Methods for treating influenza infection

Abstract

Provided herein are methods for inhibiting, reducing, or shortening influenza virus replication or infection in a human subject, and methods for treating influenza infection or a condition associated therewith in a human subject in need thereof, comprising administering a zanamivir-Fc conjugate having the structure of formula (I) at a dose of 10 to 900 mg.

Description

[Background technology]

[0001] The need for novel antiviral treatments for influenza is particularly significant in the medical field. The influenza virus, the causative agent of influenza, or the flu, causes 3 to 5 million severe cases and approximately 500,000 deaths worldwide annually. While most people fully recover from influenza within approximately one to two weeks, some develop life-threatening complications, such as pneumonia. Thus, influenza can be particularly deadly for the young, elderly, or those with chronic illnesses. People with weakened or compromised immune systems, such as those with advanced HIV infection or transplant patients whose immune systems are medically suppressed to prevent rejection of transplanted organs, are at increased risk for influenza-related complications. Pregnant women and young children are also at increased risk for complications.

[0002] The development of antiviral therapeutics against influenza remains an ongoing challenge. Several antiviral agents have been approved for clinical use, and these play an important role in modulating disease severity and controlling pandemics while vaccines are being developed. However, drug-resistant strains have emerged against the most commonly used inhibitors.

[0003] Antiviral agents primarily target proteins displayed on the surface of influenza virus particles. The influenza virus envelope contains two immunodominant glycoproteins, hemagglutinin and neuraminidase, which play important roles in viral infection and spread. Hemagglutinin attaches the virus to host cells through interaction with surface sialic acid, thereby initiating entry. Neuraminidase is an exoglycosidase enzyme that cleaves sialic acid (terminal neuraminic acid residues) from glycan structures on the surface of infected host cells, releasing progeny viruses and allowing the virus to spread from the host cell to uninfected neighboring cells. Therefore, inhibition of neuraminidase is a pharmacological target for antiviral drugs. Viral neuraminidase inhibitors used to reduce viral spread have been identified, including oseltamivir (Tamiflu™), zanamivir (Relenza™), and peramivir (Rapivab™).

[0004] Influenza in transplant recipients remains characterized by prolonged viral shedding, increasing the likelihood of developing drug-resistant strains. New, more effective therapies for treating influenza are needed. Summary of the Invention

[0005] In a first aspect, the present disclosure provides a method for inhibiting, reducing, or shortening influenza virus replication or infection in a human subject, the method comprising or consisting of subcutaneously or intramuscularly administering to the human subject a dose of 10 mg to 900 mg (e.g., 10 mg, 15 mg, 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 45 mg, 50 mg, 150 mg, 300 mg, 450 mg, or 900 mg) of a conjugate of formula (I), wherein the conjugate of formula (I) has the following structure: [ka] wherein each E is an Fc domain monomer; n is 2, T is an integer from 3 to 6, and The wavy line indicates the covalent bond of E to the nitrogen atom of a solvent-exposed lysine or the sulfur atom of a solvent-exposed cysteine.

[0006] In another aspect, the present disclosure provides a method for treating a human subject having or at risk of influenza infection, the method comprising or consisting of subcutaneously or intramuscularly administering to the human subject a dose of a conjugate of Formula (I) in an amount of 10 mg to 900 mg.

[0007] In another aspect, the present disclosure provides methods for reducing the area under the influenza viral load time curve (VL-AUC), reducing peak influenza viral load time, reducing time to confirmed negative influenza test, reducing total clinical symptom score (TSS-AUC), reducing peak TSS, or reducing time to symptom resolution in a human subject (e.g., a human subject having an influenza infection), which methods comprise or consist of subcutaneously or intramuscularly administering to the human subject an amount of 10 mg to 900 mg of a conjugate of Formula (I).

[0008] In another aspect, the disclosure provides a method for inhibiting, reducing, or shortening influenza virus replication or infection in a human subject, the method comprising or consisting of maintaining a minimum plasma concentration of the conjugate of Formula (I) of at least 300 ng / mL for 4 to 6 months. In some embodiments, the method comprises or consists of maintaining a minimum plasma concentration of the conjugate of at least 1 μg / mL. In some embodiments, a minimum plasma concentration of the conjugate of at least 300 ng / mL is maintained in the subject for 6 months. In some embodiments, a minimum plasma concentration of the conjugate of at least 300 ng / mL is maintained in the subject for 4 months. In some embodiments, a minimum plasma concentration of the conjugate of at least 1 μg / mL is maintained in the subject for 6 months. In some embodiments, a minimum plasma concentration of the conjugate of at least 1 μg / mL is maintained in the subject for 4 months.

[0009] In some embodiments of any of the above methods, the method comprises administering a single dose of the conjugate to a human subject. In some embodiments, the human subject is administered a single dose of 10 mg, 15 mg, 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 45 mg, 50 mg, 150 mg, 300 mg, 450 mg, or 900 mg of the conjugate.

[0010] In some embodiments of any of the above methods, the method comprises or consists of two or more (e.g., three or four) administrations of the conjugate to the human subject. In some embodiments, the method comprises or consists of two administrations of the conjugate to the human subject, the two administrations being separated by 60 to 120 days (e.g., between 75 and 105 days).

[0011] In some embodiments, the dose for administration comprises 10 mg, 15 mg, 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 45 mg, 50 mg, 150 mg, 300 mg, 450 mg, or 900 mg of the conjugate.

[0012] In some embodiments of any of the above methods, the conjugate is conjugate A having a structure provided herein.

[0013] In another aspect, the disclosure provides a pharmaceutical composition in unit dosage form, comprising a conjugate of Formula (I) (e.g., conjugate A) in an amount of 10 mg to 900 mg. In some embodiments, the pharmaceutical composition contains 10 mg, 15 mg, 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 45 mg, 50 mg, 150 mg, 300 mg, 450 mg, or 900 mg of the conjugate.

[0014] In another aspect, the present disclosure provides a kit comprising the pharmaceutical composition described above and instructions for use in any of the methods described herein.

[0015] In some embodiments of any of the above aspects, the conjugate comprises an Fc domain comprising a protein having an amino acid sequence at least 95% identical to any one of SEQ ID NO:62, SEQ ID NO:63, SEQ ID NO:64, SEQ ID NO:67, SEQ ID NO:68, SEQ ID NO:72, SEQ ID NO:73, SEQ ID NO:76, SEQ ID NO:77, SEQ ID NO:80, SEQ ID NO:81, SEQ ID NO:82, SEQ ID NO:85, SEQ ID NO:86, SEQ ID NO:90, SEQ ID NO:91, SEQ ID NO:94, or SEQ ID NO:95. In some embodiments, the conjugate comprises an Fc domain comprising a protein having an amino acid sequence of any one of SEQ ID NO:63, SEQ ID NO:64, SEQ ID NO:67, SEQ ID NO:68, SEQ ID NO:72, SEQ ID NO:73, SEQ ID NO:76, or SEQ ID NO:77.

[0016] In some embodiments, the influenza virus is an influenza A virus. In some embodiments, the influenza virus is an influenza B virus. In some embodiments, the influenza virus is an influenza C virus.

[0017] definition To facilitate understanding of the present invention, several terms are defined below. Terms defined herein have meanings commonly understood by one of ordinary skill in the art relevant to the present invention. Terms such as "a," "an," and "the" are not intended to refer to a singular entity only, but include general classifications for which specific examples may be used for purposes of illustration. While terms herein are used to describe particular embodiments of the present invention, their use does not delimit the scope of the invention except as outlined in the claims.

[0018] "Influenza infection" refers to the pathogenic proliferation of influenza virus in a host organism (e.g., a human subject). Influenza infection can be any situation in which the presence of influenza virus populations is damaging to the host's body. Thus, a subject "suffers" from an influenza infection when excessive influenza populations are present in or on the subject's body, or when the presence of virus population(s) is damaging to the subject's cells or other tissues.

[0019] As used herein, the term "Fc domain monomer" refers to a monomer that contains at least a hinge domain and a second and third antibody constant domain (C H 2 and C H3) or a functional fragment thereof (e.g., a fragment that is (i) capable of dimerizing with another Fc domain monomer to form an Fc domain, and (ii) capable of binding to an Fc receptor). The Fc domain monomer can be of any immunoglobulin antibody isotype, including IgG, IgE, IgM, IgA, or IgD (e.g., IgG). Additionally, the Fc domain monomer can be of an IgG subtype (e.g., IgG1, IgG2a, IgG2b, IgG3, or IgG4) (e.g., IgG1). The Fc domain monomer does not contain any portion of an immunoglobulin that can act as an antigen recognition region, e.g., a variable domain or a complementarity-determining region (CDR). The Fc domain monomer in the conjugates described herein can include one or more changes (e.g., 1-10, 1-8, 1-6, 1-4 amino acid substitutions, additions, or deletions) from the wild-type Fc domain monomer sequence that alter the interaction between the Fc domain and an Fc receptor. Examples of suitable alterations are known in the art. In certain embodiments, a human Fc domain monomer (e.g., an IgG heavy chain such as IgG1) comprises a region extending from any of Asn208, Glu216, Asp221, Lys222, or Cys226 to the carboxyl terminus of the heavy chain at Lys447. The C-terminal Lys447 of the Fc region may be present or absent and does not affect the structure or stability of the Fc region. The C-terminal Lys447 may be proteolytically cleaved upon expression of the polypeptide. In some embodiments of any of the Fc domain monomers described herein, the C-terminal Lys447 is optionally present or absent. The present disclosure expressly contemplates any of SEQ ID NOs: 1-4, 11, 16, 19, 20, 32-37, 48-53, and 60-68 that do not include the C-terminal Lys corresponding to Lys447. The N-terminal N(Asn) of the Fc region (e.g., any one of SEQ ID NOs: 60 to 77) may or may not be present and does not affect the structural stability of the Fc region. The N-terminal Asn can be deamidated upon expression of the polypeptide.In some embodiments of any of the Fc domain monomers described herein, the N-terminal Asn is optionally present or absent. The present disclosure expressly contemplates any of SEQ ID NOS: 60-77 without the N-terminal Asn. Unless otherwise specified herein, the numbering of amino acid residues in an IgG or Fc domain monomer is according to the EU numbering system for antibodies, also known as the Kabat EU index, as described, e.g., in Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD, 1991.

[0020] As used herein, the term "Fc domain" refers to a dimer of two Fc domain monomers capable of binding to an Fc receptor. In a wild-type Fc domain, the two Fc domain monomers are linked together by two C H The interactions between the three antibody constant domains result in dimerization, and in some embodiments, one or more disulfide bonds are formed between the hinge domains of the two dimerized Fc domain monomers.

[0021] As used herein, a "surface-exposed amino acid" or "solvent-exposed amino acid" (e.g., a surface-exposed cysteine ​​or a surface-exposed lysine) refers to an amino acid that is accessible to the solvent surrounding a protein. A surface-exposed amino acid can be a naturally occurring or engineered variant (e.g., a substitution or insertion) of a protein. In some embodiments, a surface-exposed amino acid is an amino acid that, when substituted, does not substantially alter the three-dimensional structure of a protein.

[0022] As used herein, the term "treating" refers to administering a pharmaceutical composition for prophylactic and / or therapeutic purposes. "Preventing disease" refers to prophylactic treatment of a subject who is not yet sick, but who is susceptible to or otherwise at risk for a particular disease. "Treating" a human subject with influenza infection means administering treatment to a subject who already has an influenza infection to improve or stabilize the subject's condition. Thus, in the claims and embodiments, treating is administration to a subject for either therapeutic or prophylactic purposes.

[0023] As used herein, the term "T" refers to the number of neuraminidase inhibitor dimers conjugated to an Fc domain in a population of conjugates. In some embodiments, the average number of neuraminidase inhibitor dimers conjugated to an Fc domain monomer in a population of conjugates (i.e., the average / mean value of T) can be 1 to 20 (e.g., the average value of T is 3 to 6 or 3.5 to 5.5). In some embodiments, the average value of T is 4.5.

[0024] As used herein, the term "pharmaceutical composition" refers to a medicinal or pharmaceutical preparation comprising a conjugate of formula (I) and one or more excipients and diluents to make the active ingredient suitable for the method of administration. The pharmaceutical composition of the present disclosure comprises pharmaceutically acceptable ingredients that are compatible with the conjugate of formula (I).

[0025] As used herein, the term "pharmaceutically acceptable carrier" refers to an excipient or diluent in a pharmaceutical composition. For example, a pharmaceutically acceptable carrier may be a vehicle capable of suspending or dissolving the conjugate of formula (I). A pharmaceutically acceptable carrier must be compatible with other ingredients of the formulation and not harmful to the recipient. In the present disclosure, a pharmaceutically acceptable carrier must provide reasonable pharmaceutical stability for the conjugate described herein.

[0026] As used herein, the term "pharmaceutically acceptable salt" refers to a salt of the conjugate of Formula (I) that is suitable, within the scope of sound medical judgment, for use in the methods described herein without undue toxicity, irritation, and / or allergic response. Pharmaceutically acceptable salts are well known in the art. For example, pharmaceutically acceptable salts are described in Pharmaceutical Salts: Properties, Selection, and Use (Eds. P.H. Stahl and C.G. Wermuth), Wiley-VCH, 2008. Salts can be prepared in situ during the final isolation and purification of the conjugates described herein, or separately by reacting the free base group with a suitable organic acid.

[0027] The term "drug-to-antibody ratio" or "DAR" refers to the average number of small molecule drug moieties (e.g., the average number of small molecule drug monomers or dimers) conjugated to an Fc domain. In some embodiments described herein, the DAR is represented by "T" (e.g., in Formula (I)). As used herein, each dimer moiety conjugated to an Fc domain corresponds to a DAR value of 1.0 (e.g., a "T" value of 1.0). For example, an Fc domain conjugated to four zanamivir dimers has a DAR of 4.0 (e.g., a "T" of 4.0). The DAR can also be calculated as the average DAR for a population of molecules, such as a population of Fc domains. The DAR value can affect the efficacy, potency, pharmacokinetics, or toxicity of a drug.

[0028] "Conjugate of formula (I)" means [ka] where each E comprises an Fc domain monomer, n is 2, T is an integer from 3 to 6, and the wavy line indicates the covalent bond of E to the nitrogen atom of a solvent-exposed lysine or the sulfur atom of a solvent-exposed cysteine. In this conjugate, conjugation of zanamivir dimers is at various solvent-exposed lysines on the Fc domain monomers.

[0029] "Conjugate A" means a conjugate of formula (I) in which the average value of T is about 4.5 and each E has the sequence of SEQ ID NO:76. [Brief explanation of the drawings]

[0030] [Figure 1] A is a summary of the clinical trial protocol, and B is a summary of the clinical trial details. [Figure 2] Summary of the demographics of human subjects in clinical trials. [Figure 3] Box plot of viral load-area under the curve (VL-AUC) data for placebo and conjugate A. [Figure 4] 1 is a graph of the 8-day mean viral load for placebo and conjugate A by qRT-PCR. [Figure 5] Box plots of peak viral load data for placebo and conjugate A. [Figure 6] FIG. 16 is a graph showing time to confirmed negative influenza test by qRT-PCR for placebo and conjugate A. [Figure 7] 1 is a box plot of the area under the curve of the total clinical symptom score (TSS-AUC) over time for placebo and conjugate A. [Figure 8] 1 is a graph of the mean Total Clinical Symptom Score (TSS) over time for placebo and conjugate A. [Figure 9] 1 is a box plot of peak Total Clinical Symptom Score (TSS) for placebo and conjugate A. [Figure 10]1 is a graph of time to symptom resolution for placebo and conjugate A. [Figure 11] 1 is a graph of the mean plasma concentration of a single dose of conjugate A versus time for 50 mg of conjugate A administered by intramuscular or subcutaneous injection. [Figure 12] 1 is a graph of the mean plasma concentration of a single dose of conjugate A versus time for 150 mg of conjugate A administered by intramuscular or subcutaneous injection. [Figure 13] 1 is a graph of the mean plasma concentration of a single dose of conjugate A versus time for 50 or 150 mg of conjugate A administered by intramuscular or subcutaneous injection on a semi-logarithmic scale. [Figure 14] 1 is a graph of the mean plasma concentration of a single dose of conjugate A versus time for 50, 150, or 450 mg of conjugate A administered by intramuscular or subcutaneous injection on a semi-logarithmic scale. [Figure 15] 1 is a graph showing the mean AUC of conjugate A as a function of dose (mg) of conjugate A when conjugate A is administered intramuscularly and subcutaneously. [Figure 16] Graph showing predicted median (line) and 10th and 90th percentiles (shaded gray areas) of conjugate A concentration versus individual Cmax. Day 5 is shown as a vertical dotted line. The gray rectangle is the in-hospital period for participants in the human challenge study. [Figure 17] 1 is a graph showing model-predicted plasma concentrations for Conjugate A administered as one 150 mg dose, one 300 mg dose, or two 150 mg doses. DETAILED DESCRIPTION OF THE INVENTION

[0031] influenza The conjugates and pharmaceutical compositions described herein can be used to treat influenza infections, such as influenza A, B, or C infections.

[0032] Influenza viruses are associated with serious human illnesses and cause annual epidemics in the fall and winter. While most people recover from seasonal influenza within 1–2 weeks without needing medical treatment, millions of people are hospitalized worldwide each year, and approximately 650,000 influenza-related deaths occur, particularly among infants, the elderly, and those with chronic illnesses. In the United States (US), the 2018–2019 influenza season saw an estimated 37.4–42.9 million cases of symptomatic influenza-related illness, 17.3–20.1 million influenza-related medical visits, 531,000–647,000 influenza-related hospitalizations, and 63,400–61,200 deaths. Comparable mortality and morbidity rates have been reported in European countries. Because no effective drugs are currently available to prevent influenza, these figures remain high year after year. Currently, influenza A viruses, H1N1 and H3N2, along with influenza B viruses, circulate in humans. H3N2 viruses have dominated most seasons, causing more deaths and hospitalizations than H1N1 and influenza B viruses.

[0033] Progress toward a drug, also known as a "universal vaccine," that would provide protection against a wide range of influenza strains and provide extended duration of protection has been disappointing. Monoclonal antibody therapeutics developed to date suffer from limited spectrum and commercial limitations due to the need for high doses and / or multiple antibody cocktails to achieve the desired spectrum and efficacy. Thus, there is a significant unmet need for long-acting universal protective agents.

[0034] Influenza human challenge models have been established to aid in understanding influenza disease and transmission, as well as to evaluate the efficacy of antivirals, immunomodulators, and vaccines. The influenza H3N2 A / Perth / 16 / 2009 challenge strain has been used in the majority of trials to date. This challenge virus has been shown to induce a measurable disease profile that is clearly distinct from uninfected participants, with approximately a 60% to 75% chance of study participants becoming infected after administration of this virus. Typical influenza illness is characterized by a sudden onset of rhinitis, nasal congestion, fever, fatigue, myalgia (muscle pain), and sore throat. In healthy adults, the disease typically resolves without any treatment, with symptom relief occurring spontaneously within 3 to 5 days. The disease profile of the challenge agent is consistent with the mild to moderate disease profile expected with wild-type challenge viruses in healthy adult participants. In summary, the influenza H3N2 A / Perth / 16 / 2009 challenge virus is safe, well tolerated, and induces disease pathology appropriate to be an effective viral challenge agent in human viral challenge (HVC) studies.

[0035] Conjugate A - Long-acting antiviral Fc conjugate Conjugate A is a conjugate of zanamivir and a crystallizable fragment (Fc). Zanamivir is a small molecule antiviral neuraminidase inhibitor. The targeting group of this small molecule selectively engages a small conserved pocket on the viral surface, which cannot be achieved with monoclonal antibodies.

[0036] Conjugate A differs from conventional antibody-drug conjugate molecules in the following aspects: In conventional antibody-drug conjugates, the drug is conjugated to full-length human IgG (Fc and antigen-binding fragment). In contrast, the zanamivir dimer in Conjugate A is conjugated to the Fc fragment of human IgG1 (not full-length IgG1). In this conjugate, conjugation of the zanamivir dimer occurs at different solvent-exposed lysines on each Fc domain monomer. In some embodiments, the zanamivir dimer is conjugated to one or more solvent-exposed lysines, such as Lys205, Lys213, Lys218, Lys246, Lys317, Lys326, Lys334, Lys392, or a combination thereof.

[0037] In conventional antibody-drug conjugates, a protease-cleavable linker is used to conjugate the drug to human IgG, allowing for release of the drug inside target cells. The linker between zanamivir and Fc in conjugate A is not a substrate for proteases, which allows it to exert its antiviral activity in the extracellular space.

[0038] While traditional antibody-drug conjugates are used to treat cancer by rapidly releasing and delivering a cytotoxic payload to target cells, Conjugate A is designed to treat and prevent infectious diseases using a long-acting, stable conjugate of a non-cytotoxic small molecule to the Fc fragment of IgG1.

[0039] Methods for making conjugate A are described in US Pat. No. 11,510,992, which is incorporated herein by reference in its entirety.

[0040] Fc domain The Fc domain is a dimer of two Fc domain monomers, which generally consist of a hinge domain, a C domain, and a C domain. H 2 antibody constant domains, and C HThe Fc domain monomer comprises three antibody constant domains. The Fc domain monomer can be of the immunoglobulin antibody isotype IgG, IgE, IgM, IgA, or IgD. The Fc domain monomer can be of any immunoglobulin antibody isotype (e.g., IgG1, IgG2a, IgG2b, IgG3, or IgG4). The Fc domain monomer can be of any immunoglobulin antibody allotype (e.g., IGH1, IGH2a, IGH3, IGH4, IGH5, IGH6, IGH7, IGH8, IGH9, IGH10, IGH11, IGH12, IGH13, IGH14, IGH15, IGH16, IGH17, IGH18, IGH19, IGH20, IGH21, IGH22, IGH23, IGH24, IGH25, IGH26, IGH27, IGH28, IGH29, IGH30, IGH31, IGH32, IGH33, IGH34, IGH35, IGH36, IGH37, IGH38, IGH40, IGH41, IGH42, IGH43, IGH44, IGH45, IGH46, IGH47, IGH48, IGH49, IGH50, IGH51, IGH52, IGH53, IGH54, IGH55, IGH56, IGH57, IGH58, IGH59, IGH60, IGH61, IGH62, IGH63, IGH64, IGH65, IGH70, IGH71, IGH72, IGH73, IGH74, IGH75, IGH76, IGH77, IGH78, IGH79, IGH71, IGH71, IGH72, IGH72, IGH73, IGH74, IGH75 * 01 (i.e., G1m(za)), IGHG1 * 07 (i.e., G1m(zax)), IGHG1 * 04 (i.e., G1m(zav)), IGHG1 * 03(G1m(f)), IGHG1 * 08 (i.e., G1m(fa)), IGHG2 * 01, IGHG2 * 06, IGHG2 * 02, IGHG3 * 01, IGHG3 * 05, IGHG3 * 10.IGHG3 * 04, IGHG3 * 09, IGHG3 * 11.IGHG3 * 12, IGHG3 * 06, IGHG3 * 07, IGHG3 * 08, IGHG3 * 13.IGHG3 * 03, IGHG3 * 14, IGHG3 * 15, IGHG3 * 16, IGHG3 * 17, IGHG3 * 18, IGHG3 * 19, IGHG2 * 04, IGHG4 * 01, IGHG4 * 03, or IGHG4 *02) (e.g., as described in Vidarsson et al. IgG subclasses and allotypes: from structure to effector function. Frontiers in Immunology. 5(520):1-17(2014)). The Fc domain monomer can be from any species (e.g., human, murine, or mouse). A dimer of Fc domain monomers (i.e., Fc domains) can bind to an Fc receptor, a receptor on the surface of a white blood cell.

[0041] In some embodiments, an Fc domain monomer in a conjugate described herein can include one or more amino acid substitutions, additions, and / or deletions relative to an Fc domain monomer having the sequence of any one of SEQ ID NOS: 1-138. In some embodiments, an Asn in an Fc domain monomer in a conjugate described herein can be replaced by Ala to prevent N-linked glycosylation (e.g., an Asn→Ala substitution can be * In some embodiments, the Fc domain monomers in the conjugates described herein may also include an additional Cys addition (e.g., a Cys addition * (See SEQ ID NOS: 9, 10, and 11, which are represented by the formula:

[0042] In some embodiments, the Fc domain monomer in the conjugates described herein comprises an additional moiety attached to the N-terminus or C-terminus of the Fc domain monomer, such as an albumin binding peptide, a purification peptide (e.g., a hexa-histidine peptide (HHHHHH (SEQ ID NO: 146)), or a signal sequence (e.g., the IL2 signal sequence MYRMQLLSCIALSLALVTNS (SEQ ID NO: 147)). In some embodiments, the Fc domain monomer in the conjugate comprises any type of antibody variable region, e.g., a V H , V L , complementarity determining regions (CDRs), or hypervariable regions (HVRs).

[0043] In some embodiments, an Fc domain monomer in a conjugate described herein may have a sequence that is at least 95% identical (e.g., 97%, 99%, or 99.5% identical) to any one of SEQ ID NOs: 1-138 shown below. In some embodiments, an Fc domain monomer in a conjugate described herein may have the sequence of any one of SEQ ID NOs: 1-138 shown below.

[0044] SEQ ID NO: 1: Murine Fc-IgG2a containing an N-terminal IL2 signal sequence (bold)

[0045] TIFF2026508522000003.tif32170

[0046] SEQ ID NO: 2: Mature murine Fc-IgG2a PRGPTIKPCPPCKCPAPNLLGGPSVFIFPPKIKDVLMISLSPIVTCVVVDVSEDDPDVQISWFVNNVEVHTAQTQTHREDYNSTLRVVSALPIQHQDWMSGKEFKCKVNNKDLPAP IERTISKPKGSVRAPQVYVLPPPEEEMTKKQVTLTCMVTDFMPEDIYVEWTNNGKTELNYKNTEPVLDSDGSYFMYSKLRVEKKNWVERNSYSCSVVHEGLHNHHTTKSFSRTPGK

[0047] SEQ ID NO: 3: Human Fc-IgG1 containing an N-terminal IL2 signal sequence (bold) and additional N-terminal MVRS amino acid residues (underlined)

[0048] TIFF2026508522000004.tif32170

[0049] SEQ ID NO: 4: Mature human Fc-IgG1 containing added N-terminal MVRS amino acid residues (underlined)

[0050] TIFF2026508522000005.tif32170

[0051] SEQ ID NO: 5: Murine Fc-IgG2a containing an N-terminal IL2 signal sequence (bold) and a C-terminal hexa-histidine peptide (italics)

[0052] TIFF2026508522000006.tif32170

[0053] SEQ ID NO: 6: Mature murine Fc-IgG2a containing a C-terminal hexa-histidine peptide (italics)

[0054] TIFF2026508522000007.tif32170

[0055] SEQ ID NO: 7: Human Fc-IgG1 with an IL2 signal sequence at the N-terminus (bold), additional N-terminal MVRS amino acid residues (underlined), and a hexa-histidine peptide at the C-terminus (italics)

[0056] TIFF2026508522000008.tif32170

[0057] SEQ ID NO: 8: Mature human Fc-IgG1 with a C-terminal hexa-histidine peptide (italics) and additional N-terminal MVRS amino acid residues (underlined)

[0058] TIFF2026508522000009.tif32170

[0059] SEQ ID NO: 9: IL2 signal sequence at the N-terminus (bold), additional N-terminal MVRS amino acid residues (underlined), two additional cysteines in the hinge region ( * ), and human Fc-IgG1 with a hexa-histidine peptide (italics) at the C-terminus

[0060] TIFF2026508522000010.tif31170

[0061] SEQ ID NO: 10: Added N-terminal MVRS amino acid residues (underlined), two additional cysteines in the hinge region ( * ), and mature human Fc-IgG1 with a hexa-histidine peptide (italics) at the C-terminus

[0062] TIFF2026508522000011.tif33170

[0063] SEQ ID NO: 11: Additional N-terminal MVRS amino acid residues (underlined) and two additional cysteines in the hinge region ( * ) mature human Fc-IgG1

[0064] TIFF2026508522000012.tif34170

[0065] SEQ ID NO: 12: IL2 signal sequence at the N-terminus (bold), Asn→Ala substitution ( * ), and murine Fc-IgG2a with a hexa-histidine peptide (italics) at the C-terminus

[0066] TIFF2026508522000013.tif32170

[0067] SEQ ID NO: 13: Asn → Ala substitution ( * ) and mature murine Fc-IgG2a with a hexa-histidine peptide (italics) at the C-terminus

[0068] TIFF2026508522000014.tif33170

[0069] SEQ ID NO: 14: IL2 signal sequence at the N-terminus (bold), added N-terminal MVRS amino acid residues (underlined), Asn→Ala substitution ( * ), and human Fc-IgG1 with a hexa-histidine peptide (italics) at the C-terminus

[0070] TIFF2026508522000015.tif32170

[0071] SEQ ID NO: 15: Asn → Ala substitution ( * ), mature human Fc-IgG1 with added N-terminal MVRS amino acid residues (underlined), and a hexa-histidine peptide (italicized) at the C-terminus.

[0072] TIFF2026508522000016.tif32170

[0073] SEQ ID NO: 16: Human IgG1 Fc with N-terminal human serum albumin signal sequence (bold) and additional N-terminal ISAMVRS amino acid residues (underlined)

[0074] TIFF2026508522000017.tif32170

[0075] SEQ ID NO: 17: Human IgG1 Fc with an N-terminal human serum albumin signal sequence (bold), additional N-terminal ISAMVRS amino acid residues (underlined), a C-terminal G4S linker (italicized), and a C-terminal c-Myc tag (underlined, italicized).

[0076] TIFF2026508522000018.tif32170

[0077] SEQ ID NO: 18: Mature human IgG1 Fc with added N-terminal ISAMVRS amino acid residues (underlined), a C-terminal G4S linker (italicized), and a C-terminal c-Myc tag (underlined, italicized).

[0078] TIFF2026508522000019.tif32170

[0079] SEQ ID NO: 19: Human serum albumin signal sequence (bold), added N-terminal ISAMVRS amino acid residues (underlined), and lysine to serine modification () to prevent lysine conjugation at this site * ) containing human IgG1 Fc

[0080] TIFF2026508522000020.tif32170

[0081] SEQ ID NO: 20: Added N-terminal ISAMVRS amino acid residue (underlined) and lysine to serine modification ( ) to prevent lysine conjugation at this site * ) containing mature human IgG1 Fc

[0082] TIFF2026508522000021.tif32170

[0083] SEQ ID NO: 21: Human serum albumin signal sequence at the N-terminus (bold), added N-terminal ISAMVRS amino acid residues (underlined), lysine to serine modification ( ) to prevent lysine conjugation at this site * ), a C-terminal G4S linker (italics), and a C-terminal C-Myc tag (underlined, italics).

[0084] TIFF2026508522000022.tif32170

[0085] SEQ ID NO: 22: Added N-terminal ISAMVRS amino acid residue (underlined), lysine to serine modification ( ) to prevent lysine conjugation at this site * ), a mature human IgG1 Fc containing a C-terminal G4S linker (italics), and a C-terminal C-Myc tag (underlined, italics).

[0086] TIFF2026508522000023.tif32170

[0087] SEQ ID NO: 23: Human serum albumin signal sequence at the N-terminus (bold), added N-terminal ISAMVRS amino acid residues (underlined), Asn→Ala substitution ( * ), a C-terminal G4S linker (italics), and a C-terminal C-myc tag (underlined, italics).

[0088] TIFF2026508522000024.tif32170

[0089] SEQ ID NO: 24: Added N-terminal ISAMVRS amino acid residue (underlined), Asn→Ala substitution ( * ), a mature human IgG1 Fc containing a C-terminal G4S linker (italics), and a C-terminal C-myc tag (underlined, italics).

[0090] TIFF2026508522000025.tif32170

[0091] SEQ ID NO: 25: Human serum albumin signal sequence at the N-terminus (bold), added N-terminal ISAMVRS amino acid residues (underlined), H310A ( * ) and H435A( * ) mutation, a C-terminal G4S (italicized), and a C-terminal C-myc tag (underlined, italicized).

[0092] TIFF2026508522000026.tif38170

[0093] SEQ ID NO: 26: Human serum albumin signal sequence at the N-terminus (bold), added N-terminal ISAMVRS amino acid residues (underlined), H310A ( * ) and H435A( * ) mutation, a C-terminal G4S (italicized), and a C-terminal C-myc tag (underlined, italicized).

[0094] TIFF2026508522000027.tif32170

[0095] SEQ ID NO: 27: Human IgG1 Fc with an N-terminal human serum albumin signal sequence (bold), additional N-terminal ISAMVRS amino acid residues (underlined), a C-terminal G4S linker (italicized), and a C-terminal mutated (lysine to phenylalanine, bolded) C-myc tag (underlined, italicized).

[0096] TIFF2026508522000028.tif32170

[0097] SEQ ID NO: 28: Mature human IgG1 Fc containing added N-terminal ISAMVRS amino acid residues (underlined), a C-terminal G4S linker (italicized), and a C-terminal mutated (lysine to phenylalanine, bold) C-myc tag (underlined, italicized).

[0098] TIFF2026508522000029.tif32170

[0099] SEQ ID NO: 29: Human serum albumin signal sequence at the N-terminus (bold), added N-terminal ISAMVRS amino acid residues (underlined), Asn→Ala substitution ( * ), a C-terminal G4S linker (italics), and a C-terminal mutated (lysine to phenylalanine, bold) C-myc tag (underlined, italics).

[0100] TIFF2026508522000030.tif34170

[0101] SEQ ID NO: 30: Added N-terminal MVRS amino acid residue (underlined), Asn→Ala substitution ( * ), a C-terminal G4S linker (italics), and a C-terminal mutated (lysine to phenylalanine, bold) C-myc tag (underlined, italics).

[0102] TIFF2026508522000031.tif32170

[0103] SEQ ID NO: 31: Human IgG1 Fc with N-terminal human serum albumin signal sequence (bold), allotype G1m(fa) (bold italics), C-terminal G4S linker (italics), and C-terminal mutated (lysine to phenylalanine, bold) C-myc tag (underlined).

[0104] TIFF2026508522000032.tif34170

[0105] SEQ ID NO: 32: Human IgG1 Fc with human serum albumin signal sequence at the N-terminus (bold), allotype G1m(fa) (bold italics)

[0106] TIFF2026508522000033.tif32170

[0107] SEQ ID NO: 33: Mature human IgG1 Fc with YTE triple mutation (bold and underlined) and additional N-terminal MVRS amino acid residues (underlined)

[0108] TIFF2026508522000034.tif32170

[0109] SEQ ID NO: 34: Human IgG1 Fc with residues EPKSS (underlined), Cys→Ser substitution (#), allotype G1m(fa) (bold italics), containing the complete hinge region at the N-terminus of mature human IgG1 Fc, with a human serum albumin signal sequence (bold) at the N-terminus.

[0110] TIFF2026508522000035.tif32170

[0111] SEQ ID NO: 35: Human IgG1 Fc with murine IgG signal sequence at the N-terminus (bold), EPKSSD hinge residues removed from the N-terminus of mature human IgG1 Fc, and allotype G1m(fa) (bold italics)

[0112] TIFF2026508522000036.tif32170

[0113] SEQ ID NO: 36: Mature human IgG1 Fc with the YTE triple mutation (bold and underlined), removal of the EPKSSD hinge residues from the N-terminus of mature human IgG1 Fc, and allotype G1m(fa) (bold italics)

[0114] TIFF2026508522000037.tif43170

[0115] SEQ ID NO: 37: Mature human IgG1 Fc with LS double mutation (bold and underlined), removal of EPKSSD hinge residues from the N-terminus of mature human IgG1 Fc, and allotype G1m(fa) (bold italics)

[0116] TIFF2026508522000038.tif38170

[0117] SEQ ID NO: 38: Mature human IgG1 Fc with N-terminal human serum albumin signal sequence (bold), YTE triple mutation (bold and underlined), allotype G1m(fa) (bold italics), C-terminal G4S linker (italics), and C-terminal C-myc tag (underlined).

[0118] TIFF2026508522000039.tif32170

[0119] SEQ ID NO: 39: Mature human Fc IgG1, X1 is Met or Tyr, X2 is Ser or Thr, X3 is Thr or Glu, X4 is Asp or Glu, X5 is Leu or Met, X6 is Met or Leu, X7 is Asn or Ser

[0120] TIFF2026508522000040.tif32170

[0121] SEQ ID NO: 40: Mature human Fc IgG1, X4 is Asp or Glu, X5 is Leu or Met

[0122] TIFF2026508522000041.tif31170

[0123] SEQ ID NO: 41: Mature human Fc IgG1 with YTE triple mutation (bold and underlined), X4 is Asp or Glu, X5 is Leu or Met

[0124] TIFF2026508522000042.tif32170

[0125] SEQ ID NO: 42: Mature human Fc IgG1 with YTE triple mutation (bold and underlined), allotype G1m(fa) (bold italics)

[0126] TIFF2026508522000043.tif32170

[0127] SEQ ID NO: 43: Mature human Fc IgG1 with YTE triple mutation (bold and underlined), allotype G1m(f) (bold italics)

[0128] TIFF2026508522000044.tif32170

[0129] SEQ ID NO: 44: Mature human Fc IgG1 with LS double mutation (bold and underlined), X4 is Asp or Glu, X5 is Leu or Met

[0130] TIFF2026508522000045.tif32170

[0131] SEQ ID NO: 45: Mature human Fc IgG1 with LS double mutation (bold and underlined), allotype G1m(fa) (bold italics)

[0132] TIFF2026508522000046.tif32170

[0133] SEQ ID NO: 46: Mature human Fc IgG1 with LS double mutation (bold and underlined), allotype G1m(f) (bold italics)

[0134] TIFF2026508522000047.tif32170

[0135] SEQ ID NO: 47: Mouse heavy chain MIgG Vh signal sequence (bold), mature human Fc IgG1 with Cys→Ser substitution (#), X1 is Met or Tyr, X2 is Ser or Thr, X3 is Thr or Glu, X4 is Asp or Glu, X5 is Leu or Met, X6 is Met or Leu, X7 is Asn or Ser

[0136] TIFF2026508522000048.tif38170

[0137] SEQ ID NO: 48: Mature human IgG1 Fc with mouse heavy chain MIgG Vh signal sequence (bold), Cys→Ser substitution (#), allotype G1m(fa) (bold italics)

[0138] TIFF2026508522000049.tif38170

[0139] SEQ ID NO: 49: Mature human IgG1 Fc with mouse heavy chain MIgG Vh signal sequence (bold), Cys→Ser substitution (#), allotype G1m(f) (bold italics)

[0140] TIFF2026508522000050.tif38170

[0141] SEQ ID NO: 50: Mature human IgG1 Fc with mouse heavy chain MIgG Vh signal sequence (bold), Cys→Ser substitution (#), M428L, N434S mutation (bold / underlined), allotype G1m(fa) (bold italics)

[0142] TIFF2026508522000051.tif37170

[0143] SEQ ID NO: 51: Mature human IgG1 Fc with mouse heavy chain MIgG Vh signal sequence (bold), Cys→Ser substitution (#), M428L, N434S mutation (bold / underlined), allotype G1m(f) (bold italics)

[0144] TIFF2026508522000052.tif38170

[0145] SEQ ID NO: 52: Mature human IgG1 Fc with mouse heavy chain MIgG Vh signal sequence (bold), Cys→Ser substitution (#), YTE triple mutation (bold and underlined), allotype G1m(fa) (bold italics)

[0146] TIFF2026508522000053.tif39170

[0147] SEQ ID NO: 53: Mature human IgG1 Fc with mouse heavy chain MIgG Vh signal sequence (bold), Cys→Ser substitution (#), YTE triple mutation (bold and underlined), allotype G1m(f) (bold italics)

[0148] TIFF2026508522000054.tif38170

[0149] SEQ ID NO: 54: Mature human IgG1 Fc with mouse heavy chain MIgG Vh signal sequence (bold), added N-terminal ISAMVRS amino acid residues (italics), M428L, N434S mutations (bold / underlined), G4S linker (italics), and C-terminal C-myc tag (underlined), allotype G1m(f) (bold italics).

[0150] TIFF2026508522000055.tif33170

[0151] SEQ ID NO: 55: Mature human IgG1 Fc with mouse heavy chain MIgG Vh signal sequence (bold), added N-terminal ISAMVRS amino acid residues (italics), M428L, N434S mutations (bold / underlined), G4S linker (italics), C-terminal C-myc tag (underlined), allotype G1m(fa) (bold italics).

[0152] TIFF2026508522000056.tif34170

[0153] SEQ ID NO: 56: Mature human IgG1 Fc with mouse heavy chain MIgG Vh signal sequence (bold), added N-terminal ISAMVRS amino acid residues (italics), YTE triple mutant (bold / underlined), G4S linker (italics), and C-terminal C-myc tag (underlined), allotype G1m(f) (bold italics).

[0154] TIFF2026508522000057.tif32170

[0155] SEQ ID NO: 57: Mature human IgG1 Fc with mouse heavy chain MIgG Vh signal sequence (bold), added N-terminal ISAMVRS amino acid residues (italics), YTE triple mutant (bold / underlined), G4S linker (italics), C-terminal C-myc tag (underlined), allotype G1m(fa) (bold italics).

[0156] TIFF2026508522000058.tif32170

[0157] SEQ ID NO: 58: Mature human IgG1 with mouse heavy chain MIgG1 signal sequence (bold), Cys to Ser substitution (#), C-terminal G4S (italics), and C-terminal IgA peptide (underlined), allotype G1m(fa) (bold italics)

[0158] TIFF2026508522000059.tif43170

[0159] SEQ ID NO: 59: Mature human IgG1 with mouse heavy chain MIgG1 signal sequence (bold), Cys to Ser substitution (#), M428L, N434S mutation (bold / underlined), C-terminal G4S (italics), and C-terminal IgA peptide (underlined), allotype G1m(fa) (bold italics).

[0160] TIFF2026508522000060.tif43170

[0161] SEQ ID NO: 60: Mature human Fc IgG1, Z1 is Cys or Ser, X1 is Met or Tyr, X2 is Ser or Thr, X3 is Thr or Glu, X4 is Asp or Glu, X5 is Leu or Met, X6 is Met or Leu, X7 is Asn or Ser

[0162] TIFF2026508522000061.tif34170

[0163] SEQ ID NO: 61: Mature human Fc IgG1, Cys→Ser substitution (#), X1 is Met or Tyr, X2 is Ser or Thr, X3 is Thr or Glu, X4 is Asp or Glu, X5 is Leu or Met, X6 is Met or Leu, X7 is Asn or Ser

[0164] TIFF2026508522000062.tif32170

[0165] SEQ ID NO: 62: Mature human IgG1 Fc, Cys→Ser substitution (#), X4 is Asp or Glu, X5 is Leu or Met

[0166] TIFF2026508522000063.tif32170

[0167] SEQ ID NO: 63: Mature human IgG1 Fc, Cys→Ser substitution (#), allotype G1m(f) (bold italics)

[0168] TIFF2026508522000064.tif32170

[0169] SEQ ID NO: 64: Mature human IgG1 Fc, Cys→Ser substitution (#), allotype G1m(fa) (bold italics)

[0170] TIFF2026508522000065.tif33170

[0171] SEQ ID NO: 65: Mature human IgG1 Fc, Cys→Ser substitution (#), M428L, N434S mutation (bold / underlined), allotype G1m(fa) (bold italics)

[0172] TIFF2026508522000066.tif33170

[0173] SEQ ID NO: 66: Mature human IgG1 Fc, Cys→Ser substitution (#), M428L, N434S mutation (bold / underlined), allotype G1m(f) (bold italics)

[0174] TIFF2026508522000067.tif32170

[0175] SEQ ID NO: 67: Mature human IgG1 Fc, Cys→Ser substitution (#), YTE triple mutation (bold and underlined), allotype G1m(fa) (bold italics)

[0176] TIFF2026508522000068.tif33170

[0177] SEQ ID NO: 68: Mature human IgG1 Fc, Cys→Ser substitution (#), YTE triple mutation (bold and underlined), allotype G1m(f) (bold italics)

[0178] TIFF2026508522000069.tif33170

[0179] SEQ ID NO: 69: Mature human Fc IgG1, Z1 is Cys or Ser, X1 is Met or Tyr, X2 is Ser or Thr, X3 is Thr or Glu, X4 is Asp or Glu, X5 is Leu or Met, X6 is Met or Leu, X7 is Asn or Ser

[0180] TIFF2026508522000070.tif34170

[0181] SEQ ID NO: 70: Mature human Fc IgG1, Cys→Ser substitution (#), X1 is Met or Tyr, X2 is Ser or Thr, X3 is Thr or Glu, X4 is Asp or Glu, X5 is Leu or Met, X6 is Met or Leu, X7 is Asn or Ser

[0182] TIFF2026508522000071.tif35170

[0183] SEQ ID NO: 71: Mature human IgG1 Fc, Cys→Ser substitution (#), X4 is Asp or Glu, X5 is Leu or Met

[0184] TIFF2026508522000072.tif34170

[0185] SEQ ID NO: 72: Mature human IgG1 Fc, Cys→Ser substitution (#), allotype G1m(f) (bold italics)

[0186] TIFF2026508522000073.tif32170

[0187] SEQ ID NO: 73: Mature human IgG1 Fc, Cys→Ser substitution (#), allotype G1m(fa) (bold italics)

[0188] TIFF2026508522000074.tif34170

[0189] SEQ ID NO: 74: Mature human IgG1 Fc, Cys→Ser substitution (#), M428L, N434S mutation (bold / underlined), allotype G1m(fa) (bold italics)

[0190] TIFF2026508522000075.tif33170

[0191] SEQ ID NO: 75: Mature human IgG1 Fc, Cys→Ser substitution (#), M428L, N434S mutation (bold / underlined), allotype G1m(f) (bold italics)

[0192] TIFF2026508522000076.tif33170

[0193] SEQ ID NO: 76: Mature human IgG1 Fc, Cys→Ser substitution (#), YTE triple mutation (bold and underlined), allotype G1m(fa) (bold italics)

[0194] TIFF2026508522000077.tif33170

[0195] SEQ ID NO: 77: Mature human IgG1 Fc, Cys→Ser substitution (#), YTE triple mutation (bold and underlined), allotype G1m(f) (bold italics)

[0196] TIFF2026508522000078.tif33170

[0197] SEQ ID NO: 78: Mature human Fc IgG1, Z1 is Cys or Ser, X1 is Met or Tyr, X2 is Ser or Thr, X3 is Thr or Glu, X4 is Asp or Glu, X5 is Leu or Met, X6 is Met or Leu, X7 is Asn or Ser

[0198] TIFF2026508522000079.tif33170

[0199] SEQ ID NO: 79: Mature human Fc IgG1, Cys→Ser substitution (#), X1 is Met or Tyr, X2 is Ser or Thr, X3 is Thr or Glu, X4 is Asp or Glu, X5 is Leu or Met, X6 is Met or Leu, X7 is Asn or Ser

[0200] TIFF2026508522000080.tif34170

[0201] SEQ ID NO: 80: Mature human IgG1 Fc, Cys→Ser substitution (#), X4 is Asp or Glu, X5 is Leu or Met

[0202] TIFF2026508522000081.tif33170

[0203] SEQ ID NO: 81: Mature human IgG1 Fc, Cys→Ser substitution (#), allotype G1m(f) (bold italics)

[0204] TIFF2026508522000082.tif33170

[0205] SEQ ID NO: 82: Mature human IgG1 Fc, Cys→Ser substitution (#), allotype G1m(fa) (bold italics)

[0206] TIFF2026508522000083.tif33170

[0207] SEQ ID NO: 83: Mature human IgG1 Fc, Cys→Ser substitution (#), M428L, N434S mutation (bold / underlined), allotype G1m(fa) (bold italics)

[0208] TIFF2026508522000084.tif32170

[0209] SEQ ID NO: 84: Mature human IgG1 Fc, Cys→Ser substitution (#), M428L, N434S mutation (bold / underlined), allotype G1m(f) (bold italics)

[0210] TIFF2026508522000085.tif32170

[0211] SEQ ID NO: 85: Mature human IgG1 Fc, Cys→Ser substitution (#), YTE triple mutation (bold and underlined), allotype G1m(fa) (bold italics)

[0212] TIFF2026508522000086.tif33170

[0213] SEQ ID NO: 86: Mature human IgG1 Fc, Cys→Ser substitution (#), YTE triple mutation (bold and underlined), allotype G1m(f) (bold italics)

[0214] TIFF2026508522000087.tif34170

[0215] SEQ ID NO: 87: Mature human Fc IgG1, Z1 is Cys or Ser, X1 is Met or Tyr, X2 is Ser or Thr, X3 is Thr or Glu, X4 is Asp or Glu, X5 is Leu or Met, X6 is Met or Leu, X7 is Asn or Ser

[0216] TIFF2026508522000088.tif33170

[0217] SEQ ID NO: 88: Mature human Fc IgG1, Cys→Ser substitution (#), X1 is Met or Tyr, X2 is Ser or Thr, X3 is Thr or Glu, X4 is Asp or Glu, X5 is Leu or Met, X6 is Met or Leu, X7 is Asn or Ser

[0218] TIFF2026508522000089.tif33170

[0219] SEQ ID NO: 89: Mature human IgG1 Fc, Cys→Ser substitution (#), X4 is Asp or Glu, X5 is Leu or Met

[0220] TIFF2026508522000090.tif33170

[0221] SEQ ID NO: 90: Mature human IgG1 Fc, Cys→Ser substitution (#), allotype G1m(f) (bold italics)

[0222] TIFF2026508522000091.tif34170

[0223] SEQ ID NO: 91: Mature human IgG1 Fc, Cys→Ser substitution (#), allotype G1m(fa) (bold italics)

[0224] TIFF2026508522000092.tif34170

[0225] SEQ ID NO: 92: Mature human IgG1 Fc, Cys→Ser substitution (#), M428L, N434S mutation (bold / underlined), allotype G1m(fa) (bold italics)

[0226] TIFF2026508522000093.tif33170

[0227] SEQ ID NO: 93: Mature human IgG1 Fc, Cys→Ser substitution (#), M428L, N434S mutation (bold / underlined), allotype G1m(f) (bold italics)

[0228] TIFF2026508522000094.tif32170

[0229] SEQ ID NO: 94: Mature human IgG1 Fc, Cys→Ser substitution (#), YTE triple mutation (bold and underlined), allotype G1m(fa) (bold italics)

[0230] TIFF2026508522000095.tif34170

[0231] SEQ ID NO: 95: Mature human IgG1 Fc, Cys→Ser substitution (#), YTE triple mutation (bold and underlined), allotype G1m(f) (bold italics)

[0232] TIFF2026508522000096.tif33170

[0233] SEQ ID NO: 96: Mature human Fc IgG1, J1 is Asn or absent, J2 is Lys or absent, Z1 is Cys or Ser, X1 is Met or Tyr, X2 is Ser or Thr, X3 is Thr or Glu, X4 is Asn or Ala, X5 is Leu or Asp, X6 is Gln or His, X7 is Asp or Glu, X8 is Leu or Met, X9 is Met or Leu, X 10 is Asn or Ser

[0234] TIFF2026508522000097.tif34170

[0235] SEQ ID NO: 97: Mature human Fc IgG1, Cys→Ser substitution (#), J1 is Asn or absent, J2 is Lys or absent, X4 is Asn or Ala, X5 is Leu or Asp, X6 is Gln or His, X7 is Asp or Glu, X8 is Leu or Met, and X 10 is Asn or Ser

[0236] TIFF2026508522000098.tif32170

[0237] SEQ ID NO: 98: Mature human Fc IgG1, Cys→Ser substitution (#), DHS triple mutation (bold and underlined), J1 is Asn or absent, J2 is Lys or absent, X4 is Asn or Ala, X7 is Asp or Glu, X8 is Leu or Met

[0238] TIFF2026508522000099.tif33170

[0239] SEQ ID NO: 99: Mature human Fc IgG1, Cys→Ser substitution (#), DHS triple mutation (bold and underlined), X4 is Asn or Ala, X7 is Asp or Glu, X8 is Leu or Met

[0240] TIFF2026508522000100.tif32170

[0241] SEQ ID NO: 100: Mature human Fc IgG1, Cys→Ser substitution (#), DHS triple mutation (bold and underlined), X7 is Asp or Glu, X8 is Leu or Met

[0242] TIFF2026508522000101.tif34170

[0243] SEQ ID NO: 101: Mature human Fc IgG1, Cys→Ser substitution (#), DHS triple mutation (bold and underlined), allotype G1m(fa) (bold italics)

[0244] TIFF2026508522000102.tif34170

[0245] SEQ ID NO: 102: Mature human Fc IgG1, Cys→Ser substitution (#), DHS triple mutation (bold and underlined), allotype G1m(f) (bold italics)

[0246] TIFF2026508522000103.tif33170

[0247] SEQ ID NO: 103: Mature human Fc IgG1, Cys→Ser substitution (#), DHS triple mutation (bold and underlined), allotype G1m(fa) (bold italics)

[0248] TIFF2026508522000104.tif34170

[0249] SEQ ID NO: 104: Mature human Fc IgG1, Cys→Ser substitution (#), DHS triple mutation (bold and underlined), allotype G1m(f) (bold italics)

[0250] TIFF2026508522000105.tif34170

[0251] SEQ ID NO: 105: Mature human Fc IgG1, Cys→Ser substitution (#), DHS triple mutation (bold and underlined), allotype G1m(fa) (bold italics)

[0252] TIFF2026508522000106.tif33170

[0253] SEQ ID NO: 106: Mature human Fc IgG1, Cys→Ser substitution (#), DHS triple mutation (bold and underlined), allotype G1m(f) (bold italics)

[0254] TIFF2026508522000107.tif34170

[0255] SEQ ID NO: 107: Mature human Fc IgG1, Cys→Ser substitution (#), DHS triple mutation (bold and underlined), allotype G1m(fa) (bold italics)

[0256] TIFF2026508522000108.tif32170

[0257] SEQ ID NO: 108: Mature human Fc IgG1, Cys→Ser substitution (#), DHS triple mutation (bold and underlined), allotype G1m(f) (bold italics)

[0258] TIFF2026508522000109.tif33170

[0259] SEQ ID NO: 109: Mature human Fc IgG1, Cys→Ser substitution (#), Asn→Ala substitution ( * ), DHS triple mutation (bold and underlined), X7 is Asp or Glu, X8 is Leu or Met

[0260] TIFF2026508522000110.tif32170

[0261] SEQ ID NO: 110: Mature human Fc IgG1, Cys→Ser substitution (#), Asn→Ala substitution ( * ), DHS triple mutation (bold and underlined), allotype G1m(fa) (bold italics)

[0262] TIFF2026508522000111.tif34170

[0263] SEQ ID NO: 111: Mature human Fc IgG1, Cys→Ser substitution (#), Asn→Ala substitution ( * ), DHS triple mutation (bold and underlined), allotype G1m(f) (bold italics)

[0264] TIFF2026508522000112.tif33170

[0265] SEQ ID NO: 112: Mature human Fc IgG1, Cys→Ser substitution (#), Asn→Ala substitution ( * ), DHS triple mutation (bold and underlined), allotype G1m(fa) (bold italics)

[0266] TIFF2026508522000113.tif34170

[0267] SEQ ID NO: 113: Mature human Fc IgG1, Cys→Ser substitution (#), Asn→Ala substitution ( * ), DHS triple mutation (bold and underlined), allotype G1m(f) (bold italics)

[0268] TIFF2026508522000114.tif33170

[0269] SEQ ID NO: 114: Mature human Fc IgG1, Cys→Ser substitution (#), Asn→Ala substitution ( * ), DHS triple mutation (bold and underlined), allotype G1m(fa) (bold italics)

[0270] TIFF2026508522000115.tif35170

[0271] SEQ ID NO: 115: Mature human Fc IgG1, Cys→Ser substitution (#), Asn→Ala substitution ( * ), DHS triple mutation (bold and underlined), allotype G1m(f) (bold italics)

[0272] TIFF2026508522000116.tif33170

[0273] SEQ ID NO: 116: Mature human Fc IgG1, Cys→Ser substitution (#), Asn→Ala substitution ( * ), DHS triple mutation (bold and underlined), allotype G1m(fa) (bold italics)

[0274] TIFF2026508522000117.tif34170

[0275] SEQ ID NO: 117: Mature human Fc IgG1, Cys→Ser substitution (#), Asn→Ala substitution ( * ), DHS triple mutation (bold and underlined), allotype G1m(f) (bold italics)

[0276] TIFF2026508522000118.tif32170

[0277] SEQ ID NO: 118: Mature human Fc IgG1, J1 is Asn or absent, J2 is Lys or absent, X4 is Asn or Ala, X5 is Leu or Asp, X6 is Gln or His, X7 is Asp or Glu, X8 is Leu or Met, and X 10 is Asn or Ser

[0278] TIFF2026508522000119.tif34170

[0279] SEQ ID NO: 119: Mature human Fc IgG1, DHS triple mutation (bold and underlined), J1 is Asn or absent, J2 is Lys or absent, X4 is Asn or Ala, X7 is Asp or Glu, X8 is Leu or Met

[0280] TIFF2026508522000120.tif36170

[0281] SEQ ID NO: 120: Mature human Fc IgG1, DHS triple mutation (bold and underlined), X4 is Asn or Ala, and X7 is Asp or Glu, X8 is Leu or Met

[0282] TIFF2026508522000121.tif34170

[0283] SEQ ID NO: 121: Mature human Fc IgG1, DHS triple mutation (bold and underlined), X7 is Asp or Glu, X8 is Leu or Met

[0284] TIFF2026508522000122.tif33170

[0285] SEQ ID NO: 122: Mature human Fc IgG1, DHS triple mutation (bold and underlined), allotype G1m(fa) (bold italics)

[0286] TIFF2026508522000123.tif34170

[0287] SEQ ID NO: 123: Mature human Fc IgG1, DHS triple mutation (bold and underlined), allotype G1m(f) (bold italics)

[0288] TIFF2026508522000124.tif34170

[0289] SEQ ID NO: 124: Mature human Fc IgG1, DHS triple mutation (bold and underlined), allotype G1m(fa) (bold italics)

[0290] TIFF2026508522000125.tif31170

[0291] SEQ ID NO: 125: Mature human Fc IgG1, DHS triple mutation (bold and underlined), allotype G1m(f) (bold italics)

[0292] TIFF2026508522000126.tif34170

[0293] SEQ ID NO: 126: Mature human Fc IgG1, DHS triple mutation (bold and underlined), allotype G1m(fa) (bold italics)

[0294] TIFF2026508522000127.tif32170

[0295] SEQ ID NO: 127: Mature human Fc IgG1, DHS triple mutation (bold and underlined), allotype G1m(f) (bold italics)

[0296] TIFF2026508522000128.tif33170

[0297] SEQ ID NO: 128: Mature human Fc IgG1, DHS triple mutation (bold and underlined), allotype G1m(fa) (bold italics)

[0298] TIFF2026508522000129.tif33170

[0299] SEQ ID NO: 129: Mature human Fc IgG1, DHS triple mutation (bold and underlined), allotype G1m(f) (bold italics)

[0300] TIFF2026508522000130.tif34170

[0301] SEQ ID NO: 130: Mature human Fc IgG1, Asn→Ala substitution ( * ), DHS triple mutation (bold and underlined), X7 is Asp or Glu, X8 is Leu or Met

[0302] TIFF2026508522000131.tif33170

[0303] SEQ ID NO: 131: Mature human Fc IgG1, Asn→Ala substitution ( * ), DHS triple mutation (bold and underlined), allotype G1m(fa) (bold italics)

[0304] TIFF2026508522000132.tif34170

[0305] SEQ ID NO: 132: Mature human Fc IgG1, Asn→Ala substitution ( * ), DHS triple mutation (bold and underlined), allotype G1m(f) (bold italics)

[0306] TIFF2026508522000133.tif34170

[0307] SEQ ID NO: 133: Mature human Fc IgG1, Asn→Ala substitution ( * ), DHS triple mutation (bold and underlined), allotype G1m(fa) (bold italics)

[0308] TIFF2026508522000134.tif33170

[0309] SEQ ID NO: 134: Mature human Fc IgG1, Asn→Ala substitution ( * ), DHS triple mutation (bold and underlined), allotype G1m(f) (bold italics)

[0310] TIFF2026508522000135.tif34170

[0311] SEQ ID NO: 135: Mature human Fc IgG1, Asn→Ala substitution ( * ), DHS triple mutation (bold and underlined), allotype G1m(fa) (bold italics)

[0312] TIFF2026508522000136.tif33170

[0313] SEQ ID NO: 136: Mature human Fc IgG1, Asn→Ala substitution ( * ), DHS triple mutation (bold and underlined), allotype G1m(f) (bold italics)

[0314] TIFF2026508522000137.tif31170

[0315] SEQ ID NO: 137: Mature human Fc IgG1, Asn→Ala substitution ( * ), DHS triple mutation (bold and underlined), allotype G1m(fa) (bold italics)

[0316] TIFF2026508522000138.tif34170

[0317] SEQ ID NO: 138: Mature human Fc IgG1, Asn→Ala substitution ( * ), DHS triple mutation (bold and underlined), allotype G1m(f) (bold italics)

[0318] TIFF2026508522000139.tif33170

[0319] As defined herein, an Fc domain is a C H The Fc domain comprises two Fc domain monomers that are dimerized by interactions between three antibody constant domains and one or more disulfide bonds formed between the hinge domains of the two dimerized Fc domain monomers. The Fc domain forms a minimal structure that binds to an Fc receptor, e.g., an Fc-gamma receptor (i.e., an Fcγ receptor (FcγR)), an Fc-alpha receptor (i.e., an Fcα receptor (FcαR)), an Fc-epsilon receptor (i.e., an Fcε receptor (FcεR)), and / or an embryonic Fc receptor (FcRn). In some embodiments, the Fc domains of the present invention bind to an Fcγ receptor (e.g., FcRn, FcγRI (CD64), FcγRIIa (CD32), FcγRIIb (CD32), FcγRIIIa (CD16a), FcγRIIIb (CD16b)), and / or FcγRIV and / or fetal Fc receptor (FcRn).

[0320] In some embodiments, the Fc domain of the invention is an aglycosylated Fc domain (e.g., an Fc domain that maintains engagement with an Fc receptor (e.g., FcRn)). For example, the Fc domain is an aglycosylated IgG1 variant that maintains engagement with an Fc receptor (e.g., an IgG1 with amino acid substitutions at glycosylation motifs N297 and / or T299). Exemplary aglycosylated Fc domains and methods for making aglycosylated Fc domains are known in the art, for example, as described in Sazinsky SLet et al., Aglycosylated immunoglobulin G1 variants productively engage activating Fc receptors, PNAS, 2008, 105(51):20167-20172 (incorporated herein by reference in its entirety).

[0321] In some embodiments, the Fc domain of the present invention is engineered to enhance binding to the fetal Fc receptor (FcRn). For example, the Fc domain may comprise a triple mutation corresponding to M252Y / S254T / T256E (YTE) (e.g., a human or humanized IgG1 with a YTE mutation, e.g., an IgG1 such as SEQ ID NO:33, SEQ ID NO:36, SEQ ID NO:38, SEQ ID NO:39, SEQ ID NO:41, SEQ ID NO:42, SEQ ID NO:43, SEQ ID NO:52, SEQ ID NO:53, SEQ ID NO:56, or SEQ ID NO:57). The Fc domain may comprise a double mutant corresponding to M428L / N434S (LS) (e.g., an IgG1 such as a human or humanized IgG1 with a LS mutation, e.g., SEQ ID NO:37, SEQ ID NO:39, SEQ ID NO:44, SEQ ID NO:45, SEQ ID NO:46, SEQ ID NO:50, SEQ ID NO:51, SEQ ID NO:54, SEQ ID NO:55, or SEQ ID NO:59). The Fc domain may comprise a single mutant corresponding to N434H (e.g., an IgG1 such as a human or humanized IgG1 with an N434H mutation). The Fc domain may comprise a single mutation corresponding to C220S (e.g., an IgG1 such as a human or humanized IgG1 with a C220S mutation such as SEQ ID NO:34, SEQ ID NO:47, SEQ ID NO:48, SEQ ID NO:49, SEQ ID NO:50, SEQ ID NO:51, SEQ ID NO:52, SEQ ID NO:53, SEQ ID NO:58, SEQ ID NO:59, SEQ ID NO:61, SEQ ID NO:62, SEQ ID NO:63, SEQ ID NO:64, SEQ ID NO:65, SEQ ID NO:66, SEQ ID NO:67, SEQ ID NO:68, SEQ ID NO:69, SEQ ID NO:70, SEQ ID NO:71, SEQ ID NO:72, SEQ ID NO:73, SEQ ID NO:74, SEQ ID NO:75, SEQ ID NO:76, SEQ ID NO:77, SEQ ID NO:78, SEQ ID NO:79, SEQ ID NO:80, SEQ ID NO:81, SEQ ID NO:82, SEQ ID NO:83, SEQ ID NO:84, SEQ ID NO:85, SEQ ID NO:86, SEQ ID NO:87, SEQ ID NO:88, SEQ ID NO:89, SEQ ID NO:90, SEQ ID NO:91, SEQ ID NO:92, SEQ ID NO:93, SEQ ID NO:94, and SEQ ID NO:95).The Fc domain can comprise a quadruple mutant corresponding to C220S / L309D / Q311H / N434S (CDHS) (e.g., an IgG1 such as a human or humanized IgG1 with CDHS mutations such as SEQ ID NO:96, SEQ ID NO:97, SEQ ID NO:98, SEQ ID NO:99, SEQ ID NO:100, SEQ ID NO:101, SEQ ID NO:102, SEQ ID NO:103, SEQ ID NO:104, SEQ ID NO:105, SEQ ID NO:106, SEQ ID NO:107, SEQ ID NO:108, SEQ ID NO:109, SEQ ID NO:110, SEQ ID NO:111, SEQ ID NO:112, SEQ ID NO:113, SEQ ID NO:114, SEQ ID NO:115, SEQ ID NO:116, and SEQ ID NO:117). The Fc domain may comprise a triple mutant corresponding to L309D / Q311H / N434S (DHS) (e.g., an IgG1, such as a human or humanized IgG1, with DHS mutations such as SEQ ID NO:118, SEQ ID NO:119, SEQ ID NO:120, SEQ ID NO:121, SEQ ID NO:122, SEQ ID NO:123, SEQ ID NO:124, SEQ ID NO:125, SEQ ID NO:126, SEQ ID NO:127, SEQ ID NO:128, SEQ ID NO:129, SEQ ID NO:130, SEQ ID NO:131, SEQ ID NO:132, SEQ ID NO:133, SEQ ID NO:134, SEQ ID NO:135, SEQ ID NO:136, SEQ ID NO:137, and SEQ ID NO:138). The Fc domain may comprise a combination of one or more of the above-mentioned mutations that enhance binding to FcRn. Enhanced binding to FcRn may increase the half-life of the Fc domain-containing conjugate. For example, by incorporating one or more amino acid mutations that increase binding to FcRn (e.g., a YTE mutation, an LS mutation, or an N434H mutation), the half-life of the conjugate may be increased by 5%, 10%, 15%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 200%, 300%, 400%, 500%, or more, compared to a conjugate having a corresponding Fc domain without the mutations that enhance FcRn binding.Exemplary Fc domains with enhanced FcRN binding and methods for producing Fc domains with enhanced FcRN binding are known in the art, as described, for example, in Maeda, A. et al., Identification of human IgG1 variant with enhanced FcRn binding and without increased binding to rheumatoid factor autoantibody, MABS, 2017, 9(5):844-853 (incorporated herein in its entirety).

[0322] As used herein, an amino acid "corresponding to" a particular amino acid residue (e.g., in a particular SEQ ID NO:) should be understood to include any amino acid residue that would be understood by one of skill in the art to align with that particular residue (e.g., in a particular sequence). For example, any one of SEQ ID NOs: 1-138 may be mutated to include a YTE mutation, an LS mutation, and / or an N434H mutation by mutating the "corresponding residue" in the amino acid sequence.

[0323] As used herein, a sulfur atom "corresponding to" a particular cysteine ​​residue in a particular SEQ ID NO: includes the sulfur atom of any cysteine ​​residue that would be understood by one of skill in the art to align with the particular cysteine ​​in that particular sequence. A protein sequence alignment of human IgG1 (UniProtKB:P01857, SEQ ID NO:142), human IgG2 (UniProtKB:P01859, SEQ ID NO:143), human IgG3 (UniProtKB:P01860, SEQ ID NO:144), and human IgG4 (UniProtKB:P01861, SEQ ID NO:145) is shown below (aligned using Clustal Omega multiple pairwise alignment). This alignment shows cysteine ​​residues (e.g., sulfur atoms of cysteine ​​residues) that "correspond" to each other (indicated by boxes and a circle). By performing such an alignment with an IgG variant of the invention, one skilled in the art would readily be able to determine the sulfur atom of the cysteine ​​that corresponds to any of the sulfur atoms of a particular cysteine ​​in a particular SEQ ID NO: 1 described herein (e.g., any one of SEQ ID NOs: 1-138). For example, one skilled in the art would readily be able to determine that Cys10 of SEQ ID NO: 10 (the first cysteine ​​of the conserved CPPC motif in the hinge region of the Fc domain) corresponds to, for example, Cys109 of IgG1, Cys106 of IgG2, Cys156 of IgG3, Cys29 of SEQ ID NO: 1, Cys9 of SEQ ID NO: 2, Cys30 of SEQ ID NO: 3, or Cys10 of SEQ ID NO: 10.

[0324] In some embodiments, an Fc domain of the invention has the sequence of any one of SEQ ID NOs: 39-138 and can further comprise an additional amino acid (Xaa)x at the N-terminus and / or an additional amino acid (Xaa)z at the C-terminus, where Xaa is any amino acid and x and z are integers equal to or greater than zero, generally less than 100, preferably less than 10, and more preferably 0, 1, 2, 3, 4, or 5. In some embodiments, the additional amino acid is at least 70% (e.g., 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100%) identical to one or more consecutive amino acids of SEQ ID NO: 81. For example, the additional amino acid can be a single amino acid on the C-terminus corresponding to Lys330 of IgG1 (SEQ ID NO: 119).

[0325] As used herein, a nitrogen atom "corresponding to" a particular lysine residue in a particular SEQ ID NO includes the nitrogen atom of any lysine residue that would be understood by one of skill in the art to align with the particular lysine in that particular sequence. A protein sequence alignment of human IgG1 (UniProtKB:P01857, SEQ ID NO:142), human IgG2 (UniProtKB:P01859, SEQ ID NO:143), human IgG3 (UniProtKB:P01860, SEQ ID NO:144), and human IgG4 (UniProtKB:P01861, SEQ ID NO:145) is shown below (aligned using Clustal Omega multiple pairwise alignment). This alignment shows lysine residues (e.g., nitrogen atoms of lysine residues) that "correspond" to each other (boxed, *(Indicated by a symbol). By performing such an alignment with an IgG variant of the present invention, one skilled in the art would readily be able to determine the lysine nitrogen atom that corresponds to any of the nitrogen atoms of a specific lysine in a particular SEQ ID NO: 1 described herein (e.g., any one of SEQ ID NOs: 1-138). For example, one skilled in the art would readily be able to determine that Lys35 in SEQ ID NO: 10 corresponds to, for example, Lys129 in IgG1, Lys126 in IgG2, Lys176 in IgG3, Lys51 in SEQ ID NO: 1, Lys31 in SEQ ID NO: 2, Lys50 in SEQ ID NO: 3, or Lys30 in SEQ ID NO: 10.

[0326] TIFF2026508522000140.tif219170

[0327] Pharmaceutical Compositions and Preparations The conjugates described herein can be formulated into pharmaceutical compositions for use in the methods described herein. In some embodiments, the conjugates described herein can be formulated alone in a pharmaceutical composition. In some embodiments, the conjugates described herein can be formulated in a pharmaceutical composition in combination with an antiviral agent or antiviral vaccine. In some embodiments, the pharmaceutical composition comprises a conjugate of Formula (I) and a pharmaceutically acceptable carrier and excipient.

[0328] Acceptable carriers and excipients in pharmaceutical compositions are nontoxic to recipients at the dosages and concentrations employed. Acceptable carriers and excipients can include buffers (e.g., phosphate, citric acid, HEPES, and TAE), antioxidants (e.g., ascorbic acid and methionine), preservatives (e.g., hexamethonium chloride, octadecyldimethylbenzylammonium chloride, resorcinol, and benzalkonium chloride), proteins (e.g., human serum albumin, gelatin, dextran, and immunoglobulins), hydrophilic polymers (e.g., polyvinylpyrrolidone), amino acid residues (e.g., glycine, glutamine, histidine, and lysine), and carbohydrates (e.g., glucose, mannose, sucrose, and sorbitol).

[0329] Examples of other excipients include, but are not limited to, anti-adherents, binders, coatings, compression aids, disintegrants, dyes, emollients, emulsifiers, fillers (diluents), film formers or coatings, flavors, fragrances, flow agents (flow enhancers), lubricants, adsorbents, suspending or dispersing agents, or sweeteners. Exemplary excipients include, but are not limited to, butylated hydroxytoluene (BHT), calcium carbonate, calcium phosphate (dibasic), calcium stearate, croscarmellose, cross-linked polyvinylpyrrolidone, citric acid, crospovidone, cysteine, ethylcellulose, gelatin, hydroxypropyl cellulose, hydroxypropylmethylcellulose, lactose, magnesium stearate, maltitol, mannitol, methionine, methylcellulose, methylparaben, microcrystalline cellulose, polyethylene glycol, povidone, pregelatinized starch, propylparaben, retinyl palmitate, shellac, silicon dioxide, sodium carboxymethylcellulose, sodium citrate, sodium starch glycolate, sorbitol, starch (corn), stearic acid, stearic acid, sucrose, talc, titanium dioxide, vitamin A, vitamin E, vitamin C, and xylitol.

[0330] The pharmaceutical composition can be formed into a unit dosage form as needed. The amount of the active ingredient, for example, the conjugate of formula (I), contained in the unit dosage form is 10 to 900 mg (for example, 10 mg to 20 mg, 15 mg to 30 mg, 10 mg to 25 mg, 15 mg to 25 mg, 20 mg to 30 mg, 25 mg to 35 mg, 25 mg to 35 mg, 30 mg to 40 mg, 35 mg to 50 mg, 30 mg to 45 mg, 40 mg to 50 mg, 45 mg to 50 mg, 50 mg to 75 mg, 50 mg to 100 mg, 50 mg to 125 mg, 50 mg to 150 mg, 50 mg to 175 mg, 50 mg to 200 mg, 50 mg to 225 mg). mg, 50mg~250mg, 60mg~75mg, 60mg~100mg, 60mg~125mg, 60mg~150mg, 60mg~175mg, 60mg~200mg, 60mg~225mg, 60mg~250mg, 70mg~75mg, 70mg~100mg , 70mg~125mg, 70mg~150mg, 70mg~175mg, 70mg~200mg, 70mg~225mg, 70mg~250mg, 80mg~100mg, 80mg~125mg, 80mg~150mg, 80mg~175mg, 80mg~200mg, 80mg~225mg, 80mg~250mg, 90mg~100mg, 90mg~125mg, 90mg~150mg, 90mg~175mg, 90mg~200mg, 90mg~225mg, 90mg~250mg, 100mg~125mg, 100mg~150m g, 100mg~175mg, 100mg~200mg, 100mg~225mg, 100mg~250mg, 150mg~175mg, 150mg~200mg, 150mg~225mg, 150mg~250mg, 200mg~225mg, 200mg~250mg , 200mg to 275mg, 200mg to 300mg, 200mg to 325mg, and 200mg to 350mg, 300mg to 325mg, 300mg to 350mg, 300mg to 375mg, 300mg to 400mg, 300mg to 425mg, 300mg to 450mg, 400mg to 425mg, 400mg to 450mg, 400mg to 475mg, 400mg to 500mg, 400mg to 525mg, 400mg to 550mg, 500mg to 525mg, 500mg to 550mg, 500mg to 575mg, 500mg to 600mg,In some embodiments, the unit dosage form comprises 150 mg of a conjugate of Formula (I) (e.g., Conjugate A). In some embodiments, the unit dosage form comprises 300 mg of a conjugate of Formula (I) (e.g., Conjugate A). In some embodiments, the unit dosage form comprises 900 mg of a conjugate of Formula (I) (e.g., Conjugate A).

[0331] Route of administration and dosage In any of the methods described herein, the conjugates herein can be administered by any suitable route to treat or protect against influenza infection, or to prevent, stabilize, or inhibit the growth or spread of influenza virus. In some embodiments, administering includes intramuscular, intravenous (e.g., as a sterile solution and in a solvent system suitable for intravenous use), or subcutaneous administration of any of the conjugates of Formula (I).

[0332] The dosage of the conjugates described herein may range from 10 to 900 mg (e.g., 10 mg to 20 mg, 15 mg to 30 mg, 10 mg to 25 mg, 15 mg to 25 mg, 20 mg to 30 mg, 25 mg to 35 mg, 25 mg to 35 mg, 30 mg to 40 mg, 35 mg to 50 mg, 30 mg to 45 mg, 40 mg to 50 mg, 45 mg to 50 mg, 50 mg to 75 mg, 50 mg to 100 mg, 50 mg to 125 mg, 50 mg to 150 mg, 50 mg to 175 mg, 50 mg to 200 mg, 50 mg to 225 mg, 50 mg to 250 mg, 60 mg to 75 mg, 70 mg to 80 mg, 80 mg to 90 mg, 90 mg to 100 mg, 100 mg to 125 mg, 100 mg to 150 mg, 100 mg to 175 mg, 100 mg to 200 mg, 100 mg to 225 mg, 100 mg to 250 ... 5mg, 60mg~100mg, 60mg~125mg, 60mg~150mg, 60mg~175mg, 60mg~200mg, 60mg~225mg, 60mg~250mg, 70mg~75mg, 70mg~100mg, 70mg~125mg, 70mg~150mg, 70mg~175mg, 70mg~200mg, 70mg~225mg, 70mg~250mg, 80mg~100mg, 80mg~125mg, 80mg~150mg, 80mg~175mg, 80mg~200mg, 80mg~225mg, 80mg~250mg, 90m g~100mg, 90mg~125mg, 90mg~150mg, 90mg~175mg, 90mg~200mg, 90mg~225mg, 90mg~250mg, 100mg~125mg, 100mg~150mg, 100mg~175mg, 100mg~200mg, 10 0mg~225mg, 100mg~250mg, 150mg~175mg, 150mg~200mg, 150mg~225mg, 150mg~250mg, 200mg~225mg, 200mg~250mg, 200mg~275mg, 200mg~300mg, 200mg~ 325mg, and 200mg to 350mg, 300mg to 325mg, 300mg to 350mg, 300mg to 375mg, 300mg to 400mg, 300mg to 425mg, 300mg to 450mg, 400mg to 425mg, 400mg to 450mg, 400mg to 475mg, 400mg to 500mg, 400mg to 525mg, 400mg to 550mg, 500mg to 525mg, 500mg to 550mg, 500mg to 575mg, 500mg to 600mg, 500mg to 625mg, 500mg to 650mg, 600mg to 625mg,In some embodiments, the dosage administered to a human subject is 150 mg of the conjugate of Formula (I) (e.g., Conjugate A). In some embodiments, the dosage administered to a human subject is 300 mg of the conjugate of Formula (I) (e.g., Conjugate A). In some embodiments, the dosage administered to a human subject is 900 mg of the conjugate of Formula (I) (e.g., Conjugate A). The conjugate of Formula (I) or a pharmaceutical composition thereof may be administered once to a human subject, for example, prior to the flu season. [Example]

[0333] The following examples are presented to provide one of ordinary skill in the art with a description of how the compositions and methods described herein can be used, made, and evaluated, and are intended to be merely exemplary of the invention and are not intended to limit the scope of what the inventors regard as their invention.

[0334] Example 1: Clinical Trial Protocol We conducted a proof-of-concept, randomized, double-blind, placebo-controlled, phase 2a study to evaluate the prophylactic antiviral activity, safety, tolerability, and pharmacokinetics of conjugate A against influenza H3N2 A / Perth / 16 / 2009 in a human virus challenge model. Conjugate A used in this study has a DAR of 4.5.

[0335] To evaluate the prophylactic efficacy of conjugate A in terms of reduction of the viral load-time area under the curve (VL-AUC) after influenza virus challenge compared with placebo, we determined the viral load-time area under the curve (VL-AUC) of the influenza challenge virus using quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR) of nasal samples starting the day after virus challenge (day 1, afternoon) through day 8 (morning) (Table 1). The study protocol is summarized in Figure 1A.

[0336] A summary of the study breakdown is shown in Figure 1B. The demographics of the human volunteers are shown in Figure 2. Box plots of the VL-AUC data from Table 1 are shown in Figure 3. The 8-day mean viral load by qRT-PCR is shown in Figure 4. [Table 1]

[0337] To evaluate the efficacy of conjugate A compared with placebo in reducing or shortening viral replication after influenza virus challenge, we determined the peak influenza viral load, defined by the maximum viral load, using quantifiable qRT-PCR measurements in nasal samples from day 1 (afternoon) through day 8 (morning) (Table 2). A boxplot of the peak viral load by qRT-PCR in Table 2 is shown in Figure 5. [Table 2]

[0338] To evaluate the efficacy of conjugate A compared with placebo in reducing the incidence of influenza infection resulting from influenza virus challenge, we used qRT-PCR to detect the incidence of influenza infection, which is summarized in Table 3. qRT-PCR-confirmed influenza infection was defined as: two quantifiable (at or above the lower limit of quantification [LLOQ]) qRT-PCR measurements (reported in two or more independent samples over two days) between Day 1 (afternoon) and Day 8 (morning); and the occurrence of at least one positive quantitative (at or above the LLOQ) cell culture measurement in a nasal sample between Day 1 (afternoon) and Day 8 (morning). qRT-PCR-confirmed symptomatic influenza infection was defined as: two quantifiable [at or above the LLOQ] qRT-PCR measurements (reported in two or more independent samples over two days) between Day 1 (afternoon) and Day 8 (morning), and two or more symptoms at a single time point. Moderate to severe symptomatic qRT-PCR-confirmed influenza infection was defined as follows: qRT-PCR-confirmed influenza infection (two quantifiable [≥ LLOQ] qRT-PCR measurements [reported on two or more independent samples over two days]) between Day 1 (afternoon) and Day 8 (morning) and Grade 2 or greater symptoms at a single time point. Culture-confirmed symptomatic influenza infection was defined as: laboratory-confirmed cultureable influenza infection (one quantifiable [≥ LLOQ] cell culture measurement) between Day 1 (afternoon) and Day 8 (morning) and two or more symptoms at a single time point. [Table 3]

[0339] To evaluate the effect of conjugate A compared to placebo in reducing or shortening viral shedding after influenza virus challenge, we measured the time to test-negative confirmation (in hours) using quantifiable qRT-PCR measurements in nasal samples from day 1 (afternoon) to the first confirmed undetectable assessment after the peak measurement. The results are summarized in Table 4 and Figure 6. [Table 4]

[0340] To evaluate the efficacy of conjugate A compared with placebo in reducing clinical symptoms resulting from influenza virus challenge, we used a graded symptom scoring system to measure the area under the curve over time (TSS-AUC) of the total clinical symptom score collected three times daily from day 1 (morning) through day 8 (morning). The TSS-AUC data are summarized in Table 5 and Figure 7. [Table 5]

[0341] To evaluate the efficacy of conjugate A compared to placebo in reducing clinical symptoms resulting from influenza virus challenge, we measured total clinical symptom scores (TSS). Peak symptom diary card scores consisted of peak TSS measured by a graded symptom scoring system collected three times daily from Day 1 (morning) through Day 8 (morning). Peak daily symptom scores were the maximum individual daily sum of symptom scores from Day 1 through Day 8. TSS results are summarized in Table 6, and mean TSS over time is shown in Figure 8. A boxplot of the peak TSS data is shown in Figure 9. [Table 6]

[0342] To further evaluate the effect of conjugate A compared to placebo in reducing clinical symptoms resulting from influenza virus challenge, we measured time to symptom resolution, measured from the time of peak daily symptom score to the time to return to baseline score, using a graded daily symptom scoring system. The time to symptom resolution data are summarized in Table 7 and Figure 10. [Table 7]

[0343] Rationale for the test This study was designed to determine the prophylactic antiviral activity of a single subcutaneous dose of conjugate A against experimental influenza infection and to confirm its safety, tolerability, and PK in a healthy adult population vaccinated with the influenza H3N2 A / Perth / 16 / 2009 challenge strain. In the current epidemiological situation, where influenza incidence is very low worldwide, it is difficult to demonstrate proof-of-concept for the therapeutic or prophylactic efficacy of anti-influenza compounds in the setting of natural infection. This HVC model provides a setting independent of the number of influenza cases present at any given time. This model also has a proven safety track record and has demonstrated proof-of-concept efficacy for numerous antiviral compounds and vaccines, which have subsequently been found to be effective in large-scale clinical trials in the setting of natural infection. Establishing the proof-of-concept and minimum effective concentration in this study is important to better design studies to demonstrate the safety and efficacy of conjugate A.

[0344] Influenza challenge strains have been used for over 20 years to aid in the evaluation of numerous antiviral, immunomodulatory, and vaccine therapies.

[0345] Challenge Agent The challenge agent used in this study is influenza H3N2 A / Perth / 16 / 2009. This challenge stock was manufactured under current Good Manufacturing Practice (cGMP). This challenge stock has undergone quality testing (identity, appearance, sterility, infectivity, and contaminants) during manufacturing according to established specifications and has subsequently passed an extensive panel of adventitious agent tests. This challenge agent is stored in a secure -80°C freezer (normal temperature range is -60°C to -90°C).

[0346] Example 2: Single-dose pharmacokinetics of conjugate A in humans This is a first-in-human, phase 1, single-center, prospective, randomized, double-blind study in which healthy adult subjects received single escalating doses of conjugate A intramuscularly (IM) or subcutaneously (SQ), followed by another single dose of conjugate A administered by the same route in the mid- and high-dose groups 3 months or 5 effective half-lives after the first dose, whichever was longer (dosing may be adjusted based on tolerability). Conjugate A used in this study has a DAR of 4.5. For subjects randomized to receive conjugate A, plasma concentrations of conjugate A were determined using a validated hybrid immunoassay LC-MS method with a lower limit of quantitation (LLOQ) of 0.1 μg / mL. The objective is to evaluate the plasma pharmacokinetics of conjugate A in humans.

[0347] Dose levels of Conjugate A followed an ascending single-dose design with a starting dose based on findings from 3-month toxicity studies in rats and monkeys. For each route (IM and SQ administration, route assignment was open-label), subjects were randomized to receive a single dose of Conjugate A injection or saline placebo (treatment assignment was blinded) according to the design in Table 8. [Table 8]

[0348] Each cohort was divided into two groups identified as the "sentinel" group (randomized 1:1) and the "main" group (randomized 7:2). Low dose (50mg) level: Cohort 1A (IM) and Cohort 1B (SQ) - Sentinel n=2 and Main n=9 for each route Intermediate dose (150 mg) level: Cohort 2A (IM) and Cohort 2B (SQ) - Sentinel n=2 and Main n=9 for each route High dose (450mg) level: Cohort 3A (IM) and Cohort 3B (SQ) - Sentinel n=2 and Main n=9 for each route Highest dose (900mg): Cohort 4B (SQ) - Sentinel n=2 and Main n=9

[0349] For each route, the sentinel group for each dose level, i.e., the first two subjects (one conjugate A and one placebo), received blinded study drug and were closely monitored for safety for at least one week. The principal investigator (PI) and sponsor reviewed the blinded safety data after the first sentinel group subjects for each dose level completed dosing and were followed for at least one week. Because no drug-related serious adverse events occurred, the corresponding main group subjects were dosed one week after dosing in the sentinel group. If the dose / route of administration was determined to be safe and well-tolerated for at least 14 days after dosing, the next cohort of subjects was enrolled and randomized to receive the next higher dose level of conjugate A injection or placebo.

[0350] Subjects in Cohort 4B received a single injection of conjugate A or placebo (SQ). Subjects in Cohorts 2A and 2B and Cohorts 3A and 3B received a second single dose of conjugate A or placebo after a five-effective half-life washout from the first dose if the safety and tolerability of the first dose were deemed acceptable upon review of cumulative safety data. For each dose and route, dropouts in the sentinel group immediately prior to the second single dose were randomly replaced with subjects in the main group while maintaining their original treatment assignment (i.e., subjects assigned to placebo remained on placebo, and subjects assigned to conjugate A remained on conjugate A; this may require unblinded oversight). The PI and sponsor reviewed blinded safety data after the first sentinel group subjects at each dose level completed dosing and were followed for the minimum observation period (7 days) determined during the first single dose. Because no drug-related serious adverse events occurred, subjects in the sentinel group were dosed one week after the corresponding main group.

[0351] Pharmacokinetics (PK) was determined by analyzing the concentration of conjugate A in plasma samples obtained from subjects receiving conjugate A injections in each cohort at various time points after administration of the first and second single doses of study drug. Plasma samples, nasopharyngeal swabs, and nasal washes are analyzed for conjugate A concentrations. At specific time points, anti-drug antibodies (ADAs) were also measured by a validated ELISA method.

[0352] In the PK population, PK parameters assessed using noncompartmental analysis in Phoenix WinNonlin (Certara) and nominal protocol-specified times included peak plasma concentration (C max ), time to reach maximum plasma concentration (T max ), terminal elimination half-life (t 1 / 2 ), apparent clearance (CL / F), apparent volume of distribution (V Z / F), the area under the plasma concentration-time curve from time 0 to the last quantifiable sample time point (AUC 0-t ), and the area under the plasma concentration-time curve extrapolated from time 0 to infinity (AUC 0-∞ ) is included.

[0353] PK parameters for each cohort are summarized in Tables 8A-8G. Figures 11-14 show the mean plasma concentrations of conjugate A over time following intramuscular versus subcutaneous administration of conjugate A. Figure 15 compares the mean AUC of conjugate A following intramuscular versus subcutaneous administration. [Table 9] [Table 10] [Table 11] [Table 12] [Table 13] [Table 14] [Table 15]

[0354] Example 3: Population Pharmacokinetic Model of Conjugate A A Phase 1 single ascending dose (SAD) study is being conducted in healthy volunteers to evaluate the safety, tolerability, and pharmacokinetics of four dose levels of conjugate A administered subcutaneously (SQ) or intramuscularly (IM). Conjugate A used in this study has a DAR of 4.5. Preliminary results from the SAD study were analyzed by nonlinear mixed-effects modeling.

[0355] A total of 66 healthy volunteers dosed with placebo (N=3 per cohort) or conjugate A (N=8 per cohort, 50 mg, 150 mg, or 450 mg of conjugate A administered SQ or IM) were included in the pharmacokinetic analysis.

[0356] Conjugate A concentrations in plasma were sampled and quantified using a hybrid LCMS method using a polyclonal antibody against conjugate A with low cross-reactivity to human Fc alone. Preliminary results were analyzed using nonlinear mixed-effects modeling. The overall procedure for developing a population pharmacokinetic model of conjugate A included exploratory data analysis and the development of a basic structural model. NONMEM version 7.3 software was used throughout the model development process, applying a conditional first-order estimation method that took interactions into account. The observed PK was adequately described by a linear one-compartment pharmacokinetic model with first-order absorption (Table 9). The half-life of conjugate A was 6 to 8 weeks. [Table 16]

[0357] All participants achieved 90% C on the fifth day after administration. max(Figure 16). Model-predicted plasma concentrations following one or two 150 mg doses (as 300 mg) on ​​day 1 show a gradual decline from peak to trough concentration of approximately 1 Log over the course of the flu season, but accumulation occurs when subjects given a 150 mg dose on day 1 receive a second 150 mg dose midway through the flu season, reducing the peak-to-trough fluctuations (Figure 17).

[0358] Thus, conjugate A exhibits an extended half-life of 6-8 weeks and may provide seasonal protection with one or two doses per season.

[0359] Example 4: Temporal pharmacokinetics of conjugate A in influenza-infected humans In this study, to evaluate the pharmacokinetics of conjugate A as a function of time in influenza-infected human subjects, a single dose of 50 mg or 150 mg of conjugate A was administered subcutaneously (SQ) to human subjects followed by influenza virus challenge. Conjugate A used in this study has a DAR of 4.5.

[0360] Following single SQ administration of 50 mg and 150 mg conjugate A to human subjects, quantifiable conjugate A plasma concentrations were observed in all participants at the first sampling time point post-dose (day -5 [viral inoculation basis], 2 hours [conjugate A dosing basis]) (Tables 10A and 10B). Human subjects were then challenged with influenza virus inoculation on day 0. Mean conjugate A plasma concentrations gradually increased for both 50 mg and 150 mg doses, reaching peak levels on day 0 (viral inoculation basis) and 120 hours (conjugate A dosing basis). Thereafter, plasma concentrations gradually declined, fluctuating in some subjects, and additional conjugate A plasma peaks could be observed. Notably, a single SQ administration of 50 mg conjugate A maintained a mean plasma concentration of 387 ng / mL 180 days after administration of conjugate A (Table 10B). In summary, the mean plasma concentrations of conjugate A at the 150 mg dose were higher across the pharmacokinetic profile compared to the mean plasma concentrations of conjugate A at the 50 mg dose (Table 11). [Table 17] [Table 18] [Table 19]

[0361] Other embodiments While the invention has been described in terms of specific embodiments thereof, it will be understood that further modifications are possible, and this application is intended to cover any variations, uses, or applications of the invention which generally follow the principles of the invention, come within the scope of known or customary practice in the art to which this invention pertains, and which are applicable to the essential features defined above, including departures therefrom as fall within the scope of the claims. All publications, patents, and patent applications mentioned in the above specification are herein incorporated by reference to the same extent as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated by reference in its entirety.

[0362] A detailed description of one or more preferred embodiments has been provided herein. However, it should be understood that the present invention can be embodied in various forms. As such, the specific details disclosed herein should not be construed as limiting, but rather as a basis for the claims and as a representative basis for teaching one skilled in the art to use the present invention in any suitable manner.

Claims

1. 1. A method for inhibiting, reducing, or shortening influenza virus replication or infection in a human subject comprising or consisting of subcutaneously or intramuscularly administering to said human subject a dose of a conjugate of formula (I) in an amount of 50 mg to 900 mg, wherein said conjugate of formula (I) has the following structure: 【Chemistry 1】 wherein each E is an Fc domain monomer; n is 2, T is an integer from 3 to 6, and The wavy line indicates the covalent bond of E to the nitrogen atom of a solvent-exposed lysine or the sulfur atom of a solvent-exposed cysteine.

2. 1. A method for treating a human subject comprising or consisting of subcutaneously or intramuscularly administering a dose of a conjugate of formula (I) in an amount of 50 mg to 900 mg, wherein said conjugate of formula (I) has the following structure:

3. 3. The method of claim 2, wherein the human subject has an influenza infection.

4. 1. A method for reducing the time to symptom resolution in a human subject, or for reducing the area under the influenza viral load curve (VL-AUC) in a human subject with influenza infection, or for reducing the time to peak influenza viral load in a human subject with influenza infection, comprising or consisting of subcutaneously or intramuscularly administering a dose of a conjugate of formula (I) in an amount of 50 mg to 900 mg.

5. 1. A method for reducing the time to a confirmed negative influenza test in a human subject with influenza infection, comprising or consisting of subcutaneously or intramuscularly administering a dose of a conjugate of formula (I) in an amount of 50 mg to 900 mg.

6. 1. A method for reducing the Total Clinical Symptom Score (TSS-AUC) in a human subject with influenza infection, comprising or consisting of subcutaneously or intramuscularly administering a dose of a conjugate of formula (I) in an amount of 50 mg to 900 mg.

7. 1. A method for reducing the peak of TSS in a human subject with influenza infection, comprising or consisting of subcutaneously or intramuscularly administering a dose of a conjugate of formula (I) in an amount of 50 mg to 900 mg.

8. 1. A method for inhibiting, reducing, or shortening influenza virus replication or infection in a human subject, said method comprising: (a) subcutaneously or intramuscularly administering to said human subject a first dose of a conjugate of formula (I) in an amount of 50 mg to 900 mg; and (b) administering a second dose of the conjugate of Formula (I) subcutaneously or intramuscularly to said human subject in an amount of 50 mg to 900 mg. comprising or consisting of The method, wherein the first dose and the second dose are administered 60 to 120 days apart.

9. 1. A method for treating a human subject, said method comprising: (a) administering a first dose of a conjugate of formula (I) subcutaneously or intramuscularly in an amount of 50 mg to 900 mg; and (b) administering a second dose of the conjugate of formula (I) subcutaneously or intramuscularly in an amount of 50 mg to 900 mg. comprising or consisting of The method, wherein the first dose and the second dose are administered 60 to 120 days apart.

10. 10. The method of claim 9, wherein the human subject has an influenza infection.

11. 1. A method for reducing the time to symptom resolution in a human subject, or for reducing the area under the influenza viral load curve (VL-AUC) in a human subject with influenza infection, or for reducing the time to peak influenza viral load in a human subject with influenza infection, said method comprising: (a) administering a first dose of a conjugate of formula (I) subcutaneously or intramuscularly in an amount of 50 mg to 900 mg; and (b) administering a second dose of the conjugate of formula (I) subcutaneously or intramuscularly in an amount of 50 mg to 900 mg. comprising or consisting of The method, wherein the first dose and the second dose are administered 60 to 120 days apart.

12. 1. A method for reducing the time to a confirmed negative influenza test in a human subject with influenza infection, said method comprising: (a) administering a first dose of a conjugate of formula (I) subcutaneously or intramuscularly in an amount of 50 mg to 900 mg; and (b) administering a second dose of the conjugate of formula (I) subcutaneously or intramuscularly in an amount of 50 mg to 900 mg. comprising or consisting of The method, wherein the first dose and the second dose are administered 60 to 120 days apart.

13. 1. A method for reducing Total Clinical Symptom Score (TSS-AUC) in a human subject with influenza infection, said method comprising: (a) administering a first dose of a conjugate of formula (I) subcutaneously or intramuscularly in an amount of 50 mg to 900 mg; and (b) administering a second dose of the conjugate of formula (I) subcutaneously or intramuscularly in an amount of 50 mg to 900 mg. comprising or consisting of The method, wherein the first dose and the second dose are administered 60 to 120 days apart.

14. 1. A method for reducing a TSS peak in a human subject having an influenza infection, said method comprising: (a) administering a first dose of a conjugate of formula (I) subcutaneously or intramuscularly in an amount of 50 mg to 900 mg; and (b) administering a second dose of the conjugate of formula (I) subcutaneously or intramuscularly in an amount of 50 mg to 900 mg. comprising or consisting of The method, wherein the first dose and the second dose are administered 60 to 120 days apart.

15. 1. A method for inhibiting, reducing or shortening influenza virus replication or infection in a human subject, comprising or consisting of maintaining a minimum plasma concentration of a conjugate of formula (I) of at least 300 ng / mL for a period of 4 to 6 months.

16. 16. The method of claim 15, wherein the minimum plasma concentration of the conjugate of at least 300 ng / mL is maintained in the subject for 6 months.

17. 16. The method of claim 15, wherein the minimum plasma concentration of the conjugate of at least 300 ng / mL is maintained in the subject for 4 months.

18. 18. The method of any one of claims 1 to 17, wherein the conjugate is conjugate A and E has the sequence of SEQ ID NO:

76.

19. 19. The method of any one of claims 1 to 18, wherein each dose comprises 150 mg of the conjugate.

20. 19. The method of any one of claims 1 to 18, wherein each dose comprises 300 mg of the conjugate.

21. 19. The method of any one of claims 1 to 18, wherein each dose comprises 450 mg of the conjugate.

22. 19. The method of any one of claims 1 to 18, wherein each dose comprises 900 mg of the conjugate.

23. A pharmaceutical composition in unit dosage form, comprising the conjugate of formula (I) in an amount of 50 mg to 900 mg.

24. 24. The pharmaceutical composition of claim 23, comprising 50 mg of the conjugate.

25. 24. The pharmaceutical composition of claim 23, comprising 150 mg of the conjugate.

26. 24. The pharmaceutical composition of claim 23, comprising 300 mg of the conjugate.

27. 24. The pharmaceutical composition of claim 23, comprising 450 mg of the conjugate.

28. 24. The pharmaceutical composition of claim 23, comprising 900 mg of the conjugate.

29. The pharmaceutical composition according to any one of claims 23 to 28, wherein the conjugate is conjugate A and E has the sequence of SEQ ID NO:

76.

30. A kit comprising the pharmaceutical composition of any one of claims 23 to 29 and instructions for use in the method of any one of claims 1 to 17.

31. 31. The method of any one of claims 1 to 17, the pharmaceutical composition of any one of claims 23 to 29, or the kit of claim 30, wherein the conjugate comprises an Fc domain comprising an amino acid sequence at least 95% identical to any one of SEQ ID NO:62, SEQ ID NO:63, SEQ ID NO:64, SEQ ID NO:67, SEQ ID NO:68, SEQ ID NO:72, SEQ ID NO:73, SEQ ID NO:76, SEQ ID NO:77, SEQ ID NO:80, SEQ ID NO:81, SEQ ID NO:82, SEQ ID NO:85, SEQ ID NO:86, SEQ ID NO:90, SEQ ID NO:91, SEQ ID NO:94, or SEQ ID NO:

95.

32. 31. The method of any one of claims 1 to 17, the pharmaceutical composition of any one of claims 23 to 29, or the kit of claim 30, wherein the conjugate comprises an Fc domain comprising the amino acid sequence of any one of SEQ ID NO:63, SEQ ID NO:64, SEQ ID NO:67, SEQ ID NO:68, SEQ ID NO:72, SEQ ID NO:73, SEQ ID NO:76, or SEQ ID NO:77.

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