Combination therapy and uses thereof for HIV infection - Patents.com

Abstract

The present disclosure relates to a method of treatment or treatment, clearing, prevention or cure of human immunodeficiency virus (HIV) infection. The present disclosure provides a combination of at least one agent selected from the group consisting of fostemsavir and temsavir, or a pharma- ceutical acceptable salt thereof, and a CD4 binding site (CD4bs) binding protein for use in the treatment of HIV and / or the clearance of HIV-infected cells.

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of U.S. Provisional Patent Application No. 63 / 290,758, filed December 17, 2021, the disclosure of which is incorporated herein in its entirety.

[0002] Sequence Listing This application contains a Sequence Listing, which has been submitted electronically in computer readable form in XML file format and is incorporated by reference in its entirety. The XML file, created on December 15, 2022, is named "70090WO01_SeqList_Updated_5Dec2022" and is 15,605 bytes in size.

[0003] FIELD OF THEINVENTION The present invention relates to a method for treating, preventing or curing human immunodeficiency virus (HIV) infection. In particular, the present invention relates to a method for treating, clearing, preventing or curing HIV-infected cells using a combination of drugs. In some embodiments, the combination comprises a drug comprising at least one drug selected from the group consisting of fostemsavir and temsavir, or a pharma- ceutically acceptable salt thereof, and a drug comprising a CD4 binding site-binding protein or a broadly neutralizing antibody, in therapeutic or prophylactic use for treating, clearing, preventing and / or curing HIV infection. [Background technology]

[0004] 2. Background of the Invention Human immunodeficiency virus (HIV), the virus that causes acquired immune deficiency syndrome (AIDS), is one of the world's most serious public health challenges. HIV remains a significant medical problem, with 79.3 million people infected with HIV and 36.3 million people dying from HIV since the beginning of the epidemic. As of the end of 2020, 37.7 million people worldwide were living with HIV. An estimated 0.7% of adults aged 15-49 years worldwide are living with HIV, yet the burden of the epidemic continues to vary widely across countries and regions.

[0005] The retrovirus HIV replicates by the assembly of new virus particles (virions) within infected host cells. These new virions leave the infected host cell (now known as producer cells) and spread the infection to other susceptible host cells. Virion morphogenesis within the producer cell can be divided into three stages: assembly, budding, and maturation. During the assembly stage, the virion packages all of the components required for infectivity, including the gp160 viral envelope (ENV) protein. ENV is an integral membrane protein that allows the virion to fuse with the target cell. As part of the packaging process, the ENV protein is co-translationally inserted into the endoplasmic reticulum (ER) membrane of the infected producer cell and then traffics through the cellular secretory pathway, where it is glycosylated, assembled into a trimeric complex, processed by the cellular protease furin into transmembrane (gp41) and surface (gp120) subunits, and delivered to the plasma membrane via vesicular trafficking (Sundquist, WI, & Kraeusslich, HG (2012). HIV-1 assembly, budding, and maturation. Cold Spring Harbor perspectives in medicine, 2(7), a006924). Thus, as the assembly process progresses, the infected host cell displays the HIV-specific ENV on the cell surface. Summary of the Invention [Problem to be solved by the invention]

[0006] ENV displayed on the surface of infected cells may be an attractive target for clearance by antibody recognition, which may result in antibody-mediated cell death. However, ENV on the surface of infected cells may exhibit multiple conformations influenced by interactions with the cis-expressed host CD4 receptor, which may result in reduced antibody recognition. Therefore, it is necessary to improve antibody recognition of ENV expressed on infected host cells. [Means for solving the problem]

[0007] Summary of the Invention In one aspect, a method for treating or preventing human HIV is provided, comprising administering therapeutically effective amounts of two or more agents selected from the following group: CD4bs binding protein; gp120 binding protein; a broadly neutralizing antibody or antigen-binding fragment thereof; fostemsavir or a pharmaceutically acceptable salt thereof; temsavir or a pharmaceutically acceptable salt thereof; and an integrase inhibitor or a pharmaceutically acceptable salt thereof.

[0008] In one aspect, a method is provided for treating or preventing HIV infection in a human in need thereof, comprising administering therapeutically effective amounts of: (a) a first agent comprising at least one agent selected from the group consisting of fostemsavir and temsavir, or a pharma- ceutically acceptable salt thereof; and (b) a second agent comprising at least one broadly neutralizing antibody or antigen-binding fragment thereof.

[0009] In one aspect, a method is provided for treating or preventing HIV infection in a human in need thereof, comprising administering therapeutically effective amounts of: (a) a first agent comprising at least one agent selected from the group consisting of fostemsavir and temsavir, or a pharmaceutically acceptable salt thereof; (b) a second agent comprising at least one broadly neutralizing antibody or antigen-binding fragment thereof; and (c) a third agent comprising at least one integrase inhibitor or a pharmaceutically acceptable salt thereof.

[0010] In one aspect, a combination for use in the treatment or prevention of HIV is provided, comprising administering to a human a first pharmaceutical composition comprising a first agent comprising at least one agent selected from the group consisting of fostemsavir and temsavir, or a pharmaceutically acceptable salt thereof, and a second pharmaceutical composition comprising a second agent comprising at least one broadly neutralizing antibody or an antigen-binding fragment thereof. In some embodiments, the combination for use further comprises administering a third pharmaceutical composition comprising a third agent comprising at least one integrase inhibitor, or a pharmaceutically acceptable salt thereof.

[0011] In one aspect, a kit is provided comprising a first pharmaceutical composition comprising a first agent comprising at least one agent selected from the group consisting of fostemsavir and temsavir, or a pharma- ceutically acceptable salt thereof; a second pharmaceutical composition comprising a second agent comprising at least one broadly neutralizing antibody or antigen-binding fragment thereof; and, optionally, a third pharmaceutical composition comprising a third agent comprising at least one integrase inhibitor or a pharma- ceutically acceptable salt thereof. [Brief description of the drawings]

[0012] [Figure 1] FIG. 1 shows the enhancement of mean fluorescent intensity (MFI) of bnAb binding to HIV-infected cells by temsavir. [Diagram 2] FIG. 1 shows the enhancement by temsavir of the percentage of HIV-infected cells bound by bnAb. [Diagram 3] FIG. 1 shows dose-dependent enhancement of antibody-dependent effector cell-mediated cytotoxicity (ADCC) by temsavir against HIV-infected primary cells. [Figure 4] FIG. 1 shows that temsavir does not cause cell death of HIV-infected primary cells in the absence of effector cells. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0013] definition Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. Although methods and materials similar or equivalent to those described herein can be used in the practice of the present invention, suitable methods and materials are described below.

[0014] All publications, patent applications, patents and other references mentioned herein are incorporated by reference in their entirety.

[0015] Further, the materials, methods, and examples are illustrative only and not intended to be limiting. The details of one or more embodiments of the invention are set forth in the accompanying description below. Other features, objects, and advantages of the invention will become apparent from the description and claims herein.

[0016] HIV Human immunodeficiency virus (HIV) is intended to mean HIV-1 unless further specified. The present invention may also be effective against HIV-2 or against patients with HIV-1 / HIV-2 co-infection. Human immunodeficiency virus type 1 (HIV-1) infection and the resulting acquired immune deficiency syndrome (AIDS) remain a global public health threat despite extensive efforts to develop anti-HIV-1 therapeutics. The enveloped virus HIV-1 evades humoral recognition by hiding behind a wide range of defense mechanisms. The major HIV-1 envelope protein (HIV-1 Env) is a glycoprotein (gp160) of approximately 160 kD. During infection, host cell proteases cleave gp160 into gp120 and gp41. gp41 is an integral membrane protein, whereas gp120 protrudes from the mature virus. gp120 binds to the host cell receptor CD4 by a CD4 binding site. Together, gp120 and gp41 form the HIV-1 envelope spike, which is the target of neutralizing antibodies. Broadly neutralizing antibodies that bind to HIV-1 Env have been identified, including the N6LS antibody, which specifically binds to the CD4 binding site of gp120 and can neutralize a high percentage of HIV-1 strains.

[0017] Binding Proteins The term "CD4 binding site (CD4bs) binding protein" as used herein refers to antibodies and other protein constructs, e.g., domains, that can bind to the CD4 binding site of the HIV envelope glycoprotein gp120. The terms "CD4bs binding protein", "CD4bs binding domain" and "antigen binding protein" are used interchangeably herein. This term does not include natural cognate ligands or receptors. In some embodiments, provided herein are monoclonal antibodies and antigen-binding fragments thereof that bind to the CD4 binding site on gp120 and neutralize HIV-1.

[0018] antibody The term "antibody" is used herein in the broadest sense to refer to a molecule having an immunoglobulin-like domain (e.g., IgG, IgM, IgA, IgD or IgE) and includes monoclonal antibodies, recombinant antibodies, polyclonal antibodies, chimeric antibodies, human antibodies, humanized antibodies, multispecific antibodies including bispecific antibodies and heteroconjugate antibodies; single variable domains (e.g., domain antibodies (DABs)), antigen-binding antibody fragments, Fab, F(ab')2, Fv, disulfide-linked Fvs, single chain Fvs, disulfide-linked scFvs, diabodies, TANDABS, etc., as well as modified versions of any of the foregoing (for a review of alternative "antibody" formats, see Holliger and Hudson, Nature Biotechnology, 2005, Vol 23, No. 9, 1126-1136).

[0019] The terms "full antibody or immunoglobulin", "whole antibody or immunoglobulin", or "intact antibody or immunoglobulin" are used interchangeably herein and refer to a heterotetrameric glycoprotein with a molecular weight of approximately 150,000 daltons. An intact antibody is composed of two identical heavy chains (HC) and two identical light chains (LC) linked by covalent disulfide bonds. This H2L2 structure folds to form three functional domains, including two antigen-binding fragments known as "Fab" fragments, and an "Fc" crystallizable fragment. The Fab fragment is composed of the variable heavy (VH) or variable light (VL) variable domains at the amino terminus, and the constant domains CH1 (heavy chain) and CL (light chain) at the carboxy terminus. The Fc fragment is composed of two domains formed by dimerization of paired CH2 and CH3 regions. Fc can trigger effector functions by binding to receptors on immune cells or by binding to C1q, the first component of the classical complement pathway. The five classes of antibodies, IgM, IgA, IgG, IgE and IgD, are defined by the amino acid sequences of different heavy chains, called μ, α, γ, ε and δ, respectively, and each heavy chain can pair with either K or λ light chains. The majority of antibodies in serum belong to the IgG class, and there are four isotypes of human IgG (IgG1, IgG2, IgG3 and IgG4), whose sequences differ mainly in their hinge region.

[0020] Fully human antibodies can be obtained using a variety of methods, for example, using yeast-based libraries or transgenic animals (e.g., mice) that can produce a repertoire of human antibodies. Yeast that display human antibodies on their surface that bind to the antigen of interest can be selected using FACS (Fluorescence-Activated Cell Sorting)-based methods or by capture onto beads using labeled antigen. Transgenic animals modified to express human immunoglobulin genes can be immunized with the antigen of interest, and antigen-specific human antibodies can be isolated using B cell sorting techniques. Human antibodies produced using these techniques can then be characterized for desirable properties, such as affinity, developability, and selectivity.

[0021] Alternative antibody formats include alternative scaffolds, in which one or more CDRs of an antigen binding protein can be placed on a suitable non-immunoglobulin protein scaffold or framework, such as an affibody, an SpA scaffold, an LDL receptor class A domain, an avimer (see, e.g., U.S. Patent Application Publication Nos. 2005 / 0053973, 2005 / 0089932, 2005 / 0164301), or an EGF domain.

[0022] The term "broadly neutralizing antibody" (bnAb) is defined as an antibody that inhibits viral attachment and cell entry via binding to HIV envelope glycoproteins (Env) (e.g., gp160, gp120, gp41), including, but not limited to, inhibiting in vitro infection by 50% in greater than 50%, 60% or more, 70% or more, 80% or more, 90% or more, 95% or more, 99% or more of a large panel (>100) of HIV-1 enveloped pseudotyped viruses and viral isolates. See, e.g., U.S. Published Patent Application No. 20120121597; Burton et al., Broadly Neutralizing Antibodies to HIV and Their Role in Vaccine Design. Annu Rev Immunol. May 20, 2016; 34:635-59.

[0023] For example, broadly neutralizing antibodies can include, but are not limited to, 2G12, 2F5, 3BC176, 3BNC60, 3BNC117, 4E10, 8ANC131, 8ANC195, 10E8, 10-1074, 12A12, 35022, b12, B2530, CH01-04, CH103, CH31, HJ16, M66.6, N6, N6LS, NIH45-46, PG9, PG16, PGDM1400, PGT121, PGT128, PGT135, PGT141-PGT145, PGT151, PGV04, VRC01, VRC01-LS, VRC07, VRC07-523, VRC07-LS, Z13, or any other broadly neutralizing antibody disclosed herein. In some embodiments, the broadly neutralizing antibody binds to an HIV envelope glycoprotein. In some embodiments, the broadly neutralizing antibody binds to an HIV envelope glycoprotein selected from the group consisting of gp160, gp120, and gp41. In one embodiment, the broadly neutralizing antibody binds to the HIV envelope glycoprotein gp41. In some embodiments, the broadly neutralizing antibody binds to gp120. In some embodiments, the broadly neutralizing antibody binds to gp120 and neutralizes HIV-1. An example of VRC07-523 is described in J. Virol, 88(21):12669-12682 (November 2014). An example of 3BNC1 17 is described in U.S. Publication No. 20140212458. An example of NIH45-46 is described in U.S. Publication No. 20150274813. An example of PGV04 is described in US Publication No. 20130251726. An example of b12 is described in US Publication No. 20160009789. An example of CH31 is described in US Publication No. 20130251726. An example of CH103 is described in US Publication No. 20140212458. In another embodiment, an antibody is provided that binds to HIV envelope glycoproteins at the gp120-gp41 interface. Such antibodies include, but are not limited to, antibodies selected from 8ANC195, 35022, and PGT151. An example of 8ANC195 is described in US Publication No. 20150361160. An example of 35022 is described in US Publication No. 20160022803.An example of PGT151 is described in US Publication No. 20150152167. In another embodiment, an antibody that binds to gp41 MPER (membrane-proximal external region) is provided, including, but not limited to, 4E10, 10E8, 2F5, and Z13e1. An example of 4E10 is described in US Publication No. 20160009789. An example of 10E8 is described in PCT Published Application No. WO2013070776. An example of 2F5 is described in US Publication No. 20150158934. An example of Z13e1 is described in US Publication No. 20120269821.

[0024] In some embodiments, the broadly neutralizing antibody is selected from the group consisting of VRC01, VRC01-LS, N6, N6LS, N6-DE, N6-LAGA, VRC07 and VRC07-523. An example of the disclosure of VRC01 is described in U.S. Patent No. 8,637,036. An example of the disclosure of VRC01-LS is described in WO2012 / 106578. An example of the disclosure of N6 and N6LS is described in WO2016 / 196975. An example of the disclosure of VRC07 and VRC07-523 is described in U.S. Patent No. 8,637,036, U.S. Patent Publication No. 2014 / 0322163A1, WO2016 / 196975 and WO2017 / 79479.

[0025] antigen binding site An antigen-binding site refers to the site on an antigen-binding protein that can specifically bind to an antigen and may be a single variable domain or, alternatively, a paired VH / VL domain such as may be found in a standard antibody. Alternatively, a single chain Fv (ScFv) domain may provide the antigen-binding site.

[0026] CDR "CDR" is defined as the amino acid sequence of the complementarity determining region of an antigen-binding protein. These are the hypervariable regions of the heavy and light chains of immunoglobulins. In the variable part of an immunoglobulin, there are three heavy chain CDRs (or CDR regions) and three light chain CDRs (or CDR regions). Thus, as used herein, "CDR" refers to all three heavy chain CDRs, all three light chain CDRs, all heavy and light chain CDRs, or at least two CDRs.

[0027] Throughout this specification, amino acid residues in variable domain sequences and variable domain regions within a full-length antigen-binding sequence, e.g., within an antibody heavy chain sequence or an antibody light chain sequence, are numbered according to the Kabat numbering convention. Similarly, the terms "CDR", "CDRL1", "CDRL2", "CDRL3", "CDRH1", "CDRH2", "CDRH3" used in the examples follow the Kabat numbering convention. For further information, see Kabat et al., Sequences of Proteins of Immunological Interest, 4th ed., USDepartment of Health and Human Services, National Institutes of Health (1987).

[0028] It will be apparent to one skilled in the art that there are alternative numbering conventions for amino acid residues in variable domain sequences and full-length antibody sequences. There are also alternative numbering conventions for CDR sequences, such as those described in Chothia et al. (1989) Nature 342:877-883. Depending on the structure of the antigen-binding protein and protein folding, other residues may be considered as part of the CDR sequence and will be understood as such by one skilled in the art.

[0029] Other numbering conventions for CDR sequences available to those skilled in the art include the "AbM" (University of Bath) and "contact" (University College London) methods.

[0030] Table 1 below shows one definition using each numbering convention for each CDR or binding unit. In Table 1, the Kabat numbering scheme is used to number the amino acid sequence of the variable domain. Please note that some of the CDR definitions may differ depending on the individual publication used.

[0031] [Table 1]

[0032] CDR variants The CDRs may be modified by the substitution, deletion or addition of one or more amino acids, wherein the variant antigen binding protein substantially retains the biological characteristics of the unmodified protein.

[0033] It will be understood that each of CDRs H1, H2, H3, L1, L2, L3 can be modified alone or in combination with any other CDR in any variation or combination.In one embodiment, the CDR is modified by substitution, deletion or addition of up to 3 amino acids, for example 1 or 2 amino acids, for example 1 amino acid.Typically, the modification is a substitution, particularly a conservative substitution, for example as shown in Table 2 below.

[0034] [Table 2]

[0035] For example, in a variant CDR, the flanking residues that make up a CDR as part of an alternative definition, eg, Kabat or Chothia, may be replaced with conservative amino acid residues.

[0036] Such antigen binding proteins comprising the above-described variant CDRs may be referred to herein as "functional CDR variants."

[0037] Epitope The term "epitope" as used herein refers to the portion of an antigen that contacts a specific binding domain (also known as a paratope) of an antigen-binding protein. An epitope may be a linear epitope or a conformational / discontinuous epitope. A conformational or discontinuous epitope comprises amino acid residues that come together in the tertiary folding of a polypeptide chain that are separated by other sequences in the primary sequence of the antigen (i.e., are not in a continuous sequence). These residues may be from different regions of the polypeptide chain, but are adjacent in the three-dimensional structure of the antigen. In the case of a multimeric antigen, a conformational or discontinuous epitope may include residues from different peptide chains. The specific residues contained within an epitope can be determined by computer modeling programs or by methods known in the art, such as the three-dimensional structure obtained by X-ray crystallography. Epitope mapping can be performed using a variety of techniques known to those skilled in the art, as described in publications such as Methods in Molecular Biology "Epitope Mapping Protocols", Mike Schutkowski and Ulrich Reineke (vol. 524, 2009) and Johan Rockberg and Johan Nilvebrant (vol. 1785, 2018). Exemplary methods include peptide-based approaches, such as pep-scanning, where binding of a series of overlapping peptides is screened using techniques such as ELISA, or in vitro display of large libraries of peptide or protein variants, such as on phage. Detailed epitope information can be determined by structural techniques, including X-ray crystallography, solution nuclear magnetic resonance (NMR) spectroscopy, and cryogenic-electron microscopy (cryo-EM). Mutagenesis, such as alanine scanning, is an effective approach where loss of binding analysis is used for epitope mapping.Another method combines hydrogen / deuterium exchange (HDX) with proteolysis and liquid-chromatography mass spectrometry (LC-MS) analysis to characterize discontinuous or conformational epitopes.

[0038] Percent Identity "Percent identity" or "% identity" between a query nucleic acid sequence and a subject nucleic acid sequence is an "identity" value expressed as a percentage, which is calculated using an appropriate algorithm (e.g., BLASTN, FASTA, Needleman-Wunsch, Smith-Waterman, LALIGN, or GenePAST / KERR) or software (e.g., DNASTAR Lasergene, GenomeQuest, EMBOSS needle, or EMBOSS infoalign) over the entire length of the query sequence after a pairwise global sequence alignment is performed using an appropriate algorithm (e.g., Needleman-Wunsch or GenePAST / KERR) or software (e.g., DNASTAR Lasergene, GenomeQuest, EMBOSS needle, or EMBOSS infoalign). Importantly, the query nucleic acid sequence may be described by a nucleic acid sequence disclosed herein, particularly one or more claims.

[0039] The query sequence may be 100% identical to the subject sequence, or may contain up to a certain integer number of amino acid or nucleotide changes relative to the subject sequence, resulting in a percent identity (%) less than 100%. For example, the query sequence is at least 50%, at least 60%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the subject sequence. In the case of nucleic acid sequences, such changes include deletion, substitution, or insertion of one or more nucleotide residues, which may occur at the 5' or 3' terminal positions of the query sequence, or may occur anywhere between these terminal positions, either individually within the nucleotide residues of the query sequence or scattered in one or more contiguous groups within the query sequence. In the case of amino acid sequences, such alterations include the deletion, substitution (including conservative and non-conservative substitutions) or insertion of one or more amino acid residues, which may occur at the amino- or carboxy-terminal positions of the query sequence, or at any position between these terminal positions, either individually within the amino acid residues of the query sequence, or scattered in one or more contiguous groups within the query sequence.

[0040] For antibody sequences, the percent identity can be determined over the entire length of the query sequence, including the CDRs. Alternatively, the percent identity can exclude one or more or all CDRs, for example, when the CDR sequences are fixed and intact, with all CDRs being 100% identical to the subject sequence, and the percent identity variation being in the remainder of the query sequence, e.g., framework sequences. The variant sequence substantially retains the biological characteristics of the unmodified protein.

[0041] Sequence variation The VH or VL (or HC or LC) sequence may be a variant sequence having up to 10 amino acid substitutions, additions or deletions. For example, the variant sequence may have up to 9, 8, 7, 6, 5, 4, 3, 2 or 1 amino acid substitutions, additions or deletions.

[0042] The HC sequence may be a variant sequence having up to 20% sequence variation.

[0043] The LC sequence may be a variant sequence having up to 20% sequence variation.

[0044] The sequence variations can exclude one or more or all CDRs, for example where the CDRs are the same in the VH or VL (or HC or LC) sequence and the variations are in the remainder of the VH or VL (or HC or LC) sequence such that the CDR sequences are fixed and intact.

[0045] Typically, the variations are substitutions, particularly conservative substitutions, such as those shown in Table 2. The variant sequence substantially retains the biological characteristics of the unmodified protein, e.g., N6.

[0046] Fc modification Fc engineering methods can be applied to modify the functional or pharmacokinetic properties of antibodies. Effector functions can be altered by making mutations in the Fc region that increase or decrease binding to C1q or Fcγ receptors, modifying antibody-mediated complement activation, including complement-dependent cytotoxicity (CDC) activity, or antibody-dependent cell-mediated cytotoxicity (ADCC) activity. Effector functions can also be altered by modifying the glycosylation pattern of the antibody. The in vivo half-life of the antibody can be altered by making mutations that affect the binding of Fc to FcRn (fetal Fc receptor).

[0047] Effector Function The term "effector function," as used herein, refers to one or more of antibody-mediated effects including antibody-dependent cell-mediated cytotoxicity (ADCC), antibody-mediated complement activation, e.g., complement-dependent cytotoxicity (CDC), complement-dependent cell-mediated phagocytosis (CDCP), antibody dependent complement-mediated cell lysis (ADCML), and Fc-mediated phagocytosis (Fe-mediated phagocytosis) or antibody-dependent cellular phagocytosis (ADCP).

[0048] The interaction between the Fc region of an antigen-binding protein or antibody and various Fc receptors (FcRs), including FcγRI (CD64), FcγRII (CD32), FcγRIII (CD16), FcRn, C1q, and type II Fc receptors, is believed to mediate the effector functions of the antigen-binding protein or antibody. Important biological effects can be the result of effector functions. Usually, binding of an antigen-binding protein or antibody to an antigen is required to mediate effector functions, and not all antigen-binding proteins or antibodies mediate all effector functions.

[0049] Effector function can be assessed in a number of ways, including, for example, assessing ADCC effector function of antibodies coated on target cells, mediated by natural killer (NK) cells via FcγRIII or monocytes / macrophages via FcγRI, or assessing CDC effector function of antibodies coated on target cells, mediated by the complement cascade via C1q. For example, ADCC effector function of antigen binding proteins of the invention can be assessed in a natural killer cell assay. Examples of such assays can be found in Shields et al., 2001, The Journal of Biological Chemistry, vol. 276, pp. 6591-6604; Chappel et al., 1993, The Journal of Biological Chemistry, vol. 268, pp. 25124-25131; Lazar et al., 2006, PNAS, 103; 4005-4010.

[0050] Exemplary assays for determining CDC function include those described in J Imm Meth, 1995, 184:29-38. The effect of mutations on effector function (e.g., FcRn binding, FcγRs and C1q binding, CDC, ADCML, ADCC, ADCP) can be assessed, for example, as described in Grevys et al., J Immunol. 2015 Jun 1;194(11):5497-5508, or Tam et al., Antibodies 2017, 6(3); Monnet et al., 2014 mAbs, 6:2, 422-436.

[0051] Throughout this specification, amino acid residues in an Fc region, an antibody sequence or a full length antigen binding protein sequence are numbered according to the EU index numbering convention.

[0052] Enhancement of effector functions Human IgG1 constant regions containing certain mutations have been shown to enhance binding to Fc receptors. In some cases, these mutations have also been shown to enhance effector functions, such as ADCC and CDC, as described below. The antigen-binding proteins of the present invention may include any of the following mutations.

[0053] Enhanced CDC: Fc engineering can be used to enhance complement-based effector functions. For example (with reference to IgG1), K326W / E333S; S267E / H268F / S324T; and IgG1 / IgG3 cross-subclasses can increase C1q binding; E345R (Diebolder et al., Science 2014;343:1260-1293) and E345R / E430G / S440Y result in preformed IgG hexamers (Wang et al., Protein Cell. 2018 January;9(1):63-73).

[0054] Enhancement of ADCC: Fc engineering can be used to enhance ADCC. For example (with reference to IgG1), F243L / R292P / Y300L / V305I / P396L; S239D / I332E; and S298A / E333A / K334A increase FcγRIIIa binding; S239D / I332E / A330L increase FcγRIIIa binding and decrease FcγRIIb binding; G236A / S239D / I332E improves binding to FcγRIIa, improves the FcγRIIa / FcγRIIb binding ratio (activation / inhibition ratio), and enhances phagocytosis of antibody-coated target cells by macrophages. An asymmetric Fc containing L234Y / L235Q / G236W / S239M / H268D / D270E / S298A mutations in one heavy chain and D270E / K326D / A330M / K334E in the opposite heavy chain increases affinity for FcγRIIIa F158 (low affinity allele) and FcγRIIIa V158 (high affinity allele) but does not increase binding affinity for the inhibitory FcγRIIb (Mimoto et al., 2013).

[0055] Enhancing ADCP: Fc engineering can be used to enhance ADCP. For example (with reference to IgG1), G236A / S239D / I332E increases FcγRIIa binding and increases FcγRIIIa binding (Richards J et al., Mol. Cancer Ther. 2008;7:2517-2527).

[0056] Increased simultaneous binding: Fc engineering can be used to increase simultaneous binding to FcRs. For example (with reference to IgG1), S267E / L328F increases FcγRIIb binding; N325S / L328F increases FcγRIIa binding and decreases FcγRIIIa binding (Wang et al., 2018).

[0057] Reduced effector function Some isotypes of human constant regions, particularly IgG4 and IgG2 isotypes, are essentially deficient in the following functions: a) activation of complement by the classical pathway; and b) ADCC. Various modifications to the constant region of the heavy chain of the antigen-binding protein can be performed to alter the effector function depending on the desired effector property. IgG1 constant regions containing specific mutations that reduce binding to Fc receptors and decrease effector functions, e.g., ADCC and CDC, have been described (Duncan et al., Nature 1988, 332; 563-564; Lund et al., J. Immunol. 1991, 147; 2657-2662; Chappel et al., PNAS 1991, 88; 9036-9040; Burton and Woof, Adv. Immunol. 1992, 51; 1-84; Morgan et al., Immunology 1995, 86; 319-324; Hezareh et al., J. Virol. 2001, 75(24); 12161-12168).

[0058] In one embodiment of the present invention, an antigen binding protein is provided that comprises a constant region such that the effector function of the antigen binding protein, e.g., ADCC and / or CDC, is reduced. In such an embodiment, the constant region of the heavy chain may comprise a naturally disabled constant region of an IgG2 or IgG4 isotype or a mutated IgG1 constant region. Examples of suitable modifications are described in EP0307434. One example includes substitutions with alanine at positions 235 and 237 (EU index numbering), i.e., L235A and G237A (commonly referred to as "LAGA" mutations). Another example includes substitutions with alanine at positions 234 and 235 (EU index numbering), i.e., L234A and L235A (commonly referred to as "LALA" mutations). Further examples described in EP2691417 and US8969526 include, in IgG1 Fc, P329G or P329R in combination with a LALA mutation (EU index numbering), and in IgG4 Fc (EU index numbering), P329G or P329R in combination with S228P and L235E.

[0059] Mutations that increase half-life by adding or increasing FcRn binding "Half-life" refers to the time required for the serum concentration of an antigen-binding protein to reach half of its original value. The serum half-life of a protein can be measured by pharmacokinetic studies according to the method described in Kim et al., 1994, Eur.J.of Immuno.24:542-548. According to this method, a radiolabeled protein is injected intravenously into mice and the plasma concentration versus time is measured periodically, for example, from about 3 minutes to about 72 hours after injection. Other methods for pharmacokinetic analysis and half-life determination of molecules will be familiar to those skilled in the art.

[0060] The antigen binding proteins of the invention may have amino acid modifications that increase the affinity of the constant domain or fragment thereof for FcRn. Increasing the half-life (i.e., serum half-life) of therapeutic and diagnostic IgG antibodies and other bioactive molecules has many advantages, including reducing the dosage and / or frequency of administration of these molecules. In one embodiment, the antigen binding proteins of the invention comprise all or a portion of an IgG constant domain (FcRn binding portion) with one or more of the following amino acid modifications:

[0061] For example, with reference to IgG1, M252Y / S254T / T256E (commonly referred to as the "YTE" mutation) and M428L / N434S (commonly referred to as the "LS" mutation) increase FcRn binding at pH 6.0 (Wang et al., 2018).

[0062] Half-life can also be enhanced by T250Q / M428L, V259I / V308F / M428L, N434A, and T307A / E380A / N434A mutations (according to IgG1 and Kabat numbering) (Monnet et al.).

[0063] Half-life and FcRn binding can also be extended by the introduction of H433K and N434F mutations (commonly referred to as "HN" or "NHance" mutations) (with reference to IgG1) (WO2006 / 130834).

[0064] WO 00 / 42072 discloses polypeptides comprising a variant Fc region with altered FcRn binding affinity, the Fc region comprising one or more amino acid modifications at amino acid positions 238, 252, 253, 254, 255, 256, 265, 272, 286, 288, 303, 305, 307, 309, 311, 312, 317, 340, 356, 360, 362, 376, 378, 380, 386, 388, 400, 413, 415, 424, 433, 434, 435, 436, 439 and 447 (EU index numbering).

[0065] WO02 / 060919 discloses modified IgGs comprising an IgG constant domain which contains one or more amino acid modifications relative to the wild-type IgG constant domain, which have an increased half-life compared to the half-life of an IgG having the wild-type IgG constant domain, wherein the one or more amino acid modifications are present at one or more of positions 251, 253, 255, 285-290, 308-314, 385-389 and 428-435.

[0066] Shields et al. (2001, J Biol Chem;276:6591-604) used alanine scanning mutagenesis to alter residues within the Fc region of a human IgG1 antibody and then assessed binding to human FcRn. Positions that effectively abrogated binding to FcRn when changed to alanine included I253, S254, H435 and Y436. Other positions that showed less pronounced reductions in binding were: E233-G236, R255, K288, L309, S415 and H433. Several amino acid positions showed improved FcRn binding when changed to alanine; notable among them were P238, T256, E272, V305, T307, Q311, D312, K317, D376, E380, E382, S424 and N434. Many other amino acid positions showed little improvement (D265, N286, V303, K360, Q362 and A378) or no change in FcRn binding (S239, K246, K248, D249, M252, E258, T260, S267, H268, S269, D270, K274, N276, Y278, D280, V282, E283, H285, T289, K290, R292, E293, E294, Q295, Y296, N297, S298, R301, N315, E318, K320, (K322, S324, K326, A327, P329, P331, E333, K334, T335, S337, K338, K340, Q342, R344, E345, Q345, Q347, R356, M358, T359, K360, N361, Y373, S375, S383, N384, Q386, E388, N389, N390, K392, L398, S400, D401, K414, R416, Q418, Q419, N421, V422, E430, T437, K439, S440, S442, S444 and K447).

[0067] The most significant effect in terms of improved FcRn binding was found for the combination variant. At pH 6.0, the E380A / N434A variant showed over 8-fold better binding to FcRn compared to native IgG1, compared to 2-fold for E380A and 3.5-fold for N434A. Addition of T307A to this variant improved binding by 12-fold compared to native IgG1. In one embodiment, the antigen binding protein of the invention comprises the E380A / N434A mutations and has increased binding to FcRn.

[0068] Dall'Acqua et al. (2002, J Immunol.;169:5171-80) described random mutagenesis and screening of a phage display library of hinge-Fc fragments of human IgG1 against mouse FcRn. They disclosed random mutagenesis of positions 251, 252, 254-256, 308, 309, 311, 312, 314, 385-387, 389, 428, 433, 434 and 436. The stability of the IgG1-human FcRn complex is largely improved by substitution of residues located in the belt-like region that crosses the Fc-FcRn interface (M252, S254, T256, H433, N434 and Y436) and to a lesser extent by substitution of surrounding residues such as V308, L309, Q311, G385, Q386, P387 and N389. The variant with the highest affinity for human FcRn was obtained by combining the M252Y / S254T / T256E mutations ("YTE" mutations) with the H433K / N434F / Y436H mutations, and showed a 57-fold increase in affinity compared to wild-type IgG1. The in vivo behavior of such mutant human IgG1 showed an almost 4-fold increase in serum half-life in cynomolgus monkeys compared to wild-type IgG1.

[0069] Thus, the present invention provides antigen-binding proteins with optimized binding to FcRn. In a preferred embodiment, the antigen binding protein comprises one or more amino acid modifications in the Fc region of said antigen binding protein, said modifications being 226, 227, 228, 230, 231, 233, 234, 239, 241, 243, 246, 250, 252, 256, 259, 264, 265, 267, 269, 270, 276, 284, 285, 288, 289, 290, 291, 292, 294, 297, 298, 299, 301, 302, 303, 305, 307, 308, 309, 311, 315, 317, 320, 322, 325, 327, 330, 332, 334, 335, 338, 340, 341, 342, 343, 344, 345, 346, 347, 348, 349, 350, 351, 352, 353, 354, 355, 356, 357, 358, 359, 360, 361, 362, 363, 364, 365, 366, 367, 368, 369, 370, 371, 372, 373, 374, 375, 376, 377, 378, 379, 380, 381, 382, ​​383, 384, 42, 343, 345, 347, 350, 352, 354, 355, 356, 359, 360, 361, 362, 369, 370, 371, 375, 378, 380, 382, ​​384, 385, 386, 387, 389, 390, 392, 393, 394, 395, 396, 397, 398, 399, 400 , 401, 403, 404, 408, 411, 412, 414, 415, 416, 418, 419, 420, 421, 422, 424, 426, 428, 433, 434, 438, 439, 440, 443, 444, 445, 446 and 447.

[0070] Furthermore, various publications describe methods for obtaining bioactive molecules with modified half-lives by introducing FcRn-binding polypeptides into the molecule (WO97 / 43316, US5869046, US5747035, WO96 / 32478 and WO91 / 14438) or by fusing the molecule to antibodies with preserved FcRn-binding affinity but greatly reduced affinity for other Fc receptors (WO99 / 43713) or to the FcRn-binding domain of an antibody (WO00 / 09560, US4703039).

[0071] FcRn affinity-enhancing Fc variants to improve both antibody cytotoxicity and half-life were identified in a screen at pH 6.0. Selected IgG variants can be produced as hypofucosylated molecules. The resulting variants show improved serum persistence and preserved and enhanced ADCC in hFcRn mice (Monnet et al.). Exemplary variants include (according to IgG1 and Kabat numbering): P230T / V303A / K322R / N389T / F404L / N434S;P228R / N434S;Q311R / K334R / Q 342E / N434Y;C226G / Q386R / N434Y;T307P / N389T / N434Y;P230S / N434S;P23 0T / V305A / T307A / A378V / L398P / N434S;P23OT / P387S / N434S;P230Q / E269 D / N434S;N276S / A378V / N434S;T307A / N315D / A330V / 382V / N389T / N434Y;T 256N / A378V / S383N / N434Y;N315D / A330V / N361D / A387V / N434Y;V259I / N315D / M428L / N434Y;P230S / N315D / M428L / N434Y;F241L / V264E / T307P / A378V / H433R;T250A / N389K / N434Y;V305A / N315D / A330V / P395A / N434Y;V264E / Q386R / P396L / N434S / K439R;E294del / T307P / N434Y (where "del" indicates a deletion).

[0072] domain The term "domain" refers to a folded polypeptide structure that retains its tertiary structure independently of the rest of the polypeptide. Generally, domains are responsible for distinct functional properties of the polypeptide and can often be added, removed, or moved to other polypeptides without impairing the function of the remainder of the protein and / or the domain.

[0073] Single variable domains The term "single variable domain" refers to a folded polypeptide domain that contains sequences characteristic of antibody variable domains. Single variable domains therefore include complete antibody variable domains such as VH, VHH and VL, and modified antibody variable domains, such as those in which one or more loops have been replaced with sequences not characteristic of antibody variable domains, or antibody variable domains that are shortened or contain N-terminal or C-terminal extensions, as well as folded fragments of variable domains that retain at least the binding activity and specificity of the full-length domain. Single variable domains as defined herein can bind to an antigen or epitope independently of different variable regions or domains. A "domain antibody" or "DAB" can be considered the same as a human "single variable domain". Single variable domains can be human single variable domains, but also include single variable domains from other species, such as rodents (e.g., those disclosed in WO00 / 29004), nurse sharks and camelid VHHs. Camelid VHHs are immunoglobulin single variable domain polypeptides from species including camel, llama, alpaca, dromedary, and guanaco, which naturally produce heavy chain-only antibodies devoid of light chains. Such VHH domains can be humanized according to standard techniques available in the art, and such domains are considered to be "single variable domains".

[0074] Protein Scaffolds An antigen-binding fragment can be provided by placing one or more CDRs on a non-antibody protein scaffold. "Protein scaffold" as used herein includes, but is not limited to, immunoglobulin (Ig) scaffolds, such as IgG scaffolds that can be four-chain or two-chain antibodies, IgG scaffolds that can include only the Fc region of an antibody, IgG scaffolds that can include one or more constant regions of an antibody, IgG scaffolds where the constant region can be human or primate derived, or IgG scaffolds that can be artificial chimeras of human and primate constant regions.

[0075] The protein scaffold can be an Ig scaffold, e.g., an IgG, or an IgA scaffold. The IgG scaffold can include some or all of the domains (i.e., CH1, CH2, CH3, VH, VL) of an intact antibody. The antigen-binding protein can include an IgG scaffold selected from IgG1, IgG2, IgG3, IgG4, or IgG4PE. For example, the scaffold can be an IgG1. The scaffold can consist of or include an Fc region or a portion thereof of an antibody.

[0076] The protein scaffold may be a derivative of a scaffold selected from the group consisting of CTLA-4, lipocalin, protein A derived molecules such as Z domain (affibody, SpA), A domain (avimer / maxibody) of protein A; heat shock proteins such as GroEl and GroES; transferrin (trans-body); ankyrin repeat proteins (DARPins); peptide aptamers; C-type lectin domain (tetranectin); human □-crystallin and human ubiquitin (affilin); PDZ domain; scorpion venom Kunitz-type domain of human protease inhibitor; and fibronectin / adnectin, which have been subjected to protein engineering to obtain binding to an antigen, e.g., the CD4 binding site of gp120.

[0077] Attachment inhibitor As used herein, "attachment inhibitors" refers to a class of small molecule drugs that bind to the gp120 protein on the outer surface of HIV and prevent HIV from binding to and entering CD4+ T lymphocytes (CD4 cells) and / or that bind to the gp120 protein expressed on the outer surface of HIV-infected CD4 cells. Examples of attachment inhibitors include temsavir and fostemsavir.

[0078] Temsa Building As used herein, temsavir refers to 1-(4-benzoylpiperazin-1-yl)-2-[4-methoxy-7-(3-methyl-1,2,4-triazol-1-yl)-1H-pyrrolo[2,3-c]pyridin-3-yl]ethane-1,2-dione or compound (I) disclosed in US Pat. No. 7,354,924.

[0079] [ka]

[0080] Hostemsa Building As used herein, fostemsavir refers to 1-benzoyl-4-[2-[4-methoxy-7-(3-methyl-1H-1,2,4-triazol-1-yl)-1-[(phosphonooxy)methyl]-1H-pyrrolo[2,3-c]pyridin-3-yl]-1,2-dioxoethyl]-piperazine or compound (II) disclosed in US 7,745,625; US 8,168,615; and US 8,461,333.

[0081] [ka]

[0082] Fostemsavir is a prodrug (methyl phosphate replaced by H) that is metabolized to temsavir. The non-phosphonated compound is 1-benzoyl-4-[2-[4-methoxy-7-(3-methyl-1H-1,2,4-triazol-1yl)-1H-pyrrolo[2,3-c]pyridin-3-yl]-1,2-dioxoethyl]-piperazine.

[0083] Producing cells As used herein, "producer cells" refers to the cell population that is targeted and infected by HIV. This cell population then promotes the replication of HIV. CD4+ T lymphocytes and macrophages fall into this category of producer cells.

[0084] Prodrug As used herein, "prodrug" refers to a bioreversible derivative of a drug molecule which undergoes enzymatic and / or chemical transformation in vivo to release the active drug, which then exerts a desired pharmacological effect.

[0085] Pharmaceutical Compositions As used herein, the term "pharmaceutical composition" refers to a composition suitable for pharmaceutical use. The antigen binding proteins, antibodies, inhibitors and integrase inhibitors described herein can be incorporated into pharmaceutical compositions for use in the treatment of human diseases described herein. In one embodiment, the pharmaceutical composition comprises an antigen binding protein and one or more pharma- ceutically acceptable carriers and / or excipients.

[0086] Pharmaceutically acceptable salts As used herein, the term "pharmaceutical acceptable salts" refers to salts that retain the desired biological activity of the subject compound and exhibit minimal undesired toxicological effects. These pharmaceutical acceptable salts may be prepared in situ during the final isolation and purification of the compounds, or may be prepared by separately reacting the purified compounds in the form of a free acid or free base with a suitable base or acid, respectively.

[0087] Pharmaceutically acceptable salts include, inter alia, those described in Berge, J. Pharm. Sci., 1977, 66, 1-19, or those listed in PH Stahl and CG Wermuth, editors, Handbook of Pharmaceutical Salts; Properties, Selection and Use, 2nd ed. Stahl / Wermuth:Wiley- VCH / VHCA, 2011.

[0088] Suitable pharma- ceutically acceptable salts may include acid or base addition salts.Suitable pharma- ceutically acceptable salts of the present invention include base addition salts.

[0089] Representative pharma- ceutically acceptable base addition salts include aluminum, 2-amino-2-(hydroxymethyl)-1,3-propanediol (TRIS, tromethamine), arginine, benethamine (N-benzylphenethylamine), benzathine (N,N'-dibenzylethylenediamine), bis-(2-hydroxyethyl)amine, bismuth, calcium, chloroprocaine, choline, clemizole (1-chlorobenzyl-2-pyrrolildine-1'-ylmethyl benzimidazole), cyclohexylamine, dibenzylethylenediamine, diethylamine, diethyltriamine, dimethylamine, dimethylethanolamine, dopamine, ethanolamine, ethylenediamine, L-histidine, iron, isoquinoline, lepidine, lithium, lysine, magnesium, meglumine (N-methylglucamine), piperazine, piperidine, potassium, procaine, quinine, quinoline, sodium, strontium, t-butylamine, and zinc.

[0090] Therapeutically Effective Dose As used herein, the term "therapeutically effective amount" or "effective amount" refers to an amount of compound administered that prevents a pathology or relieves to some extent one or more symptoms of the disorder being treated. Pharmaceutical compositions suitable for use herein include compositions in which the active ingredient is contained in an amount sufficient to achieve the intended purpose. Determining a therapeutically effective amount is well within the capabilities of those skilled in the art, especially in light of the detailed description provided herein.

[0091] Treatment As used herein, the terms "treatment", "treating" or "treat" in the context of a method of therapy refer to alleviating a particular condition, eliminating or reducing the symptoms of a condition, slowing or eliminating the progression, infiltration or spread of a condition, as well as reducing or delaying the recurrence of a condition in a previously affected human. The invention further provides for the use of a compound or composition of the invention for the preparation of a medicament in a mammal (e.g., a human) in need thereof for the treatment of some conditions.

[0092] Prevention As used herein, the terms "prevention," "preventing," or "prevent" in the context of therapy refer to preventing a particular condition or symptoms of a condition, or preventing the recurrence of a condition in a prior outbreak of infection. The invention further provides the use of a compound of the invention for the preparation of a medicament in a mammal (e.g., a human) in need thereof for the prevention of some condition.

[0093] Neutraliza As used herein, the terms "neutralize" or "neutralizing" refer to reducing the biological activity of an HIV antigen, e.g., ENV or gp120, in the presence of an antigen-binding protein, in vitro or in vivo, compared to the activity of the antigen in the absence of the antigen-binding protein. Neutralization of HIV can be by preventing viral entry through interaction with the HIV receptor.

[0094] HIV antigen recognition As used herein, the terms "HIV antigen recognition," "antigen recognition," or "ENV detection" refer to binding to ENV on the surface of an infected cell by the CD4bs binding protein.

[0095] Clearance As used herein, the terms "clearance", "clearing" or "clearing" refer to the killing of HIV-infected cells. As an example, in one embodiment, "clearance" refers to the therapeutic administration or combination of administrations, alone or in combination with one or more other compounds, inducing the removal, elimination, or killing of cells carrying replication-competent HIV genomes capable of producing virus. Clearance can be reflected, for example, as a reduction in the level of HIV DNA as measured by polymerase chain reaction (PCR) testing. Clearance can also be reflected as a delay in the detection of HIV virus when therapeutic intervention is discontinued.

[0096] Cure As used herein, the term "cure", "curing" or "viral remission" in the context of a therapeutic method refers to eradicating, halting, pausing or terminating the human immunodeficiency virus or symptoms, or the progression of the symptoms or the virus, for a defined period of time. As an example, in one embodiment, "cure" or "curing" refers to a therapeutic administration or combination of administrations, alone or in combination with one or more other compounds, that induces and maintains (after a minimum of two years) sustained viral control of the human immunodeficiency virus (e.g., no detectable plasma viremia by polymerase chain reaction (PCR) test, bDNA (branched chain DNA) test or nucleic acid sequence-based amplification (NASBA) test) without other therapeutic intervention. The above PCR, bDNA and NASBA tests are performed using techniques known and well known to those skilled in the art. As an example, eradication, halting, pausing or terminating the human immunodeficiency virus or symptoms, or the progression of the symptoms or the virus, may last for a minimum of two years.

[0097] Parenteral As used herein, the terms "parenteral" or "parenterally" in the context of therapeutic methods refer to a route of administration other than oral administration of a pharmaceutical compound or composition. Parenteral routes of administration suitable for use herein include injection, infusion, implantation, or any other route other than the digestive tract. Parenteral injection routes of administration include intravenous, intramuscular, and subcutaneous injection routes of administration.

[0098] Closed Conformation As used herein, the terms "closed conformation", "state 1 conformation", "prefusion conformation" or "pre-triggered state" refer to the state of the HIV ENV trimer assembly prior to CD4 receptor binding, which exposes an outer surface heavily covered with N-linked glycans that confers evasion from neutralizing antibody recognition. Interaction with the CD4 receptor triggers a structural rearrangement of ENV and a conformational transition to an intermediate open state, thus exposing co-receptor binding sites and epitopes that are hidden in the closed conformation.

[0099] Description of the Invention How to Treat HIV The method of the present invention may be used to treat, clear, neutralize, prevent or cure human immunodeficiency virus (HIV). In one aspect, the present invention provides a method of treating or preventing HIV, comprising administering a therapeutically effective amount of two or more agents selected from the following group: CD4bs binding protein; gp120 binding protein; broadly neutralizing antibody or antigen-binding fragment thereof; at least one agent selected from the group consisting of fostemsavir and temsavir, or a pharmaceutically acceptable salt thereof; and an integrase inhibitor or a pharmaceutically acceptable salt thereof.

[0100] In one aspect, a method is provided for treating or preventing HIV infection in a human in need thereof, comprising administering therapeutically effective amounts of: (a) a first agent comprising at least one agent selected from the group consisting of fostemsavir and temsavir, or a pharma- ceutically acceptable salt thereof; and (b) a second agent comprising at least one broadly neutralizing antibody or an antigen-binding fragment thereof.

[0101] In one aspect, a method is provided for treating or preventing HIV infection in a human in need thereof, comprising administering therapeutically effective amounts of: (a) a first agent comprising at least one agent selected from the group consisting of fostemsavir and temsavir, or a pharmaceutically acceptable salt thereof; (b) a second agent comprising at least one broadly neutralizing antibody or antigen-binding fragment thereof; and (c) a third agent comprising at least one integrase inhibitor or a pharmaceutically acceptable salt thereof.

[0102] In some embodiments, a method for treating or preventing HIV infection in a human in need thereof is provided, comprising administering a therapeutically effective amount of: (a) a first agent comprising at least one agent selected from the group consisting of fostemsavir and temsavir, or a pharmaceutically acceptable salt thereof; and (b) a second agent comprising at least one broadly neutralizing antibody or an antigen-binding fragment thereof. In some embodiments, the first agent is fostemsavir or a pharmaceutically acceptable salt thereof. In some embodiments, the first agent is temsavir or a pharmaceutically acceptable salt thereof.

[0103] In some embodiments, the second agent is at least one broadly neutralizing antibody or antigen-binding fragment thereof that binds to at least one HIV glycoprotein selected from the group consisting of HIV gp160, HIV gp120, and HIV gp41. In some embodiments, the at least one broadly neutralizing antibody binds to HIV gp160. In some embodiments, the at least one broadly neutralizing antibody binds to HIV gp120. In some embodiments, the at least one broadly neutralizing antibody binds to HIV gp41. In some embodiments, the second agent is at least one antibody selected from the group consisting of 2G12, 2F5, 3BC176, 3BNC60, 3BNC1-17, 4E10, 8ANC131, 8ANC195, 10E8, 10-1074, 12A12, 35022, b12, B2530, CH01-04, CH103, CH31, HJ16, M66.6, N6, N6LS, N6-DE, N6-LAGA, NIH45-46, PG9, PG16, PGDM1400, PGT121, PGT128, PGT135, PGT141-PGT145, PGT151, PGV04, VRC01, VRC01-LS, VRC07, VRC07-523, VRC07-LS, and Z13. In some embodiments, the second agent is at least one antibody selected from the group consisting of N6, N6LS, N6-DE, N6-LAGA, VRC01, VRC01-LS, VRC07, VRC07-523, and VRC07-LS. In some embodiments, the second agent is at least one antibody selected from the group consisting of N6, N6LS, N6-DE, VRC01, VRC01-LS, VRC07, VRC07-523, and VRC07-LS. In some embodiments, the second agent is at least one antibody selected from the group consisting of N6, N6LS, and N6 or N6LS having any one of the ADCC mutations disclosed herein.In some embodiments, the second agent comprises a heavy chain complementarity determining region (CDRH) having an amino acid sequence of CDRH1 that comprises a sequence at least 95%, at least 98%, at least 99% or 100% identical to SEQ ID NO:1, an amino acid sequence of CDRH2 that comprises a sequence at least 95%, at least 98%, at least 99% or 100% identical to SEQ ID NO:2, and an amino acid sequence of CDRH3 that comprises a sequence at least 95%, at least 98%, at least 99% or 100% identical to SEQ ID NO:3, and and a CDRH3 amino acid sequence that is at least 95%, at least 98%, at least 99% or 100% identical to SEQ ID NO: 6. In one embodiment, the second agent is an isolated monoclonal antibody or antigen-binding fragment thereof, comprising a light chain complementarity determining region (CDRL) having an amino acid sequence of CDRL1 that comprises a sequence that is at least 95%, at least 98%, at least 99% or 100% identical to SEQ ID NO: 5, an amino acid sequence of CDRL2 that comprises a sequence that is at least 95%, at least 98%, at least 99% or 100% identical to SEQ ID NO: 6. In one embodiment, the second agent is a heavy chain variable region (V) having at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO: 7. H In some embodiments, the second agent is an isolated monoclonal antibody or antigen-binding fragment thereof comprising a light chain variable region (V) having at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO:8. L In some embodiments, the isolated monoclonal antibody further comprises a recombinant constant domain comprising an M428L mutation and an N434S mutation. In some embodiments, the isolated monoclonal antibody further comprises a recombinant constant domain comprising an S239D mutation and an I332E mutation. In some embodiments, the isolated monoclonal antibody further comprises a recombinant constant domain comprising an L235A mutation and a G237A mutation.

[0104] In some embodiments, the second agent is an isolated N6 monoclonal antibody or an antigen-binding fragment thereof comprising a heavy chain complementarity determining region (CDRH) having the amino acid sequence of CDRH1 of SEQ ID NO: 1, the amino acid sequence of CDRH2 of SEQ ID NO: 2, and the amino acid sequence of CDRH3 of SEQ ID NO: 3, and a light chain complementarity determining region (CDRL) having the amino acid sequence of CDRL1 of SEQ ID NO: 4, the amino acid sequence of CDRL2 of SEQ ID NO: 5, and the amino acid sequence of CDRH3 of SEQ ID NO: 6. In some embodiments, the second agent is an isolated N6LS monoclonal antibody or an antigen-binding fragment thereof, comprising: a heavy chain complementarity determining region (CDRH) having the amino acid sequence of CDRH1 of SEQ ID NO: 1, the amino acid sequence of CDRH2 of SEQ ID NO: 2, and the amino acid sequence of CDRH3 of SEQ ID NO: 3; a light chain complementarity determining region (CDRL) having the amino acid sequence of CDRL1 of SEQ ID NO: 4, the amino acid sequence of CDRL2 of SEQ ID NO: 5, and the amino acid sequence of CDRH3 of SEQ ID NO: 6; and a recombinant constant domain comprising an M428L mutation and an N434S mutation. In some embodiments, the second agent is an isolated N6-DE monoclonal antibody or an antigen-binding fragment thereof, comprising: a heavy chain complementarity determining region (CDRH) having an amino acid sequence for CDRH1 of SEQ ID NO: 1, an amino acid sequence for CDRH2 of SEQ ID NO: 2, and an amino acid sequence for CDRH3 of SEQ ID NO: 3; a light chain complementarity determining region (CDRL) having an amino acid sequence for CDRL1 of SEQ ID NO: 4, an amino acid sequence for CDRL2 of SEQ ID NO: 5, and an amino acid sequence for CDRH3 of SEQ ID NO: 6; and a recombinant constant domain comprising an S239D mutation and an I332E mutation. In some embodiments, the second agent is an isolated N6-LAGA monoclonal antibody or an antigen-binding fragment thereof, comprising: a heavy chain complementarity determining region (CDRH) having the amino acid sequence of CDRH1 of SEQ ID NO: 1, the amino acid sequence of CDRH2 of SEQ ID NO: 2, and the amino acid sequence of CDRH3 of SEQ ID NO: 3; a light chain complementarity determining region (CDRL) having the amino acid sequence of CDRL1 of SEQ ID NO: 4, the amino acid sequence of CDRL2 of SEQ ID NO: 5, and the amino acid sequence of CDRH3 of SEQ ID NO: 6; and a recombinant constant domain comprising the L235A and G237A mutations.In some embodiments, the antigen-binding fragment is an Fv, Fab, F(ab')2, scFv or scFV2 ​​fragment.

[0105] In some embodiments, the method further comprises administering a therapeutically effective amount of a third agent comprising at least one integrase inhibitor or a pharma- ceutically acceptable salt thereof. In some embodiments, the third agent is at least one agent selected from the group consisting of raltegravir, elvitegravir, dolutegravir, bictegravir and cabotegravir. In some embodiments, the third agent is raltegravir or cabotegravir. In some embodiments, the third agent is cabotegravir. In some embodiments, the second agent is N6 with one or more Fc modifications in the constant domain disclosed herein. In some embodiments, the second agent is N6LS. In some embodiments, the second agent is N6-DE. In some embodiments, the first agent is temsavir, the second agent is selected from the group consisting of N6LS and N6-DE, and the third agent is cabotegravir. In some embodiments, the first drug is temsavir, the second drug is N6LS, and the third drug is cabotegravir. In some embodiments, the first drug is temsavir, the second drug is N6-DE, and the third drug is cabotegravir. In some embodiments, the first drug is fostemsavir, the second drug is N6LS, and the third drug is cabotegravir. In some embodiments, the first drug is fostemsavir, the second drug is N6-DE, and the third drug is cabotegravir.

[0106] In some embodiments, the method according to any one of the above embodiments, each of the first agent, the second agent, and the third agent is in the form of a pharmaceutical composition. In some embodiments, the first agent is administered before administering the second agent. In some embodiments, the method comprises orally administering to a human about 1 mg / kg body weight to about 100 mg / kg body weight of the first agent once a day, twice a day, or three times a day. In some embodiments, the method comprises parenterally administering to a human about 1 mg / kg body weight to about 100 mg / kg body weight of the first agent once a day, twice a day, or three times a day.

[0107] In some embodiments, the human is diagnosed with human immunodeficiency virus 1 (HIV-1) infection. In some embodiments, the human has been previously treated with one or more different HIV treatment modalities. In some embodiments, the human has been treated with a treatment regimen of a combination of antiretroviral drugs termed highly active antiretroviral therapy (HAART). In some embodiments, the human has been treated with state-of-the-art antiretroviral therapy (ART).

[0108] In one aspect, the present invention provides a method of treating or preventing HIV in a human in need thereof, comprising administering to the human therapeutically effective amounts of: (a) a first agent comprising at least one agent selected from the group consisting of fostemsavir and temsavir, or a pharmaceutically acceptable salt thereof; and (b) a second agent selected from the group consisting of a CD4bs binding protein, a gp120 binding protein, and an integrase inhibitor, or a pharmaceutically acceptable salt thereof.

[0109] In one aspect, the present invention provides a method of treating or preventing HIV in a human in need thereof, comprising administering to the human therapeutically effective amounts of: (a) a first agent comprising at least one agent selected from the group consisting of fostemsavir and temsavir, or a pharmaceutically acceptable salt thereof; (b) a second agent comprising at least one CD4bs binding protein; and (c) a third agent comprising at least one integrase inhibitor, or a pharmaceutically acceptable salt thereof.

[0110] According to one aspect, the present invention provides a method of treating HIV infection in a human in need thereof, comprising administering to the human a therapeutically effective amount of at least one agent selected from the group consisting of fostemsavir and temsavir, or a pharmaceutically acceptable salt thereof, and sequentially administering a therapeutically effective amount of a CD4bs binding protein, wherein the CD4bs binding protein is administered after the at least one agent selected from the group consisting of fostemsavir and temsavir, or a pharmaceutically acceptable salt thereof.

[0111] According to another aspect, the present invention provides a method for clearing HIV-infected cells from a human in need thereof, comprising administering a therapeutically effective amount of at least one agent selected from the group consisting of fostemsavir and temsavir, or a pharmaceutically acceptable salt thereof, and a therapeutically effective amount of a CD4bs binding protein. In one embodiment of the method for clearing HIV-infected cells, the administration of at least one agent selected from the group consisting of fostemsavir and temsavir, or a pharmaceutically acceptable salt thereof, and the CD4bs binding protein is simultaneous. In one embodiment of the method for clearing HIV-infected cells, the CD4bs binding protein is administered after the administration of at least one agent selected from the group consisting of fostemsavir and temsavir, or a pharmaceutically acceptable salt thereof.

[0112] According to another aspect, the present invention provides a method of preventing HIV infection in a human in need thereof, comprising administering to the human a therapeutically effective amount of at least one agent selected from the group consisting of fostemsavir and temsavir, or a pharmaceutically acceptable salt thereof, and sequentially administering a therapeutically effective amount of a CD4 binding site (CD4bs) binding protein, wherein the CD4bs binding protein is administered after the at least one agent selected from the group consisting of fostemsavir and temsavir, or a pharmaceutically acceptable salt thereof.

[0113] According to another aspect, the present invention provides a method of neutralizing HIV infection in a human in need thereof, comprising administering to the human a therapeutically effective amount of at least one agent selected from the group consisting of fostemsavir and temsavir, or a pharmaceutically acceptable salt thereof, and sequentially administering a therapeutically effective amount of a CD4 binding site (CD4bs) binding protein, wherein the CD4bs binding protein is administered following administration of the at least one agent selected from the group consisting of fostemsavir and temsavir, or a pharmaceutically acceptable salt thereof.

[0114] According to another aspect, the present invention provides a method of curing HIV infection in a human in need thereof, comprising administering to the human a therapeutically effective amount of at least one agent selected from the group consisting of fostemsavir and temsavir, or a pharmaceutically acceptable salt thereof, and sequentially administering a therapeutically effective amount of a CD4 binding site (CD4bs) binding protein, wherein the CD4bs binding protein is administered following administration of the at least one agent selected from the group consisting of fostemsavir and temsavir, or a pharmaceutically acceptable salt thereof.

[0115] In some embodiments, the at least one agent is temsavir or fostemsavir. In one embodiment, the at least one agent is temsavir. In some embodiments, the at least one agent is fostemsavir. Without being bound by any theory or mechanism of action, temsavir or fostemsavir may act by blocking the gp120 viral receptor, preventing the virus from first attaching to the host CD4+ cell and thus preventing entry into the host CD4+ cell. In some embodiments, the administration of at least one agent selected from the group consisting of fostemsavir and temsavir, or a pharmaceutically acceptable salt thereof, is oral or parenteral. In one embodiment of the method, the administration of temsavir is parenteral. In some embodiments, the administration of fostemsavir is oral. In some embodiments, the at least one agent selected from the group consisting of fostemsavir and temsavir, or a pharmaceutically acceptable salt thereof, is administered in a dose range of 1 mg / kg body weight to 100 mg / kg body weight. In some embodiments, the dose of at least one agent selected from the group consisting of fostemsavir and temsavir, or a pharmaceutically acceptable salt thereof, is 100 mg to 1200 mg. In one embodiment of the method, the dose of fostemsavir is 600 mg. In some embodiments, the administration of at least one agent selected from the group consisting of fostemsavir and temsavir, or a pharmaceutically acceptable salt thereof, is a daily dose, a twice-daily dose, or a three times daily dose. In some embodiments, the administration is a twice-daily dose. In some embodiments, the administration is a once-daily dose. In one embodiment of the method, the administration is a three times daily dose.

[0116] In some embodiments, at least one agent selected from the group consisting of fostemsavir and temsavir, or a pharmaceutically acceptable salt thereof, locks gp120 on the host cell membrane surface. In some embodiments, at least one agent selected from the group consisting of fostemsavir and temsavir, or a pharmaceutically acceptable salt thereof, locks gp120 in a closed conformation. In some embodiments, at least one agent selected from the group consisting of fostemsavir and temsavir, or a pharmaceutically acceptable salt thereof, prevents cell membrane-bound CD4 from interacting with gp120 expressed in the host cell. In some embodiments, at least one agent selected from the group consisting of fostemsavir and temsavir, or a pharmaceutically acceptable salt thereof, enables or promotes binding of CD4bs binding protein. In some embodiments, binding of CD4bs binding protein to HIV-infected host cells is measured by flow cytometry using mean fluorescence intensity (MFI).

[0117] In some embodiments, the method includes a gp120 binding protein. The gp120 binding protein binds to gp120 found on HIV virions or infected host cells. In some embodiments, the gp120 binding protein includes a CD4bs binding protein. In some embodiments, the CD4bs binding protein is at least one CD4bs binding protein. In some embodiments, the CD4bs binding protein is an antibody or a binding fragment thereof. In some embodiments, the antibody is a broadly neutralizing antibody or a binding fragment thereof. In some embodiments, the broadly neutralizing antibody is selected from the group consisting of N6, N6LS, N6-DE and N6-LAGA.

[0118] In some embodiments, the CD4bs binding protein comprises a heavy chain complementarity determination having an amino acid sequence for CDRH1 that comprises a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical to SEQ ID NO:1, an amino acid sequence for CDRH2 that comprises a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical to SEQ ID NO:2, and an amino acid sequence for CDRH3 that comprises a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical to SEQ ID NO:3. and a light chain complementarity determining region (CDRL) having an amino acid sequence for CDRL1 that comprises a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical to SEQ ID NO:4, an amino acid sequence for CDRL2 that comprises a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical to SEQ ID NO:5, and an amino acid sequence for CDRL3 that comprises a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical to SEQ ID NO:6.

[0119] In some embodiments, the CD4bs binding protein comprises an amino acid sequence of a heavy chain variable region (VH) comprising a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical to a sequence selected from the group consisting of SEQ ID NO: 7. In some embodiments, the CD4bs binding protein comprises an amino acid sequence of a light chain variable region (VL) comprising a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical to a sequence selected from the group consisting of SEQ ID NO: 8. In some embodiments, the CD4bs binding protein comprises an amino acid sequence of a heavy chain (HC) comprising a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical to a sequence selected from the group consisting of SEQ ID NO: 9, 11, 13 and 14. In some embodiments, the CD4bs binding protein comprises an amino acid sequence of a light chain (LC) comprising a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical to a sequence selected from the group consisting of SEQ ID NOs: 10 and 12.

[0120] In some embodiments, the CD4bs binding protein antibody comprises a full-length immunoglobulin CD4bs binding protein. In one embodiment of the method, the full-length immunoglobulin CD4bs binding protein comprises a fragment crystallizable region or "Fc" region or "constant region." The Fc of any of the defined antibodies may contain one or more amino acid substitutions to optimize the in vivo half-life of the antibody. The serum half-life of IgG Abs is regulated by the fetal Fc receptor (FcRn). In some embodiments, the antibody contains an amino acid substitution that increases binding to FcRn. Some such substitutions are known to those of skill in the art, such as T250Q and M428L in the IgG constant region (see, e.g., Hinton et al., J Immunol, 176:346-356, 2006); M428L and N434S ("LS" mutations, see, e.g., Zalevsky et al., Nature 1999:131-135, 2006); Biotechnology, 28:157-159, 2010); N434A (see, e.g., Petkova et al., Int. Immunol, 18:1759-1769, 2006); T307A, E380A and N434A (see, e.g., Petkova et al., Int. Immunol, 18:1759-1769, 2006); and substitutions in M252Y, S254T and T256E (see, e.g., Dall'Acqua et al., J. Biol. Chem., 281:23514-23524, 2006). The disclosed antibodies and antigen-binding fragments can be linked to an Fc polypeptide that includes any of the substitutions listed above, for example, the Fc polypeptide can include an M428L substitution and an N434S substitution. In some embodiments, the antibody comprises a recombinant constant domain comprising a modification that improves binding to a fetal Fc receptor compared to an unmodified constant domain, and the recombinant domain is an IgG1 constant domain comprising an M428L mutation and an N434S mutation.

[0121] In some embodiments, the constant region of any of the defined antibodies comprises one or more amino acid substitutions to optimize antibody-dependent cell-mediated cytotoxicity (ADCC). ADCC is primarily mediated through a set of closely related Fc gamma receptors (FcyR). In some embodiments, the antibody comprises one or more amino acid substitutions that increase binding to FcyRIIIa. Some such substitutions are known to those skilled in the art, such as S239D and I332E in the IgG constant region (see, e.g., Lazar et al., Proc. Natl, Acad. Sci. USA, 103:4005-4010, 2006); S239D, A330L and I332E (see, e.g., Lazar et al., Proc. Natl, Acad. Sci. USA, 103:4005-4010, 2006).

[0122] In some embodiments, combinations of the above substitutions are also included to generate IgG constant regions with increased binding to FcRn and FcyRIIIa. The combinations increase the half-life and ADCC of the antibody. For example, such combinations include antibodies with the following amino acid substitutions in the Fc region: (1) S239D / I332E and T250Q / M428L; (2) S239D / I332E and M428L / N434S; (3) S239D / I332E and N434A; (4) S239D / I332E and T307A / E380A / N434A; (5) S239D / I332E and M252Y / S254T / T256 (6) S239D / A330L / I332E and 250Q / M428L; (7) S239D / A330L / I332E and M428L / N434S; (8) S239D / A330L / I332E and N434A; (9) S239D / A330L / I332E and T307A / E380A / N434A; or (10) S239D / A330L / I332E and M252Y / S254T / T256E. In some embodiments, the antibody or antigen-binding fragment thereof is modified to be directly cytotoxic to infected cells or to exploit natural defenses, such as complement, antibody-dependent cellular cytotoxicity (ADCC) or phagocytosis by macrophages.

[0123] In some embodiments, the antibody is an antibody fragment. In some embodiments, the antibody fragment is selected from a Fab fragment, a F(ab')2 fragment, a single chain Fv protein ("scFv"), a disulfide stabilized Fv protein ("dsFv"), a diabody, and a TANBAB. In one embodiment of this method, the antibody fragment is a scFv.

[0124] In some embodiments, the CD4bs binding protein comprises a recombinant constant domain comprising a modification that improves binding to a fetal Fc receptor compared to an unmodified constant domain, and the recombinant domain is an IgG1 constant domain comprising an M428L mutation and an N434S mutation (the "LS" mutation).

[0125] In some embodiments, the CD4bs binding protein comprises a recombinant constant domain that comprises a modification that affects antibody-dependent cell-mediated cytotoxicity (ADCC) compared to an unmodified constant domain, and the recombinant domain is an IgG1 constant domain that comprises S239D / I332E mutations ("DE" mutations) or L235A / G237A mutations ("LAGA" mutations).

[0126] In some embodiments, the CD4bs binding protein is selected from the group consisting of N6, N6LS, N6-LAGA and N6-DE. In some embodiments, the CD4bs binding protein is N6. In some embodiments, the CD4bs binding protein is N6LS. In some embodiments, the CD4bs binding protein is N6-LAGA. In some embodiments, the CD4bs binding protein is N6-DE.

[0127] In some embodiments, the CD4bs binding protein is administered consecutively at least 30 minutes, at least 1 hour, at least 2 hours, at least 4 hours, or at least 6 hours after administration of at least one agent selected from the group consisting of fostemsavir and temsavir, or a pharmaceutically acceptable salt thereof. In some embodiments, the CD4bs binding protein is administered at least 30 minutes after administration of at least one agent selected from the group consisting of fostemsavir and temsavir, or a pharmaceutically acceptable salt thereof. In some embodiments, the CD4bs binding protein is administered at least 1 hour after administration of at least one agent selected from the group consisting of fostemsavir and temsavir, or a pharmaceutically acceptable salt thereof. In one embodiment of the method, the CD4bs binding protein is administered at least 2 hours after administration of at least one agent selected from the group consisting of fostemsavir and temsavir, or a pharmaceutically acceptable salt thereof. In some embodiments, the CD4bs binding protein is administered at least 4 hours after administration of at least one agent selected from the group consisting of fostemsavir and temsavir, or a pharmaceutically acceptable salt thereof. In one embodiment of the method, the CD4bs binding protein is administered at least 6 hours after administration of at least one agent selected from the group consisting of fostemsavir and temsavir, or a pharmaceutically acceptable salt thereof. In some embodiments, the CD4bs binding protein is administered at least 24 hours after administration of at least one agent selected from the group consisting of fostemsavir and temsavir, or a pharmaceutically acceptable salt thereof. In one embodiment of the method, the CD4bs binding protein is administered at least 48 hours after administration of at least one agent selected from the group consisting of fostemsavir and temsavir, or a pharmaceutically acceptable salt thereof. In some embodiments, the CD4bs binding protein is administered at least 72 hours after administration of at least one agent selected from the group consisting of fostemsavir and temsavir, or a pharmaceutically acceptable salt thereof.In some embodiments, the CD4bs binding protein is administered at least 96 hours after administration of at least one agent selected from the group consisting of fostemsavir and temsavir, or a pharmaceutically acceptable salt thereof. Without being bound by theory, administration of at least one agent selected from the group consisting of fostemsavir and temsavir, or a pharmaceutically acceptable salt thereof, to a human prior to administration of a CD4 binding site (CD4bs) binding protein enhances binding of the CD4bs binding protein to HIV ENV. This is believed to be because administration of at least one agent selected from the group consisting of fostemsavir and temsavir, or a pharmaceutically acceptable salt thereof, to a human alters intracellular glycosylation and / or processing of the ENV trimer, facilitating binding and HIV antigen recognition of the CD4bs binding protein. It is also believed that administration of at least one drug selected from the group consisting of fostemsavir and temsavir, or a pharma- ceutical acceptable salt thereof, to a human locks or fixes the ENV trimer to the surface membrane of the host cell, preventing shedding of ENV and promoting engagement of bnAb.

[0128] In some embodiments, the method further comprises at least a second administration, after the sequential administration, of simultaneously administering at least one agent selected from the group consisting of fostemsavir and temsavir, or a pharma- ceutically acceptable salt thereof, and a CD4bs binding protein.

[0129] In various embodiments, the method comprises administering at least one integrase inhibitor. Integrase inhibitors, i.e., INSTI (integrase strand transfer inhibitor), are active substances that function by preventing retroviral integration. Exemplary compounds include, but are not limited to, raltegravir, elvitegravir, dolutegravir, bictegravir and cabotegravir. In some embodiments, the at least one integrase inhibitor comprises cabotegravir.

[0130] Combinations for use in the treatment of HIV In one aspect, the present invention provides a pharmaceutical composition for use in the treatment of HIV, comprising two or more agents selected from the following group: CD4bs binding protein; gp120 binding protein; at least one agent selected from the group consisting of fostemsavir and temsavir, or a pharma- ceutically acceptable salt thereof; and an integrase inhibitor, or a pharma- ceutically acceptable salt thereof.

[0131] In one aspect, the present invention provides a combination for use in the treatment or prevention of HIV, comprising administering to a human a first pharmaceutical composition comprising a first agent comprising at least one agent selected from the group consisting of fostemsavir and temsavir, or a pharmaceutically acceptable salt thereof, and a second pharmaceutical composition comprising a second agent comprising at least one broadly neutralizing antibody or antigen-binding fragment thereof. In some embodiments, the combination for use in the treatment or prevention of HIV further comprises administering a third pharmaceutical composition comprising a third agent comprising at least one integrase inhibitor, or a pharmaceutically acceptable salt thereof.

[0132] In some embodiments, the first agent is temsavir or a pharmaceutically acceptable salt thereof. In yet other embodiments, the first agent is fostemsavir or a pharmaceutically acceptable salt thereof.

[0133] In some embodiments, the second agent binds to the HIV envelope glycoprotein or HIV gp120. In some embodiments, the second agent is at least one agent selected from the group consisting of 2G12, 2F5, 3BC176, 3BNC60, 3BNC1-17, 4E10, 8ANC131, 8ANC195, 10E8, 10-1074, 12A12, 35022, b12, B2530, CH01-04, CH103, CH31, HJ16, M66.6, N6, N6LS, N6-DE, N6-LAGA, NIH45-46, PG9, PG16, PGDM1400, PGT121, PGT128, PGT135, PGT141-PGT145, PGT151, PGV04, VRC01, VRC01-LS, VRC07, VRC07-523, VRC07-LS, and Z13. In some embodiments, the second agent is at least one antibody selected from the group consisting of N6, N6LS, N6-DE, N6-LAGA, VRC01, VRC01-LS, VRC07, VRC07-523, and VRC07-LS. In some embodiments, the second agent is at least one antibody selected from the group consisting of N6, N6LS, N6-DE, VRC01, VRC01-LS, VRC07, VRC07-523, and VRC07-LS. In some embodiments, the second agent is at least one antibody selected from the group consisting of N6, N6LS, and N6 or N6LS having any one of the ADCC mutations disclosed herein.In some embodiments, the second agent comprises a heavy chain complementarity determining region (CDRH) having an amino acid sequence of CDRH1 that comprises a sequence at least 95%, at least 98%, at least 99% or 100% identical to SEQ ID NO:1, an amino acid sequence of CDRH2 that comprises a sequence at least 95%, at least 98%, at least 99% or 100% identical to SEQ ID NO:2, and an amino acid sequence of CDRH3 that comprises a sequence at least 95%, at least 98%, at least 99% or 100% identical to SEQ ID NO:3, and and a CDRH3 amino acid sequence that is at least 95%, at least 98%, at least 99% or 100% identical to SEQ ID NO: 6. In one embodiment, the second agent is an isolated monoclonal antibody or antigen-binding fragment comprising a light chain complementarity determining region (CDRL) having an amino acid sequence of CDRL1 that comprises a sequence that is at least 95%, at least 98%, at least 99% or 100% identical to SEQ ID NO: 4, an amino acid sequence of CDRL2 that comprises a sequence that is at least 95%, at least 98%, at least 99% or 100% identical to SEQ ID NO: 5, and an amino acid sequence of CDRH3 that comprises a sequence that is at least 95%, at least 98%, at least 99% or 100% identical to SEQ ID NO: 6. In one embodiment, the second agent is an isolated monoclonal antibody or antigen-binding fragment comprising a light chain complementarity determining region (CDRL) having an amino acid sequence of CDRL1 that comprises a sequence that is at least 95%, at least 98%, at least 99% or 100% identical to SEQ ID NO: 7. H In some embodiments, the second agent is an isolated monoclonal antibody or antigen-binding fragment comprising a light chain variable region (V) having at least 95%, at least 98%, at least 99% or 100% sequence identity to SEQ ID NO:8. L In some embodiments, the isolated monoclonal antibody further comprises a recombinant constant domain comprising an M428L mutation and an N434S mutation. In some embodiments, the isolated monoclonal antibody further comprises a recombinant constant domain comprising an S239D mutation and an I332E mutation. In some embodiments, the isolated monoclonal antibody further comprises a recombinant constant domain comprising an L235A mutation and a G237A mutation.

[0134] In some embodiments, the second agent is an isolated N6 monoclonal antibody or antigen-binding fragment comprising a heavy chain complementarity determining region (CDRH) having the amino acid sequence of CDRH1 of SEQ ID NO: 1, the amino acid sequence of CDRH2 of SEQ ID NO: 2, and the amino acid sequence of CDRH3 of SEQ ID NO: 3, and a light chain complementarity determining region (CDRL) having the amino acid sequence of CDRL1 of SEQ ID NO: 4, the amino acid sequence of CDRL2 of SEQ ID NO: 5, and the amino acid sequence of CDRH3 of SEQ ID NO: 6. In some embodiments, the second agent is an isolated N6LS monoclonal antibody or antigen-binding fragment comprising: a heavy chain complementarity determining region (CDRH) having the amino acid sequence of CDRH1 of SEQ ID NO: 1, the amino acid sequence of CDRH2 of SEQ ID NO: 2, and the amino acid sequence of CDRH3 of SEQ ID NO: 3; a light chain complementarity determining region (CDRL) having the amino acid sequence of CDRL1 of SEQ ID NO: 4, the amino acid sequence of CDRL2 of SEQ ID NO: 5, and the amino acid sequence of CDRH3 of SEQ ID NO: 6; and a recombinant constant domain comprising the M428L and N434S mutations. In some embodiments, the second agent is an isolated N6-DE monoclonal antibody or antigen-binding fragment comprising: a heavy chain complementarity determining region (CDRH) having the amino acid sequence of CDRH1 of SEQ ID NO: 1, the amino acid sequence of CDRH2 of SEQ ID NO: 2, and the amino acid sequence of CDRH3 of SEQ ID NO: 3; a light chain complementarity determining region (CDRL) having the amino acid sequence of CDRL1 of SEQ ID NO: 4, the amino acid sequence of CDRL2 of SEQ ID NO: 5, and the amino acid sequence of CDRH3 of SEQ ID NO: 6; and a recombinant constant domain comprising the S239D and I332E mutations. In some embodiments, the second agent is an isolated N6-LAGA monoclonal antibody or antigen-binding fragment comprising: a heavy chain complementarity determining region (CDRH) having the amino acid sequence of CDRH1 of SEQ ID NO: 1, the amino acid sequence of CDRH2 of SEQ ID NO: 2, and the amino acid sequence of CDRH3 of SEQ ID NO: 3; a light chain complementarity determining region (CDRL) having the amino acid sequence of CDRL1 of SEQ ID NO: 4, the amino acid sequence of CDRL2 of SEQ ID NO: 5, and the amino acid sequence of CDRH3 of SEQ ID NO: 6; and a recombinant constant domain comprising the L235A and G237A mutations.In some embodiments, the antigen-binding fragment is an Fv, Fab, F(ab')2, scFv or scFV2 ​​fragment.

[0135] In some embodiments, the combination for use further comprises administering a therapeutically effective amount of a third pharmaceutical composition comprising a third agent comprising at least one integrase inhibitor or a pharma- ceutically acceptable salt thereof. In some embodiments, the third agent is at least one agent selected from the group consisting of raltegravir, elvitegravir, dolutegravir, bictegravir and cabotegravir. In some embodiments, the third agent is raltegravir or cabotegravir. In some embodiments, the third agent is cabotegravir. In some embodiments, the first agent is temsavir, the second agent is selected from the group consisting of N6LS and N6-DE, and the third agent is cabotegravir. In some embodiments, the second agent is N6 with one or more ADCC mutations in the constant domain disclosed herein. In some embodiments, the second agent is N6LS. In some embodiments, the second agent is N6-DE.

[0136] In some embodiments, the combination for use includes administering the first pharmaceutical composition to a human before administering the second pharmaceutical composition. In some embodiments, the first pharmaceutical composition is orally administered to a human at about 1 mg / kg body weight to about 100 mg / kg body weight once a day, twice a day, or three times a day. In some embodiments, the first pharmaceutical composition is parenterally administered to a human at about 1 mg / kg body weight to about 100 mg / kg body weight once a day, twice a day, or three times a day.

[0137] In one aspect, the present invention provides a pharmaceutical composition for use in the treatment of HIV, comprising: (a) a first agent comprising at least one agent selected from the group consisting of fostemsavir and temsavir, or a pharmaceutically acceptable salt thereof; and (b) a second agent selected from the group consisting of CD4bs binding proteins, gp120 binding proteins, and integrase inhibitors, or a pharmaceutically acceptable salt thereof.

[0138] In one aspect, the present invention provides a pharmaceutical composition for use in the treatment of HIV, comprising: (a) a first agent comprising at least one agent selected from the group consisting of fostemsavir and temsavir, or a pharma- ceutical acceptable salt thereof; (b) a second agent comprising at least one CD4bs binding protein; and (c) a third agent comprising at least one integrase inhibitor, or a pharma- ceutical acceptable salt thereof.

[0139] According to another aspect, the present invention provides a combination for use in the treatment of HIV, comprising a therapeutically effective amount of at least one agent selected from the group consisting of fostemsavir and temsavir, or a pharmaceutically acceptable salt thereof, and a therapeutically effective amount of a CD4bs binding protein. In some embodiments, for use in the treatment of HIV, a human is administered at least one agent selected from the group consisting of fostemsavir and temsavir, or a pharmaceutically acceptable salt thereof, followed by sequential administration of a CD4bs binding protein.

[0140] According to another aspect, the present invention provides a combination for use in the clearance of HIV-infected cells, comprising a therapeutically effective amount of at least one agent selected from the group consisting of fostemsavir and temsavir, or a pharmaceutically acceptable salt thereof, and a therapeutically effective amount of a CD4bs binding protein, wherein at least one agent selected from the group consisting of fostemsavir and temsavir, or a pharmaceutically acceptable salt thereof, and the CD4bs binding protein are administered to a human. In one embodiment of the combination for use in the clearance of HIV-infected cells, the administration of at least one agent selected from the group consisting of fostemsavir and temsavir, or a pharmaceutically acceptable salt thereof, and the administration of the CD4bs binding protein are simultaneous. In one embodiment of the combination for use in the clearance of HIV-infected cells, the CD4bs binding protein is administered after the administration of at least one agent selected from the group consisting of fostemsavir and temsavir, or a pharmaceutically acceptable salt thereof.

[0141] According to another aspect, the present invention provides a combination for use in the prevention of HIV comprising therapeutically effective amounts of at least one agent selected from the group consisting of fostemsavir and temsavir, or a pharmaceutically acceptable salt thereof, and a CD4bs binding protein, wherein a human is administered at least one agent selected from the group consisting of fostemsavir and temsavir, or a pharmaceutically acceptable salt thereof, followed by sequential administration of the CD4bs binding protein.

[0142] According to another aspect, the present invention provides a combination for use in neutralizing HIV comprising therapeutically effective amounts of at least one agent selected from the group consisting of fostemsavir and temsavir, or a pharmaceutically acceptable salt thereof, and a CD4bs binding protein, wherein a human is administered at least one agent selected from the group consisting of fostemsavir and temsavir, or a pharmaceutically acceptable salt thereof, followed by sequential administration of the CD4bs binding protein.

[0143] According to another aspect, the present invention provides a combination for use in curing HIV, comprising therapeutically effective amounts of at least one agent selected from the group consisting of fostemsavir and temsavir, or a pharmaceutically acceptable salt thereof, and a CD4bs binding protein, wherein a human is administered at least one agent selected from the group consisting of fostemsavir and temsavir, or a pharmaceutically acceptable salt thereof, and then sequentially administered the CD4bs binding protein.

[0144] In some embodiments, at least one agent is temsavir or a pharmaceutically acceptable salt thereof. In one embodiment of the combination for use, at least one agent is fostemsavir or a pharmaceutically acceptable salt thereof.

[0145] In some embodiments, the administration of at least one agent selected from the group consisting of fostemsavir and temsavir or a pharmaceutically acceptable salt thereof is oral or parenteral.In some embodiments, the administration of temsavir is parenteral.In some embodiments, the administration of fostemsavir is oral.

[0146] In some embodiments, at least one agent selected from the group consisting of fostemsavir and temsavir or a pharmaceutically acceptable salt thereof is administered at a dose ranging from 1 mg / kg body weight to 100 mg / kg body weight. In some embodiments, the dose of at least one agent selected from the group consisting of fostemsavir and temsavir or a pharmaceutically acceptable salt thereof is 100 mg to 1000 mg. In some embodiments, the dose of fostemsavir is 600 mg.

[0147] In some embodiments, the administration of at least one agent selected from the group consisting of fostemsavir and temsavir, or a pharma- ceutically acceptable salt thereof, is a daily dose, a twice-daily dose, or a three times daily dose. In some embodiments, the administration is a two-times daily dose. In some embodiments, the administration is a one-time daily dose. In some embodiments, the administration is a three-times daily dose.

[0148] In some embodiments, at least one agent selected from the group consisting of fostemsavir and temsavir, or a pharmaceutically acceptable salt thereof, locks gp120 on the host cell membrane surface. In some embodiments, at least one agent selected from the group consisting of fostemsavir and temsavir, or a pharmaceutically acceptable salt thereof, locks gp120 in a closed conformation. In one embodiment of the combination for use, at least one agent selected from the group consisting of fostemsavir and temsavir, or a pharmaceutically acceptable salt thereof, prevents cell membrane-bound CD4 from interacting with gp120 expressed in the host cell. In some embodiments, at least one agent selected from the group consisting of fostemsavir and temsavir, or a pharmaceutically acceptable salt thereof, enables or promotes binding of CD4bs binding protein. In some embodiments, binding of CD4bs binding protein to HIV-infected host cells is measured by flow cytometry using mean fluorescence intensity (MFI).

[0149] In some embodiments, the combination for use comprises a gp120 binding protein. The gp120 binding protein binds to gp120 found on HIV virions or infected host cells. In some embodiments, the gp120 binding protein comprises a CD4bs binding protein. In some embodiments, the CD4bs binding protein is at least one CD4bs binding protein. In some embodiments, the CD4bs binding protein is an antibody or a binding fragment thereof. In some embodiments, the antibody is a broadly neutralizing antibody or a binding fragment thereof. In some embodiments, the broadly neutralizing antibody is selected from the group consisting of N6, N6LS, N6-DE and N6-LAGA.

[0150] In some embodiments, the CD4bs binding protein comprises a heavy chain complementarity determination having an amino acid sequence for CDRH1 that comprises a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical to SEQ ID NO:1, an amino acid sequence for CDRH2 that comprises a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical to SEQ ID NO:2, and an amino acid sequence for CDRH3 that comprises a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical to SEQ ID NO:3. and a light chain complementarity determining region (CDRL) having an amino acid sequence of CDRL1 that comprises a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical to SEQ ID NO:4, an amino acid sequence of CDRL2 that comprises a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical to SEQ ID NO:5, and an amino acid sequence of CDRL3 that comprises a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical to SEQ ID NO:6.

[0151] In some embodiments, the CD4bs binding protein comprises an amino acid sequence of a heavy chain variable region (VH) comprising a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical to a sequence selected from the group consisting of SEQ ID NO:7.

[0152] In some embodiments, the CD4bs binding protein comprises an amino acid sequence of a light chain variable region (VL) comprising a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical to a sequence selected from the group consisting of SEQ ID NO:8.

[0153] In some embodiments, the CD4bs binding protein comprises an amino acid sequence of a heavy chain (HC) comprising a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical to a sequence selected from the group consisting of SEQ ID NOs: 9, 11, 13 and 14.

[0154] In some embodiments, the CD4bs binding protein comprises an amino acid sequence of a light chain (LC) comprising a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical to a sequence selected from the group consisting of SEQ ID NOs: 10 and 12.

[0155] In some embodiments, the CD4bs binding protein antibody comprises a full-length immunoglobulin CD4bs binding protein. In some embodiments, the full-length immunoglobulin CD4bs binding protein comprises a fragment crystallizable region or "Fc" region or "constant region". The Fc of any of the defined antibodies may contain one or more amino acid substitutions to optimize the in vivo half-life of the antibody. The serum half-life of IgG Abs is regulated by the fetal Fc receptor (FcRn). In one embodiment of the combination for use, the antibody comprises an amino acid substitution that increases binding to FcRn. Some such substitutions are known to those of skill in the art, for example, T250Q and M428L in the IgG constant region (see, e.g., Hinton et al., J Immunol, 176:346-356, 2006); M428L and N434S ("LS" mutations, see, e.g., Zalevsky et al., Nature 1999:131-135, 2006); Biotechnology, 28:157-159, 2010); N434A (see, e.g., Petkova et al., Int. Immunol, 18:1759-1769, 2006); T307A, E380A and N434A (see, e.g., Petkova et al., Int. Immunol, 18:1759-1769, 2006); and M252Y, S254T and T256E (see, e.g., Dall'Acqua et al., J. Biol. Chem., 281:23514-23524, 2006). The disclosed antibodies and antigen-binding fragments can be linked to an Fc polypeptide that includes any of the substitutions listed above, for example, an Fc polypeptide can include an M428L substitution and an N434S substitution. In one embodiment of the combination for use, the antibody comprises a recombinant constant domain comprising a modification that improves binding to a fetal Fc receptor compared to an unmodified constant domain, the recombinant domain being an IgG1 constant domain comprising the M428L and N434S mutations.

[0156] In some embodiments, any constant region of the defined antibodies contains one or more amino acid substitutions to optimize ADCC. ADCC is primarily mediated through a set of closely related Fcy receptors. In one embodiment of the combination for use, the antibody contains one or more amino acid substitutions that increase binding to FcyRIIIa. Some such substitutions are known to those skilled in the art, such as S239D and I332E in the IgG constant region (see, e.g., Lazar et al., Proc. Natl, Acad. Sci. USA, 103:4005-4010, 2006); S239D, A330L and I332E (see, e.g., Lazar et al., Proc. Natl, Acad. Sci. USA, 103:4005-4010, 2006).

[0157] In some embodiments, combinations of the above substitutions are also included to generate IgG constant regions with increased binding to FcRn and FcyRIIIa. The combinations increase the half-life and ADCC of the antibody. For example, such combinations include antibodies with the following amino acid substitutions in the Fc region: (1) S239D / I332E and T250Q / M428L; (2) S239D / I332E and M428L / N434S; (3) S239D / I332E and N434A; (4) S239D / I332E and T307A / E380A / N434A; (5) S239D / I332E and M252Y / S254T / T256 (6) S239D / A330L / I332E and 250Q / M428L; (7) S239D / A330L / I332E and M428L / N434S; (8) S239D / A330L / I332E and N434A; (9) S239D / A330L / I332E and T307A / E380A / N434A; or (10) S239D / A330L / I332E and M252Y / S254T / T256E. In some examples, the antibody or antigen-binding fragment thereof is modified to be directly cytotoxic to infected cells or to exploit natural defenses, such as complement, ADCC, or phagocytosis by macrophages.

[0158] In some embodiments, the antibody is an antibody fragment. In one embodiment of the combination for use, the antibody fragment is selected from a Fab fragment, a F(ab')2 fragment, a single chain Fv protein ("scFv"), a disulfide stabilized Fv protein ("dsFv"), a diabody and a TANBAS. In one embodiment of the combination for use, the antibody fragment is a scFv.

[0159] In some embodiments, the CD4bs binding protein comprises a recombinant constant domain comprising a modification that improves binding to a fetal Fc receptor compared to an unmodified constant domain, and the recombinant domain is an IgG1 constant domain comprising an M428L mutation and an N434S mutation (the "LS" mutation).

[0160] In some embodiments the CD4bs binding protein comprises a recombinant constant domain that comprises a modification that affects ADCC compared to an unmodified constant domain, and the recombinant domain is an IgG1 constant domain that comprises S239D / I332E mutations ("DE" mutations).

[0161] In some embodiments, the CD4bs binding protein is selected from the group consisting of N6, N6LS, N6-LAGA and N6-DE. In some embodiments, the CD4bs binding protein is N6. In some embodiments, the CD4bs binding protein is N6LS. In some embodiments, the CD4bs binding protein is N6-LAGA. In some embodiments, the CD4bs binding protein is N6-DE.

[0162] In some embodiments, the CD4bs binding protein is administered continuously at a time selected from the group consisting of at least 30 minutes, at least 1 hour, at least 2 hours, at least 4 hours and at least 6 hours after administration of at least one agent selected from the group consisting of fostemsavir and temsavir or a pharmaceutically acceptable salt thereof. In some embodiments, the CD4bs binding protein is administered at a time selected from the group consisting of at least one agent selected from the group consisting of fostemsavir and temsavir or a pharmaceutically acceptable salt thereof. In one embodiment of the combination for use, the CD4bs binding protein is administered at a time at least 1 hour after administration of at least one agent selected from the group consisting of fostemsavir and temsavir or a pharmaceutically acceptable salt thereof. In some embodiments, the CD4bs binding protein is administered at a time at least 2 hours after administration of at least one agent selected from the group consisting of fostemsavir and temsavir or a pharmaceutically acceptable salt thereof. In one embodiment of the combination for use, the CD4bs binding protein is administered at least 4 hours after administration of at least one agent selected from the group consisting of fostemsavir and temsavir, or a pharmaceutically acceptable salt thereof. In some embodiments, the CD4bs binding protein is administered at least 6 hours after administration of at least one agent selected from the group consisting of fostemsavir and temsavir, or a pharmaceutically acceptable salt thereof. In some embodiments, the CD4bs binding protein is administered at least 24 hours after administration of at least one agent selected from the group consisting of fostemsavir and temsavir, or a pharmaceutically acceptable salt thereof. In some embodiments, the CD4bs binding protein is administered at least 48 hours after administration of at least one agent selected from the group consisting of fostemsavir and temsavir, or a pharmaceutically acceptable salt thereof. In one embodiment of the combination for use, the CD4bs binding protein is administered at least 72 hours after administration of at least one agent selected from the group consisting of fostemsavir and temsavir, or a pharmaceutically acceptable salt thereof.In some embodiments, the CD4bs binding protein is administered at least 96 hours after administration of at least one agent selected from the group consisting of fostemsavir and temsavir, or a pharma- ceutically acceptable salt thereof.

[0163] In some embodiments, the combination for use further comprises, after sequential administration, at least a second administration of simultaneously administering at least one agent selected from the group consisting of fostemsavir and temsavir, or a pharma- ceutically acceptable salt thereof, and a CD4bs binding protein.

[0164] In various embodiments, the combination of the present invention can also include an integrase inhibitor. An integrase inhibitor, i.e., INSTI (integrase strand transfer inhibitor), is an active agent that functions by preventing retroviral integration. Exemplary compounds include, but are not limited to, raltegravir, elvitegravir, dolutegravir, bictegravir and cabotegravir.

[0165] Other Aspects According to another aspect, the present invention provides the use of a therapeutically effective amount of at least one agent selected from the group consisting of fostemsavir and temsavir, or a pharma- ceutically acceptable salt thereof, and a therapeutically effective amount of a CD4bs binding protein, as defined above, in the manufacture of a medicament for use in the treatment of HIV, wherein for use in the treatment of HIV, a human is administered at least one agent selected from the group consisting of fostemsavir and temsavir, or a pharma- ceutically acceptable salt thereof, followed by sequential administration of a CD4bs binding protein.

[0166] According to another aspect, the present invention provides the use of a therapeutically effective amount of at least one agent selected from the group consisting of fostemsavir and temsavir, or a pharma- ceutically acceptable salt thereof, and a therapeutically effective amount of a CD4bs binding protein, as defined above, in the manufacture of a medicament for use in the treatment of HIV, wherein for use in the prophylaxis of HIV, a human is administered at least one agent selected from the group consisting of fostemsavir and temsavir, or a pharma- ceutically acceptable salt thereof, followed by sequential administration of a CD4bs binding protein.

[0167] According to another aspect, the present invention provides the use of a therapeutically effective amount of at least one agent selected from the group consisting of fostemsavir and temsavir, or a pharma- ceutically acceptable salt thereof, and a therapeutically effective amount of a CD4bs binding protein, as defined above, in the manufacture of a medicament for use in the treatment of HIV, wherein a human is administered at least one agent selected from the group consisting of fostemsavir and temsavir, or a pharma- ceutically acceptable salt thereof, followed by sequential administration of a CD4bs binding protein, for use in neutralizing HIV.

[0168] According to another aspect, the present invention provides the use of a therapeutically effective amount of at least one agent selected from the group consisting of fostemsavir and temsavir, or a pharma- ceutically acceptable salt thereof, and a therapeutically effective amount of a CD4bs binding protein, as defined above, in the manufacture of a medicament for use in the treatment of HIV, wherein a human is administered at least one agent selected from the group consisting of fostemsavir and temsavir, or a pharma- ceutically acceptable salt thereof, followed by sequential administration of a CD4bs binding protein, for use in curing HIV.

[0169] According to another aspect, the present invention provides the use of a therapeutically effective amount of at least one agent selected from the group consisting of fostemsavir and temsavir, or a pharma- ceutically acceptable salt thereof, and a therapeutically effective amount of a CD4bs binding protein, as defined above, in the manufacture of a medicament for use in the clearance of HIV-infected cells.

[0170] According to another aspect, the present invention provides a kit comprising a therapeutically effective amount of at least one agent selected from the group consisting of fostemsavir and temsavir, or a pharma- ceutically acceptable salt thereof, and a therapeutically effective amount of a CD4bs binding protein.

[0171] In some embodiments, the present invention provides a method of treating HIV infection in a human in need thereof, comprising administering to the human a therapeutically effective amount of at least one agent selected from the group consisting of fostemsavir and temsavir, or a pharmaceutically acceptable salt thereof, and a therapeutically effective amount of a CD4 binding site (CD4bs) binding protein, wherein the CD4bs binding protein is administered following administration of at least one agent selected from the group consisting of fostemsavir and temsavir, or a pharmaceutically acceptable salt thereof.

[0172] In some embodiments, the present invention provides a method for clearing HIV infected cells from a human in need thereof, comprising administering a therapeutically effective amount of at least one agent selected from the group consisting of fostemsavir and temsavir, or a pharma- ceutically acceptable salt thereof, and a therapeutically effective amount of a CD4bs binding protein.

[0173] In some embodiments, the present invention provides a combination for use in the treatment of HIV, comprising a therapeutically effective amount of at least one agent selected from the group consisting of fostemsavir and temsavir, or a pharmaceutically acceptable salt thereof, and a therapeutically effective amount of a CD4bs binding protein, wherein a human is administered at least one agent selected from the group consisting of fostemsavir and temsavir, or a pharmaceutically acceptable salt thereof, and then sequentially administered with a CD4bs binding protein. In an embodiment, a combination for use in the clearance of HIV-infected cells comprises a therapeutically effective amount of at least one agent selected from the group consisting of fostemsavir and temsavir, or a pharmaceutically acceptable salt thereof, and a therapeutically effective amount of a CD4bs binding protein, wherein a human is administered at least one agent selected from the group consisting of fostemsavir and temsavir, or a pharmaceutically acceptable salt thereof, and a CD4bs binding protein.

[0174] In some embodiments, at least one agent selected from the group consisting of fostemsavir and temsavir or a pharmaceutically acceptable salt thereof, and the CD4bs binding protein are administered concurrently, simultaneously, separately, or sequentially. In some embodiments, at least one agent selected from the group consisting of fostemsavir and temsavir or a pharmaceutically acceptable salt thereof, and the CD4bs binding protein is administered sequentially, and the CD4bs binding protein is administered after the administration of at least one agent selected from the group consisting of fostemsavir and temsavir or a pharmaceutically acceptable salt thereof. In some embodiments, at least one agent is temsavir or a pharmaceutically acceptable salt thereof. In other embodiments, at least one agent is fostemsavir or a pharmaceutically acceptable salt thereof.

[0175] In some embodiments, at least one agent selected from the group consisting of fostemsavir and temsavir or a pharmaceutically acceptable salt thereof is administered orally or parenterally.In some embodiments, the administration of temsavir is parenteral.In other embodiments, the administration of fostemsavir is oral.

[0176] In some embodiments, at least one agent selected from the group consisting of fostemsavir and temsavir or a pharmaceutically acceptable salt thereof is administered at a dose ranging from 1 mg / kg body weight to 100 mg / kg body weight. In some embodiments, the dose of at least one agent selected from the group consisting of fostemsavir and temsavir or a pharmaceutically acceptable salt thereof is 100 mg to 1200 mg. In some embodiments, at least one agent selected from the group consisting of fostemsavir and temsavir or a pharmaceutically acceptable salt thereof is administered once a day, twice a day, or three times a day. In one embodiment, at least one agent selected from the group consisting of fostemsavir and temsavir or a pharmaceutically acceptable salt thereof is administered twice a day.

[0177] In some embodiments, at least one agent selected from the group consisting of fostemsavir and temsavir, or a pharmaceutically acceptable salt thereof, promotes binding of a CD4bs binding protein. In some embodiments, at least one agent selected from the group consisting of fostemsavir and temsavir, or a pharmaceutically acceptable salt thereof, locks the gp120 envelope glycoprotein (gp120 ENV) in a closed conformation.

[0178] In some embodiments, the CD4bs binding protein comprises an antibody or binding fragment thereof that binds to CD4bs. In some embodiments, the antibody is a broadly neutralizing antibody or binding fragment thereof. In some embodiments, the CD4bs binding protein comprises a heavy chain complementarity determination having an amino acid sequence for CDRH1 that comprises a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical to SEQ ID NO:1, an amino acid sequence for CDRH2 that comprises a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical to SEQ ID NO:2, and an amino acid sequence for CDRH3 that comprises a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical to SEQ ID NO:3. and a light chain complementarity determining region (CDRL) having an amino acid sequence of CDRL1 that comprises a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical to SEQ ID NO:4, an amino acid sequence of CDRL2 that comprises a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical to SEQ ID NO:5, and an amino acid sequence of CDRL3 that comprises a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical to SEQ ID NO:6.

[0179] In some embodiments the CD4bs binding protein comprises an amino acid sequence of a heavy chain variable region (VH) that comprises a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical to SEQ ID NO: 7. In some embodiments the CD4bs binding protein comprises an amino acid sequence of a light chain variable region (VL) that comprises a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical to SEQ ID NO:8.

[0180] In some embodiments the CD4bs binding protein comprises an amino acid sequence of a heavy chain (HC) comprising a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical to a sequence selected from the group consisting of SEQ ID NOs: 9, 11, 13 and 14. In some embodiments the CD4bs binding protein comprises an amino acid sequence of a light chain (LC) comprising a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% identical to a sequence selected from the group consisting of SEQ ID NOs: 10 and 12.

[0181] In certain embodiments, the CD4bs binding protein comprises a full-length immunoglobulin CD4bs binding protein. In some embodiments, the CD4bs binding protein is a CD4bs binding protein antibody fragment. In some embodiments, the CD4bs binding protein antibody fragment is a Fab fragment, a F(ab')2 fragment, a single chain Fv protein ("scFv"), a disulfide stabilized Fv protein ("dsFv"), a diabody or a TANDABS. In some embodiments, the CD4bs binding protein antibody fragment is a scFv.

[0182] In some embodiments, the CD4bs binding protein comprises a recombinant constant domain comprising a modification that improves binding to fetal Fc receptors compared to an unmodified constant domain, the recombinant domain being an IgG1 constant domain comprising an M428L mutation and an N434S mutation. In some embodiments, the CD4bs binding protein comprises a recombinant constant domain comprising a modification that affects antibody-dependent cell-mediated cytotoxicity (ADCC) compared to an unmodified constant domain, the recombinant domain being an IgG1 constant domain comprising an S239D / I332E mutation or an L235A / G237A mutation. In some embodiments, the CD4bs binding protein is a monoclonal antibody selected from the group consisting of N6, N6LS, N6-DE and N6-LAGA. In some embodiments, the CD4bs binding protein is N6. In some embodiments, the CD4bs binding protein is N6LS.

[0183] In some embodiments, the CD4bs binding protein is administered at least 30 minutes, at least 1 hour, at least 2 hours, at least 4 hours, and at least 6 hours after administration of at least one agent selected from the group consisting of fostemsavir and temsavir, or a pharmaceutically acceptable salt thereof. In some embodiments, the method further comprises administering at least one additional agent selected from the group consisting of fostemsavir and temsavir, or a pharmaceutically acceptable salt thereof. In some embodiments, the method or combination further comprises a third agent comprising at least one integrase inhibitor, or a pharmaceutically acceptable salt thereof.

[0184] In some embodiments, a therapeutically effective amount of at least one agent selected from the group consisting of fostemsavir and temsavir, or a pharmaceutically acceptable salt thereof, and a therapeutically effective amount of a CD4bs binding protein are provided for use in the manufacture of a medicament for use in the treatment of HIV. In some embodiments, at least one agent selected from the group consisting of fostemsavir and temsavir, or a pharmaceutically acceptable salt thereof, and a CD4bs binding protein are administered to a human concurrently, simultaneously, separately, or sequentially. In some embodiments, a therapeutically effective amount of at least one agent selected from the group consisting of fostemsavir and temsavir, or a pharmaceutically acceptable salt thereof, and a therapeutically effective amount of a CD4bs binding protein are provided for use in the manufacture of a medicament for use in the clearance of HIV-infected cells according to any of the embodiments disclosed herein.

[0185] In one aspect, the present invention provides a kit comprising a first pharmaceutical composition comprising a first agent comprising at least one agent selected from the group consisting of fostemsavir and temsavir or a pharmaceutically acceptable salt thereof, a second pharmaceutical composition comprising a second agent comprising at least one broadly neutralizing antibody or an antigen-binding fragment thereof, and optionally a third pharmaceutical composition comprising a third agent comprising at least one integrase inhibitor or a pharmaceutically acceptable salt thereof. In some embodiments, the present invention provides a kit comprising a therapeutically effective amount of at least one agent selected from the group consisting of fostemsavir and temsavir or a pharmaceutically acceptable salt thereof and a therapeutically effective amount of a CD4bs binding protein. EXAMPLES

[0186] Example 1: Antiviral activity overview Human CD4 using the HIV-1 laboratory strain HxB2 +The effect of temsavir on the binding of bnAbs N6, N6-DE and N6-LAGA to HIV-1 envelope expressed on the cell surface upon productive infection of T cells was assessed by flow cytometry.

[0187] procedure Preparation of target cells and HIV-1 infection Peripheral blood mononuclear cells (PBMCs) were obtained from healthy donors by continuous-flow leukapheresis, isolated by ficoll-hypaque density gradient centrifugation, and cryopreserved in the vapor phase of liquid nitrogen. PBMCs were rapidly thawed to room temperature in a 37°C water bath and cultured in assay medium (RPMI 1640 medium supplemented with 10% fetal bovine serum + 30 U / mL recombinant IL-2) containing 2 μg / mL phytohemagglutinin-P (Sigma) for 3 days in a humidified incubator at 37°C with 5% CO2. CD4 + T cells were isolated by negative selection (Stemcell Technologies), washed twice, and titrated to a final density of 1–2 × 10 using assay medium. 6 The cells were then resuspended at 5 × 10 cells / mL and incubated overnight at 37 °C, 5% CO2. The cells were centrifuged at 500 × g for 10 min and diluted with assay medium at 5 × 10 6 Resuspend cells / mL in 200 µL (total 1 x 10 6 HIV-1 laboratory strain HxB2 virus previously grown in human PBMC cells was diluted in assay medium containing 50 μg / mL DEAE-dextran and 200 μL was added to the cells in the 24-well plate. The plate was centrifuged at 1,200 × g for 1.5 h, and the infected cells were harvested and pooled in a 50 mL conical tube. The infected cells were washed twice and incubated at a final density of 1–2 × 10 6 The cells were resuspended in assay medium to 200 cells / mL and incubated at 37° C., 5% CO2 for 3 days.

[0188] Treatment of HIV-infected target cells with temsavir and N6 bnAbHIV-infected cells were centrifuged at 500 × g for 10 min and diluted to 3 × 10 in assay medium. 6 The antibodies were resuspended at 1000x1000 cells / mL and 50 μL (total of 150,000 cells) were dispensed into 96-well U-bottom tissue culture treated plates. Infected cell cultures were then treated with temsavir dissolved in DMSO at a final concentration of 0.5 μM or with DMSO alone as a vehicle control and assay plates were incubated at 37°C, 5% CO2 for 16-24 hours. N6 bnAbs (N6, N6-DE and N6-LAGA) were produced from HEK293 cells by ChemPartner according to the sequence described. Antibodies were eluted and diluted in a buffer consisting of 150 mM arginine, 50 mM sodium acetate (pH 5.5, 150 mM NaCl), titrated stepwise in assay medium and added to the temsavir treated cells at a final concentration range of 50 μg / mL to 0.01 μg / mL. Plates were incubated at 37°C, 5% CO2 for 30 minutes.

[0189] Flow cytometry: The assay plate was removed from the incubator and the cells were pelleted by centrifugation at 500×g for 2 min at 25° C. The cells were washed twice, mixing with 200 μL ice-cold wash buffer (phosphate-buffered saline with 2% fetal bovine serum) for each wash cycle and centrifuging at 500×g for 2 min at 25° C. The cells were resuspended in 100 μL ice-cold wash buffer containing 1:2,000 LIVE / DEAD™ fixable aqua dead cell stain (Invitrogen), 1:400 BV786 mouse anti-human CD4 mAb (clone OKT4, BD Biosciences) and 1:200 R-phycoerythrin F(ab′)2 fragment goat anti-human IgG (Jackson Immuno Research) and incubated at 4° C. in the dark for 30 min. Stained cells were washed twice with wash buffer as described above, fixed and permeabilized with BD cytofix / cytoPerm™, and then stained intracellularly with 50 μL of FITC-anti-HIV-1 core antigen (KC57, Beckman Coulter) prepared 1:200 in BD Perm / Wash™ buffer for 30 minutes at 4° C. Stained cells were washed twice with wash buffer as described above, data acquisition was performed on a Fortessa cytometer (BD Biosciences), and data were analyzed by mean fluorescence intensity (MFI) using FlowJo v10.8 software (Tree Star).

[0190] result Treatment of HIV-infected cells with 0.5 μM temsavir for 24 hours significantly increased CD4 + / p24 + Binding of bnAbs N6, N6-DE and N6-LAGA to the cell surface of cells was significantly increased (Fig. 1), and treatment with 0.5 μM temsavir significantly increased the total percentage of cells containing bound bnAbs compared to cells treated with DMSO alone (Fig. 2).

[0191] Example 2: Antibody-dependent cellular cytotoxicity overview Temsavir enhancement of HIV-infected cell killing mediated by the three CD4 binding site bnAbs, N6, N6-DE and N6-LAGA, was assessed in an ADCC assay.

[0192] procedure Isolation and preparation of target and effector cells Peripheral blood mononuclear cells (PBMCs) were obtained from healthy donors by continuous flow leukapheresis, isolated by ficoll-hypaque density gradient centrifugation, and cryopreserved in the vapor phase of liquid nitrogen. PBMCs were rapidly thawed to room temperature in a 37°C water bath and resuspended in RPMI 1640 medium supplemented with 10% fetal bovine serum. Target cells were CD4 + T lymphocytes were purified using immunomagnetic negative selection beads according to the manufacturer's instructions (StemCell Technologies). Cells were activated with phytohemagglutinin-P (Sigma; PHA-P; 5 μg / mL) for 72 h at 37°C, 5% CO2 and then maintained under the same incubation conditions in RPMI 1640 medium supplemented with 10% fetal bovine serum + 50 U / mL recombinant IL-2. Natural killer (NK) effector cells were purified from autologous PBMCs using immunomagnetic negative selection beads according to the manufacturer's instructions (StemCell Technologies). Cells were maintained in RPMI 1640 medium supplemented with 10% fetal bovine serum and incubated for 24 h at 37°C, 5% CO2 before the ADCC assay.

[0193] Infection of target cells : HIV-1 laboratory strain IIIB was used to immunize activated CD4 + T lymphocytes were infected. Infected cells were harvested, washed, resuspended in RPMI 1640 medium supplemented with 10% fetal bovine serum + 50 U / mL recombinant IL-2, and incubated at 37°C, 5% CO2 for 3 days until the infection rate exceeded 20%.

[0194] ADCC assay :HIV infection CD4 + T cells (targets) were incubated with a cell proliferation dye (eFluor670; eBioscience) according to the manufacturer's instructions and plated in 96-well U-bottom plates. Temsavir was dissolved in DMSO, diluted in RPMI 1640 medium supplemented with 10% fetal bovine serum, and titrated five-fold in the same medium to give a dose range of 50 nM to 0.08 nM. To exclude the possibility that the reduction in the percentage of infected cells was the result of an antiviral effect of N6 and temsavir, raltegravir was added to the culture medium to prevent viral spread, thus allowing the analysis of the elimination of infected cells to be performed independently of the blocking effect of temsavir and N6 on viral infection. Temsavir titrations were added to the dye-loaded target cells and the assay plates were incubated for 16–24 h at 37 °C and 5% CO2. Antibodies were added to the appropriate wells of the assay plate at a final concentration of 5 μg / mL and incubated for 15 min at room temperature. NK effector cells, purified according to the target and effector cell isolation and preparation described above, were added at a 3:1 effector to target cell ratio and the assay plates were incubated at 37° C., 5% CO 2 for 24 hours.

[0195] Flow cytometry Following the ADCC part of the assay, cells were incubated with a viability dye (LiveDeadAqua; ThermoFisher), anti-CD3-BV421 (clone SP34-2; BD Biosciences, 1:200 final concentration) and anti-CD4-BV786 (clone OKT4; BD Bioscience, 1:400 final concentration) for 20-30 min at room temperature. Intracellular staining of HIV-1 p24 on cells was then performed by use of a Cytofix / Cytoperm Fixation / Permeabilization Kit (BD Biosciences) followed by the addition of anti-p24 mAb (FITC anti-p24, clone KC57; Beckman Coulter / Immunotech; 1:200 final concentration) for 20-30 min at room temperature. Cells were incubated at 5 × 104 Flow cytometry particles / mL (AccuCount blank particles; 5.3 μm; Spherotech) were resuspended in Pharmingen Stain Buffer (BSA) (BD Biosciences). Samples were acquired on a Fortessa cytometer (BD Biosciences) and data were analyzed using FlowJo v10.8 software (Tree Star). Percentage specific killing was calculated by gating on live target cells using the formula: (p24 + p3 + p4 + p5 + p6 + p1 + p2 ... + Percentage (%) of cells) - (p24 in target cells + effector cells + Ab + Percentage of cells (%) / (p24 in target cells alone, without antibody or temsavir + Percentage of cells (%)).

[0196] result Temsavir enhances NK killing of HIV-infected cells mediated by CD4-binding site bnAbs (Figure 3). The Fc-binding enhanced version, N6-DE, promoted the highest killing rate (up to 75%), whereas the Fc-binding reduced version, N6-LAGA, showed the lowest killing (up to 25%), indicating that NK-mediated killing of target cells is directly related to FcγRIIIB receptor function. There was no evidence of killing when effector cells were excluded from the assay (Figure 4).

[0197] Sequence Listing

[0198] [Table 3-1]

[0199] [Table 3-2]

[0200]

Table 3-3

Claims

1. 1. A pharmaceutical combination for treating human immunodeficiency virus (HIV) infection in a human in need thereof, comprising: (a) a first agent comprising at least one agent selected from the group consisting of fostemsavir and temsavir, or a pharmaceutically acceptable salt thereof; and (b) a second agent comprising at least one broadly neutralizing antibody or antigen-binding fragment thereof. A combination medicine comprising:

2. 2. The pharmaceutical combination of claim 1, wherein the first drug is fostemsavir or a pharmaceutically acceptable salt thereof.

3. 2. The pharmaceutical combination of claim 1, wherein the first drug is temsavir or a pharmaceutically acceptable salt thereof.

4. 2. The pharmaceutical combination of claim 1, wherein the second agent binds to at least one HIV envelope glycoprotein selected from the group consisting of HIV gp160, HIV gp120, and HIV gp41.

5. 2. The pharmaceutical combination of claim 1, wherein the second agent binds to HIV gp120.

6. The second drug was 2G12, 2F5, 3BC176, 3BNC60, 3BNC1-17, 4E10, 8ANC131, 8ANC195, 10E8, 10-1074, 12A12, 35022, b12, B2530, CH01-04, CH103, CH31, HJ16, M66.6, N6, N6LS, N6-DE, N6-LAGA, NIH45- 46, PG9, PG16, PGDM1400, PGT121, PGT128, PGT135, PGT141-PGT145, PGT151, PGV04, VRC01, VRC01-LS, VRC07, VRC07-523, VRC07-LS and Z13.

7. The second drug is a heavy chain complementarity determining region (CDRH) having a CDRH1 amino acid sequence that comprises a sequence at least 95%, at least 98%, at least 99%, or 100% identical to SEQ ID NO:1; a CDRH2 amino acid sequence that comprises a sequence at least 95%, at least 98%, at least 99%, or 100% identical to SEQ ID NO:2; and a CDRH3 amino acid sequence that comprises a sequence at least 95%, at least 98%, at least 99%, or 100% identical to SEQ ID NO:3; and a light chain complementarity determining region (CDRL) having a CDRL1 amino acid sequence that comprises a sequence that is at least 95%, at least 98%, at least 99%, or 100% identical to SEQ ID NO:4; a CDRL2 amino acid sequence that comprises a sequence that is at least 95%, at least 98%, at least 99%, or 100% identical to SEQ ID NO:5; and a CDRH3 amino acid sequence that comprises a sequence that is at least 95%, at least 98%, at least 99%, or 100% identical to SEQ ID NO:

6. The pharmaceutical combination of claim 1, which is an isolated monoclonal antibody or an antigen-binding fragment thereof comprising:

8. The second agent comprises a heavy chain variable region (V) having at least 95%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO:

7. Ｈ 2. The pharmaceutical combination of claim 1, which is an isolated monoclonal antibody or an antigen-binding fragment thereof comprising:

9. The second agent comprises a light chain variable region (V) having at least 95%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO:

8. Ｌ 2. The pharmaceutical combination of claim 1, which is an isolated monoclonal antibody or an antigen-binding fragment thereof comprising:

10. 8. The pharmaceutical combination of claim 7, wherein the isolated monoclonal antibody further comprises a recombinant constant domain comprising the M428L and N434S mutations.

11. 8. The pharmaceutical combination of claim 7, wherein the isolated monoclonal antibody further comprises a recombinant constant domain comprising the S239D and I332E mutations.

12. 8. The pharmaceutical combination of claim 7, wherein the isolated monoclonal antibody further comprises a recombinant constant domain comprising the L235A and G237A mutations.

13. The second drug is a heavy chain complementarity determining region (CDRH) having a CDRH1 amino acid sequence of SEQ ID NO: 1, a CDRH2 amino acid sequence of SEQ ID NO: 2, and a CDRH3 amino acid sequence of SEQ ID NO: 3; and a light chain complementarity determining region (CDRL) having a CDRL1 amino acid sequence of SEQ ID NO: 4, a CDRL2 amino acid sequence of SEQ ID NO: 5, and a CDRH3 amino acid sequence of SEQ ID NO: 6; 2. The pharmaceutical combination of claim 1, which is an isolated monoclonal antibody (N6) or an antigen-binding fragment thereof comprising:

14. The second drug is a heavy chain complementarity determining region (CDRH) having a CDRH1 amino acid sequence of SEQ ID NO: 1, a CDRH2 amino acid sequence of SEQ ID NO: 2, and a CDRH3 amino acid sequence of SEQ ID NO: 3; a light chain complementarity determining region (CDRL) having a CDRL1 amino acid sequence of SEQ ID NO: 4, a CDRL2 amino acid sequence of SEQ ID NO: 5, and a CDRH3 amino acid sequence of SEQ ID NO: 6; and Recombinant constant domain containing the M428L and N434S mutations 2. The pharmaceutical combination of claim 1, which is an isolated monoclonal antibody (N6LS) or an antigen-binding fragment thereof comprising:

15. The second drug is a heavy chain complementarity determining region (CDRH) having a CDRH1 amino acid sequence of SEQ ID NO: 1, a CDRH2 amino acid sequence of SEQ ID NO: 2, and a CDRH3 amino acid sequence of SEQ ID NO: 3; a light chain complementarity determining region (CDRL) having a CDRL1 amino acid sequence of SEQ ID NO: 4, a CDRL2 amino acid sequence of SEQ ID NO: 5, and a CDRH3 amino acid sequence of SEQ ID NO: 6; and Recombinant constant domain containing S239D and I332E mutations The pharmaceutical combination according to claim 1, which is an isolated monoclonal antibody (N6-DE) or an antigen-binding fragment thereof comprising:

16. The second drug is a heavy chain complementarity determining region (CDRH) having a CDRH1 amino acid sequence of SEQ ID NO: 1, a CDRH2 amino acid sequence of SEQ ID NO: 2, and a CDRH3 amino acid sequence of SEQ ID NO: 3; a light chain complementarity determining region (CDRL) having a CDRL1 amino acid sequence of SEQ ID NO: 4, a CDRL2 amino acid sequence of SEQ ID NO: 5, and a CDRH3 amino acid sequence of SEQ ID NO: 6; and Recombinant constant domain containing the L235A and G237A mutations The pharmaceutical combination according to claim 1, which is an isolated monoclonal antibody (N6-LAGA) or an antigen-binding fragment thereof comprising:

17. The antigen-binding fragment may be Fv, Fab, F(ab') ２ , scFv or scFV ２ The pharmaceutical combination according to claim 7, which is a fragment.

18. 10. The pharmaceutical combination of claim 1, further comprising a third agent comprising at least one integrase inhibitor or a pharmaceutically acceptable salt thereof.

19. 19. The pharmaceutical combination of claim 18, wherein the third agent comprises at least one agent selected from the group consisting of raltegravir, elvitegravir, dolutegravir, bictegravir, and cabotegravir.

20. 19. The pharmaceutical combination of claim 18, wherein the third agent is raltegravir or cabotegravir.

21. 19. The pharmaceutical combination of claim 18, wherein the third agent is cabotegravir.

22. 19. The pharmaceutical combination of claim 18, wherein the first drug is temsavir, the second drug is N6LS, and the third drug is cabotegravir.

23. 19. The pharmaceutical combination of claim 18, wherein the first drug is temsavir, the second drug is N6-DE, and the third drug is cabotegravir.

24. 20. The pharmaceutical combination of claim 18, wherein each of the first agent, the second agent, and the third agent is in the form of a pharmaceutical composition.

25. The pharmaceutical combination of any one of claims 1 to 24, wherein the first agent is administered before the second agent is administered.

26. 25. The pharmaceutical combination of any one of claims 1 to 24, wherein about 1 mg / kg body weight to about 100 mg / kg body weight of the first agent is orally administered to a human once daily, twice daily, or three times daily.

27. 25. The pharmaceutical combination of any one of claims 1 to 24, wherein about 1 mg / kg body weight to about 100 mg / kg body weight of the first agent is parenterally administered to a human once daily, twice daily, or three times daily.

28. The pharmaceutical combination of any one of claims 1 to 24, wherein the human is diagnosed with human immunodeficiency virus 1 (HIV-1) infection.

29. The pharmaceutical combination of any one of claims 1 to 24, wherein the human has previously been treated with one or more different HIV treatment modalities.

30. A combination pharmaceutical described in any one of claims 1 to 24, wherein the first drug is in the form of a first pharmaceutical composition comprising the first drug, and the second drug is in the form of a second pharmaceutical composition comprising the second drug.

31. 31. The pharmaceutical combination of claim 30, wherein the first pharmaceutical composition is administered to a human before the second pharmaceutical composition is administered to a human.

32. 31. The pharmaceutical combination of claim 30, wherein about 1 mg / kg to about 100 mg / kg of body weight of the first pharmaceutical composition is orally administered to a human once daily, twice daily, or three times daily.

33. 31. The pharmaceutical combination of claim 30, wherein about 1 mg / kg to about 100 mg / kg of body weight of the first pharmaceutical composition is parenterally administered to a human once daily, twice daily, or three times daily.

34. (a) a first drug comprising at least one drug selected from the group consisting of fostemsavir and temsavir, or a pharmaceutically acceptable salt thereof; (b) a second agent comprising at least one broadly neutralizing antibody or antigen-binding fragment thereof; and Optionally, (c) a third agent comprising at least one integrase inhibitor or a pharmaceutically acceptable salt thereof. Includes a kit.

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