequence and the search sequence and a percentage of sequence identity (see, for example, Pearson, WR, Methods Mol Biol, Vol. 132: pp. 185–219, 2000, incorporated herein by reference). When comparing the sequences of this invention with databases containing a large number of sequences from different organisms, another preferred algorithm is the computer program BLAST using default parameters, especially BLASTP or TBLASTN. See, for example, Altschul et al., J Mol Biol, Vol. 215: pp. 403-410, 1990, and Altschul et al., Nucleic Acids Res, Vol. 25: pp. 3389-3402, 1997, each of which is incorporated herein by reference.

[0233] Specific binding

[0234] As used herein, the term "specific binding" refers to the formation of a complex between an antigen-specific binding protein or antigen-specific binding domain and a specific antigen, characterized by a dissociation constant (K). D The concentration is 50 nM or less, and it does not bind to other irrelevant antigens under normal detection conditions. "Irrelevant antigens" are proteins, peptides, or polypeptides that have less than 95% amino acid identity with each other. Methods for determining whether two molecules specifically bind to each other are well known in the art and include, for example, equilibrium dialysis, surface plasmon resonance, etc. For example, as used in the context of this invention, an antigen-specific binding protein or antigen-specific binding domain includes a molecule that binds a specific antigen (e.g., a specific epitope of the SARS-CoV-2 spike protein, the SARS-CoV-2 spike protein RBD, or the SARS-CoV-2 spike protein RBD) or a portion thereof, wherein the K of the specific antigen or a portion thereof... DLess than about 50 nM, less than about 40 nM, less than about 30 nM, less than about 20 nM, less than about 10 nM, less than about 5 nM, less than about 4 nM, less than about 3 nM, less than about 2 nM, or less than about 1 nM, as measured in surface plasmon resonance measurements.

[0235] As used herein, the term "surface plasmon resonance" refers to an optical phenomenon that allows for the analysis of real-time interactions by detecting changes in protein concentration within a biosensor matrix, for example, using BIAcore. TM System (Biacore LifeSciences division of GE Healthcare, Piscataway, NJ).

[0236] As used in this article, the term "K" D "K" refers to the equilibrium dissociation constant of a specific protein-protein interaction (e.g., antibody-antigen interaction). Unless otherwise stated, the K values ​​disclosed herein are not necessarily the same. D The value refers to K, which is determined by surface plasmon resonance at 25°C. D value.

[0237] Antibodies containing heavy chain constant region variants

[0238] According to certain embodiments of the present invention, antibodies against the SARS-CoV-2 spike protein comprising an Fc domain are provided, wherein the Fc domain contains one or more mutations, for example, that enhance or weaken the binding of the antibody to the FcRn receptor at acidic pH compared to neutral pH. For example, the present invention includes C in the Fc domain... H Zone 2 or C HRegion 3 contains a mutated anti-SARS-CoV-2 spike protein antibody, wherein the mutation increases the affinity of the Fc domain for FcRn in an acidic environment (e.g., in the endosome with a pH range of about 5.5 to about 6.0). When administered to animals, this type of mutation can lead to an increased serum half-life of the antibody. Non-limiting examples of such Fc modifications include, for example, modifications at position 250 (e.g., E or Q); modifications at positions 250 and 428 (e.g., L or F); modifications at position 252 (e.g., L / Y / F / W or T), modifications at position 254 (e.g., S or T), and modifications at position 256 (e.g., S / R / Q / E / D or T); or modifications at positions 428 and / or 433 (e.g., H / L / R / S / P / Q or K) and / or modifications at position 434 (e.g., A, W, H, F, or Y [N434A, N434W, N434H, N434F, or N434Y]); or modifications at positions 250 and / or 428; or modifications at positions 307 or 308 (e.g., 308F, V308F) and modifications at position 434. In one embodiment, the modifications include 428L (e.g., M428L) and / or 434S (e.g., N434S) modifications; 428L, 259I (e.g., V259I) and 308F (e.g., V308F) modifications; 433K (e.g., H433K) and 434 (e.g., 434Y) modifications; 252, 254 and 256 (e.g., 252Y, 254T and 256E) modifications; 250Q and 428L modifications (e.g., T250Q and M428L); and 307 and / or 308 modifications (e.g., 308F or 308P). In another embodiment, the modifications include 265A (e.g., D265A) and / or 297A (e.g., N297A) modifications.

[0239] For example, the present invention includes an anti-SARS-CoV-2 spike protein antibody comprising an Fc domain comprising one or more pairs of mutations or mutation sets selected from the group consisting of: 250Q and 248L (e.g., T250Q and M248L); 252Y, 254T and 256E (e.g., M252Y, S254T and T256E); 428L and 434S (e.g., M428L and N). 434S); 257I and 311I (e.g., P257I and Q311I); 257I and 434H (e.g., P257I and N434H); 376V and 434H (e.g., D376V and N434H); 307A, 380A and 434A (e.g., T307A, E380A and N434A); and 433K and 434F (e.g., H433K and N434F). All possible combinations of the aforementioned Fc domain mutations and other mutations within the antibody variable domains disclosed herein are within the scope of this invention.

[0240] In various implementations, the anti-SARS-CoV-2 spike protein antibody includes a heavy chain constant region that binds to sequences derived from more than one immunoglobulin isotype. For example, the chimeric heavy chain constant region may include sequences derived from human IgG1, human IgG2, or human IgG4 C. H C in Zone 2 H 2. Part or all of the sequence and C derived from human IgG1, human IgG2 or human IgG4 H 3. Part or all of the sequence. The chimeric heavy chain constant region may also contain a chimeric hinge region. For example, the chimeric hinge may include an “upper hinge” sequence derived from the hinge region of human IgG1, human IgG2, or human IgG4, combined with a “lower hinge” sequence derived from the hinge region of human IgG1, human IgG2, or human IgG4. Specific examples of chimeric heavy chain constant regions that may be included in any of the antibodies shown herein include, from the N-terminus to the C-terminus: [IgG4 C H 1]-[IgG4 upper hinge]-[IgG2 lower hinge]-[IgG4 CH2]-[IgG4 CH3]. Another example of a chimeric heavy chain constant region that may be included in any of the antibodies shown herein includes, from the N-terminus to the C-terminus: [IgG1 C H 1]-[IgG1 upper hinge]-[IgG2 lower hinge]-[IgG4 CH2]-[IgG1 CH3]. These and other examples of chimeric heavy chain constant regions that may be included in any of the antibodies of the present invention are described in WO 2014 / 121087 (8550-WO). Chimeric heavy chain constant regions having these general structural arrangements and their variants may have altered Fc receptor binding, which in turn affects Fc effector function.

[0241] In various implementations, the anti-SARS-CoV-2 spike protein antibody comprises a heavy chain constant region including a hinge domain, wherein positions 233-236 within the hinge domain can be G, G, G and unoccupied; G, G, unoccupied and unoccupied; G, unoccupied, unoccupied and unoccupied; or all unoccupied, wherein the positions are numbered according to EU designations. Optionally, the heavy chain constant region includes a hinge domain, a CH2 domain, and a CH3 domain from the N-terminus to the C-terminus. Optionally, the heavy chain constant region includes a CH1 domain, a hinge domain, a CH2 domain, and a CH3 domain from the N-terminus to the C-terminus. Optionally, the CH1 region (if present), the remainder of the hinge region (if present), the CH2 region, and the CH3 region are the same human isotype. Optionally, the CH1 region (if present), the remainder of the hinge region (if present), the CH2 region, and the CH3 region are human IgG1. Optionally, the CH1 region (if present), the remainder of the hinge region (if present), the CH2 region, and the CH3 region are human IgG2. Optionally, the CH1 region (if present), the remainder of the hinge region (if present), the CH2 region, and the CH3 region are human IgG4. Optionally, the constant region has a CH3 domain modified to reduce binding to protein A. These and other examples of modified heavy chain constant regions that may be included in any of the antibodies of the present invention are described in WO 2016 / 161010 (10140WO01).

[0242] Epitope plotting and related techniques

[0243] This invention includes an anti-SARS-CoV-2 spike protein antibody that interacts with one or more amino acids found within the SARS-CoV-2 spike protein (e.g., within the spike protein RBD). The antibody-binding epitope may consist of a single, continuous sequence of three or more amino acids (e.g., 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or more) located within the spike protein RBD. Alternatively, the epitope may consist of multiple non-continuous amino acids (or amino acid sequences) within the spike protein RBD.

[0244] Various techniques known to those skilled in the art can be used to determine whether an antibody "interacts with one or more amino acids" within a peptide or protein. Exemplary techniques include, for example, conventional cross-blocking assays (such as those used in...). AntibodiesCross-blocking assays described in Harlow and Lane (Cold Spring Harbor Press, Cold Spring Harb., NY), alanine scanning mutation analysis, peptide blotting analysis (Reineke, Methods Mol Biol, Vol. 248: pp. 443-463, 2004), and peptide cleavage analysis can be used. Additionally, methods such as epitope excision, epitope extraction, and chemical modification of the antigen can be employed (Tomer, Protein Science, Vol. 9: pp. 487-496, 2000). Another method that can be used to identify amino acids within peptides interacting with antibodies is hydrogen / deuterium exchange detected by mass spectrometry. Generally, hydrogen / deuterium exchange methods involve deuterating the protein of interest and then binding the antibody to the deuterated protein. The protein / antibody complex is then transferred to water to allow hydrogen-deuterium exchange to occur at all residues except those protected by the antibody (which remain deuterated). After dissociation of the antibody, the target protein is subjected to protease cleavage and mass spectrometry analysis to reveal deuterated residues corresponding to specific amino acids that interact with the antibody. See, for example, Ehring, Analytical Biochemistry, Vol. 267, No. 2: pp. 252-259, 1999; Engen and Smith, Anal. Chem., Vol. 73: pp. 256A-265A, 2001.

[0245] The present invention also includes an anti-SARS-CoV-2 spike protein antibody that binds to the same epitope as any of the exemplary antibodies described above (e.g., mAb10933, mAb10987, or mAb10989). Similarly, the present invention also includes an anti-SARS-CoV-2 spike protein antibody that competes with any of the specific exemplary antibodies described herein (e.g., mAb10933, mAb10987, or mAb10989) for binding to the SARS-CoV-2 spike protein.

[0246] Using conventional methods known in the art and illustrated herein, it can be readily determined whether an antibody binds to the same epitope as or competitively binds to a reference anti-SARS-CoV-2 spike protein antibody. For example, to determine whether a test antibody binds to the same epitope as a reference anti-SARS-CoV-2 spike protein antibody discussed herein, the reference antibody is allowed to bind to the SARS-CoV-2 spike protein. Next, the ability of the test antibody to bind to the SARS-CoV-2 spike protein is evaluated. If the test antibody is able to bind to the SARS-CoV-2 spike protein after saturation binding with the reference anti-SARS-CoV-2 spike protein antibody, it can be concluded that the test antibody binds to a different epitope than the reference anti-SARS-CoV-2 spike protein antibody. On the other hand, if the test antibody is unable to bind to the SARS-CoV-2 spike protein after saturation binding with the reference anti-SARS-CoV-2 spike protein antibody, the test antibody may bind to the same epitope as defined by the reference anti-SARS-CoV-2 spike protein antibody discussed herein. Further routine experiments (e.g., peptide mutation and binding assays) can then be performed to confirm whether the observed lack of binding of the test antibody is actually due to binding to the same epitope as the reference antibody or whether the observed lack of binding is caused by steric hindrance (or other phenomena). Such experiments can be performed using ELISA, RIA, Biacore, flow cytometry, or any other quantitative or qualitative antibody binding assay available in the art. According to certain embodiments of the invention, if, for example, an excess of 1, 5, 10, 20, or 100 times of one antibody inhibits the binding of another antibody by at least 50%, but preferably 75%, 90%, or even 99%, then the two antibodies bind to the same (or overlapping) epitope, as measured in a competitive binding assay (see, for example, Junghans et al., Cancer Res., Vol. 50: pp. 1495-1502, 1990). Alternatively, if substantially all amino acid mutations in the antigen that reduce or eliminate the binding of one antibody reduce or eliminate the binding of the other antibody, then the two antibodies are considered to bind to the same epitope. If a subset of amino acid mutations that reduce or eliminate the binding of one antibody reduces or eliminates the binding of another antibody, then the two antibodies are considered to have "overlapping epitopes".

[0247] Preparation of human antibodies

[0248] Methods for generating monoclonal antibodies (including fully human monoclonal antibodies) are known in the art. Any such known methods can be used in the context of this invention for preparing human antibodies that specifically bind to the SARS-CoV-2 spike protein.

[0249] For example, using VELOCIMMUNE TMThe technique, or any other known method for producing fully human monoclonal antibodies, initially isolates a high-affinity chimeric antibody containing a human variable region and a mouse constant region of the SARS-CoV-2 spike protein. The antibody is characterized and desired properties, including affinity, selectivity, epitopes, etc., are selected. If desired, the mouse constant region is replaced with the desired human constant region (e.g., wild-type or modified IgG1 or IgG4) to produce a fully human anti-SARS-CoV-2 spike protein antibody. While the selected constant region can vary depending on the specific application, high-affinity antigen binding and target-specific properties are present in the variable region. In some cases, fully human anti-SARS-CoV-2 spike protein antibodies are isolated directly from antigen-positive B cells.

[0250] bioequivalent

[0251] The anti-SARS-CoV-2 spike protein antibodies and antibody fragments of the present invention encompass proteins having an amino acid sequence different from that of the antibodies but retaining the ability to bind the SARS-CoV-2 spike protein. When compared with parental sequences, such variant antibodies or antibody fragments contain one or more additions, deletions, or substitutions of amino acids but exhibit biological activity substantially equivalent to that of the antibodies. Similarly, when compared with disclosed sequences, the DNA sequences encoding anti-SARS-CoV-2 spike protein antibodies of the present invention encompass sequences containing one or more additions, deletions, or substitutions of nucleotides but encoding anti-SARS-CoV-2 spike protein antibodies or antibody fragments substantially bioequivalent to the anti-SARS-CoV-2 spike protein antibodies or antibody fragments of the present invention.

[0252] If, for example, two antibodies are pharmaceutical equivalents or substitutes that do not show significant differences in absorption rate and extent when administered at the same molar dose (single or multiple doses) under similar experimental conditions, then the two antibodies are considered bioequivalent. If some antibodies are equivalent in their extent of absorption but not in their rate of absorption, then the antibodies are considered equivalents or substitutes, and can be considered bioequivalent because such intentional differences in absorption rate reflected in the label are not necessary for achieving effective bodily drug concentrations, for example, with prolonged use, and are considered medically irrelevant to the specific pharmaceutical product under investigation.

[0253] In one implementation, the two antibodies are bioequivalent if there are no clinically significant differences in their safety, purity, and potency.

[0254] In one implementation, two antibodies are bioequivalent if a patient can switch between the reference product and the biologic product once or multiple times without an expected increased risk of side effects, including clinically significant changes in immunogenicity or reduced efficacy, compared to continued treatment without such switching.

[0255] In one implementation, the two antibodies are bioequivalent if they both function through one or more co-operating mechanisms against one or more conditions of use (provided those mechanisms are known).

[0256] Bioequivalence can be demonstrated through in vivo and in vitro methods. Bioequivalence measurements include, for example, (a) in vivo studies in humans or other mammals, in which the concentration of the antibody or its metabolites in blood, plasma, serum, or other biological fluids is measured as a function of time; (b) in vitro studies that correlate with and reasonably predict in vivo bioavailability data; (c) in vivo studies in humans or other mammals, in which the appropriate acute pharmacological effect of the antibody (or its target) is measured as a function of time; and (d) in clinical trials that establish good controls for the safety, efficacy, or bioavailability or bioequivalence of the antibody.

[0257] Bioequivalent variants of the anti-SARS-CoV-2 spike protein antibodies of the present invention can be constructed, for example, by various substitutions or deletions of terminal or internal residues or sequences that are not biologically essential. For example, cysteine ​​residues that are not biologically essential can be deleted or substituted with other amino acids to prevent the formation of unwanted or incorrect intramolecular disulfide bonds during renaturation. In other contexts, bioequivalent antibodies may include anti-SARS-CoV-2 spike protein antibody variants that incorporate amino acid changes that modify the glycosylation properties of the antibody, such as mutations that eliminate or remove glycosylation.

[0258] In some embodiments, the antibodies disclosed herein lack fucose in their constant-region glycosylation. Methods for measuring fucose in antibody compositions have been described in the art, for example, U.S. Patent No. 8,409,838 (Regeneron Pharmaceuticals), which is incorporated herein by reference. In some embodiments, fucose is not detected in the composition comprising a population of antibody molecules. In some embodiments, the fucose-deficient antibody exhibits enhanced ADCC activity.

[0259] In some implementations, antibodies lacking fucose can be generated using cell lines that lack the ability to reduce or eliminate fucosylated proteins. Fucosylation of glycans requires the synthesis of GDP-fucose via a de novo or salvage pathway, both involving the sequential function of several enzymes that result in the addition of the fucose molecule to the first N-acetylglucosamine (GlcNAc) moiety at the reducing end of the glycan. Two key enzymes responsible for the de novo pathway of GDP-fucose production are GDP-D-mannose-4,6-dehydratase (GMD) and GDP-keto-6-deoxymannose-3,5-episomerase,4-reductase (FX). In the absence of fucose, these two de novo pathway enzymes (GMD and FX) convert mannose and / or glucose to GDP-fucose, which is then transported to the Golgi complex. In the Golgi complex, nine fucosyltransferases (FUT1-9) work synergistically to fucosylate the first GlcNAc molecule of the glycan. However, in the presence of fucose, the salvage pathway enzymes, fucokinase and GDP-fucosylationase, convert fucose to GDP-fucose.

[0260] Cell lines lacking the ability to produce fucosylated proteins have been described in the art. In some embodiments, the cell lines lacking the ability to produce fucosylated proteins are mammalian cell lines (e.g., CHO cell lines, such as CHOK1, DXB-11CHO, Veggie-CHO) that contain mutations or genetic modifications in one or more of the endogenous FUT1 to 9 genes, resulting in the lack of one or more functional fucosylated transferases. In some embodiments, the mammalian cell line contains a mutation in the endogenous FUT8 gene (e.g., a FUT8 knockout cell line, wherein the FUT8 gene has been disrupted, resulting in a lack of functional α1,6-fucosyltransferase in the cell line, as described in the following patents incorporated herein by reference: U.S. Patent No. 7,214,775 (Kyowa Hakko Kogyo Co., Ltd.) and U.S. Patent No. 7,737,725 (Kyowa Hakko Kirin Co., Ltd.). In some embodiments, the mammalian cell line contains a mutation or genetic modification in the endogenous GMD gene, resulting in a lack of functional GMD in the cell line, e.g., a GMD knockout cell line wherein the GMD gene has been disrupted, as described in, for example, U.S. Patent No. 7,737,725 (Kyowa Hakko Kirin Co., Ltd.), incorporated herein by reference. In some embodiments, the mammalian cell line contains a mutation or genetic modification of the endogenous Fx gene, resulting in a lack of functional Fx protein. In some embodiments, the mammalian cell line is an Fx knockout cell line in which the endogenous Fx gene has been disrupted (see, for example, U.S. Patent 7,737,725 (Kyowa Hakko Kirin Co., Ltd.), which is incorporated herein by reference). In some embodiments, the mammalian cell line contains a mutation of the endogenous Fx mutation that confers a temperature-sensitive phenotype (as described in U.S. Patent No. 8,409,838 (Regeneron Pharmaceuticals), which is incorporated herein by reference). In some embodiments, the mammalian cell line lacking the ability to fucosylate proteins is selected based on resistance to certain lectins (e.g., lentil lectin). See, for example, U.S. Patent No. 8,409,838 (Regeneron Pharmaceuticals), which is incorporated herein by reference.

[0261] Therapeutic preparations and administration

[0262] The anti-SARS-CoV-2 spike protein antibody or antigen-binding fragment used in the methods and uses of this invention can be formulated into pharmaceutical compositions for administration with one or more pharmaceutically acceptable carriers, excipients, or diluents. These pharmaceutical compositions are formulated with suitable carriers, excipients, and other agents that provide improved transfer, delivery, tolerability, etc. Many suitable formulations can be found in all formularies known to medicinal chemists, such as Remington's Pharmaceutical Sciences, Mack Publishing Company, Easton, PA. These formulations include, for example, powders, pastes, ointments, gels, waxes, oils, lipids, and vesicle-containing lipids (cationic or anionic) (such as LIPOFECTIN). TM Life Technologies (Carlsbad, California), DNA conjugates, anhydrous absorbent pastes, oil-in-water and water-in-oil emulsions, polyethylene glycol emulsions (polyethylene glycol of various molecular weights), semi-solid gels, and semi-solid mixtures containing polyethylene glycol. See also Powell et al., “Compendium of excipients for parenteral formulations,” PDA, J Pharm SciTechnol, Vol. 52: pp. 238–311, 1998.

[0263] mAb10933 and mAb10987 are human IgG1 mAbs that simultaneously bind to different non-overlapping epitopes on the spike (S) glycoprotein of SARS-CoV-2. mAb10933 and mAb10987, in combination, can be found in antibody mixtures called REGN-COV2 or REGEN-COV, which are produced via recombinant DNA technology in Chinese hamster ovary (CHO) cell suspension cultures and have molecular weights of approximately 145.23 kDa and 144.14 kDa, respectively. The antibodies described herein (e.g., mAb10933 and mAb10987) can be formulated alone or in combination. For example, co-formulated compositions can be used to simplify administration (e.g., intravenous or subcutaneous), while individual formulations offer greater flexibility in dosing. In a particular embodiment, the two antibodies (mAb10933 and mAb10987) referred to as REGEN-COV in the composition may be co-formulated, or the two antibodies may be formulated separately and combined prior to administration.

[0264] In some embodiments, mAb10933 and mAb10987 are sterile, preservative-free, clear to slightly milky white and colorless to pale yellow solutions with a pH of 6.0. In some embodiments, each of mAb10933 and mAb10987 may be formulated as: 120 mg / mL antibody, 10 mM histidine, 8% (w / v) sucrose, and 0.1% (w / v) polysorbate 80, pH 6.0. Each antibody is available in two concentrations: 300 mg / 2.5 mL and 1332 mg / 11.1 mL. In some embodiments, mAb10933 and mAb10987 are each available as vials containing 300 mg of antibody (e.g., in a 2.5 mL solution) or 1332 mg of antibody (e.g., in an 11.1 mL solution). Exemplary contents of each vial are shown below:

[0265] 300mg vial

[0266] • mAb10933: Each 2.5 mL solution contains 300 mg of mAb10933, L-histidine (1.9 mg), L-histidine monohydrochloride monohydrate (2.7 mg), polysorbate 80 (2.5 mg), sucrose (200 mg), and water for injection, USP. pH 6.0.

[0267] • mAb10987: Each 2.5 mL solution contains 300 mg of mAb10987, L-histidine (1.9 mg), L-histidine monohydrochloride monohydrate (2.7 mg), polysorbate 80 (2.5 mg), sucrose (200 mg), and water for injection, USP. pH 6.0.

[0268] 1332mg vial

[0269] • mAb10933: Each 11.1 mL solution contains 1332 mg of mAb10933, L-histidine (8.3 mg), L-histidine monohydrochloride monohydrate (12.1 mg), polysorbate 80 (11.1 mg), sucrose (888 mg), and water for injection, USP. pH 6.0.

[0270] • mAb10987: Each 11.1 mL solution contains 1332 mg of mAb10987, L-histidine (8.3 mg), L-histidine monohydrochloride monohydrate (12.1 mg), polysorbate 80 (11.1 mg), sucrose (888 mg), and water for injection, USP. pH 6.0.

[0271] The dosage of the antibody administered to a patient can vary depending on the patient's age and body size, symptoms, route of administration, etc. A preferred dosage is typically calculated based on body weight or body surface area. When the antibody of the present invention is used to treat adult patients, it may be advantageous to administer the antibody of the present invention intravenously in a single dose of about 0.01 mg / kg to about 20 mg / kg body weight, more preferably about 0.02 mg / kg to about 7 mg / kg body weight, about 0.03 mg / kg to about 5 mg / kg body weight, or about 0.05 mg / kg to about 3 mg / kg body weight. The frequency and duration of treatment can be adjusted according to the severity of the symptoms. The effective dosage and regimen for administering the anti-SARS-CoV-2 spike protein antibody can be determined empirically; for example, patient progression can be monitored through periodic assessments, and the dosage adjusted accordingly. Furthermore, interspecies scaling of the dosage can be performed using methods well known in the art (e.g., Mordenti et al., Pharmaceut Res, Vol. 8: p. 1351, 1991).

[0272] Various delivery systems are known and can be used to administer the pharmaceutical compositions of the present invention, such as encapsulation in liposomes, microparticles, microcapsules, recombinant cells capable of expressing the antibodies or other therapeutic proteins of the present invention, and receptor-mediated endocytosis (see, for example, Wu et al., J Biol Chem, Vol. 262: pp. 4429-4432, 1987). The antibodies and other therapeutically active components of the present invention can also be delivered via gene therapy techniques. Methods of administration include, but are not limited to, intradermal, intramuscular, intraperitoneal, intravenous, subcutaneous, intranasal, epidural, and oral routes. The compositions can be administered via any convenient route, such as by infusion or rapid concentration, absorption through the epithelial or mucosal lining (e.g., oral mucosa, rectal and intestinal mucosa), and can be administered with other bioactive agents. Administration can be systemic or local.

[0273] The drug composition can be delivered subcutaneously or intravenously using standard needles and syringes. Additionally, for subcutaneous delivery, pen delivery devices are readily applicable for delivering the drug compositions of this invention. Such pen delivery devices can be reusable or disposable. Reusable pen delivery devices typically utilize a replaceable cartridge containing the drug composition. Once the entire drug composition in the cartridge has been administered and the cartridge is empty, the empty cartridge can be easily discarded and replaced with a new cartridge containing the drug composition. The pen delivery device can then be reused. In disposable pen delivery devices, there is no replaceable cartridge. Instead, the disposable pen delivery device is pre-loaded with the drug composition held in a reservoir within the device. Once the drug composition in the reservoir is emptied, the entire device is discarded.

[0274] Many reusable pen delivery devices and autoinjector delivery devices are used for subcutaneous delivery of the pharmaceutical compositions discussed herein. Examples include, but are not limited to, AUTOPEN. TM (Owen Mumford, Inc., Woodstock, UK), DISETRONIC TM Pen (Disetronic Medical Systems, Bodolf, Switzerland), HUMALOGMIX 75 / 25 TM Pen, HUMALOG TM Pen, HUMALIN 70 / 30 TM Pen (Eli Lilly and Co., Indianapolis, Indiana), NOVOPEN TM I, II and III (Novo Nordisk, Copenhagen, Denmark), NOVOPEN JUNIOR TM (Novo Nordisk, Copenhagen, Denmark), BD TM Pen (Becton Dickinson, Franklin Lake, New Jersey), OPTIPEN TM OPTIPEN PRO TM OPTIPEN STARLET TM And OPTICLIK TM (Sanofi-Aventis, Frankfurt, Germany), to name just a few. Examples of disposable pen delivery devices for the pharmaceutical compositions of the present invention used in subcutaneous delivery include, but are not limited to, SOLOSTAR. TM Pen (Sanofi), FLEXPEN TM (Novo Nordisk) and KWIKPEN TM (Lilly), SURECLICK TM Self-injector (Amgen, Thousand Oaks, California), PENLET TM (Haselmeier, Stuttgart, Germany), EPIPEN (Dey GmbH (Dey, LP)) and HUMIRA TM Pens (Abbott Labs, Abbott Science Park IL), to name just a few.

[0275] In some cases, drug compositions can be delivered in controlled-release systems. In one embodiment, a pump can be used (see Langer, ibid.; Sefton, CRC Crit. Ref. Biomed. Eng., Vol. 14: p. 201, 1987). In another embodiment, polymeric materials can be used; see Medical Applications of Controlled Release, Langer and Wise (eds.), 1974, CRC Pres., Boca Raton, Florida. In yet another embodiment, the controlled-release system can be placed near the target of the composition, thus requiring only a fraction of the systemic dose (see, for example, Goodson, 1984, Controlled Release in Medicine, ibid., Vol. 2, pp. 115–138). Other controlled-release systems are discussed in the following review: Langer, Science, Vol. 249: pp. 1527–1533, 1990.

[0276] Injectable formulations can comprise dosage forms for intravenous, subcutaneous, intradermal, and intramuscular injection, infusion, etc. These injectable formulations can be prepared by known methods. For example, injectable formulations can be prepared by, for instance, dissolving, suspending, or emulsifying the aforementioned antibody or its salts in a sterile aqueous medium or an oily medium conventionally used for injection. Aqueous media for injection include, for example, physiological saline, isotonic solutions containing glucose, and other adjuvants.

[0277] Combination therapy

[0278] In some cases, anti-SARS-CoV-2 spike protein antibodies may be administered in conjunction with additional therapeutic agents. In some embodiments, the additional therapeutic agent is an antiviral drug or a vaccine. In some embodiments, the additional therapeutic agent is selected from the group consisting of: anti-inflammatory agents, antimalarial agents, antibodies that specifically bind to TMPRSS2 or their antigen-binding fragments, and antibodies or antigen-binding fragments that specifically bind to the SARS-CoV-2 spike protein. In some cases, the antimalarial agent is chloroquine or hydroxychloroquine. In some cases, the anti-inflammatory agent is an antibody, such as thalidomide, tocilizumab, or gemcitabine. In some embodiments, the additional therapeutic agent is a second antibody or antigen-binding fragment comprising the sequences HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 of Table 1.

[0279] Additional therapeutic agents may be administered to the subject or used prior to the administration of the anti-SARS-CoV-2 spike protein antibody of the present invention. For example, if the first component is administered / used 1 week, 72 hours, 60 hours, 48 ​​hours, 36 hours, 24 hours, 12 hours, 6 hours, 5 hours, 4 hours, 3 hours, 2 hours, 1 hour, 30 minutes, 15 minutes, 10 minutes, 5 minutes, or less than 1 minute prior to the administration / use of the second component, the first component may be considered to have been administered / used "before" the second component. In other embodiments, additional therapeutic agents may be administered to the subject or used after the administration of the anti-SARS-CoV-2 spike protein antibody of the present invention. For example, if the first component is administered / used 1 minute, 5 minutes, 10 minutes, 15 minutes, 30 minutes, 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 12 hours, 24 hours, 36 hours, 48 ​​hours, 60 hours, or 72 hours after the second component is administered / used, then the first component can be considered to be administered / used "after" the second component. In other embodiments, an additional therapeutic agent may be administered to the subject or used concurrently with the administration of the anti-SARS-CoV-2 spike protein antibody of the present invention. For the purposes of the present invention, "concurrent" administration includes, for example, administering the anti-SARS-CoV-2 spike protein antibody and the additional therapeutically active component to the subject in a single dosage form, or administering separate dosage forms to the subject at intervals of approximately 30 minutes or less. If administered in separate dosage forms, each dosage form may be administered via the same route (e.g., the anti-SARS-CoV-2 spike protein and the additional therapeutically active component may be administered intravenously, subcutaneously, etc.). In any event, for the purposes of this disclosure, administration of the components in a single dosage form, in a single dosage form via the same route, or in a single dosage form via a different route is considered "simultaneous administration." For the purposes of this disclosure, administration of an additional therapeutic agent "before," "simultaneously with," or "after" administration of the anti-SARS-CoV-2 spike protein antibody (as defined above) is considered administration of the anti-SARS-CoV-2 spike protein antibody "in combination" with the additional therapeutic agent.

[0280] dose

[0281] The amount of active ingredient (e.g., an anti-SARS-CoV-2 spike protein antibody or other therapeutic agents administered in combination with an anti-SARS-CoV-2 spike protein antibody) that can be administered to a subject is typically a therapeutically effective amount, as discussed elsewhere in this document.

[0282] In some implementations, the therapeutically effective dose can be from about 0.05 mg to about 20 g; for example, 0.05 mg, about 0.1 mg, about 1.0 mg, about 1.5 mg, about 2.0 mg, about 10 mg, about 20 mg, about 30 mg, about 40 mg, about 50 mg, about 60 mg, about 70 mg, about 80 mg, about 90 mg, about 100 mg, about 110 mg, about 120 mg, about 130 mg, about 140 mg, about 150 mg, about 160 mg, about 170 mg, about 180 mg, about 190 mg, about 200 mg, about 210 mg, about 220 mg, about 230 mg, about 240 mg, about 250 mg, about 260 mg, about 270 mg, about... 280mg, approximately 290mg, approximately 300mg, approximately 310mg, approximately 320mg, approximately 330mg, approximately 340mg, approximately 350mg, approximately 360mg, approximately 370mg, approximately 380mg, approximately 390mg, approximately 400mg, approximately 410mg, approximately 420mg, approximately 430mg, approximately 440mg, approximately 450mg, approximately 460mg, approximately 470mg, approximately 480mg, approximately 490mg, approximately 500mg, approximately 510mg, approximately 520mg, approximately 530mg, approximately 540mg, approximately 550mg, approximately 560mg, approximately 570mg, approximately 580mg, approximately 590mg, approximately 600mg, approximately 610mg, approximately 620mg, approximately 630mg g, approximately 640mg, approximately 650mg, approximately 660mg, approximately 670mg, approximately 680mg, approximately 690mg, approximately 700mg, approximately 710mg, approximately 720mg, approximately 730mg, approximately 740mg, approximately 750mg, approximately 760mg, approximately 770mg, approximately 780mg, approximately 790mg, approximately 800mg, approximately 810mg, approximately 820mg, approximately 830mg, approximately 840mg, approximately 850mg, approximately 860mg, approximately 870mg, approximately 880mg, approximately 890mg, approximately 900mg, approximately 910mg, approximately 920mg, approximately 930mg, approximately 940mg, approximately 950mg, approximately 960mg, approximately 970mg, approximately 980mg, approximately 9 90mg, approximately 1g, approximately 1.1g, approximately 1.2g, approximately 1.3g, approximately 1.4g, 1.5g, approximately 1.6g, approximately 1.7g, approximately 1.8g, approximately 1.9g, approximately 2g, approximately 2.1g, approximately 2.2g, approximately 2.3g, approximately 2.4g, approximately 2.5g, approximately 2.6g, approximately 2.7g, approximately 2.8g, approximately 2.9g, approximately 3g Approximately 3.1g, approximately 3.2g, approximately 3.3g, approximately 3.4g, approximately 3.5g, approximately 3.6g, approximately 3.7g, approximately 3.8g, approximately 3.9g, approximately 4g, approximately 4.1g, approximately 4.2g, approximately 4.3g, approximately 4.4g, approximately 4.5g, approximately 4.6g, approximately 4.7g, approximately 4.8g, approximately 4.9g, approximately 5g, approximately 5.1g, approximately 5g.2g, approx. 5.3g, approx. 5.4g, approx. 5.5g, approx. 5.6g, approx. 5.7g, approx. 5.8g, approx. 5.9g, approx. 6g, approx. 6.1g, approx. 6.2g, approx. 6.3g, approx. 6.4g, approx. 6.5g, approx. 6.6g, approx. 6.7g, approx. 6.8g, approx. 6.9g, approx. 7g, approx. 7.1g, approx. 7.2g, approx. 7.3g, approx. 7.4g, approx. 7.5g, approx. 7.6g, approx. 7.7g, approx. 7.8g, approx. 7.9g, approx. 8g, approx. 8g. The corresponding antibodies are available in doses of approximately 1g, 8.2g, 8.3g, 8.4g, 8.5g, 8.6g, 8.7g, 8.8g, 8.9g, 9g, 9.1g, 9.2g, 9.3g, 9.4g, 9.5g, 9.6g, 9.7g, 9.8g, 9.9g, 10g, 11g, 12g, 13g, 14g, 15g, 16g, 17g, 18g, 19g, or 20g. In some cases, the therapeutically effective dose is 0.1g to 3.5g. In some cases, the therapeutically effective dose is 0.5g to 2g. In some cases, the therapeutically effective dose is 0.8g to 1.6g. In some cases, the therapeutically effective dose is 1.0g to 1.4g. In some cases, the effective therapeutic dose is 1g to 7g. In some cases, the effective therapeutic dose is 3g to 5g. In some cases, the effective therapeutic dose is 3.5g to 4.5g. In any of these embodiments, the dose may represent the dose of a single antibody or the total dose of a combination of antibodies. For example, two different anti-SARS-CoV-2 spike glycoprotein antibodies may be administered co-administered, wherein the dose of each antibody represents half of the total dose administered.

[0283] In some embodiments, mAb10933 and mAb10987 are administered intravenously or subcutaneously in a total dose of 300 mg to 2400 mg. In some cases, the total dose is 100 mg to 5000 mg. In some embodiments, the total dose is 200 mg to 400 mg, 500 mg to 700 mg, 1000 mg to 1400 mg, or 2000 mg to 2800 mg. In some embodiments, the total dose is 250 mg to 350 mg, 550 mg to 650 mg, 1150 mg to 1250 mg, or 2300 mg to 2500 mg. In some cases, the total dose is 300 mg, 600 mg, 1200 mg, or 2400 mg. In some cases, the total dose is 100mg, 150mg, 200mg, 250mg, 300mg, 350mg, 400mg, 450mg, 500mg, 550mg, 600mg, 650mg, 700mg, 750mg, 800mg, 850mg, 900mg, 1000mg, 1050mg, 1100mg, 1150mg, 1200mg, 1250mg, 1300mg, 1350mg, 1400mg, 1450mg, 1500mg, 1550mg, 160mg. 0 mg, 1650 mg, 1700 mg, 1750 mg, 1800 mg, 1850 mg, 1900 mg, 1950 mg, 2000 mg, 2050 mg, 2100 mg, 2150 mg, 2200 mg, 2250 mg, 2300 mg, 2350 mg, 2400 mg, 2450 mg, 2500 mg, 2550 mg, 2600 mg, 2650 mg, 2700 mg, 2750 mg, 2800 mg, 2850 mg, 2900 mg, 2950 mg, or 3000 mg. In some embodiments, the total dose is 2400 mg, and each of mAb10933 and mAb10987 is administered intravenously at a dose of 1200 mg. In some embodiments, the total dose is 1200 mg, and each of mAb10933 and mAb10987 is administered intravenously at a dose of 600 mg. In some embodiments, the total dose is 600 mg, and each of mAb10933 and mAb10987 is administered intravenously at a dose of 300 mg. In some embodiments, the total dose is 300 mg, and each of mAb10933 and mAb10987 is administered intravenously at a dose of 150 mg. In some embodiments, the total dose is 1200 mg, and each of mAb10933 and mAb10987 is administered subcutaneously at a dose of 600 mg. In some embodiments, the total dose is 600 mg, and each of mAb10933 and mAb10987 is administered subcutaneously at a dose of 300 mg.In some embodiments, each individual antibody is administered at a dose of 100 mg to 200 mg, 200 mg to 400 mg, 500 mg to 700 mg, or 2300 mg to 2500 mg. In some cases, each individual antibody is administered at a dose of 124 mg to 175 mg, 250 mg to 350 mg, 550 mg to 650 mg, or 1150 mg to 1250 mg.

[0284] The amount of anti-SARS-CoV-2 spike protein antibody or other therapeutic agent contained in each dose can be expressed in milligrams of antibody per kilogram of patient body weight (i.e., mg / kg). For example, anti-SARS-CoV-2 spike protein antibody can be administered to patients at doses from approximately 0.0001 mg / kg to approximately 200 mg / kg of patient body weight (e.g., 0.1 mg / kg, 0.5 mg / kg, 1.0 mg / kg, 1.5 mg / kg, 2.0 mg / kg, 2.5 mg / kg, 3.0 mg / kg, 3.5 mg / kg, 4.0 mg / kg, 4.5 mg / kg, 5.0 mg / kg, 5.5 mg / kg, 6.0 mg / kg, 6.5 mg / kg, 7.0 mg / kg, 7.5 mg / kg, 8.0 mg / kg, 8.5 mg / kg, 9.0 mg / kg, 9.5 ... mg / kg, 10.0mg / kg, 10.5mg / kg, 11.0mg / kg, 11.5mg / kg, 12.0mg / kg, 12.5mg / kg, 13.0mg / kg, 13.5mg / kg, 14.0mg / kg, 14.5mg / kg, 15.0mg / kg, 15.5mg / kg, 16.0mg / kg, 16.5mg / kg, 17.0mg / kg, 17.5mg / kg, 18.0mg / kg, 18.5mg / kg, 19.0mg / kg, 19.5mg / kg, 20.0mg / kg, 20.5mg / kg, 21.0mg / kg, 21.5mg / kg, 22.0mg / kg, 22.5mg / kg, 23.0mg / kg, 23.5mg / kg, 24.0mg / kg, 24.5mg / kg, 25.0mg / kg, 25.5mg / kg, 26.0mg / kg, 26.5mg / kg, 27. 0mg / kg, 27.5mg / kg, 28.0mg / kg, 28.5mg / kg, 29.0mg / kg, 29.5mg / kg, 30.0mg / kg, 30.5mg / kg, 31.0mg / kg, 31.5mg / kg, 32.0mg / kg, 32.5mg / kg, 33.0mg / kg, 33.5mg / kg, 34.0mg / kg, 34.5mg / kg, 35.0mg / kg, 35.5mg / kg, 36.0mg / kg, 36.5mg / kg, 37.0mg / kg, 37.5mg / kg, 38.0mg / kg, 38.5mg / kg, 39.0mg / kg, 39.5mg / kg, 40.0mg / kg, 40.5mg / kg, 41.0mg / kg, 41.5mg / kg, 42.0mg / kg, 42.5mg / kg, 43.0mg / kg, 43.5mg / kg, 44.0mg / kg, 44.5mg / kg、45.0mg / kg、45.5mg / kg、46.0mg / kg、46.5mg / kg、47.0mg / kg、47.5mg / kg、48.0mg / kg、48.5mg / kg、49.0mg / kg、49.5mg / kg、50.0mg / kg、50.5mg / kg、51.0mg / kg、51.5mg / kg、52.0mg / kg、52.5mg / kg、53.0mg / kg、53.5mg / kg、54.0mg / kg、54.5mg / kg、55.0mg / kg、55.5mg / kg、56.0mg / kg、56.5mg / kg、57.0mg / kg、57.5mg / kg、58.0mg / kg、58.5mg / kg、59.0mg / kg、59.5mg / kg、60.0mg / kg、60.5mg / kg、61.0mg / kg、61.5mg / kg、62.0mg / kg、62.5mg / kg、63.0mg / kg、63.5mg / kg、64.0mg / kg、64.5mg / kg、65.0mg / kg、65.5mg / kg、66.0mg / kg、66.5mg / kg、67.0mg / kg、67.5mg / kg、68.0mg / kg、68.5mg / kg、69.0mg / kg、69.5mg / kg、70.0mg / kg、70.5mg / kg、71.0mg / kg、71.5mg / kg、72.0mg / kg、72.5mg / kg、73.0mg / kg、73.5mg / kg、74.0mg / kg、74.5mg / kg、75.0mg / kg、75.5mg / kg、76.0mg / kg、76.5mg / kg、77.0mg / kg、77.5mg / kg、78.0mg / kg、78.5mg / kg、79.0mg / kg、79.5mg / kg、80.0mg / kg、80.5mg / kg、81.0mg / kg、81.5mg / kg、82.0mg / kg、82.5mg / kg、83.0mg / kg、83.5mg / kg、84.0mg / kg、84.5mg / kg、85.0mg / kg、85.5mg / kg、86.0mg / kg、86.5mg / kg、87.0mg / kg、87.5mg / kg、88.0mg / kg、88.5mg / kg、89.0mg / kg、89.5mg / kg、90.0mg / kg、90.5mg / kg、91.0mg / kg、91.5mg / kg、92.0mg / kg、92.5mg / kg、93.0mg / kg、93.5mg / kg、94.0mg / kg、94.5mg / kg、95.0mg / kg、95.5mg / kg、96.0mg / kg、96.5mg / kg、97.0mg / kg、97.5mg / kg、98.0mg / kg、98.5mg / kg、99.0mg / kg、99.5mg / kg、100.0mg / kg、100.5mg / kg、101.0mg / kg、101.5mg / kg、102.0mg / kg、102.5mg / kg、103.0mg / kg、103.5mg / kg、104.0mg / kg、104.5mg / kg、105.0mg / kg、105.5mg / kg、106.0mg / kg、106.5mg / kg、107.0mg / kg、107.5mg / kg、108.0mg / kg、108.5mg / kg、109.0mg / kg、109.5mg / kg、110.0mg / kg、110.5mg / kg、111.0mg / kg、111.5mg / kg、112.0mg / kg、112.5mg / kg、113.0mg / kg、113.5mg / kg、114.0mg / kg、114.5mg / kg、115.0mg / kg、115.5mg / kg、116.0mg / kg、116.5mg / kg、117.0mg / kg、117.5mg / kg、118.0mg / kg、118.5mg / kg、119.0mg / kg、119.5mg / kg、120.0mg / kg、120.5mg / kg、121.0mg / kg、121.5mg / kg、122.0mg / kg、122.5mg / kg、123.0mg / kg、123.5mg / kg、124.0mg / kg、124.5mg / kg、125.0mg / kg、125.5mg / kg、126.0mg / kg、126.5mg / kg、127.0mg / kg、127.5mg / kg、128.0mg / kg、128.5mg / kg、129.0mg / kg、129.5mg / kg、130.0mg / kg、130.5mg / kg、131.0mg / kg、131.5mg / kg、132.0mg / kg、132.5mg / kg、133.0mg / kg、133.5mg / kg、134.0mg / kg、134.5mg / kg、135.0mg / kg、135.5mg / kg、136.0mg / kg、136.5mg / kg、137.0mg / kg、137.5mg / kg、138.0mg / kg、138.5mg / kg、139.0mg / kg、139.5mg / kg、140.0mg / kg、140.5mg / kg、141.0mg / kg、141.5mg / kg、142.0mg / kg、142.5mg / kg、143.0mg / kg、143.5mg / kg、144.0mg / kg、144.5mg / kg、145.0mg / kg、145.5mg / kg、146.0mg / kg、146.5mg / kg、147.0mg / kg、147.5mg / kg、148.0mg / kg、148.5mg / kg、149.0mg / kg、149.5mg / kg、150.0mg / kg、150.5mg / kg、151.0mg / kg、151.5mg / kg、152.0mg / kg、152.5mg / kg、153.0mg / kg、153.5mg / kg、154.0mg / kg、154.5mg / kg、155.0mg / kg、155.5mg / kg、156.0mg / kg、156.5mg / kg、157.0mg / kg、157.5mg / kg、158.0mg / kg、158.5mg / kg、159.0mg / kg、159.5mg / kg、160.0mg / kg、160.5mg / kg、161.0mg / kg、161.5mg / kg、162.0mg / kg、162.5mg / kg、163.0mg / kg、163.5mg / kg、164.0mg / kg、164.5mg / kg、165.0mg / kg、165.5mg / kg、166.0mg / kg、166.5mg / kg、167.0mg / kg、167.5mg / kg、168.0mg / kg、168.5mg / kg、169.0mg / kg、169.5mg / kg、170.0mg / kg、170.5mg / kg、171.0mg / kg、171.5mg / kg、172.0mg / kg、172.5mg / kg、173.0mg / kg、173.5mg / kg、174.0mg / kg、174.5mg / kg、175.0mg / kg、175.5mg / kg、176.0mg / kg、176.5mg / kg、177.0mg / kg、177.5mg / kg、178.0mg / kg、178.5mg / kg、179.0mg / kg、179.5mg / kg、180.0mg / kg、180.5mg / kg、181.0mg / kg、181.5mg / kg、182.0mg / kg、182.5mg / kg、183.0mg / kg、183.5mg / kg、184.0mg / kg、184.5mg / kg、185.0mg / kg、185.5mg / kg, 186.0mg / kg, 186.5mg / kg, 187.0mg / kg, 187.5mg / kg, 188.0mg / kg, 188.5mg / kg, 189.0mg / kg, 189.5mg / kg, 190.0mg / kg, 190.5mg / kg, 191.0mg / kg, 191.5mg / kg, 192.0mg / kg, 192.5mg / kg, 19 3.0mg / kg, 193.5mg / kg, 194.0mg / kg, 194.5mg / kg, 195.0mg / kg, 195.5mg / kg, 196.0mg / kg, 196.5mg / kg, 197.0mg / kg, 197.5mg / kg, 198.0mg / kg, 198.5mg / kg, 199.0mg / kg, 199.5mg / kg or 200.0mg / kg). .

[0285] Application plan

[0286] According to certain embodiments of the invention, multiple doses of the active ingredient (e.g., an anti-SARS-CoV-2 spike protein antibody) may be administered to a subject over a predetermined time period. A method according to this aspect of the invention comprises sequentially administering multiple doses of the active ingredient of the invention to a subject. As used herein, “sequentially administering” means that each dose of the active ingredient is administered to the subject at different time points, for example, on different dates spaced apart by predetermined intervals (e.g., hours, days, weeks, or months). The invention includes a method comprising sequentially administering a single initial dose of the active ingredient to a patient, followed by one or more second doses of the active ingredient, and optionally subsequently administering one or more third doses of the active ingredient.

[0287] The terms “initial dose,” “second dose,” and “third dose” refer to the temporal sequence of administration of the active ingredient (e.g., the anti-SARS-CoV-2 spike protein antibody of the present invention) or the combination therapy of the present invention (e.g., two different anti-SARS-CoV-2 spike protein antibodies). Thus, the “initial dose” is the dose administered at the start of the treatment regimen (also known as the “baseline dose”); the “second dose” is the dose administered after the initial dose; and the “tertiary dose” is the dose administered after the second dose. The initial, second, and third doses may all contain the same amount of the active ingredient (e.g., the anti-SARS-CoV-2 spike protein antibody), but may generally differ from each other in terms of administration frequency. However, in some embodiments, the amount of the active ingredient (e.g., the anti-SARS-CoV-2 spike protein antibody) contained in the initial, second, and / or third doses during the treatment process differs from each other (e.g., appropriately increased or decreased). In some embodiments, two or more (e.g., 2, 3, 4, or 5) doses are administered at the start of the treatment regimen as a “loading dose,” followed by subsequent doses (e.g., a “maintenance dose”) on a less frequent basis.

[0288] In one exemplary embodiment of the invention, each second and / or third dose is administered 1 to 26 weeks after the previous dose (e.g., 1, 1...). 1 / 2、2、2 1 / 2, 3, 3 1 / 2、4、4 1 / 2, 5, 5 1 / 2, 6, 6 1 / 2、7、7 1 / 2、8、8 1 / 2, 9, 9 1 / 2, 10, 10 1 / 2, 11, 11 1 / 2, 12, 12 1 / 2, 13, 13 1 / 2, 14, 14 1 / 2, 15, 15 1 / 2, 16, 16 1 / 2, 17, 17 1 / 2, 18, 18 1 / 2, 19, 19 1 / 2, 20, 20 1 / 2、21、21 1 / 2、22、22 1 / 2、23、23 1 / 2、24、24 1 / 2、25、25 1 / 2, 26, 261 / 2 or more weeks). As used herein, the phrase “previous dose” means, in a sequence of multiple administrations, the dose of the active ingredient (e.g., anti-SARS-CoV-2 spike protein antibody) administered to the patient before the next dose in the sequence without intermediate doses.

[0289] Methods according to this aspect of the invention may include administering any number of second and / or third doses of the active ingredient of the invention, such as an anti-SARS-CoV-2 spike protein antibody, to a patient. For example, in some embodiments, only a single second dose is administered to the patient. In other embodiments, two or more (e.g., 2, 3, 4, 5, 6, 7, 8, or more) second doses are administered to the patient. Similarly, in some embodiments, only a single third dose is administered to the patient. In other embodiments, two or more (e.g., 2, 3, 4, 5, 6, 7, 8, or more) third doses are administered to the patient.

[0290] In embodiments involving multiple second doses, each second dose may be administered at the same frequency as the other second doses. For example, each second dose may be administered to the patient 1 to 2 weeks or 1 to 2 months after the previous dose. Similarly, in embodiments involving multiple third doses, each third dose may be administered at the same frequency as the other third doses. For example, each third dose may be administered to the patient 2 to 12 weeks after the previous dose. Alternatively, the frequency of administration of the second and / or third doses to the patient may vary throughout the treatment regimen. The frequency of administration may also be adjusted by the physician during treatment based on the individual patient's needs following clinical examination.

[0291] The present invention includes an administration regimen in which two to six loading doses are administered to a patient at a first frequency (e.g., once a week, once every two weeks, once every three weeks, once a month, once every two months, etc.), followed by two or more maintenance doses administered to the patient on a less frequent basis. For example, according to this aspect of the invention, if the loading dose is administered once a month, the maintenance dose can be administered to the patient once every six weeks, once every two months, once every three months, etc. In some embodiments, a single dose is administered to the subject as part of a prophylactic or therapeutic treatment procedure. In some embodiments, one or more doses are administered to treat adult or pediatric patients diagnosed with mild to moderate coronavirus disease (COVID-19) at high risk.

[0292] In some implementation schemes, the dosage for adult and pediatric patients (aged 12 and older, weighing at least 40 kg) is:

[0293] ο600 mg mAb10933 (cascirelimab) and 600 mg mAb10987 (edevimab) are administered together via pump or gravity as a single intravenous infusion (see Table 4A), a single subcutaneous injection, or two subcutaneous injections; or

[0294] ο 1,200 mg of cassicimarab and 1,200 mg of edevimab were administered together via a pump or gravity in a single intravenous infusion (see Table 4B); exemplary preparation instructions for mAb10933 + mAb10987 (cassicimarab and edevimab, respectively) are as follows:

[0295] 1. Remove the vials of cassicimarab and edevimab from refrigerated storage and allow them to equilibrate to room temperature for approximately 20 minutes before preparation. Do not expose directly to high temperatures. Do not shake the vials.

[0296] 2. Before administration, visually inspect the vials of cassicimarumab and edevimab for particulate matter and discoloration. If either is observed, the solution must be discarded and a fresh solution prepared. The solution in each vial should be clear to slightly milky white, colorless to pale yellow.

[0297] 3. Obtain a pre-filled IV infusion bag containing 50 mL, 100 mL, 150 mL, or 250 mL of 0.9% sodium chloride injection solution.

[0298] 4. If a 600mg / 600mg dose is administered:

[0299] Then, using two separate syringes, draw 5 mL of cassicimarab and 5 mL of edevimab from each vial (see Table 4A), and inject all 10 mL into a pre-filled infusion bag containing 0.9% sodium chloride injection solution (see Table 4A). Discard any remaining product in the vials.

[0300] or

[0301] If an alternative dose of 1,200 mg / 1,200 mg is administered:

[0302] Then, using two separate syringes, draw 10 mL of cassicimarab and 10 mL of edevimab from each vial (see Table 4B), and inject all 20 mL into a pre-filled infusion bag containing 0.9% sodium chloride injection solution (see Table 4B). Discard any remaining product in the vials.

[0303] 5. Gently invert the IV bag by hand about 10 times. Do not shake it.

[0304] 6. This product does not contain preservatives, therefore the diluted infusion solution should be administered immediately.

[0305] • If immediate administration is not possible, store the diluted cascislimab together with the edevimab infusion solution in a refrigerator at 2°C to 8°C (36°F to 46°F) for no more than 36 hours, or at room temperature to 25°C (77°F) for no more than 4 hours. If refrigerated, allow the infusion solution to equilibrate to room temperature for approximately 30 minutes before administration.

[0306] Exemplary application instructions are as follows:

[0307] 1. Collect recommended infusion materials:

[0308] a. PVC or polyurethane (PU) infusion sets lined with polyvinyl chloride (PVC) or polyethylene (PE).

[0309] b. In-line or attached 0.2-micron polyethersulfone (PES) filter

[0310] 2. Attach the infusion set to the IV bag.

[0311] 3. Infusion set.

[0312] 4. Administer via intravenous infusion over at least 60 minutes via pump or gravity through an intravenous line containing a sterile, in-line or attached 0.2-micron polyethersulfone (PES) filter (see Table 4A or Table 4B).

[0313] 5. The prepared infusion solution should not be administered concurrently with any other drugs. The compatibility of mAb10933 and mAb10987 injections with IV solutions and drugs, except for 0.9% sodium chloride injection, is currently unknown.

[0314] 6. After the infusion is completed, flush with 0.9% sodium chloride injection solution.

[0315] 7. Discard unused products.

[0316] 8. Clinically monitor the patient during administration and observe the patient for at least 1 hour after the infusion is completed.

[0317] Table 4A: Recommended dosage, dilution, and administration for intravenous infusions of 600 mg casciretumab and 600 mg edevimab illustrate

[0318]

[0319]

[0320] a Add 600 mg of cascisizumab and 600 mg of edevimab to the same infusion bag and administer together as a single intravenous infusion.

[0321] b After the infusion is completed, flush with 0.9% sodium chloride injection solution.

[0322] c When administering both cassicimab and edevimab to a patient using a 50mL pre-filled 0.9% sodium chloride infusion bag, the minimum infusion time must be at least 20 minutes to ensure safe use.

[0323] Table 4B: Recommended dosage, dilution and administration of 1,200 mg casciretumab and 1,200 mg edevimab for IV infusion Instructions for use

[0324]

[0325] a 1,200 mg of cascisizumab and 1,200 mg of edevimab were added to the same infusion bag and administered together as a single intravenous infusion.

[0326] b After the infusion is completed, flush with 0.9% sodium chloride injection solution.

[0327] c When administering both cassicimab and edevimab to a patient using a 50mL pre-filled 0.9% sodium chloride infusion bag, the minimum infusion time should be at least 20 minutes to ensure safe use.

[0328] Reagent test kit

[0329] The present invention further provides an article or kit comprising packaging material, a container, and a pharmaceutical agent contained within the container, wherein the pharmaceutical agent comprises at least one anti-SARS-CoV-2 spike glycoprotein antibody, and wherein the packaging material comprises a label or instruction manual showing indications and instructions for use. In one embodiment, the kit may comprise two anti-SARS-CoV-2 spike glycoprotein antibodies, and the two antibodies may be contained in separate containers.

[0330] Example

[0331] The following examples are provided to offer a complete disclosure and description to those skilled in the art regarding how to prepare and use the methods and compositions of the present invention, and are not intended to limit the scope of what the inventors consider to be their invention. Efforts have been made to ensure the accuracy of the figures used (e.g., amounts, temperatures, etc.), but some experimental errors and deviations should be taken into account. Unless otherwise specified, parts are parts by weight, molecular weights are average molecular weights, temperatures are degrees Celsius, and pressures are or close to atmospheric pressure.

[0332] Example 1. Clinical observation of anti-SARS-CoV-2 spike glycoprotein antibodies in hospitalized adult patients with COVID-19. evaluate .

[0333] The clinical study described below is an adaptive, phase 1 / 2 / 3, randomized, double-blind, placebo-controlled primary protocol designed to evaluate the efficacy, safety, and tolerability of mAb10933+mAb10987 in hospitalized adult patients with COVID-19. The safety, tolerability, and efficacy of mAb10989 will also be evaluated in the phase 1 portion of this study to enable further investigation in other clinical settings.

[0334] Research Objectives The following section outlines the primary and secondary objectives for each phase of the study.

[0335] Main objectives :

[0336] Phase 1

[0337] Part A

[0338] • Evaluate the safety and tolerability of mAb10933+mAb10987 compared to placebo.

[0339] • Evaluate the virological efficacy of mAb10933+mAb10987 in reducing SARS-CoV-2 viral shedding compared to placebo.

[0340] Part B

[0341] • Evaluate the safety and tolerability of mAb10989 compared to placebo.

[0342] • Evaluate the virological efficacy of mAb10989 in reducing SARS-CoV-2 viral shedding compared to placebo.

[0343] Phase 2

[0344] • Evaluate the virological efficacy of mAb10933+mAb10987 in reducing SARS-CoV-2 viral shedding compared to placebo.

[0345] • Evaluate the clinical efficacy of mAb10933+mAb10987 in improving clinical status compared to placebo.

[0346] Phase 3

[0347] The primary objective of Phase 3 is to evaluate and confirm the effect of placebo on the comparison between the two drugs.

[0348] Clinical efficacy of mAb10933+mAb10987 in improving clinical status.

[0349] Secondary objectives :

[0350] Phase 1

[0351] Part A

[0352] • Other endpoints of virological efficacy evaluation of mAb10933+mAb10987 compared to placebo.

[0353] • Evaluate the clinical efficacy of mAb10933+mAb10987 in improving clinical outcomes compared to placebo.

[0354] • Characterize the pharmacokinetic (PK) features of mAb10933 and mAb10987 in serum.

[0355] • Assess the immunogenicity of mAb10933 and mAb10987

[0356] Part B

[0357] • Other endpoints of the virological efficacy of mAb10989 compared to placebo.

[0358] • Evaluate the clinical efficacy of mAb10989 in improving clinical outcomes compared to placebo.

[0359] • Compare the results of quantitative reverse transcription polymerase chain reaction (RT-qPCR) obtained using different sample types (nasopharynx, nasal cavity, and saliva).

[0360] • Characterizing the PK properties of mAb10989 in serum

[0361] • Assess the immunogenicity of mAb10989

[0362] Phase 2

[0363] • Other endpoints of virological efficacy evaluation of mAb10933+mAb10987 compared to placebo.

[0364] Other endpoints evaluating the clinical efficacy of mAb10933+mAb10987 compared to placebo.

[0365] • Evaluate the safety and tolerability of mAb10933+mAb10987 compared to placebo.

[0366] • Characterizes the changes in serum mAb10933 and mAb10987 concentrations over time.

[0367] • Assess the immunogenicity of mAb10933 and mAb10987

[0368] Phase 3

[0369] • Evaluate the clinical efficacy of mAb10933+mAb10987 compared to placebo.

[0370] • Evaluate the safety and tolerability of mAb10933+mAb10987 compared to placebo.

[0371] • Characterizes the changes in serum mAb10933 and mAb10987 concentrations over time.

[0372] • Assess the immunogenicity of mAb10933 and mAb10987

[0373] Research Design This study is an adaptive, phase 1 / 2 / 3, randomized, double-blind, placebo-controlled primary protocol to evaluate the efficacy, safety, and tolerability of mAb10933+mAb10987 in hospitalized adult patients with COVID-19. The safety, tolerability, and efficacy of mAb10989 were evaluated in the phase 1 portion of this study to enable further investigation in other clinical settings. Eligible patients hospitalized for ≤72 hours at screening were recruited into one of four cohorts based on disease severity upon randomization. Phase 2 was initiated following approval of the phase 1 sentinel safety group by the Independent Data Monitoring Committee (IDMC), and recruitment was concurrent with phase 1 upon initiation. Once phase 2 was activated, phase 1 recruitment continued until completion, but phase 2 recruitment was not required to complete phase 1 recruitment.

[0374] Study duration Phase 1 of this study lasted 170 days. Phase 2 of this study lasted 58 days. Phase 3 of this study lasted 58 days.

[0375] research group In order to evaluate hospitalization

[0376] The study investigated and analyzed potential differential treatment effects across the spectrum of COVID-19 patients in four cohorts of hospitalized adult patients with COVID-19: Cohort 1A (patients with COVID-19 symptoms but not requiring supplemental oxygen); Cohort 1 (patients requiring low-flow oxygen supplementation); Cohort 2 (patients requiring high-intensity oxygen therapy but not mechanical ventilation); and Cohort 3 (patients requiring mechanical ventilation).

[0377] queue—Eligible patients were recruited into one of four cohorts based on disease severity at the time of randomization: Cohort 1A (patients with COVID-19 symptoms but not requiring supplemental oxygen); Cohort 1 (O2 saturation >93% via low-flow oxygen via nasal cannula, simple mask or other similar device); Cohort 2 (high-intensity oxygen therapy* but not requiring mechanical ventilation—*high-intensity oxygen therapy is defined as the use of a non-rebreathing mask with an oxygen flow rate of at least 10 L / min; the use of a high-flow device with at least 50% FiO2; or the use of non-invasive ventilation to treat hypoxemia); and Cohort 3 (mechanical ventilation).

[0378] Sample size —The Phase 1 portion of this study includes up to 100 patients from Cohort 1: Part A for mAb10933+mAb10987: approximately 20 patients per group, 60 patients in total across the 3 treatment groups; Part B for mAb10989: approximately 20 patients per group, 40 patients in total across the 2 treatment groups. The Phase 2 portion of this study includes approximately 1560 patients: Cohort 1A: approximately 130 patients per group, 390 patients in total across the 3 treatment groups; Cohort 1: approximately 130 patients per group, 390 patients in total across the 3 treatment groups; Cohort 2: approximately 130 patients per group, 390 patients in total across the 3 treatment groups; and Cohort 3: approximately 130 patients per group, 390 patients in total across the 3 treatment groups. The Phase 3 sample size is estimated to be approximately 1350 patients (each of the 3 cohorts has 3 treatment groups, with 150 patients in each group). The final determination of the sample size and patient population for Phase 3 may change and will be determined after a full review of the Phase 2 data.

[0379] Selection criteria Patients must meet the following criteria to be eligible for the study:

[0380] 1. Provide informed consent (signed by the study patient or their legal representative);

[0381] 2. During randomization, male or female adults must be ≥18 years old (or the legal age of majority in the country);

[0382] 3. A positive molecular diagnostic test for SARS-CoV-2 (by a validated SARS-CoV-2 RT-PCR or other molecular diagnostic assay, using an appropriate sample, such as NP, nasal cavity, oropharyngeal [OP], or saliva) was performed ≤72 hours prior to randomization, and there is no alternative explanation for the current clinical symptoms. A history of positive results from tests performed ≤72 hours prior to randomization is acceptable;

[0383] 4. Having symptoms consistent with COVID-19, with onset within ≤10 days prior to randomization; and

[0384] 5. Patients hospitalized for COVID-19 for less than 72 hours and meeting at least one of the following criteria during randomization will be classified into the most severely affected category:

[0385] a. Cohort 1A: Individuals with COVID-19 symptoms but not requiring supplemental oxygen.

[0386] b. Queue 1: Maintaining O2 saturation >93% via nasal cannula, simple mask, or other similar device at low flow rates.

[0387] c. Cohort 2: High-intensity oxygen therapy without mechanical ventilation, where high intensity is defined as receiving supplemental oxygen delivered by one of the following devices:

[0388] - Non-rebreathing mask (SpO2 ≤ 96%, while receiving an oxygen flow rate of at least 10 L / min)

[0389] - High-flow devices with at least 50% FiO2 (e.g., AIRVO) TM or Optiflow TM )

[0390] - Non-invasive ventilation, including continuous positive airway pressure (CPAP), for the treatment of hypoxemia (excluding sleep-disordered breathing alone).

[0391] d. Queue 3: Mechanical ventilation.

[0392] Exclusion criteria Patients meeting any of the following criteria will be excluded from this study:

[0393] 1. Stage 1 only: Patients maintain an O2 saturation level >94% in indoor air;

[0394] 2. In the researchers' view, it is unlikely that the cells will survive for more than 48 hours after screening;

[0395] 3. Receive extracorporeal membrane oxygenation (ECMO);

[0396] 4. New-onset stroke or epilepsy during hospitalization;

[0397] 5. Initiation of renal replacement therapy due to COVID-19;

[0398] 6. Patients with circulatory shock requiring vasopressors at the time of randomization (patients requiring vasopressors for reasons other than sedation-related hypotension or circulatory shock may be eligible for this study);

[0399] 7. Patients who have received convalescent plasma or IVIG within the past 5 months or who are scheduled to receive any indication-specific treatment during the study period;

[0400] 8. Participate in clinical research, including any double-blind study, evaluate the investigational drug within 30 days prior to the screening visit, and the investigational drug has a half-life of less than 5 days (allowing the use of remdesivir, hydroxychloroquine or other treatments for the treatment of COVID-19 (excluding COVID-19 convalescent plasma or IVIG) in the context of local standards of care or open-label studies or compassionate use protocols).

[0401] 9. Any medical examination results, medical history and / or accompanying medications, in the view of the researchers in the study, may confound the study results or pose additional risks to the patient due to participation in the study;

[0402] 10. Has a known allergic or hypersensitivity reaction to any component of the research drug;

[0403] 11. Pregnant women or breastfeeding women; or

[0404] 12. Women of childbearing potential (WOCBP)* or sexually active men who do not wish to continue sexual activity and do not wish to use highly effective contraception at least 6 months before the initial dose of the first treatment, during the study, and after the last dose.

[0405] Highly effective contraceptive methods for women include:

[0406] Stable use of combined hormonal contraceptives (containing estrogen and progestin) (oral, vaginal, transdermal) or progestin-only hormonal contraceptives (oral, injectable, implantable) and suppression of ovulation for the first two or more menstrual cycles prior to screening.

[0407] Intrauterine device (IUD)

[0408] Intrauterine Hormone-Releasing System (IUS)

[0409] Bilateral tubal ligation

[0410] • Partners after vasectomy and / or

[0411] Sexual abstinence ,§

[0412] Male study participants with WOCBP partners were asked to use condoms unless they underwent vasectomy. or sexual abstinence ,§.

[0413] *WOCBP is defined as a woman who remains fertile from menarche until menopause, unless permanently infertile. Postmenopausal status is defined as the absence of menstruation for 12 months without other medical cause. High follicle-stimulating hormone (FSH) levels within the postmenopausal range can be used to confirm postmenopausal status in women who are not using hormonal contraception or hormone replacement therapy. However, in the absence of 12 months of amenorrhea, a single FSH measurement is insufficient to determine the occurrence of postmenopausal status. The above definitions are based on the Clinical Trials Facilitation Group (CTFG) guidelines. Women with documented hysterectomy or tubal ligation do not require pregnancy testing or contraception. Permanent sterilization methods include hysterectomy, bilateral salpingectomy, and bilateral oophorectomy.

[0414] Partners who have undergone vasectomy or research participants who have undergone vasectomy must undergo a medical evaluation to assess the success of the surgery.

[0415] Sexual abstinence is considered an effective method only when defined as avoiding heterosexual intercourse throughout the entire risk period associated with the investigational drug. The reliability of sexual abstinence needs to be assessed regarding the duration of clinical trials and the patient's preferred and usual lifestyle.

[0416] Regular abstinence (calendar, ovulation-based, post-ovulation methods), withdrawal (coitus interruptus), spermicide-only methods, and lactogenic amenorrhea (LAM) are unacceptable methods of contraception. Female condoms and male condoms should not be used together.

[0417] Research on treatment In Phase 1, Part A, patients received a single intravenous (IV) dose of 2.4g (1.2g mAb10933 plus 1.2g mAb10987) of the co-administered mAb10933 + mAb10987 combination therapy, a single IV dose of 8.0g (4.0g mAb10933 plus 4.0g mAb10987) of the co-administered mAb10933 + mAb10987 combination therapy, or a single IV dose of placebo. In Phase 1, Part B, patients received a single IV dose of 1.2g mAb10989 monotherapy or a single IV dose of placebo. In Phase 2, patients received a single IV dose of either the co-administered mAb10933 + mAb10987 combination therapy 2.4g (1.2g mAb10933 plus 1.2g mAb10987), the co-administered mAb10933 + mAb10987 combination therapy 8.0g (4.0g mAb10933 plus 4.0g mAb10987), or a single IV dose of placebo. Phase 3 treatment groups were determined after reviewing the Phase 2 data.

[0418] endFor each period, a primary endpoint, secondary endpoint, and exploratory endpoint are specified, as defined below.

[0419] Primary endpoint

[0420] Phase 1 (Queue 1 only)

[0421] The primary endpoints for period 1 (part A and part B) are:

[0422] • The proportion of patients who experienced a serious adverse event (SAE) during treatment by day 169

[0423] • Proportion of patients experiencing infusion-related reactions (grade ≥2) by day 4

[0424] • The proportion of patients who developed hypersensitivity reactions (grade ≥2) by day 29

[0425] • Time-weighted average change in viral shedding from baseline (day 1) to day 22 (log 10 (Copies / mL), as measured in nasopharyngeal (NP) swab samples by quantitative reverse transcription polymerase chain reaction (RT-qPCR) (the time-weighted average change in viral shedding from baseline (day 1) to day 22 will be calculated for each patient using the trapezoidal rule, as the area under the curve of the change from baseline at each time point divided by the time interval of the observation period).

[0426] Phase 2

[0427] The primary endpoints for each cohort in two phases are:

[0428] Queue 1A and Queue 1

[0429] • Time-weighted average change in viral shedding from baseline (day 1) to day 22 (log 10 The proportion of patients with at least a 1-point improvement in clinical status, measured by RT-qPCR in nasopharyngeal (NP) swabs from day 1 (randomization time) to day 8, using a 7-point ordinal scale.

[0430] Queues 2 and 3

[0431] • Time-weighted average change in viral shedding from baseline (day 1) to day 22 (log 10 (Copies / mL), as measured by RT-qPCR in NP swabs. From day 1 (randomization time) to day 22, the proportion of patients with at least a 1-point improvement in clinical status using a 7-point ordinal scale.

[0432] Phase 3

[0433] The primary endpoints for the three phases in each cohort are:

[0434] Queue 1A and Queue 1

[0435] • The proportion of patients whose clinical status improved by at least 1 point using a 7-point ordinal scale from day 1 (randomization time) to day 8.

[0436] Queues 2 and 3

[0437] • The proportion of patients whose clinical status improved by at least 1 point using a 7-point ordinal scale from day 1 (randomization time) to day 22.

[0438] After reviewing the Phase 2 data, the final Phase 3 patient population (cohort 1A, cohort 1, cohort 2 and / or cohort 3) and primary clinical efficacy endpoint will be determined.

[0439] Secondary endpoint

[0440] Phase 1 (Queue 1 only)

[0441] The secondary endpoint for Phase 1 is:

[0442] • Time-weighted average change in viral shedding from baseline (day 1) to day 22 (log 10 (copy / mL), such as in saliva samples measured by RT-qPCR.

[0443] • Time-weighted average change in viral shedding from baseline (day 1) to day 22 (log 10 (copy / mL), such as that measured by RT-qPCR in nasal samples.

[0444] • Time from negative RT-qPCR in all tested samples to no positive RT-qPCR in any subsequent tested samples (NP swabs, saliva, or nasal swabs)

[0445] • Changes in SARS-CoV-2 virus shedding from baseline to day 29 at each visit, as measured by RT-qPCR in NP swabs.

[0446] • Changes in SARS-CoV-2 virus shedding from baseline to day 29 at each visit, as measured by RT-qPCR in saliva samples.

[0447] • Changes in SARS-CoV-2 virus shedding from baseline to day 29 at each visit, such as those measured by RT-qPCR in nasal swabs.

[0448] • Correlation and consistency of RT-qPCR results over time among different sample types (NP, nasal cavity, and saliva)

[0449] • Time-weighted mean change (log10 copies / mL) of viral shedding from baseline (day 1) to post-baseline study days (e.g., days 5, 7, 15, and 29), from day 1 (randomization time) to day 8, using a 7-point ordinal scale, of patients with at least a 1-point improvement in clinical status.

[0450] • The proportion of patients whose clinical status improved by at least 2 points using a 7-point ordinal scale from day 1 (randomization time) to day 8.

[0451] • The proportion of patients whose clinical status improved by at least 1 point using a 7-point ordinal scale from day 1 (randomization time) to day 29 or at discharge.

[0452] • The proportion of patients who showed at least a 2-point improvement in clinical status using a 7-point ordinal scale from day 1 (randomization time) to day 29 or at discharge.

[0453] • The period after day 29 when supplemental oxygen supply is no longer needed

[0454] • Number of days of supplemental oxygen supply used up to day 29

[0455] • The proportion of patients who started high-intensity oxygen therapy by day 29 or at discharge.

[0456] • Number of days of high-intensity oxygen therapy up to day 29

[0457] • The proportion of patients who started mechanical ventilation by day 29 or at discharge

[0458] • Number of days of mechanical ventilation up to day 29

[0459] • Number of days without a ventilator up to day 29

[0460] • Number of days hospitalized up to day 29

[0461] • The proportion of patients who were readmitted after being discharged at the end of the study

[0462] • Percentage of patients admitted to the Intensive Care Unit (ICU) by day 29

[0463] • Number of days spent in the ICU up to day 29

[0464] • All-cause mortality rate up to day 29

[0465] • All-cause mortality rate at the end of the study

[0466] Overall survival

[0467] • Proportion of patients with treatment-phase SAE by day 29

[0468] • The concentrations of mAb10987, mAb10933, and mAb10989 in serum and their corresponding pharmacokinetic parameters

[0469] • Immunogenicity, as measured by anti-drug antibodies (ADAs) against mAb10933, mAb10987, and mAb10989.

[0470] Phase 2

[0471] The secondary endpoint for Phase 2 is:

[0472] Only queue 1A and queue 1

[0473] • The proportion of patients whose clinical status improved by at least 2 points using a 7-point ordinal scale from day 1 (randomization time) to day 8.

[0474] Only queues 2 and 3

[0475] • The proportion of patients whose clinical status improved by at least 2 points using a 7-point ordinal scale from day 1 (randomization time) to day 22.

[0476] Queues 1A, 1, 2 and 3

[0477] • Time from negative RT-qPCR in NP swabs to no positive RT-qPCR results

[0478] • Changes in viral shedding from baseline to day 29 at each visit, as measured by RT-qPCR in NP swabs.

[0479] • Time-weighted average change (log) of viral shedding from baseline (day 1) to post-baseline study days (e.g., days 5, 7, 15, and 29). 10 (copy / mL)

[0480] • The proportion of patients whose clinical status improved by at least 1 point using a 7-point ordinal scale from day 1 (randomization time) to day 29 or at discharge.

[0481] • The proportion of patients who showed at least a 2-point improvement in clinical status using a 7-point ordinal scale from day 1 (randomization time) to day 29 or at discharge.

[0482] • No further oxygen supplementation is needed until day 29 (only for queues 1, 2, and 3)

[0483] • Number of days of supplemental oxygen supply used up to day 29

[0484] • The proportion of patients who started high-intensity oxygen therapy by day 29

[0485] • Number of days of high-intensity oxygen therapy up to day 29

[0486] • The proportion of patients who started mechanical ventilation by day 29 or at discharge

[0487] • Number of days of mechanical ventilation up to day 29

[0488] • Number of days without a ventilator up to day 29

[0489] • Number of days hospitalized up to day 29

[0490] • The proportion of patients who were readmitted after being discharged at the end of the study

[0491] • Proportion of patients admitted to the ICU by day 29

[0492] • Number of days spent in the ICU up to day 29

[0493] • All-cause mortality rate up to day 29

[0494] • All-cause mortality rate at the end of the study

[0495] Overall survival

[0496] • Proportion of patients with treatment-phase SAE by day 29

[0497] • The proportion of patients with treatment-stage SAE by day 57

[0498] • Proportion of patients experiencing infusion-related reactions (grade ≥2) by day 4

[0499] • The proportion of patients who developed hypersensitivity reactions (grade ≥2) by day 29

[0500] • Changes in serum mAb10933 and mAb10987 concentrations over time

[0501] • Immunogenicity, such as that measured by ADA against mAb10933 and mAb10987. Phase 3

[0502] After reviewing the Phase 2 data, the final Phase 3 patient population (cohort 1A, cohort 1, cohort 2 and / or cohort 3) and secondary clinical efficacy endpoints were determined.

[0503] Other possible secondary endpoints for Phase 3 include:

[0504] • The proportion of patients with treatment-stage SAE by day 57

[0505] • Proportion of patients experiencing infusion-related reactions (grade ≥2) by day 4

[0506] • The proportion of patients who developed hypersensitivity reactions (grade ≥2) by day 29

[0507] • Changes in serum mAb10933 and mAb10987 concentrations over time

[0508] • Immunogenicity, such as that measured by ADA against mAb10933 and mAb10987.

[0509] Exploratory endpoint

[0510] Exploratory endpoints include:

[0511] • The proportion of patients who failed treatment by day 29 with mutations in the gene encoding the SARS-CoV-2S protein.

[0512] • Changes in neutrophil-lymphocyte ratio (NLR) and percentage from each visit to day 29

[0513] • Changes and percentage changes in D-dimer levels by day 29 at each visit

[0514] • Changes and percentage changes in ferritin levels up to day 29 at each visit

[0515] • Changes and percentage changes of C-reactive protein (CRP) by day 29 at each visit

[0516] • Changes and percentage changes in lactate dehydrogenase (LDH) levels by day 29 at each visit

[0517] Procedures and assessments Efficacy—Nasopharyngeal (all phases), saliva (phase 1 only), and / or nasal swabs (phase 1 only) were used for SARS-CoV-2 RT-PCR, as well as clinical and oxygenation status; Safety—Serious adverse events and adverse events of particular concern were documented. Secretions were collected from patients using nasal swabs, saliva samples, and (in phase 1) nasopharyngeal samples to determine the presence or absence of SARS-CoV-2 virus and to measure viral shedding. Samples were used for RT-qPCR analysis. Samples may also be used for exploratory viral RNA sequencing (nasopharyngeal, nasal swabs, saliva) and / or viral culture (nasopharyngeal, nasal swabs).

[0518] Statistical Plan :

[0519] Phase 1 —The sample size is the sum of 60 patients in Part A of Phase 1 and 40 patients in Part B. The sample size allows for a preliminary estimate of the incidence of SAE, AESI, and grade 3 or 4 TEAE in the treatment group relative to placebo.

[0520] The primary efficacy endpoint for Phase 1 was the time-weighted average change (log) of viral shedding in NP swab samples from baseline (day 1) to day 22. 10(Copies / mL). Assume a standard deviation of 2.1 log [copies / mL]. 10 The sample size was measured at copies / mL. A two-sample t-test with a two-sided significance level of α = 0.05 was used. In Phase 1, a sample size of 20 patients in each group should have at least 80% influence in detecting a 1.91 log [value missing] between the treatment and placebo groups. 10 Differences in copies / mL.

[0521] Phase 2 —The sample size for Phase 2 is based on the time-weighted average change (log) of viral shedding from NP swab samples from baseline (day 1) to day 22. 10 (Copies / mL). Assuming an exit rate of approximately 23% (including missing data at baseline) and 2.1 log... 10 The standard deviation of copies / mL was determined using a two-sample t-test with two-tailed significance of α = 0.05. A sample size of 130 patients per treatment group (i.e., 100 patients per group with available data) in each of the three cohorts should have an 80% influence in detecting a 0.84 log difference between each treatment group and the placebo group in each cohort. 10 Difference in copies / mL. If the standard deviation is assumed to be 3.8 log... 10 At copies / mL, the detectable difference at 80% influence is 1.51 log [copy / mL]. 10 Copy / mL.

[0522] For the clinical endpoint of the proportion of patients whose clinical status improved by at least 1 point from baseline to day 22, the minimum detectable difference (MDD) between the treatment and placebo groups, based on a chi-square test of equal proportions, would be as follows, with a sample size of 100 patients per group (assuming a dropout rate of approximately 23%, or 130 patients per group):

[0523] • In cohorts 1A and 1, the hypothetical response rate in the placebo group was 51%, and the MDD would be 13.7% (i.e., 64.7% for anti-SARS-CoV-2S protein mAb and 51% for placebo). The hypothetical response rate in the placebo group was similar to the observed response rate with remdesivir.

[0524] In cohorts 2 and 3, the hypothetical response rate in the placebo group is 57.1%, and the MDD will be 13.3% (i.e., 70.4% for anti-SARS-CoV-2S protein mAb and 57.1% for placebo). The hypothetical response rate in the placebo group is similar to that observed in the thalidomide COVID-19 Phase 2 / 3 study (6R88-COV-2040), in which the progression population was similar to that in cohorts 2 and 3.

[0525] Phase 3—The study will continue to recruit additional patients seamlessly into the Phase 3 portion of the study until an adaptive decision is made on the primary endpoint and final sample size for Phase 3 based on a complete analysis of the Phase 2 data. The initial sample size estimate for the Phase 3 portion of the study is a total of 1350 patients (150 patients in each of the three treatment groups across three cohorts). For example, for cohort 3, a sample size of 450 patients (150 patients in each group) will be assessed using a chi-square test, providing 90% influence, to detect a 15.9% difference in the proportion of patients surviving and weaning off mechanical ventilation at day 22, assuming a rate of 68.2% in the placebo group.

[0526] result —A Phase 1 / 2 / 3 clinical trial of the antibody mixtures cassicimarab and edevimab (mAb10933 and mAb10987, respectively) in hospitalized COVID-19 patients requiring low-flow oxygen (see [link to relevant documentation]). Figure 19 The analysis focused on patients who had not yet developed an autoimmune response to SARS-CoV-2 (i.e., no antibodies at baseline: seronegative), as evidence (see Example 2) indicated these patients were at higher risk. Additionally, among subjects treated with placebo, those who developed an immune response at baseline (seropositive patients) had significantly lower viral levels at baseline compared to those who did not develop an immune response at baseline (seronegative patients), and reached viral loads below the lower limit of quantitation (“LLQ”) more quickly, even without treatment. See also Figure 16 Additionally, among hospitalized COVID-19 patients receiving low-flow supplemental oxygen, seronegative patients had a lower cumulative incidence of death or mechanical ventilation compared to seronegative patients. See also Figure 17 In cohort 1, patients who were seronegative at baseline or had high viral loads at baseline had worse clinical outcomes. See also Figure 18 The primary clinical objective of this preliminary analysis was to determine whether these patients had sufficient efficacy to justify continuing the trial (i.e., a nullification analysis). The results passed the nullification analysis (p<0.3, one-sided) because seronegative patients treated with the antibody mixture had a lower risk of death or requiring mechanical ventilation (hazard ratio (HR): 0.78; 80% CI: 0.51–1.2). Based on a post-hoc analysis, this benefit was driven by results starting one week after treatment, at which point the risk of death or requiring mechanical ventilation was reduced by approximately half in antibody mixture treatment.

[0527] The incidence of seronegativity was analyzed in cohort 1 in the full analysis set (FAS; randomized and treated patients) and the modified full analysis set (mFAS; patients who tested positive for SARS-CoV-2 at baseline via nasopharyngeal qualitative testing), and the incidence of seronegativity was similar in the FAS and mFAS groups. See also Figure 20Serologically negative patients (n=217) had significantly higher viral loads than those who had developed their own SARS-CoV-2 antibodies (serologically positive) at the time of randomization. See also Figure 16 As hypothesized, in patients who did not develop an autoimmune response (serologically negative at baseline), the antibody mixture had a stronger antiviral effect than placebo, patients treated with the antibody mixture experienced faster viral reduction compared to placebo, and the mixture reduced viral load faster than placebo across all baseline viral load thresholds. See also Figures 21 to 24 In seronegative patients, the antibody mixture reduced the time-weighted mean daily viral load by -0.54 log10 copies / mL by day 7 and by -0.63 log10 copies / mL by day 11 (nominal p = 0.002 for the combination dose). On day 5, the relative reduction was -1.1 log10 copies compared to placebo (nominal p = 0.002 for the combination dose). In seronegative patients (n = 270), the clinical and virological benefits of the antibody mixture were limited (clinical endpoint HR: 0.98; time-weighted mean viral load reduction of -0.20 log10 copies / mL on day 7 for the combination dose). Treatment with the mixture resulted in similar viral load reductions in both inpatient and outpatient settings, and the most significant difference was observed in seronegative patients between those treated with the mixture and those receiving placebo, consistent with data from Example 2. Figures 25 to 28 ).

[0528] Clinical and virological analyses included data from hospitalized patients receiving low-flow oxygen (defined as maintaining oxygen saturation >93% via nasal cannula, simple face mask, or similar device), comprising 217 seronegative patients and 270 seronegative patients at entry into the trial; although seronegative patients comprised less than half of the trial population, they accounted for approximately two-thirds of deaths in the absence of antibody mixture treatment, based on placebo ratios. Patients were randomized to receive either the antibody mixture (8000 mg high dose or 2400 mg low dose) or placebo, along with standard of care; 67% received remdesivir and 74% received systemic corticosteroids. Similar clinical and virological efficacy was observed for both high- and low-dose antibody mixtures.

[0529] Both antibody mixtures were well tolerated at both doses. The incidence of serious adverse events was 21% in the high-dose group, 20% in the low-dose group, and 24% in the placebo group across the entire trial population. Infusion reactions were more common in the high-dose antibody mixtures (2.7% high-dose, 0.9% low-dose, 1.4% placebo), and two cases of discontinuation due to infusion-related reactions occurred, both in the high-dose group.

[0530] Example 2. Clinical evaluation of anti-SARS-CoV-2 spike glycoprotein antibodies in non-bedridden patients with COVID-19 price .

[0531] The following clinical study is an adaptive, phase 1 / 2 / 3, randomized, double-blind, placebo-controlled primary protocol designed to evaluate the efficacy, safety, and tolerability of mAb10933+mAb10987 combination therapy (collectively referred to as REGN-COV2 or REGEN-COV) or mAb10989 monotherapy in adult outpatients (i.e., non-bedridden patients) with COVID-19 or asymptomatic SARS-CoV-2 infection.

[0532] Research Objectives The following section outlines the primary and secondary objectives for each phase of the study.

[0533] Exemplary uses The exemplary uses that can be authorized based on the results (including interim results) of this embodiment are as follows:

[0534] This exemplary use applies to intravenous infusion of REGEN-COV, with mAb10933 and mAb10987 administered together. REGEN-COV should be administered as soon as possible after a positive SARS-CoV-2 virus test, and within 7 days of symptom onset in adult and pediatric patients aged 12 years and older and weighing at least 40 kg, who are at high risk of developing severe COVID-19 and / or hospitalization. COVID-19 illness can range from very mild (including some unreported symptoms) to severe, including fatal illness. While information to date suggests that most COVID-19 illness is mild, severe illness can occur and may worsen some of other medical conditions. People of all ages with serious, long-term (chronic) medical conditions such as heart disease, lung disease, and diabetes, as well as other conditions including obesity, appear to have a higher risk of hospitalization due to COVID-19. Regardless of other medical conditions, the risk of hospitalization due to COVID-19 increases with age.

[0535] This exemplary authorization is for the use of REGEN-COV to treat adult and pediatric patients with mild to moderate coronavirus disease 2019 (COVID-19) who have tested positive for direct SARS-CoV-2 virus, are 12 years of age or older, weigh at least 40 kg, and are at high risk of developing severe COVID-19 and / or hospitalization.

[0536] The following medical conditions or other factors may cause adults and

[0537] Children (aged 12 to 17 years and weighing at least 40 kg) are in a developmental stage.

[0538] In the higher risk of severe COVID-19:

[0539] Older age (e.g., age ≥ 65 years)

[0540] Obese or overweight (e.g., BMI > 25 kg / m²) 2 Adults, or if aged 12 to 17 years, with a BMI ≥ 85% based on the CDC growth chart (https: / / www.cdc.gov / growthcharts / clinical_charts.htm).

[0541] pregnancy

[0542] Chronic kidney disease

[0543] diabetes

[0544] Immunosuppressive diseases or immunosuppressive therapy

[0545] Cardiovascular disease (including congenital heart disease) or hypertension

[0546] Chronic lung diseases (such as chronic obstructive pulmonary disease, asthma [moderate to severe], interstitial lung disease, cystic fibrosis, and pulmonary hypertension)

[0547] sickle cell disease

[0548] Neurodevelopmental disorders (e.g., cerebral palsy) or other symptoms that lead to medical complications (e.g., genetic or metabolic syndromes and severe congenital abnormalities).

[0549] ο Having medically relevant technical dependence (e.g., tracheostomy, gastrostomy, or positive pressure ventilation (unrelated to COVID-19))

[0550] Other medical conditions or factors (e.g., race or ethnicity) may also put an individual patient at high risk of progressing to severe COVID-19, and the REGEN-COV authorization under the EUA is not limited to the conditions mentioned above. For more information on medical conditions and factors associated with an increased risk of progressing to severe COVID-19, see the CDC website: www.cdc.gov / coronavirus / 2019-ncov / need-extra-precautions / people-with-medical-conditions.html. Healthcare providers should consider the benefit-risk profile of individual patients.

[0551] Restrictions on the use of the license:

[0552] • In this exemplary use, REGEN-COV should not be used in the following patients:

[0553] - Hospitalized due to COVID-19, or

[0554] -Requires oxygen therapy due to COVID-19, or

[0555] - COVID-19 in patients receiving long-term oxygen therapy due to underlying non-COVID-19 related comorbidities requires an increase in baseline oxygen flow rate.

[0556] However, alternative authorized uses are considered for REGEN-COV in patients who are hospitalized for COVID-19; and / or require oxygen therapy for COVID-19; and / or have COVID-19 who are receiving long-term oxygen therapy due to underlying non-COVID-19 related comorbidities and require an increase in baseline oxygen flow rate.

[0557] Main objectives :

[0558] Phase 1

[0559] Part A

[0560] • Evaluate the safety and tolerability of mAb10933+mAb10987 compared to placebo.

[0561] • Evaluate the virological efficacy of mAb10933+mAb10987 in reducing SARS-CoV-2 virus shedding compared to placebo.

[0562] Part B

[0563] • Evaluate the safety and tolerability of mAb10989 compared to placebo.

[0564] • Evaluate the virological efficacy of mAb10989 in reducing SARS-CoV-2 virus shedding compared to placebo.

[0565] Phase 2

[0566] To evaluate the virological efficacy of mAb10933+mAb10987 and mAb10989 in reducing SARS-CoV-2 viral shedding compared to placebo.

[0567] Phase 3

[0568] To evaluate the clinical efficacy of mAb10933+mAb10987 and mAb10989 compared to placebo.

[0569] Secondary objectives:

[0570] Phase 1

[0571] Part A

[0572] • Other endpoints of virological efficacy evaluation of mAb10933+mAb10987 compared to placebo.

[0573] • Evaluate the clinical efficacy of mAb10933+mAb10987 compared to placebo.

[0574] • Compare the results of quantitative reverse transcription polymerase chain reaction (RT-qPCR) obtained using different sample types (nasopharynx [NP], nasal cavity, and saliva).

[0575] • Characterize the pharmacokinetic (PK) features of mAb10933 and mAb10987 in serum.

[0576] • Assess the immunogenicity of mAb10933 and mAb10987

[0577] Part B

[0578] • Other endpoints of virological efficacy evaluation of mAb10933+mAb10987 compared to placebo.

[0579] • Evaluate the clinical efficacy of mAb10989 compared to placebo.

[0580] • Compare RT-qPCR results obtained using different sample types (NP, nasal cavity, and saliva).

[0581] • Characterizing the PK properties of mAb10989 in serum

[0582] • Assess the immunogenicity of mAb10989

[0583] Phase 2

[0584] • Other endpoints of virological efficacy evaluation of mAb10933+mAb10987 compared to placebo.

[0585] • Evaluate the clinical efficacy of mAb10933+mAb10987 and mAb10989 compared to placebo.

[0586] • Evaluate the safety and tolerability of mAb10933 + mAb10987 and mAb10989 compared to placebo.

[0587] • Characterize the concentrations of mAb10933, mAb10987, and mAb10989 in serum.

[0588] • Assess the immunogenicity of mAb10933, mAb10987, and mAb10989

[0589] Phase 3

[0590] • Evaluate the virological efficacy of mAb10933+mAb10987 and mAb10989 in reducing SARS-CoV-2 virus shedding compared to placebo.

[0591] • Evaluate the safety and tolerability of mAb10933 + mAb10987 and mAb10989 compared to placebo.

[0592] • Characterize the concentrations of mAb10933, mAb10987, and mAb10989 in serum.

[0593] • Assess the immunogenicity of mAb10933, mAb10987, and mAb10989

[0594] Research Design This is an adaptive, phase 1 / 2 / 3, randomized, double-blind, placebo-controlled primary protocol designed to evaluate the efficacy, safety, and tolerability of mAb10933+mAb10987 combination therapy and mAb10989 monotherapy in adult outpatients (i.e., non-bedridden patients) with COVID-19 or asymptomatic SARS-CoV-2 infection. To be eligible, adult patients must have laboratory-confirmed SARS-CoV-2 and COVID-19 symptoms but must not have been previously hospitalized or be currently not hospitalized. In phase 1, only patients with COVID-19 will be recruited. In phase 2, symptomatic and asymptomatic patients will be recruited into separate cohorts.

[0595] Phase 1

[0596] In Part A of Phase 1, randomization will be limited to low-dose mAb10933+mAb10987, high-dose mAb10933+mAb10987, and placebo. In Part B, randomization will be limited to mAb10989 and placebo. On day 1, eligible patients from Part A will be randomized to a single intravenous (IV) administration of mAb10933+mAb10987 (low-dose), mAb10933+mAb10987 (high-dose), mAb10989, or placebo.

[0597] Patients were then isolated for the first 48 hours after administration, during which time they were closely monitored for serious adverse events (SAEs) and adverse events of particular concern (AESIs). On day 3, patients were discharged home after completing their assessment for the day, if medically appropriate. All patients were discharged home after completing their assessment on day 7, if medically appropriate. Throughout the study, safety information (SAEs and AESIs) and any relevant information from COVID-19-related medical care visits were collected. Nasopharyngeal (NP) swabs, nasal swabs, and saliva samples were collected to assess viral shedding. The study ended on day 29, when patients underwent a final assessment in person, including NP swab, nasal swab, and / or saliva sample collection (if feasible), and blood draws for PK, anti-drug antibody (ADA), and exploratory analyses.

[0598] Phase 2

[0599] On day 1, eligible patients were randomized 1:1:1:1 to a single dose of mAb10933+mAb10987 (low dose), mAb10933+mAb10987 (high dose), mAb10989, or placebo. Patients were observed for 2 hours after infusion of the study drug and were discharged home if no SAE or AES was observed. Nasopharyngeal swabs were collected every other day for the first 2 weeks, then twice weekly thereafter. Blood samples were collected periodically. Treatment-period SAEs, AES, and medical care information related to COVID-19 were recorded throughout the study. On day 29, patients underwent final evaluation, including nasopharyngeal swab collection and blood draw for PK, ADA, and exploratory analyses.

[0600] Study duration The duration of this study was 30 days per patient.

[0601] research group This study recruited adult non-hospitalized patients who tested positive for SARS-CoV-2.

[0602] Sample size—Phase 1, recruitment up to 100 patients randomized. Phase 2, recruitment up to approximately 1300 patients randomized. Phase 3 is estimated to require 704 patients (176 patients per group).

[0603] Inclusion criteria: Patients must meet the following criteria to be eligible for inclusion in the study:

[0604] 1. During randomization, are males or females ≥18 years of age (or the legal age of majority in the country);

[0605] 2. A positive molecular diagnostic test for SARS-CoV-2 was performed ≤72 hours prior to randomization (by a validated SARS-CoV-2 RT-PCR or other molecular diagnostic assay, using an appropriate sample such as NP, nasal cavity, oropharyngeal [OP], or saliva). A history of positive results from tests performed ≤72 hours prior to randomization is acceptable;

[0606] 3. Meets one of the following two criteria:

[0607] a. Symptomatic cohort (all periods): Individuals with investigator-defined COVID-19 symptoms, with onset ≤7 days prior to randomization.

[0608] or

[0609] b. Asymptomatic cohort (Phase 2): Meets all of the following criteria:

[0610] • No symptoms consistent with COVID-19 (as determined by the investigator), occurring at any time within 2 months prior to randomization.

[0611] • Samples collected >7 days prior to randomization tested negative for SARS-CoV-2.

[0612] • No known contact (of any duration) with an individual who tested positive for COVID-19 or SARS-CoV-2 within >14 days prior to randomization.

[0613] 4. Maintain indoor air O2 saturation ≥ 93%;

[0614] 5. Willing and able to provide informed consent signed by the research patient or their legal representative; and

[0615] 6. Willing and able to comply with research procedures after discharge, including providing samples for virus shedding detection.

[0616] Exclusion criteria Patients meeting any of the following criteria will be excluded from this study:

[0617] 1. Patients who were already hospitalized for COVID-19 before randomization, or who were hospitalized at the time of randomization (inpatients);

[0618] 2. Within 3 months prior to the screening visit or within less than 5 half-lives of the investigational product (whichever is longer), have participated in or are currently participating in clinical studies evaluating COVID-19 convalescent plasma, anti-SARS-CoV-2 monoclonal antibodies, or intravenous immunoglobulin (IVIG).

[0619] 3. Within the past 30 days prior to the screening visit or within less than 5 half-lives of the investigational drug (whichever is longer), the patient has previously, is currently, or plans to use COVID-19 convalescent plasma, anti-SARS-CoV-2 mAb, intravenous immunoglobulin (IVIG) (for any indication), systemic corticosteroids (for any indication), or any emergency use authorization (EUA) approved treatment.

[0620] 4. Has a known allergic or hypersensitivity reaction to the research drug component;

[0621] 5. Those who have been discharged from the hospital or plan to be discharged to the quarantine center;

[0622] 6. Pregnant women or breastfeeding women; or

[0623] 7. Women of childbearing potential (WOCBP)* or sexually active men who do not wish to continue sexual activity and do not wish to use highly effective contraception at least 6 months before the initial dose of the first treatment, during the study, and after the last dose.

[0624] Hypersensitivity signs and symptoms, including infusion-related reactions, may include: fever, chills, nausea, headache, bronchospasm, hypotension, angioedema, throat irritation, rash (including urticaria), itching, myalgia, and dizziness.

[0625] Highly effective contraceptive methods for women include:

[0626] • Stable use of combined (estrogen and progestin) hormonal contraceptives (oral, vaginal, transdermal) or progestin-only hormonal contraceptives (oral, injectable, implantable) and suppression of ovulation for the first two or more menstrual cycles prior to screening.

[0627] Intrauterine device (IUD),

[0628] • Intrauterine hormone-releasing system (IUS)

[0629] Bilateral tubal ligation,

[0630] • Partners after vasectomy and / or

[0631] Sexual abstinence

[0632] Male study participants with WOCBP partners were asked to use condoms unless they underwent vasectomy. or sexual abstinence

[0633] *WOCBP is defined as a woman who remains fertile from menarche until menopause, unless permanently infertile. Postmenopausal status is defined as the absence of menstruation for 12 months without other medical cause. High follicle-stimulating hormone (FSH) levels within the postmenopausal range can be used to confirm postmenopausal status in women who are not using hormonal contraception or hormone replacement therapy. However, in the absence of 12 months of amenorrhea, a single FSH measurement is insufficient to determine the occurrence of postmenopausal status. The above definitions are based on the Clinical Trials Facilitation Group (CTFG) guidelines. Women with documented hysterectomy or tubal ligation do not require pregnancy testing or contraception.

[0634] Permanent sterilization methods include hysterectomy, bilateral salpingectomy, and bilateral oophorectomy.

[0635] Partners who have undergone vasectomy or research participants who have undergone vasectomy must undergo a medical evaluation to assess the success of the surgery.

[0636] Sexual abstinence is considered an effective method only when defined as avoiding heterosexual intercourse throughout the entire risk period associated with the investigational drug. The reliability of sexual abstinence needs to be assessed regarding the duration of clinical trials and the patient's preferred and usual lifestyle.

[0637] Research on treatment The following treatments are available: 1.4g (1.2g each of mAb10933 and mAb10987) IV single dose of combination therapy; 8.0g (4.0g each of mAb10933 and mAb10987) IV single dose of combination therapy; mAb10989 monotherapy, 1.2g IV single dose; or placebo IV single dose.

[0638] end For each period, a primary endpoint, secondary endpoint, and exploratory endpoint are specified, as defined below.

[0639] Primary endpoint

[0640] Phase 1

[0641] The primary endpoint for Phase 1 is:

[0642] Part A and B

[0643] • The proportion of patients who experienced a serious adverse event (SAE) during treatment by day 29

[0644] • Proportion of patients experiencing infusion-related reactions (grade ≥2) by day 4

[0645] • The proportion of patients who developed hypersensitivity reactions (grade ≥2) by day 29

[0646] • Time-weighted average change in viral shedding from baseline (day 1) to day 22 (log 10 (Copies / mL), as measured in nasopharyngeal (NP) swab samples by quantitative reverse transcription quantitative polymerase chain reaction (RT-qPCR).

[0647] Phase 2

[0648] The primary endpoint of Phase 2 was the time-weighted average change in viral shedding from baseline (day 1) to day 22 (log 10 (Copies / mL), as measured by RT-qPCR in NP swab samples.

[0649] Phase 3

[0650] The primary endpoint of Phase 3 was the proportion of patients who had ≥1 COVID-19 healthcare visit by day 29.

[0651] Secondary endpoint

[0652] Phase 1

[0653] virology

[0654] • Time-weighted average change in viral shedding from baseline (day 1) to day 22 (log 10 (copy / mL), such as the time-weighted average change in viral shedding from baseline (day 1) to day 22 as measured by RT-qPCR in saliva samples (log 10 (Copies / mL), as measured by RT-qPCR in nasal swab samples. The time from RT-qPCR negative in all tested samples and no subsequent RT-qPCR becoming positive in any tested sample (NP swab, saliva, or nasal swab).

[0655] • Changes in SARS-CoV-2 virus shedding from baseline to day 29 at each visit, as measured by RT-qPCR in NP swabs.

[0656] • Changes in SARS-CoV-2 virus shedding from baseline to day 29 at each visit, as measured by RT-qPCR in saliva samples.

[0657] • Changes in SARS-CoV-2 virus shedding from baseline to day 29 at each visit, such as those measured by RT-qPCR in nasal swabs.

[0658] • Correlation and consistency of RT-qPCR results among different sample types (NP, nasal cavity, and saliva)

[0659] • Time-weighted average change (log) of viral shedding from baseline (day 1) to post-baseline study days (e.g., days 5, 7, 15, and 29). 10 (copy / mL)

[0660] clinical

[0661] • The proportion of patients who had ≥1 COVID-19 related medical care visit by day 29; a COVID-19 related medical care visit will be defined as a visit primarily due to COVID-19 hospitalization or primarily due to COVID-19 outpatient visits (including ER, UCC, physician office, or telemedicine visits).

[0662] • Proportion of patients who had ≥2 COVID-19 related medical care visits by day 29

[0663] • Total number of COVID-19 related medical care visits by day 29

[0664] • Percentage of patients admitted to hospitals due to COVID-19 as of day 29

[0665] • Percentage of patients who had ≥1 outpatient or telemedicine visit due to COVID-19 by day 29 PK / ADA

[0666] • The concentrations of mAb10933, mAb10987, and mAb10989 in serum and their corresponding pharmacokinetic parameters

[0667] • Immunogenicity as measured by anti-drug antibodies (ADA) against mAb10933, mAb10987, and mAb10989

[0668] Phase 2

[0669] The secondary endpoint for Phase 2 is:

[0670] virology

[0671] • Time from negative RT-qPCR in NP swabs to no positive RT-qPCR results

[0672] • Changes in viral shedding from baseline to day 29 at each visit, as measured by RT-qPCR in NP swabs.

[0673] • Time-weighted average change (log) of viral shedding from baseline (day 1) to post-baseline study days (e.g., days 5, 7, 15, and 29). 10 (copy / mL)

[0674] clinical

[0675] • Proportion of patients who had ≥1 COVID-19 related medical care visit by day 29

[0676] • Proportion of patients who had ≥2 COVID-19 related medical care visits by day 29

[0677] • Total number of COVID-19 related medical care visits by day 29

[0678] • Percentage of patients admitted to hospitals due to COVID-19 as of day 29

[0679] • Percentage of patients admitted to ICU due to COVID-19 as of day 29

[0680] • Percentage of patients who had ≥1 outpatient or telemedicine visit due to COVID-19 by day 29

[0681] • Percentage of patients requiring mechanical ventilation due to COVID-19 as of day 29

[0682] ·Number of days hospitalized due to COVID-19

[0683] The proportion of patients who died from all causes by day 29

[0684] • Proportion of patients with treatment-phase SAE by day 29

[0685] • Proportion of patients experiencing infusion-related reactions (grade ≥2) by day 4

[0686] • The proportion of patients who developed hypersensitivity reactions (grade ≥2) by day 29

[0687] • The time of first onset of any COVID-19 symptoms

[0688] Duration consistent with COVID-19 symptoms

[0689] PK / ADA

[0690] • Concentrations of mAb10933, mAb10987, and mAb10989 in serum

[0691] • Immunogenicity as measured by anti-drug antibodies (ADA) against mAb10933, mAb10987, and mAb10989

[0692] Phase 3

[0693] The secondary endpoint for Phase 3 is:

[0694] virology

[0695] • Time-weighted average change in viral shedding from baseline (day 1) to day 22 (log 10 (copy / mL), as measured by RT-qPCR in NP swabs.

[0696] • Time from negative RT-qPCR in NP swabs to no positive RT-qPCR results

[0697] • Changes in SARS-CoV-2 virus shedding from baseline to day 29 at each visit, as measured by RT-qPCR in NP swabs.

[0698] • Time-weighted average change (log) of viral shedding from baseline (day 1) to post-baseline study days (e.g., days 5, 7, 15, and 29). 10 (copy / mL)

[0699] clinical

[0700] • Proportion of patients who had ≥2 COVID-19 related medical care visits by day 29

[0701] • Total number of COVID-19 related medical care visits by day 29

[0702] • Percentage of patients who had ≥1 outpatient or telemedicine visit due to COVID-19 as of day 29 • Percentage of patients admitted to hospital due to COVID-19 as of day 29

[0703] • Percentage of patients admitted to ICU due to COVID-19 as of day 29

[0704] • Percentage of patients requiring mechanical ventilation due to COVID-19 as of day 29

[0705] ·Number of days hospitalized due to COVID-19

[0706] The proportion of patients who died from all causes by day 29

[0707] • Proportion of patients with treatment-phase SAE by day 29

[0708] • Proportion of patients experiencing infusion-related reactions (grade ≥2) by day 4

[0709] • The proportion of patients who developed hypersensitivity reactions (grade ≥2) by day 29

[0710] PK / ADA

[0711] • Concentrations of mAb10933, mAb10987, and mAb10989 in serum

[0712] • Immunogenicity as measured by anti-drug antibodies against mAb10933, mAb10987, and mAb10989

[0713] Exploratory endpoint

[0714] The exploratory endpoints for Phase 1 and Phase 2 are:

[0715] • The proportion of patients who failed treatment by day 29 with mutations in the gene encoding the SARS-CoV-2S protein.

[0716] • Changes in neutrophil-lymphocyte ratio (NLR) and percentage from each visit to day 29

[0717] • Changes and percentage changes in D-dimer levels by day 29 at each visit

[0718] • Changes and percentage changes in ferritin levels up to day 29 at each visit

[0719] • Changes and percentage changes of C-reactive protein (CRP) by day 29 at each visit

[0720] • Changes and percentage changes in lactate dehydrogenase (LDH) levels by day 29 at each visit

[0721] • Changes in SE-C19 item scores over time

[0722] • Changes in PGIS scores over time

[0723] • PGIC score on day 29

[0724] • Percentage of patients admitted to ICU due to COVID-19 as of day 29 (Stage 1 only)

[0725] • Percentage of patients requiring mechanical ventilation due to COVID-19 as of day 29

[0726] Procedures and assessmentsEfficacy—Details of nasopharyngeal swabs (all phases), nasal swabs (Phase 1 only), and / or saliva samples (Phase 1 only) for SARS-CoV-2 RT-qPCR, and medical care COVID-19 visits; Safety—Documentation of serious and particular-concern adverse events, laboratory blood collection, and vital signs. Nasal swabs and saliva samples were used to collect secretions from patients to determine the presence or absence of SARS-CoV-2 virus and to measure viral shedding.

[0727] Statistical Plan :

[0728] Main therapeutic effects analysis The primary efficacy variable for both Phase 1 and Phase 2 was the time-weighted change in viral shedding from baseline (day 1) to day 22, as measured by RT-qPCR in NP swab samples. Estimates of the primary hypothesis were the mean differences in the primary efficacy variable in the FAS between each anti-SARS-CoV-2 mAb treatment group and placebo. Based on the observed data, the primary efficacy variable was calculated using the trapezoidal rule and analyzed using an analysis of covariance (ANCOVA) model with the treatment group and random stratification variable as fixed effects and baseline viral shedding as a covariate. For Phase 2, analyses were performed separately for each cohort (symptomatic and asymptomatic) and for combinations of both cohorts. Least square mean estimates of the time-weighted mean change in viral shedding from baseline for each treatment group, and the differences between each anti-spike mAb treatment group and placebo (in Phase 2, for each cohort and for combinations of both cohorts), are shown along with corresponding p-values, standard errors, and associated 95% confidence intervals. The primary efficacy variable in Phase 3 was the proportion of patients receiving medical care visits due to worsening COVID-19 symptoms and signs, and was compared between groups using a two-sided stratified Cochran-Mantel-Haenszel test at the 0.05 level. P-values ​​and 95% confidence intervals for treatment differences are shown below.

[0729] Security Analysis —The treatment team listed and summarized safety data, including serious adverse events and adverse events of particular concern, vital signs, and laboratory tests.

[0730] result —The aforementioned seamless Phase 1 / 2 / 3 trials demonstrated that, when treated with the combination of mAb10933 and mAb10987 (REGEN-COV-2), non-hospitalized patients with COVID-19 experienced a significant reduction in SARS-CoV-2 viral load and time to symptom relief. REGEN-COV also significantly reduced COVID-19-related medical care visits. This randomized, double-blind trial measured the effects of adding REGEN-COV to standard of care and compared it to adding placebo to standard of care.

[0731] The final analysis of the Phase 1 / 2 portion included 799 patients: 275 (Group 1) and 524 (Group 2). Patients were randomized (1:1:1) to a placebo group, a 2.4g mAb10933+mAb10987 antibody compound (also known as REGEN-COV), or an 8.0g REGEN-COV group, and their endogenous immune response to SARS-CoV-2 was characterized at baseline (serological antibody positive / negative). Efficacy was assessed in patients with positive baseline RT-qPCR results; safety was assessed in all patients. A pre-specified stratified analysis of virological endpoints in Group 2 was performed to confirm the previously reported descriptive analysis of Group 1. The proportion of patients in Groups 1+2 who had ≥1 COVID-19 related medical care visit (MAV) by day 29 was assessed.

[0732] By day 7, the baseline viral load was >10. 7 In patients with a pre-specified primary endpoint, the time-weighted mean reduction in viral load (log10 copies / mL) with REGEN-COV (the 2.4g + 8.0g combination dose group) was significantly greater than that with placebo: -0.68 (95% CI, -0.94 to -0.41; P < 0.0001). This change in baseline viral load was -0.73 in seronegative patients (P < 0.0001) and -0.36 in the overall population (P = 0.0003). The proportion of patients with ≥1 COVID-19-related MAV was 2.8% (12 / 434) with REGEN-COV and 6.5% (15 / 231) with placebo (P = 0.024; relative risk reduction rate = 57%), with greater relative risk reduction rates in patients with ≥1 hospitalization risk factor (72%) or seronegative patients (65%). Adverse events were similar across groups.

[0733] Trial design summary: Patients were randomly assigned (1:1:1) to receive either placebo, 2.4g REGEN-COV (1.2g each of cassicimarab and edevimab) or 8.0g REGEN-COV (4.0g each of cassicimarab and edevimab). Figure 5 The 29-day Phase 2 trial includes a screening / baseline period (day -1 to day 1), a follow-up period (day 2 to day 25), and a study-end visit (day 29). The Phase 1 and Phase 2 portions of the trial are identical, except for additional pharmacokinetic analysis in Phase 1.

[0734] Patients: Eligible patients were ≥18 years of age and not hospitalized, had a positive SARS-CoV-2 nasopharyngeal (NP) PCR test result ≤72 hours prior, and had symptom onset ≤7 days prior to randomization. Randomization was stratified by country and presence of ≥1 severe COVID-19 risk factor: age >50 years, obesity (BMI >30), immunosuppression, and chronic cardiovascular, metabolic, liver, kidney, or lung disease. All patients were assessed for the presence of anti-SARS-CoV-2 antibodies: anti-spike [S1]IgA, anti-spike [S1]IgG, and anti-nucleocapsid IgG. Because these results were not available at randomization, patients were randomized regardless of their baseline serological antibody status and then grouped into serologically negative (if all available tests were negative), serologically positive (if any test was positive), or unknown status (missing or indeterminate results). The demographic and baseline medical characteristics of the patients are shown in Table 5A below.

[0735] Table 5A. Demographic and Baseline Medical Characteristics* (N=799; Full Analysis Set)

[0736]

[0737]

[0738] SD standard deviation

[0739] *Positive and negative values ​​are the mean ± SD. Due to rounding, the sum of percentages may not equal 100. IQR represents the interquartile range, and RT-PCR represents reverse transcription polymerase chain reaction.

[0740] Patient-reported race and ethnicity.

[0741] Body Mass Index (BMI) is calculated by dividing weight in kilograms by the square of height in meters.

[0742] Obesity is defined as a body mass index (BMI) greater than 30.

[0743] Hospitalization risk factors include age over 50, obesity, cardiovascular disease (including hypertension), chronic lung disease (including asthma), chronic metabolic disease (including diabetes), chronic kidney disease (including dialysis), chronic liver disease, and immunodeficiency (immunosuppression or immunosuppressant treatment).

[0744] Intervention: At baseline (day 1), intravenous injection of mAb10933 (cascirelimab) and mAb10987 (edevi) (diluted in 250 mL of normal saline for co-administration) or normal saline placebo was administered for 1 hour.

[0745] end

[0746] The primary virological endpoint and two key secondary clinical endpoints were pre-specified in the Phase 1+2 analysis (collectively referred to as Phase 1 / 2) and tested stratified as shown in Table 5B. The primary virological endpoint was defined as the time-weighted mean change in viral load from baseline (Day 1) to Day 7 (log10 copies / mL). The key secondary clinical endpoints were the proportion of patients who had at least one COVID-19-related medical care visit (MAV) by Day 29, and the proportion of patients who had at least one COVID-19-related MAV (including only inpatient or emergency room (ER) visits or urgent care visits). MAV was defined as investigator-confirmed.

[0747] Hospitalization or ER, emergency care or physician office / telemedicine visit related to COVID-19.

[0748] Table 5B. Preliminary analysis of Phase 2 virological and clinical endpoints.

[0749]

[0750]

[0751] mFAS, Modified Full Analysis Set; MAV, Medical Care Visits.

[0752] Safety endpoints for the Phase 1 / 2 portion of the trial included adverse events that occurred or worsened during the observation period (Phase 1 only; Grade 3 and Grade 4 only), serious adverse events (SAEs), and adverse events of particular concern (AESIs): hypersensitivity or infusion-related reactions of Grade ≥2.

[0753] Statistical analysis

[0754] The statistical analysis plan for this analysis is in the database.

[0755] This was completed before the lockdown and unblinding of the Phase 2 dataset of an additional 524 patients. The full analysis set (FAS) included randomized patients with COVID-19 symptoms. Patients who were positive for SARS-CoV-2 nasopharyngeal (NP) PCR at randomization (baseline) for ≤72 hours but negative for a central laboratory qualitative PCR test at baseline (limit of detection, 714 copies / mL) were excluded from the virological and clinical endpoint analyses of the modified full analysis set (mFAS). Subgroup analyses based on serological status and baseline viral load were pre-specified in the statistical analysis plan. Safety was evaluated in FAS patients receiving the investigational drug (active or placebo).

[0756] To confirm the virological efficacy observed in analysis group 1 (patients 1 to 275), virological endpoint analyses were performed using data from patients 276 to 799 (including 524 patients; analysis group 2). However, analyses of clinical endpoints and safety utilized data from all available patients, including the first 275 patients (patients 1 to 799; analysis groups 1+2).

[0757] Virological efficacy endpoints were calculated as described below. Key secondary clinical endpoints were analyzed using Fisher's exact test. Analysis of virological and clinical endpoints was performed using a hierarchical test strategy to control for type I error at a two-sided α = 0.05. Statistical analysis was performed using SAS software version 9.4 or later (SAS Institute).

[0758] Baseline characteristics

[0759] 799 patients were randomized in phase 1 / 2 of the trial. Of the pooled 799 patients, 266, 267, and 266 patients, respectively, were assigned to receive low-dose REGEN-COV, high-dose REGEN-COV, or placebo. Figure 6 Of the 799 patients (analysis groups 1+2), 87 (10.9%) tested negative in a central laboratory SARS-CoV-2NP RT-qPCR test, and 47 (5.9%) had no central laboratory baseline viral load data; therefore, the modified full analysis (mFAS) set included 665 patients. Similarly, in analysis group 2 (primary virological efficacy analysis), the mFAS set included 437 patients out of 524.

[0760] Of the 799 randomized patients, the median age was 42.0 years, 47% were male, 9% were identified as Black or African American, and 50% were identified as Hispanic or Latino (Table 5A). 483 (60.5%) patients had ≥1 risk factor for hospitalization due to COVID-19, including obesity (37.3%), age >50 years (29.3%), cardiovascular disease (20.5%), or chronic metabolic disease (13.1%). Baseline characteristics were similar between analysis group 1 (275 patients) and analysis group 2 (524 patients) (Table 5C).

[0761] Table 5C.1 / 2 Demographic and Baseline Medical Characteristics* (N=524; Full Analysis Set)

[0762]

[0763]

[0764]

[0765] SD, standard deviation.

[0766] *Positive and negative values ​​are the mean ± SD. Due to rounding, the sum of percentages may not equal 100. IQR represents the interquartile range, and RT-PCR represents reverse transcription polymerase chain reaction.

[0767] Patient-reported race and ethnicity.

[0768] Body Mass Index (BMI) is calculated by dividing weight in kilograms by the square of height in meters.

[0769] Obesity is defined as a body mass index (BMI) greater than 30.

[0770] Hospitalization risk factors include age over 50, obesity, cardiovascular disease (including hypertension), chronic lung disease (including asthma), chronic metabolic disease (including diabetes), chronic kidney disease (including dialysis), chronic liver disease, and immunodeficiency (immunosuppression or immunosuppressant treatment).

[0771] At randomization, 408 patients (51.1%) were seronegative, 304 (38.0%) were seronegative, and 87 (10.9%) had unknown serone levels. The mean baseline viral load was 5.48 log10 copies / mL (baseline data were missing for 47 out of 799); 256 patients (32.0%) had a baseline viral load >10⁷ copies / mL. The mean time from symptom onset to randomization was 3.4 days in the entire trial population: 3.2 days in seronegative patients; 3.6 days in seronegative patients; 2.9 days in patients with a viral load >10⁷ copies / mL; and 3.8 days in patients with a viral load ≤10⁷ copies / mL. Of the 408 patients with ≥1 risk factor for hospitalization, 336 (82.3%) were seronegative or had a viral load >10⁴ copies / mL.

[0772] Natural History of Diseases

[0773] In this analysis, the COVID-19 natural history of placebo-treated patients confirmed that the presence of endogenous anti-SARS-CoV-2 antibodies at baseline is an important indicator of viral load and clinical outcomes. Compared with seropositive patients, patients in the placebo group who were seronegative at baseline had a higher median viral load at baseline (7.73 log10 copies / mL vs. 3.88 log10 copies / mL), and they required a longer time for their viral levels to reach LLQ or become undetectable. Figure 7 and Figure 8Similarly, in terms of clinical outcomes, the incidence of COVID-19-related MAV was higher in placebo patients with negative baseline serum antibodies (9.7%; 12 / 124) than in placebo patients with positive baseline serum antibodies (2.4%; 2 / 83). Since the endogenous immune response is correlated with baseline viral titer, the risk of COVID-19-related MAV was expected to be correlated with baseline viral load and the presence of risk factors: the incidence of MAV was 0% (0 / 55) in patients with a baseline viral load ≤10⁴ copies / mL, compared to 8.5% (15 / 176) in patients with a baseline viral load >10⁴ copies / mL, and 2.2% (2 / 89) in patients without risk factors, and 9.2% (13 / 142) in patients with ≥1 risk factor. Figure 9 ).

[0774] Virological treatment

[0775] In the 524 patients analyzed in Group 2, pre-specified comparisons of virological efficacy endpoints were hierarchically assessed; these patients were confirmed positive for SARS-CoV-2 by NP RT-qPCR at baseline (mFAS; n = 437) (Tables 5B and 5D). In all pre-specified virological efficacy comparisons, REGEN-COV treatment significantly reduced viral load by day 7 compared to placebo (Table 5D;). Figure 10A , Figure 10B and Figure 11 In the first comparison, the baseline viral load was >10. 7 In patients with a viral load of -0.68 log10 copies / mL, the least-squares mean difference in daily change of the time-weighted mean (TWA) of viral load up to day 7 between REGEN-COV treatment (2.4 g and 8.0 g doses in combination) and placebo was -0.68 log10 copies / mL (95% CI, -0.94 to -0.41; P < 0.0001) (Table 5D). Similarly, in patients who were serum antibody negative at baseline (n = 256), the least-squares mean difference in daily TWA of REGEN-COV treatment and placebo from baseline was -0.73 log10 copies / mL (95% CI, -0.97 to -0.48; P < 0.0001), compared to -0.36 log10 copies / mL (95% CI, -0.56 to -0.16; P = 0.0003) in the entire modified full analysis set (n = 437). These data suggest that the observed reduction in viral load after treatment with antibody compounds is primarily driven by their effect in serum antibody-negative patients, as previously observed (Table 5D). Figure 10A , Figure 10B and Figure 11In comparisons of all virological efficacy endpoints, low-dose and high-dose antibody compounds showed similar therapeutic effects (Table 5D). Table 5E, Figure 12 and Figure 13 Results from other key virological endpoints were provided.

[0776] Table 5D. Key virological and clinical endpoints

[0777]

[0778]

[0779]

[0780] *The time-weighted average change in viral load is analyzed based on a covariance model, where treatment group, risk factors, and baseline antibody status are fixed effects, and baseline viral load and treatment-to-baseline viral load are covariates. Confidence intervals are not adjusted for multiplicity.

[0781] The confidence interval for the difference (REGEN-COV minus placebo) is based on an exact methodology and is not adjusted for multiplicity.

[0782] Table 5E. Viral load at each visit for patients with no hospitalization risk factors or with ≥1 hospitalization risk factor (log 10 Change from baseline (copy / mL)

[0783]

[0784] The p-value is based on the MMRM model, where baseline serological status, country, treatment, visit, treatment by visit, treatment by baseline, and interaction by baseline visit are fixed effects, and subjects are random effects CI confidence intervals; LS, least squares; SE, standard error.

[0785]

[0786] The virological efficacy endpoint of the daily time-weighted mean (TWA) change from baseline to day 7 was calculated for each patient using the linear trapezoidal rule (area under the curve of change from baseline divided by the time interval of the observation period) and analyzed using a covariance model, in which treatment group, country, and risk factors (no risk factor versus at least one risk factor) were used as fixed effects, and baseline viral load and treatment interaction at baseline were used as covariates.

[0787] Clinical efficacy

[0788] Two clinical efficacy endpoints were pre-specified for the tiered test: the proportion of patients with at least one COVID-19-related medical visit (MAV) and the proportion of patients with at least one COVID-19-related MAV (including only inpatient or ER or emergency care visits) (Tables 5B and 5D). Both endpoints were assessed by day 29 in the pooled group of 799 patients (analysis group 1+2) who were confirmed positive for SARS-CoV-2 by NP RT-qPCR at baseline (mFAS; n = 665). Overall, 67% of COVID-19-related MAVs were inpatient or emergency room (ER) visits (30% and 37%, respectively), 26% were physician office visits / telemedicine visits, and 7% were emergency care visits. A description of COVID-19-related MAVs is included in Table 5F.

[0789] Table 5F. Description of COVID-19-related MAVs*

[0790]

[0791]

[0792]

[0793]

[0794]

[0795] *Analysis set 1+2; Modified full analysis set.

[0796] Only 12.7 mL infusions are accepted, and infusions may be stopped in the ER (Emergency Room) due to the possibility of infusion-related reactions; ICU (Intensive Care Unit); MAV (Medical Care Visit).

[0797] Compared to 6.5% (15 out of 231) in the placebo group, the proportion of patients with ≥1 COVID-19-related MAV in the REGEN-COV treatment group (combination of 2.4g and 8.0g doses) was 2.8% (12 out of 434), representing a relative reduction of 57% (absolute difference of -3.7 percentage points compared to placebo; 95% CI, -7.9% to -0.3%; P = 0.024) (Table 5D). The REGEN-COV treatment effect was more pronounced in patients with negative baseline serum antibodies (3.4% vs. 9.7% in placebo; a relative reduction of 65%) (Table 5G). For the final tiered endpoint, the proportion of patients with COVID-19-related hospitalizations or ER or emergency care visits in the REGENCOV group (compared to placebo) was numerically lower, but the difference did not reach statistical significance (Table 5D). Post-hoc analysis showed that patients receiving the antibody mixture (combination dose group) had a lower rate of hospitalization or death (0.7% [3 of 434] vs. 2.2% [5 of 231]; a relative decrease of 68%), and a lower rate of hospitalization or ER visits (1.8% [8 of 434] vs. 4.3% [10 of 231]; a relative decrease of 58%) (Table 5H).

[0798] Table 5G. Proportion of patients with ≥1 COVID-19-related MAV, broken down by baseline serum antibody status.

[0799]

[0800]

[0801] *COVID-19 related MAVs include hospitalizations, ER visits, emergency care outpatient visits, and patient / physician office / telemedicine visits.

[0802] 95% CI and p-values ​​are based on precise methods.

[0803] CI, confidence interval; ER, emergency room; MAV, medical care visit.

[0804] Table 5H. Proportion of patients hospitalized, visited ER, and / or died.

[0805]

[0806]

[0807] *95% CI and p-values ​​are based on an accurate methodology.

[0808] CI stands for confidence interval.

[0809] Additional post-hospital analysis investigated the effect of antibody combination therapy on MAV in different high-risk subgroups. In the REGEN-COV group (combination dose) versus the placebo group, the proportion of patients with ≥1 hospitalization risk factor (n=408) who developed COVID-19-related MAV was 2.6% vs. 9.2% (absolute difference compared to placebo, -6.5 percentage points; 95% CI, -17 to 4; relative reduction of 72%). Figure 14A , Figure 14B and Figure 14C (Table 5I). In the REGEN-COV group (combination dose) and the placebo group, the proportion of patients with ≥1 risk factor who were baseline serologically negative and had a viral load >104 copies / mL (n=217) and developed COVID-19-related MAV was 2.1% and 13.2% respectively (absolute difference compared to placebo, -11.0 percentage points; 95% CI, -21 to -3; 84% relative reduction) (Table 5J). Most (59%) patients who developed MAV had a viral load ≥4 log10 copies / mL at the time of medical care visit (Table 5F). Figure 15A , Figure 15B and Figure 15C Similar to the virological endpoints, no meaningful differences in clinical outcomes were observed between low-dose and high-dose treatments.

[0810] Table 5I. Patients with ≥1 COVID-19-related MAV in the absence of hospitalization risk factors or with ≥1 hospitalization risk proportion of patients with risk factors

[0811]

[0812]

[0813] *COVID-19 related MAVs include inpatient visits, ER visits, emergency care outpatient visits, and patient / physician office / telemedicine visits.

[0814] 95% CI and p-values ​​are based on precise methods.

[0815] CI, confidence interval; ER, emergency room; MAV, medical care visit.

[0816] Table 5J. Serologically antibody-negative individuals at high risk with viral load >10 4 Among the patients, there was ≥1 COVID-19 related event. The proportion of patients with MAV

[0817]

[0818] *COVID-19 related MAVs include hospitalizations, ER visits, emergency care outpatient visits, and patient / physician office / telemedicine visits.

[0819] 95% CI and p-values ​​are based on precise methods.

[0820] CI, confidence interval; ER, emergency room; MAV, medical care visit.

[0821] At a two-sided alpha level of 0.05, Fisher's exact test was used to compare the proportion of patients receiving medical care due to COVID-19 exacerbation between the REGEN-COV combination dose group and the placebo group, and between each REGEN-COV treatment group and the placebo group. Similar analyses were performed on the proportion of patients with COVID-19-related hospitalizations or emergency room or urgent care visits, and on the proportion of patients with each type of medical care visit.

[0822] Security

[0823] Serious adverse events (SAEs) occurred in 4 out of 258 patients (1.6%) in the REGEN-COV 2.4g group, 2 out of 260 patients (0.8%) in the REGEN-COV 8.0g group, and a greater number of patients (i.e., 6 out of 262 patients [2.3%]) in the placebo group (Tables 5K and 5L). All serious adverse events were considered to be due to advanced or progressive COVID-19 disease and / or associated comorbid clinical conditions and were assessed as unrelated to the treatment with the study drug.

[0824] Adverse events of particular concern (AESIs) that occurred or worsened during the safety observation period—Grade 2 infusion-related reactions and hypersensitivity reactions—were not reported in the 2.4g group, were reported in 4 patients (1.5%) in the 8.0g group, and were reported in 2 patients (0.8%) in the placebo group (Tables 5K and 5L).

[0825] Table 5K. Overview of serious and adverse events of particular concern in the safety population.

[0826]

[0827]

[0828] *The event is a grade 2 or higher hypersensitivity reaction or infusion-related reaction.

[0829] The events listed here were not present at baseline, or were exacerbations of pre-existing conditions that occurred during the observation period, defined as the time from the administration of REGEN-COV or placebo to the last follow-up visit.

[0830] Table 5L. Serious adverse events and events of particular concern reported during treatment in subjects receiving REGEN-COV. Adverse events

[0831]

[0832]

[0833] *Only serious adverse events and adverse events of particular concern (grade 2 or higher infusion-related reactions and hypersensitivity reactions) were collected.

[0834] IV. Intravenous (local).

[0835] Pharmacokinetics: The mean concentrations of both cassicimarumab and edevimab increased in a dose-proportional manner, consistent with linear pharmacokinetics following a single intravenous administration (Table 5M). The mean ± SD concentrations of cassicimarumab and edevimab in serum on day 29 were 79.7 ± 34.6 mg / L and 65.2 ± 28.1 mg / L, respectively, for the low (1.2 g) dose group, and 250 ± 97.4 mg / L and 205 ± 82.7 mg / L, respectively, for the high (4.0 g) dose group (Table 5M).

[0836] Table 5M. Mean concentrations of REGN10933 and REGN10987 in serum

[0837] *Mean average (SD)[N], where N is the number of observations.

[0838] The infusion lasted for 1 hour.

[0839] § Concentrations observed 28 days after administration (i.e., day 29)

[0840] Serum samples for drug concentration analysis were collected from all patients before administration (at screening or baseline visit), on day 1, and on day 29 after infusion. Additional serum samples were collected only on days 3, 5, 7, and 15 from stage 1 patients. Human serum concentrations of REGN10933 (casciretumab) and REGN10987 (edevimab) were measured using a validated immunoassay using streptavidin microplates from Meso Scale Discovery (MSD, Gaithersburg, MD, USA). This method utilized two anti-idiotypic monoclonal antibodies as capture antibodies, each specific for mAb10933 or mAb10987. Captured mAb10933 and mAb10987 were detected using two different non-competitive anti-idiotypic monoclonal antibodies, each also specific for mAb10933 or mAb10987. The bioanalytical method specifically quantifies the level of each anti-SARS-CoV-2 spike mAb independently, without interference from other antibodies. The limit of quantification (LLOQ) for each analyte in undiluted serum samples is 0.156 μg / mL.

[0841] discuss

[0842] This final phase 1 / 2 analysis of REGEN-COV antibody mixture treatment in outpatients with confirmed COVID-19 confirmed confirmed cases confirmed and expanded the findings of the initial 275 patients. To better understand the natural history of COVID-19 in outpatients, data from placebo patients in this trial are described. These data confirmed previous findings that patients who had not developed an autoimmune response at baseline (i.e., seronegative at baseline) had a median viral load at baseline that was almost 3 log copies / mL higher than seronegative patients and required a longer time to reach low or undetectable levels. Similar to other viral infections such as HIV, Ebola virus disease, and influenza, high viral load appears to be a predictor of COVID-19 disease progression, as evidenced by baseline viral load >10. 4 A higher incidence of COVID-19-related MAV was observed in placebo patients with a viral load of [copy / mL]. This data also suggests that risk factors for severe illness, such as older age and obesity, may help predict outpatients most likely to subsequently develop COVID-19-related MAV. For example, 9.2% (13 / 142) of placebo patients with ≥1 risk factor had MAV compared to 2.2% (2 / 89) of placebo patients with no risk factors. In this trial, >80% of patients with risk factors were seronegative or had a viral load >10 [copy / mL]. 4Copy / mL. In the absence of rapid serological tests or quantitative PCR assays to identify high-risk patients, identifying patients with hospitalization risk factors may help identify outpatients most likely to benefit from early treatment with the antibody mixture.

[0843] The anticipated and statistically significant pre-specified hierarchical analysis described in this article confirmed the virological efficacy of REGEN-COV and revealed similar virological efficacy between the 2.4g and 8.0g doses of the antibody mixture. The reduction in viral load was greatest in patients with negative seroantibodies or high viral load at baseline during the first 5 days after treatment. No additional virological benefit was observed in patients who had already developed an effective endogenous antibody response to the infection (seroantibodies positive). Following treatment with either dose of REGEN-COV, the reduction in viral load was accompanied by a significant decrease in the proportion of patients requiring subsequent COVID-19-related medical care visits, the majority of whom (67%) were hospitalized or visited by an ER. The REGEN-COV antibody mixture resulted in a relative reduction in MAV of 57% (6.5% in placebo vs. 2.8% in the combination dose group; P = 0.0240). Interestingly, the proportion of patients with MAV treated with REGEN-COV decreased only during the first week of treatment compared to placebo. One possible explanation for this finding is that, despite accelerating viral clearance, the medical visits that occurred during the first week were not altered. For example, in patients treated with the antibody mixture, all three hospitalizations occurred within the first three days after treatment while the viral load remained ≥4 log10 copies / mL, but no hospitalizations occurred after day 7 (Table 5F). Figure 15A , Figure 15B and Figure 15C In contrast, among patients receiving placebo, 3 out of 5 hospitalizations occurred after day 7, at which point viral load remained high (≥4 log10 copies / mL). These data support early identification and rapid treatment of COVID-19 outpatients to optimize the efficacy of REGEN-COV treatment.

[0844] A low incidence of serious adverse events, infusion-related reactions, and hypersensitivity reactions was observed. Similar to previously reported results, serum concentrations of each antibody on day 29 were significantly higher than the neutralization target concentrations predicted based on in vitro and preclinical data.

[0845] Clinical evidence from this trial suggests that treatment is most beneficial when administered to high-risk patients who present with symptoms early after diagnosis, as they are most likely to have a high viral load and may not yet have developed their own immune response. Furthermore, no adverse findings were observed in patients who were seropositive at baseline. Early treatment of COVID-19 outpatients is crucial, and risk-benefit assessments support treatment to prevent MAV in high-risk patients if viral load or serostatin status cannot be rapidly determined.

[0846] Phase 3 trial plan

[0847] The patient population in cohort 1—part of cohort 1 in phase 3 of this study consisted of adult male and female patients (≥18 years of age) who had the following characteristics:

[0848] • SARS-CoV-2 positive antigen or molecular diagnostic test ≤72 hours prior to randomization (by effective SARS-CoV-2 antigen, RT-PCR or other molecular diagnostic assay), and

[0849] • Symptoms defined by researchers as consistent with COVID-19, occurring within ≤7 days prior to randomization, and

[0850] • ≥1 severe COVID-19 risk factor

[0851] Risk factors are defined as follows:

[0852] a. Age > 50 years old

[0853] b. Obesity, defined as a body mass index (BMI) > 30 kg / m2

[0854] c. Cardiovascular diseases, including hypertension

[0855] d. Chronic lung diseases, including asthma

[0856] e. Type 1 or type 2 diabetes

[0857] f. Chronic kidney disease, including those undergoing dialysis.

[0858] g. Chronic liver disease

[0859] h. Pregnancy

[0860] i. Immunosuppression (e.g., including cancer treatment, bone marrow or organ transplantation, immunodeficiency, HIV (if poorly controlled or with AIDS), sickle cell anemia, thalassemia, and long-term use of immunosuppressive drugs).

[0861] Primary and critical secondary endpoints of queue 1:

[0862] For Cohort 1, the primary endpoint was COVID-19 related medical care visits (MAV) up to day 29. COVID-19 related medical care visits were defined as: inpatient, emergency room (ER) visits, urgent care visits, physician office visits, or telemedicine visits where the primary cause of the visit was COVID-19. Multiple medical care visits were considered as one event.

[0863] The key pre-specified secondary endpoint was the cumulative incidence of COVID-19-related hospitalizations or emergency room visits up to day 29.

[0864] Other key pre-specified secondary endpoints include various types of COVID-19-related MAVs and related outcomes.

[0865] Virological data collectively provided clear evidence that mAb10933+mAb10987 significantly improved SARS-CoV-2 virus clearance. Furthermore, data from the Phase 1 / 2 pooled analysis showed that the reduction in viral load translated into clinical benefit by significantly reducing COVID-19-associated MAVs, defined as those caused by COVID-19 hospitalizations, ER visits, emergency care visits, or physician office or telemedicine visits. Specifically, a pre-specified and multiple-control analysis of the Phase 1 / 2 pooled data (n=799) showed a statistically significant reduction in MAVs in the mAb10933+mAb10987 treatment group compared to placebo (2.8% in the combination dose group vs. 6.5% in placebo; p=0.0240). Most MAVs occurred in high-risk patients defined at baseline as seronegative, with high baseline viral load, or with at least one pre-existing severe COVID-19 risk factor (e.g., age >50 years, obesity, comorbidities). In the exploratory analysis, treatment with mAb10933+mAb10987 showed the greatest benefit in these high-risk groups, with a 62% reduction in the proportion of patients with MAV and a baseline viral load >104 copies / mL (3.2% with combination therapy vs. 8.5% with placebo) compared to placebo, a 65% reduction in the proportion of patients who were seronegative at baseline (3.4% with combination therapy vs. 9.7% with placebo), and a 72% reduction in the proportion of patients with at least one severe COVID-19 risk factor (2.6% with combination therapy vs. 9.2% with placebo). Given the clinical benefit observed in Phase 2, Phase 3 will focus on confirming the clinical benefit of mAb10933+mAb10987 in reducing MAV in high-risk patients, thereby demonstrating a clinical benefit in reducing viral load.

[0866] The sample size for Phase 3 Cohort 1 was estimated to be approximately 5,400 patients. Cohort 1 continued until at least 80 patients (mFAS patients with at least one risk factor) recruited in the primary analysis cohort were observed to be hospitalized or have ER visits, and the total number of patients hospitalized or had ER visits during the study period in the primary analysis cohort exceeded 120.

[0867] The primary efficacy endpoint for Phase 3 Cohort 1 was the cumulative incidence of COVID-19-related MAV in mFAS up to day 29 (randomized and treated PCR-positive patients with at least one risk factor at baseline).

[0868] Based on the time of the first hospitalization / ER visit, the key secondary endpoint of cohort 1 in phase 3, namely the cumulative incidence of COVID-19-related hospitalizations / ER visits up to day 29, was analyzed.

[0869] For Phase 3, the planned virological analysis was descriptive. Using the same methodology as the primary virological endpoint in Phase 2, the time-weighted mean change in viral load (log10 copies / mL) from baseline (day 1) to post-baseline visit time points was analyzed for both seronegative and seronegative mFAS, separately for patients receiving intensive sampling protocols. Proportional endpoints based on observed virological data were compared between groups using methods similar to those for proportional clinical endpoints. Seronegative mFAS and mFAS were analyzed.

[0870] To assess the time course of viral load treatment efficacy, repeated measures mixed-effects (MMRM) models were used to analyze changes in viral load from baseline (log10 copies / mL) at each visit for seronegative mFAS and mFAS, including baseline, random stratified variables, treatment, visits, treatment-to-baseline interaction, treatment-to-visit interaction, and treatment-to-visit interaction.

[0871] The Phase 3 portion of this study evaluated two dose levels of mAb10933+mAb10987 at a 1:1 ratio: 1200 mg and 2400 mg (600 mg / mAb and 1200 mg / mAb, respectively). In the Phase 1 and Phase 2 results, MAV assessment showed that the 2400 mg and 8000 mg doses of mAb10933+mAb10987 had similar virological and clinical efficacy, and both doses had similar and acceptable safety profiles. Given the similarity between the 2400 mg and 8000 mg doses, this Phase 3 study investigated the 2400 mg dose as the highest dose, while also investigating the lower dose.

[0872] Children aged 0 to <18 years may be included in the Phase 3 portion of this study as a separate cohort (Cohort 2) to evaluate the safety, pharmacokinetics, immunogenicity, and efficacy of mAb10933+mAb10987. Patients with COVID-19 symptoms or asymptomatic patients who were SARS-CoV-2 positive at baseline may be included in this cohort. Children with severe COVID-19 risk factors may be included in Cohort 2.

[0873] In cohort 2, pediatric patients were randomly assigned in a 1:1:1 ratio to receive a single intravenous (IV) dose of either low-dose or high-dose combination therapy of mAb10933+mAb10987 or a placebo. However, the mAb10933+mAb10987 treatment group was further classified according to body weight, as defined in Table 6 below.

[0874] Dosage selection in the pediatric population (<18 years of age) can utilize both high-dose and low-dose weight-class fixed-dose approaches. For each weight-class dose targeting the higher adult dose (2400 mg), the goal is to select a dose predicted by a population PK model to ensure that the fifth percentile (C28) of serum concentration at 28 days post-dose is similar to or greater than the fifth percentile of adult C28 observed at the 2400 mg dose. An additional consideration is to ensure that the predicted Cmax and AUC0-28 for each weight-class dose do not exceed values ​​previously achieved in adults. Both mAb10933 and mAb10987 have shown linear PK; therefore, the 50% reduction used in phase 3 selection of the lower adult dose (2400 mg to 1200 mg) also applies to each pediatric weight-class fixed-dose targeting the 1200 mg adult dose (Table 6).

[0875] Table 6: mAb10933+mAb10987IV doses for each weight group, Phase 3 Cohort 2 (age 0 to <18 years)

[0876]

[0877] 1 The dose value represents the total amount of the combination therapy of mAb10933 + mAb10987 administered in IV single doses.

[0878] In cohort 2, the primary objective for patients was safety, while MAV was a descriptive secondary objective.

[0879] The primary endpoints for cohort 2 were safety / tolerability and changes in serum drug concentrations over time.

[0880] • The proportion of patients who experienced a serious adverse event (SAE) during treatment by day 29

[0881] • Proportion of patients experiencing infusion-related reactions (grade ≥2) by day 4

[0882] • The proportion of patients who developed hypersensitivity reactions (grade ≥2) by day 29

[0883] • Changes in serum mAb10933 and mAb10987 concentrations over time

[0884] • Immunogenicity of mAb10933 and mAb10987 as measured by anti-drug antibody (ADA) and neutralizing antibody (NAb)

[0885] Up to approximately 180 pediatric patients in Cohort 2 (60 in each treatment group) will allow 45 patients to be randomized to each PK-ADA sampling protocol.

[0886] Summary of Phase 3 Adult Data

[0887] Confirmatory Phase 3 trial ( Figure 31 and Figure 32 The goal of this trial was to prospectively demonstrate a clinically significant impact of COVID-19 hospitalization or all-cause mortality risk on high-risk outpatients and to confirm safety. The trial also prospectively evaluated the potential benefit of symptom duration. A seamless design was initially used to compare 8000 mg and 2400 mg with placebo, and was modified based on the final analysis of the Phase 1 / 2 portion to evaluate 2400 mg and 1200 mg with placebo, which showed no difference between the 8000 mg and 2400 mg doses in terms of antiviral activity and clinical endpoints (and clinical events occurred primarily in high-risk patients). Data comparing 8000 mg with placebo were converted to descriptive analyses. A formal tiered analysis first evaluated the 2400 mg dose with placebo (n = approximately 2700 in patients with ≥1 risk factor in the original and revised portions), and then evaluated the 1200 mg dose with placebo (n = approximately 1500 in patients with ≥1 risk factor). A concomitant dose-range virology study conducted in outpatients further evaluated the antiviral efficacy of REGEN-COV doses ranging from 2400 mg to 300 mg IV (and 1200 mg to 600 mg subcutaneously) (Example 7). Key results are shown below.

[0888] Table 7: Key Results of the Phase 3 Outpatient Trial 1-3

[0889]

[0890] 1. Based on the modified full analysis set (mFAS) population, including all randomized patients who tested positive for SARS-CoV-2 RT-qPCR by nasopharyngeal swabs at randomization and had ≥1 severe COVID-19 risk factor.

[0891] 2. The formal stratified analysis first evaluated the 2400 mg dose compared to concurrent placebo, and then evaluated the 1200 mg dose compared to concurrent placebo.

[0892] 3. Based on the Phase 1 / 2 analysis showing no difference between 8,000 mg and 2,400 mg doses, the Phase 3 protocol was modified to compare 2,400 mg and 1,200 mg doses with placebo, and the 8,000 mg data were converted to a descriptive analysis.

[0893] In a phase 3 trial involving 4,567 high-risk patients, mAb10933+mAb10987 (REGEN-COV) significantly reduced COVID-19 hospitalizations or all-cause mortality and shortened symptom resolution time by 4 days, thus confirming a phase 1 / 2 clinical benefit. Furthermore, in patients with SARS-CoV-2 PCR-positive status at baseline and ≥1 severe COVID-19 risk factor, a single IV infusion of REGEN-COV at 1200 mg or 2400 mg significantly reduced the proportion of patients with COVID-19-related hospitalizations or all-cause mortality. The two dose levels showed similar therapeutic effects: 2400 mg versus placebo (PBO), a 71.3% reduction (1.3% vs. 4.6%; p<0.0001); and 1200 mg versus PBO, a 70.4% reduction (1.0% vs. 3.2%; p=0.0024). After day 3 of the study, the incidence of COVID-19 hospitalization or all-cause mortality was significantly reduced (89.2%, 2400 mg vs. PBO, p < 0.0001; 71.7%, 1200 mg vs. PBO, p = 0.0101); early events were less modifiable. See also Figure 35 , Figure 36 and Figure 37 The effect was more pronounced in patients with high viral load and / or seronegative status at baseline, but a significant risk reduction was observed in seronegative patients. On day 7, viral load was also significantly reduced in all subgroups, consistent between the 2400 mg and 1200 mg doses. Figure 43 , Figure 44 , Figure 52 , Figure 53 , Figure 54 , Figure 55 , Figure 56 and Figure 58 Administration of REGEN-COV resulted in faster symptom resolution at both doses: 2400 mg with PBO, median 10 days vs. 14 days; p < 0.0001; 1200 mg with PBO, median 10 days vs. 14 days; p < 0.0001. Figure 38 , Figure 39 This data summary is in Figure 33As shown in the figure. Furthermore, compared to the REGEN-COV dose group, the placebo (PBO) group had more frequent serious adverse events (including fatal events) (4.0% PBO combined with 1.4% REGEN-COV group). Figure 40 , Figure 41 , Figure 42 The demographics of this study are as follows: Figure 34 As shown in the image.

[0894] Phase 3 Adult Data: Complete Results and Discussion

[0895] In the Phase 1 / 2 portion of this adaptive phase 1–3 randomized placebo-controlled main regimen, REGEN-COV demonstrated efficacy in outpatients, showing a rapid reduction in viral load and COVID-19-related healthcare needs. In fact, on February 19, 2021, the Independent Data Monitoring Committee (IDMC) recommended discontinuing patient enrollment in the placebo group of the Phase 3 portion of this main regimen due to the proven efficacy of REGEN-COV.

[0896] This adaptive, randomized, main-protocol phase 3 portion included 4,057 COVID-19 outpatients with one or more risk factors for severe disease. Patients were randomized to either intravenous placebo or a single treatment group receiving different doses of REGEN-COV and were followed up for 29 days. A pre-specified stratified analysis compared REGEN-COV 2400 mg dose with concurrent placebo, followed by a 1200 mg dose with concurrent placebo, as endpoints for assessing hospitalization or death risk and time to symptom resolution. Safety was evaluated in all treated patients.

[0897] Compared with placebo, REGEN-COV 2400 mg and 1200 mg significantly reduced COVID-19-related hospitalizations or all-cause mortality (71% reduction, 1.0% vs. 3.2%, p<0.0024; 70% reduction, 1.3% vs. 4.6%, p<0.0001). Compared with placebo, the median time to resolution of COVID-19 symptoms was shortened by 4 days in both dose groups (10 days vs. 14 days; p<0.0001). The efficacy of REGEN-COV was consistent across subgroups, including seropositive patients. REGEN-COV reduced viral load more rapidly than placebo. Serious adverse events occurred more frequently in the placebo group (4.0% vs. 1.1% and 1.3% in the 1200 mg and 2400 mg groups, respectively), and infusion-related reactions were rare (<2 patients in all groups).

[0898] REGEN-COV treatment was well tolerated and significantly reduced COVID-19-related hospitalizations or all-cause mortality, provided rapid symptom relief, and reduced viral load.

[0899] Trial Design—This is an adaptive, multicenter, randomized, double-blind, placebo-controlled, phase 1 / 2 / 3 master protocol (NCT04425629) for outpatients with COVID-19. The phase 3 portion includes three cohorts: cohort 1 (≥18 years), cohort 2 (<18 years), and cohort 3 (pregnant at randomization). Initially, phase 3 patients were randomized 1:1:1 to receive placebo, REGEN-COV 2400 mg (cascisembazone and edevimab 1200 mg each) IV, or REGEN-COV 8000 mg (4000 mg of each antibody) IV. Figure 81 Based on Phase 1 / 2 results showing similar antiviral and clinical efficacy between the 8000 mg and 2400 mg doses, and with most clinical events occurring in high-risk patients, the trial was subsequently modified on November 14, 2020, to revise the cohort and dosage. As a result of the modification, patients subsequently recruited with ≥1 risk factor for severe COVID-19 were randomized in a 1:1:1 ratio to receive placebo, 1200 mg (600 mg per antibody) IV REGEN-COV, or 2400 mg (1200 mg per antibody) IV REGEN-COV. On February 19, 2021, following a recommendation from the IDMC, patients were no longer randomized to receive placebo. The Phase 3 analysis presented here includes Cohort 1 patients (≥18 years of age) who were randomized to receive either 2400 mg or 1200 mg REGEN-COV, with the placebo group serving as the control group.

[0900] Eligible patients (cohort 1) were ≥18 years of age and not hospitalized, had a confirmed local SARS-CoV-2 positive diagnostic test result ≤72 hours prior, and had any COVID-19 symptom onset ≤7 days prior to randomization. Randomization was performed in the initial Phase 3 portion based on country and the presence of severe COVID-19 risk factors. In the modified Phase 3 portion, only patients with ≥1 severe COVID-19 risk factor were eligible. All patients were assessed for anti-SARS-CoV-2 antibodies at baseline: anti-spike [S1]IgA, anti-spike [S1]IgG, and anti-nucleocapsid IgG. Because assay results were unavailable at randomization, patients were subsequently grouped as serologically negative (if all available tests were negative), serologically positive (if any available test was positive), or other (indeterminate / unknown results) for virological and subgroup analysis purposes.

[0901] At baseline (Day 1), REGEN-COV (diluted in saline solution for co-administration) or a saline placebo was administered intravenously. Investigators assessed hospitalization as COVID-19-related. The COVID-19 Symptom Evolution (SE-C19) instrument, an electronic diary, assessed 23 COVID-19 symptoms daily. Quantitative virological analysis and serum antibody testing of nasopharyngeal (NP) swab samples were performed in a central laboratory, as previously described.

[0902] Hierarchical tests were performed on the pre-specified primary endpoint and two key secondary endpoints. Figure 89 The primary endpoint was the proportion of patients with ≥1 COVID-19-related hospitalization or all-cause death by day 29. Two key secondary clinical endpoints were: (1) the proportion of patients with ≥1 COVID-19-related hospitalization or all-cause death from day 4 to day 29; and (2) the time to resolution of COVID-19 symptoms. The time to resolution of COVID-19 symptoms was defined as the time from randomization to day 1, during which subjects rated all symptoms except cough, fatigue, and headache as “asymptomatic” (0 points), which may be “mild / moderate symptoms” (1 point) or “asymptomatic” (0 points). Safety endpoints for the Phase 3 portion of the trial included serious adverse events (SAEs) and adverse events of particular concern (AESIs) that occurred or worsened during the observation period: grade ≥2 hypersensitivity reactions and infusion-related reactions, and treatment-related adverse events requiring medical care at a healthcare facility.

[0903] Statistical analyses for this study were planned to be completed prior to database locking and unblinding of Phase 3 cohort 1; the primary analysis excluded patients from the previously reported Phase 1 / 2 trials. The full analysis set (FAS) included all randomized symptomatic patients. Efficacy analyses were performed based on a modified FAS (mFAS), defined as all randomized patients who were positive for SARS-CoV-2 at baseline using a central laboratory-ascertained RT-qPCR test and had ≥1 risk factor for severe COVID-19. Safety was assessed in patients treated within the FAS. The proportion of patients with ≥1 COVID-19-related hospitalization or all-cause mortality was compared between the dose and placebo groups using a graded Cochran-Mantel-Haenszel (CMH) test, with country as the grading factor. The p-values ​​and 95% confidence intervals (CIs) for the graded CMH test were presented using the Farrington-Manning method. Time to resolution of COVID-19 symptoms was assessed in patients with a baseline total severity score >3 and analyzed using a graded log-rank test (with country as the grading factor). Median time and 95% CI were derived from the Kaplan-Meier method. Hazard ratios and 95% CIs were estimated using a Cox regression model. Analysis of the primary and key clinical endpoints was performed using a hierarchical test strategy to control for Type I errors at a two-sided α = 0.05. Figure 89 Statistical analysis was performed using SAS software version 9.4 or later (SAS Institute).

[0904] Results (Experimental Group)

[0905] Phase 3 patient recruitment took place between September 24, 2020, and January 17, 2021. Initially, in the initial Phase 3 portion, a total of 3088 patients with or without severe COVID-19 risk factors were randomized to receive a single dose of placebo, REGEN-COV 8000 mg, or REGEN-COV 2400 mg. Subsequently, in the revised Phase 3 portion, an additional 2519 patients with ≥1 risk factor were randomized to receive a single dose of placebo, REGEN-COV 2400 mg, or REGEN-COV 1200 mg. Figure 76 The median follow-up time for patients was 45 days, and 96.6% of patients had a follow-up time of >28 days.

[0906] The primary treatment cohort includes individuals with ≥1 severe COVID-19 risk factor and a baseline positive SARS-CoV-2 test in a central laboratory (mFAS). Figure 76 In the mFAS population (n=4057), the demographic and baseline medical characteristics of the placebo and REGEN-COV groups were balanced. Figure 78The median age was 50 years (interquartile range [IQR, 38-59]), 52% were male, 14% were ≥65 years old, 28% were Hispanic, and 61% were obese. The most common risk factors were obesity (58%), age ≥50 years (52%), and cardiovascular disease (36%); 3% of patients were immunosuppressed or receiving immunosuppressive medications. Figure 90 Similar demographic and baseline medical characteristics were observed in the overall full analysis set (n=5607) and the REGEN-COV 8000mg group. Figure 91 ).

[0907] The median NP viral load was 6.98 log. 10 The number of copies / mL (IQR 5.45-7.85) and most patients (69%) were SARS-CoV-2 serologically negative at baseline. Figure 78 These high viral loads and lack of endogenous immune response at baseline suggest that recruited individuals were in the early stages of their infection process. In the treatment groups, NP viral load and seronegative status were similar. The median time to onset of COVID-19 symptoms at randomization was 3 days (IQR 2–5), and this was well-balanced across treatment groups.

[0908] Results (Natural History)

[0909] There is an association between COVID-19-related hospitalization or all-cause mortality risk and baseline viral load: patients with high viral load (baseline viral load >10) had a higher proportion of hospitalizations / mortality compared to patients with low viral load at baseline. 6 Copy / mL: 6.3% [55 / 876] and 4.2% [20 / 471] in the placebo groups receiving 2400 mg and 1200 mg respectively; baseline viral load ≤10 6 Copy / mL: 1.3% [6 / 457] and 1.5% [4 / 273] of patients in the placebo group receiving 2400 mg and 1200 mg respectively. Figure 92 ).

[0910] Compared with patients who were seropositive, patients in the placebo group who were serone at baseline had a higher median viral load at baseline (7.45 log). 10 copies / mL and 4.96 log 10 (copy / mL), and they require a longer time to bring their viral levels below the limit of quantitation (LLQ) (copy / mL). Figure 82 ).

[0911] Baseline serological antibody status in placebo patients did not predict subsequent COVID-19-related hospitalizations or all-cause mortality, as these rates were similar in both antibody-negative and antibody-positive patients (antibody-negative: 5.3% [49 / 930] and 3.5% [18 / 519] in the 2400 mg and 1200 mg placebo groups, respectively; antibody-positive: 4.0% [12 / 297] and 3.7% [6 / 164] in the 2400 mg and 1200 mg placebo groups, respectively). However, placebo patients who were antibody-positive and subsequently required hospitalization or death had high viral loads at baseline and day 7, similar to antibody-negative patients who required hospitalization or death, suggesting that some antibody-positive patients may have an ineffective innate antibody response. Figure 93 ).

[0912] Therapeutic effect (primary endpoint)

[0913] REGEN-COV 2400mg and 1200mg similarly reduced COVID-19-related hospitalizations or all-cause mortality by 71.3% (1.3% vs. 4.6% placebo; 95% CI: 51.7%, 82.9%; p<0.0001) and 70.4% (1.0% vs. 3.2% placebo; 95% CI: 31.6%, 87.1%; p<0.0024), respectively. Figure 79 , Figure 77A , Figure 77B , Figure 94 A similar reduction in COVID-19-related hospitalizations or all-cause mortality was observed in subgroups (including patients who were seropositive at baseline). Figure 79 , Figure 83A , Figure 83B and Figure 83C ).

[0914] Therapeutic effect (key secondary endpoint)

[0915] A decrease in the proportion of patients hospitalized or dying from COVID-19 was observed approximately 1 to 3 days after treatment with REGEN-COV. Figure 77A , Figure 77B , Figure 79 One to three days prior to this, patients in the placebo group continued to experience COVID-19-related hospitalizations or deaths during the study period (46 / 1340 [3.4%]), while very few events occurred in the 2400 mg or 1200 mg REGEN-COV treatment groups (5 / 1351 [0.4%] and 5 / 735 [0.7%], respectively). Figure 79 , Figure 84A , Figure 84B ).

[0916] In both REGEN-COV dosage groups, the median time to resolution of COVID-19 symptoms was 4 days earlier than in placebo (10 days vs. 14 days; p<0.0001 for each of the 2400 mg and 1200 mg doses). Figure 79 and Figure 77C By day 3, it was clear that both doses of REGEN-COV resulted in faster resolution of COVID-19 symptoms. Both REGEN-COV doses were associated with similar improvements in symptom resolution in subgroups. Figure 85 ).

[0917] Compared with placebo, all REGEN-COV dose levels (1200 mg, 2400 mg, and 8000 mg) resulted in a similar and rapid decrease in viral load. Figure 86A , Figure 86B , Figure 86C , Figure 87 , Figure 88A , Figure 88B and Figure 88C ).

[0918] Therapeutic effects (other secondary endpoints)

[0919] REGEN-COV treatment was associated with a lower proportion of patients hospitalized for COVID-19-related reasons. Figure 95 Among patients hospitalized for COVID-19, the REGEN-COV group had shorter hospital stays and a lower rate of intensive care unit admission. Figure 96 ).

[0920] REGEN-COV treatment resulted in a lower proportion of patients with COVID-19-related hospitalizations, emergency room visits, or all-cause mortality by day 29. Figure 97 ), and requiring any medical care visit due to worsening COVID-19 (hospitalization, emergency room visit, emergency care visit, or doctor's office / telemedicine visit) or all-cause death ( Figure 95 , Figure 98 and Figure 99 The proportion of ) is relatively low.

[0921] Security

[0922] Compared with the following REGEN COV dose groups, more patients (4.0%) in the placebo group experienced serious adverse events (SAEs): 1.1% in the 1200 mg group, 1.3% in the 2400 mg group, and 1.7% in the 8000 mg group. Figure 80Compared to the following REGEN-COV dose groups, more patients (5 patients, 0.3%) in the placebo group experienced treatment-induced adverse events (TEAEs) leading to death: 1 in the 1200 mg group (0.1%), 1 in the 2400 mg group (<0.1%), and 0 in the 8000 mg group. Figure 80 and Figure 100 Most adverse events are consistent with complications of COVID-19. Figure 101 and Figure 102 Most of these reactions were considered unrelated to the study drug. Few patients experienced infusion-related reactions (0 in the placebo group; 2, 1, and 3 patients in the 1200 mg, 2400 mg, and 8000 mg groups, respectively) or hypersensitivity reactions (1 in the placebo group and 1 in the 2400 mg group). Figure 91 Similar safety profiles were observed between REGEN-COV doses, with no identifiable imbalances in the event of safety incidents. No safety signals were observed in laboratory safety parameters collected up to day 29.

[0923] Pharmacokinetics

[0924] On day 29, the mean concentrations of cascisimilab and edevimab in serum increased in a dose-proportional manner, consistent with linear pharmacokinetics. Figure 103 The mean serum concentrations of cassicimarab and edevimab on day 29 were 46.4 ± SD 22.5 mg / L and 38.3 ± SD 19.6 mg / L for a 1200 mg dose, and 73.2 ± SD 27.2 mg / L and 60.0 ± SD 22.9 mg / L for a 2400 mg dose, respectively. The mean estimated half-life of cassicimarab was 28.8 days, and that of edevimab was 25.5 days. Figure 103 ).

[0925] discuss

[0926] Previous Phase 1 / 2 data showed that REGEN-COV robustly reduced viral load, decreased the need for medical care, and, despite a small number of events, strongly suggested a reduced risk of hospitalization in outpatients with COVID-19. These clinical outcome data now definitively demonstrate that early treatment with REGEN-COV in outpatients with severe COVID-19 risk factors significantly reduces the risk of hospitalization or all-cause mortality. Both the 1200 mg IV dose and the 2400 mg IV dose of REGEN-COV reduced COVID-19 hospitalization or all-cause mortality by approximately 70% within 28 days of treatment (compared to placebo). Among those hospitalized, REGEN-COV treatment also shortened the duration of hospitalization and reduced the proportion of patients requiring intensive care. Furthermore, both doses of REGEN-COV resulted in faster resolution of COVID-19 symptoms, with a median time to resolution of 4 days. Therefore, a single dose of REGEN-COV for outpatients with COVID-19 has the potential to improve patient outcomes by reducing morbidity and mortality, and significantly alleviate the healthcare burden during the pandemic (including hospitalization and intensive care). Furthermore, REGEN-COV can significantly accelerate recovery from COVID-19, which is an additional benefit for patients, as there is increasing evidence that some patients (including those with mild symptoms) will experience varying degrees of prolonged recovery.

[0927] To avoid being bound by theory, we previously hypothesized that while host factors play a role in the disease process, SARS-CoV-2 morbidity and mortality are caused by high viral load, and that early treatment with a mixture of anti-spike monoclonal antibodies may significantly improve this risk. In the placebo group, we found that patients hospitalized or who died from all causes had significantly higher viral loads at baseline and slower viral clearance, independent of baseline serological status. Patients in the placebo group who developed an endogenous antibody response to SARS-CoV-2 (seropositive) had similar hospitalization or mortality rates compared to seronegative patients, suggesting that some seropositive patients have an ineffective immune response. Furthermore, placebo patients who were seropositive and had COVID-19-related hospitalizations or deaths also had similarly high baseline viral load levels as seronegative patients with these events, confirming that high viral load is a key driver of severe COVID-19. Additionally, this study demonstrates that REGEN-COV has clinical benefit regardless of baseline serological status, reducing the importance of serological testing at the time of COVID-19 diagnosis for clinical treatment decisions. This conclusion is important given the widespread use of vaccines, as widespread use will result in a baseline seropositivity status, which may not be effective in preventing serious infections in some patients (as in some patients with ineffective natural immunity in this trial) or serious infections due to the emergence of worrying variants (VOCs).

[0928] Both 1200 mg and 2400 mg doses of REGEN-CoV demonstrated similar antiviral and clinical efficacy, indicating that we are significantly above the lowest effective dose. Both doses rapidly reduced viral load and achieved faster viral clearance compared to placebo. In addition to providing clinical benefit to individual patients receiving REGEN-CoV, the rapid antiviral effect is likely associated with public health benefits by reducing the risk of viral transmission and inhibiting SARS-CoV-2 VOC.

[0929] A low incidence of serious adverse events, hypersensitivity reactions, and infusion-related reactions was observed. Serum concentrations of each antibody on day 29 were significantly higher than the neutralization target concentrations predicted based on in vitro and preclinical data.

[0930] The emergence of SARS-CoV-2 drug-resistant variants during antiviral treatment or circulating in global communities will continue to challenge the success of COVID-19 therapeutics and vaccines. Although in vitro studies or in vivo animal studies using recombinant viruses have shown that combinations of non-competitive ...

Claims

1. Use of a first monoclonal antibody or an antigen-binding fragment thereof and a second monoclonal antibody or an antigen-binding fragment thereof in the preparation of a composition for improving one or more clinical parameters of COVID-19 in a subject, wherein the first monoclonal antibody or an antigen-binding fragment thereof binds to a first epitope on the SARS-CoV-2 spike (S) glycoprotein and comprises a heavy chain variable region (HCVR) containing three complementarity-determining regions HCDR1, HCDR2, and HCDR3 and a light chain variable region (LCVR) containing three complementarity-determining regions LCDR1, LCDR2, and LCDR3, wherein HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 are respectively derived from SEQ ID NO: The second monoclonal antibody or its antigen-binding fragment binds to a second epitope on the SARS-CoV-2 spike (S) glycoprotein and comprises an HCVR containing three complementarity-determining regions HCDR1, HCDR2, and HCDR3 and an LCVR containing three complementarity-determining regions LCDR1, LCDR2, and LCDR3, wherein HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 are composed of the amino acid sequences of SEQ ID NO: 24, 26, 28, 32, 34, and 36, respectively.

2. The use according to claim 1, wherein the subject is a human patient suffering from laboratory-confirmed SARS-CoV-2 and one or more COVID-19 symptoms.

3. The use according to claim 2, wherein one or more COVID-19 symptoms include fever, cough, or shortness of breath.

4. The use according to claim 2, wherein the subject is selected from the group consisting of: (a) COVID-19 patients requiring low-flow oxygen supplementation; (b) COVID-19 patients requiring high-intensity oxygen therapy but not mechanical ventilation; and (c) COVID-19 patients requiring mechanical ventilation.

5. The use according to claim 1, wherein the subject is hospitalized due to one or more COVID-19 symptoms.

6. The use according to claim 1, wherein the subject is an outpatient.

7. Use of a first monoclonal antibody or an antigen-binding fragment thereof and a second monoclonal antibody or an antigen-binding fragment thereof in the preparation of a composition for the prevention of SARS-CoV-2 infection or COVID-19 in a subject, wherein the first monoclonal antibody or an antigen-binding fragment thereof binds to a first epitope on the SARS-CoV-2 spike (S) glycoprotein and comprises a heavy chain variable region (HCVR) containing three complementarity-determining regions HCDR1, HCDR2, and HCDR3 and a light chain variable region (LCVR) containing three complementarity-determining regions LCDR1, LCDR2, and LCDR3, wherein HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 are respectively derived from SEQ ID NO. The second monoclonal antibody or its antigen-binding fragment binds to the second epitope on the SARS-CoV-2 spike (S) glycoprotein and comprises an HCVR containing three complementarity-determining regions HCDR1, HCDR2, and HCDR3 and an LCVR containing three complementarity-determining regions LCDR1, LCDR2, and LCDR3, wherein HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 are composed of the amino acid sequences of SEQ ID NO: 24, 26, 28, 32, 34, and 36, respectively.

8. The use according to claim 7, wherein the subject is an uninfected individual at high risk of SARS-CoV-2 infection.

9. The use according to claim 8, wherein the subject at high risk of SARS-CoV-2 infection is a healthcare worker, a field first responder, or a family member of an individual who has tested positive for SARS-CoV-2 infection.

10. The use according to any one of claims 1-9, wherein the SARS-CoV-2 spike (S) glycoprotein consists of the amino acid sequence of SEQ ID NO:

59.

11. The use according to any one of claims 1-9, wherein the first monoclonal antibody or antigen-binding fragment comprises an HCVR / LCVR amino acid sequence pair consisting of the amino acid sequence of SEQ ID NO: 2 / 10, and the second monoclonal antibody or antigen-binding fragment comprises an HCVR / LCVR amino acid sequence pair consisting of the amino acid sequence of SEQ ID NO: 22 / 30.

12. The use according to claim 11, wherein the first monoclonal antibody and the second monoclonal antibody comprise the constant region of the human IgG heavy chain.

13. The use according to claim 12, wherein the first monoclonal antibody and the second monoclonal antibody comprise a heavy chain constant region of an IgG1 or IgG4 isotype.

14. The use according to claim 12, wherein the first monoclonal antibody comprises a heavy chain and a light chain, the heavy chain consisting of the amino acid sequence of SEQ ID NO: 18, the light chain consisting of the amino acid sequence of SEQ ID NO: 20, and the second monoclonal antibody comprises a heavy chain and a light chain, the heavy chain consisting of the amino acid sequence of SEQ ID NO: 38, the light chain consisting of the amino acid sequence of SEQ ID NO:

40.

15. The use according to any one of claims 1-9, wherein the composition comprises 1 mg to 10 g of the first monoclonal antibody or its antigen-binding fragment and the second monoclonal antibody or its antigen-binding fragment.

16. The use according to any one of claims 1-9, wherein the composition comprises 1.2 g of a first monoclonal antibody or an antigen-binding fragment thereof and 1.2 g of a second monoclonal antibody or an antigen-binding fragment thereof.

17. The use according to any one of claims 1-9, wherein the composition comprises 150 mg of a first monoclonal antibody or an antigen-binding fragment thereof and 150 mg of a second monoclonal antibody or an antigen-binding fragment thereof.

18. The use according to any one of claims 1-9, wherein the composition comprises 300 mg of a first monoclonal antibody or an antigen-binding fragment thereof and 300 mg of a second monoclonal antibody or an antigen-binding fragment thereof.

19. The use according to any one of claims 1-9, wherein the composition comprises 600 mg of a first monoclonal antibody or an antigen-binding fragment thereof and 600 mg of a second monoclonal antibody or an antigen-binding fragment thereof.

20. The use according to any one of claims 1-9, wherein the composition comprises 150 mg to 1200 mg of a first monoclonal antibody or an antigen-binding fragment thereof and 150 mg to 1200 mg of a second monoclonal antibody or an antigen-binding fragment thereof.

21. The use according to any one of claims 1-9, wherein the composition is configured to be administered to the subject by intravenous infusion or subcutaneous injection.

22. The use according to any one of claims 1-6, wherein after administration of the composition, the subject exhibits one or more therapeutic parameters selected from the group consisting of: (a) Decrease in SARS-CoV-2 virus shedding from baseline; (b) Using a 7-point ordinal scale, the clinical condition has improved by at least 1 point; (c) Reduce or eliminate the need for supplemental oxygen; (d) Reduce or eliminate the need for mechanical ventilation; (e) Preventing COVID-19-related deaths; (f) Prevention of all-cause mortality; and (g) Changes in serum concentrations of one or more disease-related biomarkers.

23. The use according to claim 22, wherein the 7-point ordinal scale is: [1] Death; [2] Hospitalization, requiring invasive mechanical ventilation or extracorporeal membrane oxygenation; [3] Hospitalization, requiring non-invasive ventilation or high-flow oxygen device; [4] Hospitalization requires supplemental oxygen supply; [5] Hospitalization, no need for supplemental oxygen supply—requires continuous medical care; [6] Hospitalization, no need for supplemental oxygen supply—no longer requiring continuous medical care; and [7] No hospitalization.

24. The use according to claim 22, wherein the one or more therapeutic parameters are measured 21 days after administration of the first dose of the composition.

25. The use according to claim 22, wherein the reduction in SARS-CoV-2 virus shedding from baseline is determined by real-time quantitative PCR (RT-qPCR) of nasopharyngeal swab samples, nasal cavity samples, or saliva samples.

26. The use according to claim 22, wherein the change in serum concentration of one or more disease-related biomarkers is a change in C-reactive protein, lactate dehydrogenase, D-dimer, or ferritin.

27. The use of claim 6, wherein, Following administration of the composition, the subjects exhibited fewer than 5 COVID-19 related medical care visits, telemedicine visits, hospital admissions, and / or intensive care unit (ICU) admissions.

28. The use according to claim 27, wherein the subject exhibits fewer than 5 COVID-19 related medical care visits, telemedicine visits, hospital admissions and / or admissions to the intensive care unit (ICU) within 29 days after administration of the first dose of the composition.

29. The use according to claim 27, wherein the subject exhibits fewer than 4, 3, 2, or 1 COVID-19 related medical care visits, telemedicine visits, hospital admissions, and / or admissions to the intensive care unit (ICU).

30. The use according to claim 22, wherein the subject tests negative for SARS-CoV-2 within 2 days to 3 weeks after the first administration of the composition.

31. The use according to claim 30, wherein a negative test for SARS-CoV-2 is determined by RT-qPCR in a nasopharyngeal swab sample, nasal cavity sample, or saliva sample.

32. The use according to any one of claims 1-6, wherein the composition is administered to the subject in combination with an additional therapeutic agent.

33. The use according to claim 32, wherein the additional therapeutic agent is an antiviral compound.

34. The use according to claim 33, wherein the antiviral compound is remdesivir.

35. The use according to claim 32, wherein the additional therapeutic agent is an IL-6 or IL-6R blocker.

36. The use according to claim 35, wherein the additional therapeutic agent is tocilizumab or thalidomide.

37. The use according to claim 32, wherein the additional therapeutic agent is a steroid.

38. The use according to claim 32, wherein the additional therapeutic agent is applied prior to the composition.

39. The use according to claim 32, wherein the additional therapeutic agent is applied after or simultaneously with the composition.

40. The use according to any one of claims 1-9, wherein the subject is serum negative for SARS-CoV-2 infection.

41. The use according to any one of claims 1-6, wherein the composition is used to more rapidly alleviate at least one symptom of SARS-CoV-2 infection when administered to a comparable seronegative subject population compared to a placebo-administered population.

42. The use according to any one of claims 1-6, wherein the composition is used to more rapidly alleviate at least one symptom of SARS-CoV-2 infection when administered to a seronegative subject population compared to a comparable seronegative subject population.

43. The use according to any one of claims 1-6, wherein the composition is used to reduce the viral load in a subject population 7 days after administration, compared to the day of administration.

44. The use according to claim 43, wherein, compared with a comparable group of subjects treated with placebo, in patients treated with 0.6 g of the first monoclonal antibody and 0.6 g of the second monoclonal antibody, the time-weighted mean change in nasopharyngeal (NP) viral load from baseline to day 7 was at least 0.86 log10 copies / mL, wherein the p-value was <0.0001.

45. The use according to claim 43, wherein, compared with a comparable group of subjects treated with placebo, the nasopharyngeal (NP) viral load in the seronegative subject group decreased by at least 1.04 log10 copies / mL from baseline to day 7 by at least 1.2 g of the first monoclonal antibody and 1.2 g of the second monoclonal antibody, wherein the p-value was <0.0001.

46. ​​The use according to claim 43, wherein, compared with a comparable group of subjects treated with placebo, the mean change in nasopharyngeal (NP) viral load from baseline to day 7 in the subject group treated with 0.6 g of the first monoclonal antibody and 0.6 g of the second monoclonal antibody was at least 0.71 log10 copies / mL less, wherein the p-value was <0.0001.

47. The use according to claim 43, wherein, compared with a comparable group of subjects treated with placebo, the mean change in nasopharyngeal (NP) viral load from baseline to day 7 in the subject group treated with 1.2 g of the first monoclonal antibody and 1.2 g of the second monoclonal antibody was greater than a decrease of 0.86 log10 copies / mL, wherein the p-value was <0.0001.

48. The use according to any one of claims 1-6, wherein the composition is used to reduce the viral load in a subject population.

49. The use according to claim 48, wherein the composition is administered in the form of 0.6 g of the first monoclonal antibody and 0.6 g of the second monoclonal antibody, and wherein the composition produces an average viral load reduction of at least 3.00 log10 copies / mL on day 7 after administration, compared with the baseline viral load measured on day 0 before administration.

50. The use according to claim 49, wherein the reduction is at least 3.50 log10 copies / mL.

51. The use according to claim 49, wherein the reduction is at least 3.90 log10 copies / mL.

52. The use according to claim 48, wherein the composition is administered in the form of 1.2 g of the first monoclonal antibody and 1.2 g of the second monoclonal antibody, and wherein the composition produces a reduction in average viral load of at least 3.50 log10 copies / mL on day 7 after administration, compared with the baseline viral load measured on day 0 before administration.

53. The use according to claim 52, wherein the reduction is at least 3.75 log10 copies / mL.

54. The use according to claim 52, wherein the reduction is at least 4.09 log10 copies / mL.

55. The use according to any one of claims 1-6, wherein the composition is used to shorten the median time to symptom relief by 4 days in a comparable group of subjects treated with 0.6g of the first monoclonal antibody and 0.6g of the second monoclonal antibody or 1.2g of the first monoclonal antibody and 1.2g of the second monoclonal antibody compared to a comparable group of subjects treated with placebo.

56. The use according to claim 48, wherein the population includes subjects who have not been hospitalized due to COVID-19.

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