Methods of identifying HIV patients susceptible to therapy with GP120 V3 glycan-directed antibodies

Abstract

The present invention provides a method for identifying a population of patients infected with HIV, which can be targeted by an antibody that binds to an HIV gl20V3 glycan region.

Description

[0001] This application is a divisional application of an invention patent application with an application date of May 18, 2020, application number 202080037770.7, and invention name “Method for identifying HIV patients sensitive to therapy using GP120 V3 glycan-directed antibodies”.

[0002] CROSS-REFERENCE TO RELATED APPLICATIONS

[0003] This patent application claims the benefit of U.S. Provisional Application No. 62 / 850,994, filed May 21, 2019, under 35 U.S.C. 119(e), which is hereby incorporated by reference in its entirety for all purposes.

[0004] Sequence Listing

[0005] This patent application contains a sequence listing filed electronically in ASCII format and is hereby incorporated by reference in its entirety. The ASCII copy was created on April 27, 2020, is named 1289PF_SL.txt, and is 199,543 bytes in size. Background Art

[0006] Human immunodeficiency virus (HIV) infection and related diseases are a major public health problem worldwide. Most currently approved therapies for HIV infection target viral reverse transcriptase, protease, and integrase, but HIV resistance to these existing drugs, long-term toxicity, and lack of patient compliance with daily dosing regimens have proven to be problems associated with these therapies. Therefore, the discovery and development of new HIV drugs is of great importance.

[0007] WO 2009 / 066702, WO 2012 / 030904, WO 2014 / 063059, WO 2016 / 149698, WO 2017 / 106346; WO 2018 / 075564 and WO 2018 / 125813, McCoy, Retrovirology, 2018, Vol. 15, p. 70; Sok and Burton, Nat Immunol., 2018, Vol. 19, No. 11, pp. 1179-1188; Possas et al., Expert Opin Ther Pat., July 2018, Vol. 28, No. 7, pp. 551-560; and Stephenson and Barouch, Curr HIV / AIDS Rep, 2016, Vol. 13, pp. 31-37 describes human anti-HIV antibodies derived from memory B cells of HIV-infected donors that target the V3 glycan region of gp120 and are able to inhibit infection by HIV-1 species from multiple clades. The therapeutic use of the antibodies may be limited by the need to identify patients infected with HIV-1 species that can be targeted by antibodies to the HIV gp120 V3 glycan region. Summary of the invention

[0008] The present invention provides methods for identifying patients most likely to benefit from therapy with antibodies targeted to the V3 glycan region of HIV gpl20.

[0009] Thus, in one aspect, the invention provides a method for treating or preventing HIV in a human subject in need thereof, the method comprising: a) identifying a human subject infected with HIV or a population of HIV expressing gp120 comprising: a glycosylated asparagine at a position corresponding to amino acid residue position 332 (N332 glycan), an aspartic acid at a position corresponding to amino acid residue position 325 (D325), and one or more amino acid residues selected from the group consisting of a threonine at a position corresponding to amino acid residue position 63 (T63), a leucine at a position corresponding to amino acid residue position 179 (L179), a threonine at a position corresponding to amino acid residue position 320 (T320), and a histidine at a position corresponding to amino acid residue position 330 (H330), wherein the amino acid positions are referenced to SEQ ID NO: 1. NO:4; and b) administering to the subject an effective amount of an antibody or antigen-binding fragment thereof that competes with or comprises the VH and VL regions that bind to an epitope of gp120 within the third variable loop (V3) and / or the high mannose patch comprising the N332 oligomannose glycan. In some embodiments, the method entails identifying a subject infected with HIV or a population of HIV that expresses a gp120 comprising the following amino acid residues: i. N332 glycan, D325, and T63; ii. N332 glycan, D325, and L179; iii. N332 glycan, D325, and T320; iv. N332 glycan, D325, and H330; v. N332 glycan, D325, T63, and L179; vi. N332 glycan, D325, T63, and T320; vii. N332 glycan, D325, T63, and H330; viii. N332 glycan, D325, L179 and T320; ix. N332 glycan, D325, L179 and H330; x. N332 glycan, D325, T320 and H330; xi. N332 glycan, D325, T63, T320 and H330; xii. N332 glycan, D325, T63, L179 and T320; xiii. N332 glycan, D325, T63, L179 and H330; xiv. N332 glycan, D325, L179, T320 and H330; or xv. N332 glycan, D325, T63, L179, T320 and H330, wherein the amino acid positions refer to SEQ ID NO:4.In some embodiments, the method entails identifying a subject infected with HIV or a population of HIV that expresses gp120 comprising the following amino acid residues: i. N332 glycan, D325 and T63; ii. N332 glycan, D325 and L179; iii. N332 glycan, D325 and T320; or iv. N332 glycan, D325 and H330, wherein the amino acid positions are referenced to SEQ ID NO:4. In some embodiments, the method entails identifying a subject infected with HIV or a population of HIV that expresses gp120 comprising the following amino acid residues: i. N332 glycan, D325, T63 and L179; ii. N332 glycan, D325, T63 and T320; iii. N332 glycan, D325, T63 and H330; iv. N332 glycan, D325, L179 and T320; v. N332 glycan, D325, L179 and H330; or vi. N332 glycan, D325, T320 and H330, wherein the amino acid positions are referenced to SEQ ID NO:4. In some embodiments, the method entails identifying a subject infected with HIV or a population of HIV that expresses gp120 comprising the following amino acid residues: i. N332 glycan, D325, L179, T320 and H330; ii. N332 glycan, D325, T63, T320 and H330; iii. N332 glycan, D325, T63, L179 and T320; or iv. N332 glycan, D325, T63, L179 and H330, wherein the amino acid positions are referenced to SEQ ID NO:4. In some embodiments, the method entails identifying a subject infected with HIV or a population of HIV that expresses gp120 comprising the following amino acid residues: i. N332 glycan, D325, T63 and H330; ii. N332 glycan, D325, T320 and H330; iii. N332 glycan, D325, L179, T320 and H330; or iv. N332 glycan, D325, T63, L179, T320 and H330, wherein the amino acid positions are referenced to SEQ ID NO:4.In some embodiments, the subject is infected with HIV or a population of HIV that expresses gp120 that further comprises one or more of the following amino acid residues: a glycan at amino acid residue 301 (glycan 301); a lysine at amino acid residue 677 (K677); an amino acid residue other than tryptophan at position 17 (e.g., A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, or Y) (not_W17); an amino acid residue other than arginine at position 747 (e.g., A, C, D, E, F, G, H, I, K, L, M, N, P, Q, S, T, V, W, or Y) (not_R747); an insertion n_321.01 (e.g., insertion of any amino acid between positions G321 and K322 (e.g., A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, or Y)); glutamic acid at position 429 (E429); glutamine at position 442 (Q442); arginine at position 335 (R335); isoleucine at position 165 (I165); serine at position 393 (S393); isoleucine at position 307 (I307); glycan at position 295 (295 glycan); and / or asparagine at position 300 (N300), wherein the amino acid positions are referenced to SEQ ID NO:4. In some embodiments, at least 90%, e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% of the HIV species in the HIV population comprise the amino acid residue. In some embodiments, the administered antibody or antigen-binding fragment thereof competes with or comprises the VH and VL regions of an antibody selected from the group consisting of GS-9722 (ipavirumab), GS-9721, PGT-121, PGT-121.66, PGT-121.414, PGT-122, PGT-123, PGT-124, PGT-125, PGT-126, PGT-128, PGT-130, PGT-133, PGT-134, PGT-135, PGT -136, PGT-137, PGT-138, PGT-139, 10-1074, 10-1074-J, VRC24, 2G12, BG18, 354BG8, 354BG18, 354BG42, 354BG33, 354BG129, 354BG188, 354BG411, 354BG426, DH270.1, DH270.6, PGDM12, VRC41.01, PGDM21, PCDN-33A, BF520.1, and VRC29.03.In some embodiments, the antibody or antigen-binding fragment thereof competes with or comprises the VH and VL regions of an antibody selected from the group consisting of GS-9722, GS-9721, PGT-121, PGT-121.66, PGT-121.414, PGT-124, PGT-134, GS-2872, 10-1074, 10-1074-J, PGT-122, and PGT-123. In some embodiments, the methods include administering an antibody comprising an Fc region comprising one or more amino acid substitutions that extend serum half-life. In some embodiments, the methods include administering an antibody comprising an Fc region comprising the following amino acids at the specified positions (EU index number): i. tyrosine at position 252, threonine at position 254, and glutamic acid (YTE) at position 256; or ii. leucine at position 428 and serine (LS) at position 434. In some embodiments, these methods include administering an antibody comprising an Fc region comprising the following amino acids at the specified positions (EU index number): i. Aspartic acid at position 239 and glutamic acid at position 332 (DE); ii. Aspartic acid at position 239, glutamic acid at position 332, and leucine at position 330 (DEL); iii. Aspartic acid at position 239, glutamic acid at position 332, alanine at position 236 (DEA); or iv. Aspartic acid at position 239, glutamic acid at position 332, alanine at position 236, and leucine at position 330 (DEAL). In some embodiments, these methods include administering an antigen binding fragment. In some embodiments, the antigen binding fragment is selected from scFv, Fab, Fab2, Fab', F(ab')2, Fv, and a bivalent antibody. In some embodiments, the antibody is one or more arms of a multispecific antibody (e.g., a bispecific antibody). In some embodiments, the human subject is acutely infected with HIV. In some embodiments, the antibody is administered to a human subject with HIV infection at Fiebig stage IV or earlier, e.g., Fiebig stage III, Fiebig stage II, or Fiebig stage I. In some embodiments, the antibody is administered to a human subject with HIV infection that has not seroconverted. In some embodiments, the human subject has recently been infected with HIV, e.g., within 1 week, 2 weeks, 3 weeks, or 4 weeks, or prior to detection, seroconversion, or symptom manifestation. In some embodiments, the antibody is administered to a human subject with HIV infection at Fiebig stage V or Fiebig stage VI. In some embodiments, the human subject is chronically infected with HIV. In some embodiments, the human subject is infected with HIV clade B virus.In some embodiments, the human subject is infected with HIV clade B virus, and these methods require identification of subjects infected with HIV or HIV populations expressing gp120 comprising the following amino acid residues: i.N332 polysaccharide, D325, T63 and H330; ii.N332 polysaccharide, D325, L179, T320 and H330; or iii.N332 polysaccharide, D325, T63, L179, T320 and H330. In some embodiments, the human subject is infected with HIV clade A virus. In some embodiments, the human subject is infected with HIV clade C virus. In some embodiments, these methods further comprise administering to the subject one or more additional therapeutic agents for treating HIV infection. In some embodiments, the subject is not receiving antiretroviral therapy (ART), or ART is discontinued before the antibody is administered. In some embodiments, ART is discontinued after one or more administrations of the antibody or its antigen-binding fragment. In some embodiments, the methods further comprise administering to the subject one or more antiretroviral therapy (ART) agents. In some embodiments, the methods further comprise administering to the subject a second antibody or antigen binding fragment thereof that binds to an epitope or region of gp120 selected from: the second variable loop (V2) and / or the Env trimer vertex; the CD4 binding site (CD4bs); the gp120 / gp41 interface; or the gp120 silent face. In some embodiments, the second antibody or antigen-binding fragment thereof binds to an epitope or region of gp120 in the second variable loop (V2) and / or the apex of the Env trimer and competes with or comprises the VH and VL regions of an antibody selected from the group consisting of PG9, PG16, PGC14, PGG14, PGT-142, PGT-143, PGT-144, PGT-145, CHO1, CH59, PGDM1400, CAP256, CAP256-VRC26.08, CAP256-VRC26.09, CAP256-VRC26.25, PCT64-24E, and VRC38.01.In some embodiments, the second antibody or antigen-binding fragment thereof binds to an epitope or region of gp120 in the CD4 binding site (CD4bs) and competes with or comprises a VH region and a VL region from an antibody selected from the group consisting of b12, F105, VRC01, VRC07, VRC07-523, VRC03, VRC06, VRC06b01VRC08, VRC0801, NIH45-46, GS-97 23, GS-5423, 3BNC117, 3BNC60, VRC-PG04, PGV04; CH103, 44-VRC13.01, 1NC9, 12A12, N6, N6LS (VRC-HIVMAB091-00-AB), N49-P7, NC-Cow1, IOMA, CH235 and CH235.12, N49P6, N49P7, N49P11, N49P9 and N60P25. In some embodiments, the second antibody or antigen-binding fragment thereof binds to an epitope or region of gp120 in the gp120 / gp41 interface and competes with or comprises the VH and VL regions from an antibody selected from PGT-151, CAP248-2B, 35022, 8ANC195, ACS202, VRC34, and VRC34.01. In some embodiments, the second antibody or antigen-binding fragment thereof binds to an epitope or region of the silent face of gp120 and competes with or comprises the VH and VL regions from the antibody VRC-PG05. In some embodiments, the second antibody or antigen-binding fragment thereof binds to an epitope or region of gp41 in the membrane proximal region (MPER) and competes with or comprises the VH and VL regions of an antibody selected from the group consisting of 10E8, 10E8v4, 10E8-5R-100cF, 4E10, DH511.11P, 2F5, 7b2, and LN01. In some embodiments, the second antibody or antigen-binding fragment thereof binds to an epitope or region of the gp41 fusion peptide and competes with or comprises the VH and VL regions of an antibody selected from the group consisting of VRC34 and ACS202. In some embodiments, the methods further comprise administering a TLR agonist to the subject. In some embodiments, the TLR agonist is a TLR2 agonist, a TLR3 agonist, a TLR7 agonist, a TLR8 agonist, or a TLR9 agonist. In some embodiments, the TLR7 agonist is selected from visammod, imiquimod, and resiquimod. In some embodiments, the methods comprise multiple administrations of the antibody or antigen-binding fragment thereof at predetermined intervals, optionally together with a TLR agonist.In some embodiments, after one or more administrations of the antibody or antigen-binding fragment thereof, in the absence of antiretroviral therapy (ART), the subject does not show symptoms of HIV or AIDS for at least 6 months, at least 1 year, at least 2 years, at least 3 years, at least 4 years, at least 5 years or longer. In some embodiments, after one or more administrations of the antibody, in the absence of antiretroviral therapy (ART), the subject has less than 500, e.g., less than 400, less than 300, less than 200, less than 100, less than 50 HIV viral load copies / ml blood for at least 6 months, at least 1 year, at least 2 years, at least 3 years or longer.

[0010] In another aspect, the invention provides a method for identifying a human subject infected with HIV or a population of HIV susceptible to an antibody or antigen binding fragment thereof that competes with or comprises VH and VL regions that bind to an epitope of gp120 within the third variable loop (V3) and / or the high mannose patch comprising the N332 oligomannose glycan. In some embodiments, the methods include identifying in a biological sample from a human subject a gp120 comprising the following amino acid residues: a glycosylated asparagine at a position corresponding to amino acid residue position 332 (N332 glycan), an aspartic acid at a position corresponding to amino acid residue position 325 (D325), and one or more amino acid residues selected from the following amino acid residues: a threonine at a position corresponding to amino acid residue position 63 (T63), a leucine at a position corresponding to amino acid residue position 179 (L179), a threonine at a position corresponding to amino acid residue position 320 (T320), and a histidine at a position corresponding to amino acid residue position 330 (H330), wherein the amino acid positions are referenced to SEQ ID NO:4. In some embodiments, the method entails identifying a subject infected with HIV or a population of HIV that expresses a gp120 comprising the following amino acid residues: i. N332 glycan, D325, and T63; ii. N332 glycan, D325, and L179; iii. N332 glycan, D325, and T320; iv. N332 glycan, D325, and H330; v. N332 glycan, D325, T63, and L179; vi. N332 glycan, D325, T63, and T320; vii. N332 glycan, D325, T63, and H330; viii. N332 glycan, D325, L179 and T320; ix. N332 glycan, D325, L179 and H330; x. N332 glycan, D325, T320 and H330; xi. N332 glycan, D325, T63, T320 and H330; xii. N332 glycan, D325, T63, L179 and T320; xiii. N332 glycan, D325, T63, L179 and H330; xiv. N332 glycan, D325, L179, T320 and H330; or xv. N332 glycan, D325, T63, L179, T320 and H330, wherein the amino acid positions refer to SEQ ID NO:4.In some embodiments, the method entails identifying a subject infected with HIV or a population of HIV that expresses gp120 comprising the following amino acid residues: i. N332 glycan, D325 and T63; ii. N332 glycan, D325 and L179; iii. N332 glycan, D325 and T320; or iv. N332 glycan, D325 and H330, wherein the amino acid positions are referenced to SEQ ID NO:4. In some embodiments, the method entails identifying a subject infected with HIV or a population of HIV that expresses gp120 comprising the following amino acid residues: i. N332 glycan, D325, T63 and L179; ii. N332 glycan, D325, T63 and T320; iii. N332 glycan, D325, T63 and H330; iv. N332 glycan, D325, L179 and T320; v. N332 glycan, D325, L179 and H330; or vi. N332 glycan, D325, T320 and H330, wherein the amino acid positions are referenced to SEQ ID NO:4. In some embodiments, the method entails identifying a subject infected with HIV or a population of HIV that expresses gp120 comprising the following amino acid residues: i. N332 glycan, D325, L179, T320 and H330; ii. N332 glycan, D325, T63, T320 and H330; iii. N332 glycan, D325, T63, L179 and T320; or iv. N332 glycan, D325, T63, L179 and H330, wherein the amino acid positions are referenced to SEQ ID NO:4. In some embodiments, the method entails identifying a subject infected with HIV or a population of HIV that expresses gp120 comprising the following amino acid residues: i. N332 glycan, D325, T63 and H330; ii. N332 glycan, D325, T320 and H330; iii. N332 glycan, D325, L179, T320 and H330; or iv. N332 glycan, D325, T63, L179, T320 and H330, wherein the amino acid positions are referenced to SEQ ID NO:4.In some embodiments, the subject is infected with HIV or a population of HIV that expresses gp120 that further comprises one or more of the following amino acid residues: a glycan at amino acid residue 301 (glycan 301); a lysine at amino acid residue 677 (K677); an amino acid residue other than tryptophan at position 17 (e.g., A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, or Y) (not_W17); an amino acid residue other than arginine at position 747 (e.g., A, C, D, E, F, G, H, I, K, L, M, N, P, Q, S, T, V, W, or Y) (not_R747); an insertion n_321.01 (e.g., insertion of any amino acid between positions G321 and K322 (e.g., A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, or Y)); glutamic acid at position 429 (E429); glutamine at position 442 (Q442); arginine at position 335 (R335); isoleucine at position 165 (I165); serine at position 393 (S393); isoleucine at position 307 (I307); glycan at position 295 (295 glycan); and / or asparagine at position 300 (N300), wherein the amino acid positions are referenced to SEQ ID NO:4. In some embodiments, the antibody or antigen-binding fragment thereof competes with or comprises a VH region and a VL region from an antibody selected from the group consisting of: GS-9722, GS-9721, PGT-121, PGT-121.66, PGT-121.414, PGT-122, PGT-123, PGT-124, PGT-125, PGT-126, PGT-128, PGT-130, PGT-133, PGT-134, PGT-135, PGT-136, PGT-137, PGT-138, PGT-139, 10-1074, 10-1074-J, VRC24, 2G12, BG18, 354BG8, 354BG18, 354BG42, 354BG33, 354BG129, 354BG188, 354BG411, 354BG426, DH270.1, DH270.6, PGDM12, VRC41.01, PGDM21, PCDN-33A, BF520.1, and VRC29.03.In some embodiments, the antibody or antigen-binding fragment thereof competes with or comprises the VH and VL regions of an antibody selected from the group consisting of GS-9722, GS-9721, PGT-121, PGT-121.66, PGT-121.414, PGT-124, PGT-134, GS-2872, 10-1074, 10-1074-J, PGT-122, and PGT-123. In some embodiments, the human subject is acutely infected with HIV. In some embodiments, the antibody is administered to a human subject with HIV infection at Fiebig stage IV or earlier. In some embodiments, the antibody is administered to a human subject who has not seroconverted. In some embodiments, the human subject has recently been infected with HIV. In some embodiments, the antibody is administered to a human subject with HIV infection at Fiebig stage V or Fiebig stage VI. In some embodiments, the human subject is chronically infected with HIV. In some embodiments, the human subject is infected with HIV clade B virus. In some embodiments, the human subject is infected with HIV clade B virus, and these methods require identifying subjects infected with HIV or HIV populations expressing gp120 comprising the following amino acid residues: i. N332 glycan, D325, T63 and H330; ii. N332 glycan, D325, L179, T320 and H330; or iii. N332 glycan, D325, T63, L179, T320 and H330. In some embodiments, the human subject is infected with HIV clade A virus. In some embodiments, the human subject is infected with HIV clade C virus.

[0011] Relative to other embodiments of the methods described herein, in some embodiments, gp120 amino acids are identified in one or more gp120 polypeptide sequences expressed in HIV or HIV populations isolated from a subject. In some embodiments, gp120 amino acids are identified in one or more gp120 polynucleotide sequences encoding gp120 polypeptides from HIV or HIV populations isolated from a subject. In various embodiments, these methods require next generation sequencing (NGS) of polynucleotide sequences encoding gp120 from HIV populations. In some embodiments, gp120 variants are detected at a frequency level of about 1% of the viral population (e.g., a frequency level of about 0.5%). In some embodiments, gp120 amino acids are identified in one or more biological samples from a subject, wherein the one or more biological samples are obtained from blood, peripheral blood mononuclear cells (PBMC), serum, plasma, semen, or lymph nodes. In some embodiments, these methods require identification of HIV RNA populations in serum or plasma samples. In some embodiments, these methods also include the step of obtaining one or more biological samples from a subject. In some embodiments, two or more biological samples are obtained from a subject. In some embodiments, two or more biological samples are obtained from the same tissue or fluid at two or more different time points. In some embodiments, two or more biological samples are obtained from different tissues or fluids or from different anatomical locations.

[0012] definition

[0013] Unless otherwise indicated, the terms "a" and "an" mean one or more.

[0014] By "about" is meant a quantity, level, value, numerical value, frequency, percentage, dimension, size, amount, weight, or length that differs by up to 30%, 25%, 20%, 15%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, or 1% from a reference quantity, level, value, numerical value, frequency, percentage, dimension, size, amount, weight, or length. In any embodiments discussed in the context of a numerical value used in conjunction with the term "about," it is specifically contemplated that the term "about" may be omitted.

[0015] Unless the context requires otherwise, throughout this specification and claims, the word "include" and variations such as "comprises" and "comprising" are to be interpreted in an open, inclusive sense, i.e., as "including but not limited to." Where the terms "includes" or "comprising" are used herein, it should be understood that the disclosure also includes embodiments in which these terms are replaced with "consisting of" or "consisting essentially of."

[0016] By "consisting of is meant including and limited to whatever follows the phrase "consisting of." Thus, the phrase "consisting of" indicates that the listed elements are required or mandatory, and other elements may not be present.

[0017] By "consisting essentially of is meant including any elements listed after the phrase and limited to other elements that do not interfere with or contribute to the activity or action of the listed elements as specified in the present disclosure. Thus, the phrase "consisting essentially of indicates that the listed elements are required or mandatory, but other elements are optional and may or may not be present, depending on whether they affect the activity or action of the listed elements.

[0018] Reference throughout this specification to "one embodiment" or "an embodiment" means that a particular feature, structure, or characteristic described in connection with the embodiment is included in at least one embodiment described herein. Thus, the phrases "in one embodiment" or "in an embodiment" appearing in various places throughout this specification are not necessarily all referring to the same embodiment. Furthermore, in one or more embodiments, the particular features, structures, or characteristics may be combined in any suitable manner.

[0019] An "increased" or "enhanced" amount is typically a "statistically significant" amount and may include an increase of 1.1 times, 1.2 times, 1.3 times, 1.4 times, 1.5 times, 1.6 times, 1.7 times, 1.8 times, 1.9 times, 2 times, 2.5 times, 3 times, 3.5 times, 4 times, 4.5 times, 5 times, 6 times, 7 times, 8 times, 9 times, 10 times, 15 times, 20 times, 30 times, 40 times or 50 times or more (e.g., 100 times, 500 times, 1000 times) of the amounts or levels described herein (including all integers and decimal points between 1 and greater than 1, e.g., 2.1, 2.2, 2.3, 2.4, etc.). It may also include an increase of at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 100%, at least 150%, at least 200%, at least 500%, or at least 1000% of the amounts or levels described herein.

[0020] A "reduced" or "decreased" or "less" amount is typically a "statistically significant" amount, and may include a decrease (including all integers and decimal points between 1 and greater than 1, e.g., 1.5, 1.6, 1.7, 1.8, 1.9, 2, 2.5, 3, 3.5, 4, 4.5, 5, 6, 7, 8, 9, 10, 15, 20, 30, 40, or 50 or more times (e.g., 100, 500, 1000 times) of the amounts or levels described herein. It may also include a decrease of at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, or at least 90%, at least 100%, at least 150%, at least 200%, at least 500%, or at least 1000% of the amounts or levels described herein.

[0021] A "composition" may include an active agent, such as a contrast agent, and an inert or active carrier, such as a pharmaceutically acceptable carrier, diluent or excipient. The composition may be a pharmaceutical composition. In certain embodiments, the composition is sterile, substantially free of endotoxin or non-toxic to the recipient at the dose or concentration employed.

[0022] "Pharmaceutically acceptable carrier, diluent or excipient" includes, but is not limited to, any adjuvant, carrier, excipient, glidant, sweetener, diluent, preservative, dye / colorant, flavor enhancer, surfactant, wetting agent, dispersant, suspending agent, stabilizer, isotonic agent, solvent or emulsifier that has been approved by the U.S. Food and Drug Administration as acceptable for use in humans or livestock.

[0023] "Biological sample" or "sample" refers to any fluid, cell, or solid tissue sample from a subject who has or is suspected of having detectable HIV.

[0024] "Subject," "individual," or "patient" refers to any mammal, including humans and non-human primates. In certain embodiments, the mammal is a human.

[0025] As used herein, the term "buffer" refers to a pharmaceutically acceptable excipient that stabilizes the pH of a pharmaceutical formulation. Suitable buffers are well known in the art. Suitable pharmaceutically acceptable buffers include, but are not limited to, acetate buffers, histidine buffers, citrate buffers, succinate buffers, tris buffers, and phosphate buffers. In certain embodiments, the concentration of the buffer is about 0.01 mM to about 1000 mM, about 0.1 mM to about 1000 mM, about 0.1 mM to about 500 mM, about 0.1 mM to about 200 mM, about 0.1 mM to about 100 mM, about 1 mM to about 1000 mM, about 1 mM to about 500 mM, about 1 mM to about 200 mM, about 1 mM to about 100 mM, about 1 mM to about 500 mM, about 2 mM to about 60 mM, about 4 mM to about 60 mM, or about 4 mM to about 40 mM, about 5 mM to about 20 mM, or about 5 mM to about 25 mM.

[0026] "Optional" or "optionally" means that the subsequently described event of circumstances may or may not occur, and that the description includes instances where said event or circumstance occurs and instances where said event or circumstance does not occur.

[0027] "Pharmaceutical composition" refers to a preparation of a compound and a medium generally accepted in the art for delivering a biologically active compound to a mammal (eg, a human). Such a medium may include any pharmaceutically acceptable carrier, diluent, or excipient.

[0028] "Effective amount" or "therapeutically effective amount" refers to an amount of an antibody or antigen-binding fragment thereof, when administered alone or in combination with another therapeutic agent to a cell, tissue or subject, sufficient to achieve a therapeutic or beneficial result in the subject. The amount constituting an "effective amount" will vary depending on the antibody or antigen-binding fragment thereof and its specific use, as well as the underlying condition and its severity, the mode of administration and the age of the subject to be treated, but can be routinely determined by a person of ordinary skill in the art with reference to his or her own knowledge and this disclosure. A therapeutically effective dose also refers to an amount of an antibody or antigen-binding fragment thereof sufficient to treat, prevent or ameliorate an infection or disease condition or infection or disease progression, and an amount sufficient to achieve an increase in the rate of treatment, cure, prevention or improvement of such conditions. When applicable to a single antibody or antigen-binding fragment thereof administered alone, a therapeutically effective dose refers to the active ingredient alone. When applicable to a combination, a therapeutically effective dose refers to the combined amount of the active ingredients that result in a therapeutic effect, whether administered in combination, sequentially or simultaneously.

[0029] As used herein, "treat", "treating" or "treatment" encompasses treating a disease, injury or condition of interest, such as HIV-1 infection, in a subject (e.g., a mammal, such as a human) having the disease or condition of interest, and includes: (i) inhibiting the progression of the disease, injury or condition, i.e., preventing its development; (ii) reducing or alleviating the disease, injury or condition, i.e., causing regression of the disease or condition; or (iii) alleviating the symptoms caused by the disease, injury or condition. As used herein, the terms "disease", "disorder" and "disorder" are used interchangeably. As used herein, "inhibit", "treatment", "treating" and "improve" are used interchangeably and refer to, for example, stagnation of symptoms, prolonged survival, partial or complete improvement of symptoms, and partial or complete eradication of a condition, disease or disorder.

[0030] As used herein, "prevent" or "prevention" includes (i) preventing or inhibiting the occurrence of a disease, injury or condition in a subject, particularly when such subject is susceptible to the condition but has not yet been diagnosed as having the condition; or (ii) reducing the likelihood of a subject developing a disease, injury or condition.

[0031] As used herein, the term "antibody" means an isolated or recombinant binding agent that contains the necessary variable region sequences that specifically bind to an antigenic epitope. Therefore, an antibody is an antibody or any form of a fragment thereof that exhibits a desired biological activity (e.g., binding to a specific target antigen). Therefore, it is used in the broadest sense, and specifically encompasses monoclonal antibodies (including full-length monoclonal antibodies), polyclonal antibodies, human antibodies, humanized antibodies, chimeric antibodies, nanobodies, bivalent antibodies, multispecific antibodies (e.g., bispecific antibodies) and antibody fragments, including but not limited to scFv, Fab and Fab2, as long as they exhibit desired biological activity.

[0032] The term "human antibody" refers to an antibody comprising sequences of human origin excluding possible non-human CDR regions, and does not imply the existence of the complete structure of an Ig molecule, only that the antibody has minimal immunogenic effects in humans.

[0033] "Antibody fragments" comprise a portion of an intact antibody, e.g., the antigen binding or variable region of an intact antibody. Examples of antibody fragments include Fab, Fab', F(ab')2, and Fv fragments; bivalent antibodies; linear antibodies (e.g., Zapata et al., Protein Eng., Vol. 8, No. 10, pp. 1057-1062, 1995); single-chain antibody molecules (e.g., scFv); and multispecific antibodies formed from antibody fragments. Papain digestion of antibodies produces two identical antigen-binding fragments, called "Fab" fragments, each with a single antigen-binding site, and a residual "Fc" fragment, a nomenclature that reflects the ability to crystallize easily. Pepsin treatment produces F(ab')2 fragments that have two antigen-binding sites and are still able to cross-link antigen.

[0034] "Fv" is the smallest antibody fragment containing a complete antigen recognition and binding site. This region is composed of a dimer of a heavy chain variable domain and a light chain variable domain that are tightly non-covalently associated. In this configuration, the three CDRs of each variable domain usually interact to define the antigen binding site on the surface of the VH-VL dimer. Generally speaking, the six CDRs together confer antibody antigen binding specificity, but when one or more CDRs in the six CDRs are missing or modified, for example, by changing the amino acid sequence of the one or more CDRs, such as by amino acid insertion, deletion or substitution, there is an example of maintaining antigen binding specificity. In addition, even a single variable domain (or half of an Fv containing only three CDRs specific for an antigen) has the ability to recognize and bind to an antigen, but the affinity is lower than the entire binding site. Residues other than those present in the CDRs may also be important or play a role in antigen binding and / or specificity, as shown by PGT121 and a closely related somatic variant that uses residues in the light chain framework 3 to interact with the gp120 antigen (Julien et al., Science, Vol. 342, pp. 1477-1483, 2013; Julien et al., PLOS Pathog., Vol. 9, p. e1003342, (2013)). These residues arise in part from an unusual three amino acid insertion that extends an otherwise short surface loop in PGT121 and related somatic variants (e.g., PGT122, PGT123, PGT124, PGT133, PGT134, 10-1074) that contacts both the N332 glycan and protein residues on HIV Env, effectively forming an additional (e.g., fourth) complementarity determining region (CDR) loop in the PGT121 light chain between LC CDR 2 and LC CDR 3.

[0035] The term "hypervariable region" refers to the amino acid residues of an antibody that are generally responsible for antigen binding. A hypervariable region generally comprises amino acid residues from a "complementarity determining region" or "CDR" (e.g., approximately residues 24-34 (L1), 50-56 (L2), and 89-97 (L3) in VL, and approximately residues 31-35 (H1), 50-65 (H2), and 95-102 (H3) in VH when numbered according to the Kabat numbering system; Kabat et al., Sequences of Proteins of Immunological Interest, 5th ed., Public Health Service, National Institutes of Health, Bethesda, Md., 1991); and / or those residues from a "hypervariable loop" (e.g., residues 24-34 (L1), 50-56 (L2), and 89-97 (L3) in VL, and residues 26-32 (H1), 52-56 (H2), and 95-101 (H3) in VH, when numbered according to the Chothia numbering system; Chothia and Lesk, J. Mol. Biol. , Vol. 196, pp. 901-917, 1987); and / or those residues from the “hypervariable loop” VCDR (e.g., residues 27-38 (L1), 56-65 (L2), and 105-120 (L3) in VL, and residues 27-38 (H1), 56-65 (H2), and 105-120 (H3) in VH when numbered according to the IMGT numbering system; Lefranc, MP et al., Nucl. Acids Res., Vol. 27, pp. 209-212, 1999; Ruiz, M. et al., Nucl. Acids Res., Vol. 28, pp. 219-221, 2000). Optionally, the antibody has a symmetric insertion at one or more of the following points 28, 36 (L1), 63, 74-75 (L2) and 123 (L3) in VL and 28, 36 (H1), 63, 74-75 (H2) and 123 (H3) in VH when numbered according to AHo; Honneger, A and Plunkthun, A., J., Mol. Biol., Vol. 309, pp. 657-670, 2001).

[0036] "Fab" fragment is the region on the antibody that binds to the antigen. It is composed of a constant domain and a variable domain of each of the heavy chain and the light chain. These domains shape the complementary position (antigen binding site) of the amino terminal of the monomer. The two variable domains bind to the epitope on its specific antigen. The difference between the Fab fragment and the Fab' fragment is that some residues are added to the carboxyl terminal of the heavy chain CH1 domain, including one or more cysteines from the hinge region of the antibody. Fab'-SH is the name of Fab' herein, in which the cysteine ​​residues of the constant domain carry free thiol groups. F(ab')2 antibody fragments were originally produced as Fab' fragment pairs, with hinge cysteines between them. Other chemical couplings of antibody fragments are also known.

[0037] The "light chains" of antibodies (immunoglobulins) from any vertebrate species can be assigned to one of two clearly distinct types, called kappa and lambda, based on the amino acid sequences of their variable or constant domains. Depending on the amino acid sequence of the constant domain of their heavy chains, immunoglobulins can be assigned to different classes. There are five major classes of immunoglobulins: IgA, IgD, IgE, IgG, and IgM, and several of these immunoglobulins can be further divided into subclasses (isotypes), e.g., IgG1, IgG2, IgG3, IgG4, IgA, and IgA2.

[0038] "Single-chain Fv" or "scFv" or "sFv" antibody fragments comprise the VH and VL domains of an antibody, wherein these domains are present in a single polypeptide chain. In some embodiments, the Fv polypeptide further comprises a polypeptide linker between the VH and VL domains that enables the sFv to form a desired antigen-binding structure.

[0039] The term "diabody" refers to small antibody fragments with two antigen binding sites, which contain a heavy chain variable domain (VH) connected to a light chain variable domain (VL) in the same polypeptide chain (VH-VL). By using a linker that is too short to allow pairing between the two domains on the same chain, the domains are forced to pair with the complementary domains of another chain and generate two antigen binding sites. Diabodies are more fully described in, for example, EP404,097; WO 93 / 11161; and Hollinger et al., Proc. Natl. Acad. Sci. USA, Vol. 90, pp. 6444-6448, 1993.

[0040] An "isolated" antibody or antigen-binding fragment thereof is an antibody or antigen-binding fragment thereof that has been identified and separated and / or recovered from a component of its natural environment. Contaminant components of its natural environment are materials that will interfere with the diagnostic or therapeutic use of the antibody, and may include enzymes, hormones, and other proteinaceous or non-proteinaceous solutes. In some embodiments, the antibody will be purified (1) to greater than 95% by weight of the antibody as determined by the Lowry method, e.g., greater than 99% by weight, (2) to a degree sufficient to obtain at least 15 N-terminal or internal amino acid sequence residues by use of a spinning cup sequencer, or (3) to homogeneity by SDS-PAGE under reducing or non-reducing conditions using Coomassie blue or silver stain. An isolated antibody includes an antibody in situ within a recombinant cell, since at least one component of the antibody's natural environment will not be present. However, typically, an isolated antibody will be prepared by at least one purification step.

[0041] An antibody or antigen-binding fragment thereof that "specifically binds to" or is "specific for" a particular polypeptide or an epitope on a particular polypeptide is an antibody or antigen-binding fragment thereof that binds to that particular polypeptide or an epitope on a particular polypeptide and does not substantially bind to any other polypeptide or polypeptide epitope. In some embodiments, the antibodies of the present disclosure specifically bind to an antigen in the form of a monoclonal antibody, scFv, Fab, or other form of an antibody measured at a temperature of about 4°C, 25°C, 37°C, or 42°C, for example, HIV-1 gp120 polypeptide with a dissociation constant Kd equal to or lower than 100 nM, optionally lower than 10 nM, optionally lower than 1 nM, optionally lower than 0.5 nM, optionally lower than 0.1 nM, optionally lower than 0.01 nM, or optionally lower than 0.005 nM. The affinity of the antibody can be easily determined using conventional techniques, such as those described by Scatchard et al. (Ann. NY, Acad. Sci. USA, Vol. 51, p. 660, 1949, ELISA assays, biolayer interferometer (BLI) assays, and surface plasmon resonance (SPR) assays. The binding properties of antibodies to antigens, cells, or their tissues can generally be determined and assessed using immunodetection methods (including, for example, immunofluorescence-based assays such as immunohistochemistry (IHC) and / or fluorescence-activated cell sorting (FACS)).

[0042] As used herein, an "internalizing" antibody is an antibody that is absorbed by (i.e., enters) a cell when it is bound to an antigen on a mammalian cell (e.g., a cell surface polypeptide or receptor). Of course, internalizing antibodies will include antibody fragments, human or chimeric antibodies, and antibody conjugates. For certain therapeutic applications, in vivo internalization is contemplated. The number of internalized antibody molecules will be sufficient or sufficient to kill cells or inhibit their growth, especially infected cells. Depending on the efficacy of the antibody or antibody conjugate, in some cases, a single antibody molecule is taken into a cell to kill the target cell to which the antibody is bound. For example, some toxins are highly effective in killing, so that the internalization of one molecule of the toxin conjugated to the antibody is sufficient to kill infected cells.

[0043] The term "antagonist" antibody is used in the broadest sense and includes antibodies that partially or completely block, inhibit, or neutralize the biological activity of an epitope, polypeptide, or cell to which it specifically binds. Methods for identifying antagonist antibodies may include contacting a polypeptide or cell specifically bound by a candidate antagonist antibody with the candidate antagonist antibody and measuring a detectable change in one or more biological activities normally associated with the polypeptide or cell.

[0044] "Antibodies that inhibit the growth of infected cells" or "growth inhibitory" antibodies are antibodies that bind to infected cells expressing or capable of expressing HIV1 epitopes bound by antibodies and cause measurable growth inhibition thereof. Compared to a suitable control (the control is usually an infected cell not treated with the tested antibody), a preferred growth inhibitory antibody inhibits the growth of infected cells by greater than 20%, preferably about 20% to about 50%, and even more preferably greater than 50% (e.g., about 50% to about 100%). Growth inhibition can be measured in cell culture at an antibody concentration of about 0.1 μg / ml to about 30 μg / ml or about 0.5 nM to about 200 nM, wherein growth inhibition is measured 1 to 10 days after the infected cells are exposed to the antibody. Infection cell growth inhibition in vivo can be determined in various ways known in the art. If the percentage of infected cells or the total number of infected cells caused by administration of the antibody at about 1 μg / kg to about 100 mg / kg body weight is reduced within about 5 days to 3 months from the first administration of the antibody, preferably within about 5 days to 30 days, then the antibody is growth inhibitory in vivo.

[0045] An antibody that "induces apoptosis" is an antibody that induces programmed cell death as determined by the binding of annexin V, DNA fragmentation, cell shrinkage, endoplasmic reticulum expansion, cell fragmentation, and / or the formation of membrane vesicles (called apoptotic bodies). Preferably, the cell is an infected cell. There are a variety of methods for evaluating cellular events associated with apoptosis. For example, phosphatidylserine (PS) translocation can be measured by annexin binding; DNA fragmentation can be evaluated by DNA gradient; and nuclear / chromatin condensation together with DNA fragmentation can be evaluated by any increase in low ploidy cells. Preferably, an antibody that induces apoptosis is an antibody that causes annexin binding in an annexin binding assay relative to untreated cells by about 2 to 50 times, preferably about 5 to 50 times, and most preferably about 10 to 50 times.

[0046] Antibody "effector function" refers to those biological activities attributable to the Fc region (native sequence Fc region or amino acid sequence variant Fc region) of an antibody, and varies with the antibody isotype. Examples of antibody effector functions include: Clq binding and complement dependent cytotoxicity; Fc receptor binding; antibody-dependent cell-mediated cytotoxicity (ADCC); phagocytosis (e.g., antibody-dependent cell-mediated phagocytosis (ADCP)); down-regulation of cell surface receptors (e.g., B cell receptors); and B cell activation.

[0047] "Antibody-dependent cell-mediated cytotoxicity" or "ADCC" refers to a form of cytotoxicity in which secreted or exogenously administered Ig binds to Fc receptors (FcRs) on certain cytotoxic cells (e.g., natural killer (NK) cells, neutrophils, and macrophages), enabling these cytotoxic effector cells to specifically bind to target cells carrying antigens and subsequently kill the target cells with cytotoxins. Antibodies "arm" cytotoxic cells and are required for such killing. Primary cell NK cells used to mediate ACC express only FcγRIII, while monocytes express FcγRI, FcγRII, and FcγRIII. Ravetch and Kinet, Annu. Rev. Immunol, Vol. 9, pp. 457-492, 1991 Table 4 on p. 464 summarizes the expression of FcRs on hematopoietic cells. In order to evaluate the ADCC activity of the molecule of interest, an in vitro ADCC assay, such as that described in U.S. Pat. No. 5,500,362 or U.S. Pat. No. 5,821,337, may be performed. Effector cells that can be used for such assays include peripheral blood mononuclear cells (PBMC) and natural killer (NK) cells. Alternatively or additionally, the ADCC activity of an antibody or its antigen-binding fragment may be assessed in vivo, for example, in an animal model, such as that disclosed in Clynes et al., Proc. Natl. Acad. Sci. (USA), Vol. 95, pp. 652-656, 1998.

[0048] "Fc receptor" or "FcR" describes a receptor that binds to the Fc region of an antibody. In certain embodiments, the FcR is a native sequence human FcR. In addition, a preferred FcR is an FcR that binds to an IgG antibody (gamma receptor), and includes receptors of FcγRI, FcγRII, and FcγRIII subclasses, including allelic variants and alternative splicing forms of these receptors. FcγRII receptors include FcγRIIA ("activating receptor") and FcγRIIB ("inhibiting receptor"), which have similar amino acid sequences that differ primarily in their cytoplasmic domains, and FcγRIIC, which includes an FcγRIIB extracellular domain fused to an activation cytoplasmic region. The activating receptor FcγRIIA contains an immunoreceptor tyrosine-based activation motif (ITAM) in its cytoplasmic domain. Inhibiting receptor FcγRIIB contains an immunoreceptor tyrosine-based inhibition motif (ITIM) in its cytoplasmic domain (see review by M. in Daeron, Annu. Rev. Immunol., Vol. 15, pp. 203-234, 1997). FcRs are reviewed in Ravept and Kinet, Annu. Rev. Immunol, Vol. 9, pp. 457-492, 1991; Capel et al., Immunomethods, Vol. 4, pp. 25-34, 1994; and de Haas et al., J. Lab. Clin. Med., Vol. 126, pp. 330-341, 1995. The term "FcR" herein encompasses other FcRs, including those to be identified in the future. The term also includes the neonatal receptor FcRn, which is responsible for the transfer of maternal IgG to the fetus (Guyer et al., J. Immunol., Vol. 117, p. 587, 1976; and Kim et al., J. Immunol., Vol. 24, p. 249, 1994), and plays a role in salvaging IgG from lysosomal degradation by FcRn-dependent recycling after endocytosis. FcRn binding after endothelial cell pinocytosis has been shown to be important for maintaining the extended pharmacokinetic half-life of antibodies. An in vitro assessment of the pH-dependent human FcRn binding of antibodies can be performed to provide a prediction of favorable clinical pharmacokinetic potential (Datta-Mannan and Wroblewski, Drug Metab. Dispos., Vol. 42, p. 1867-1872, 2014).

[0049] "Human effector cells" are leukocytes that express one or more FcRs and perform effector functions. Preferably, the cells express at least FcγRIII and perform ADCC effector functions. Examples of human leukocytes that mediate ADCC include PBMCs, NK cells, monocytes, cytotoxic T cells, and neutrophils; of which PBMCs and NK cells are preferred. Effector cells can be isolated from natural sources, for example, from blood.

[0050] "Complement dependent cytotoxicity" or "CDC" refers to the lysis of target cells in the presence of complement. Activation of the classical complement pathway is initiated by the binding of the first component of the complement system (Clq) to an antibody (of the appropriate subclass) bound to its cognate antigen. To assess complement activation, a CDC assay may be performed, e.g., as described in Gazzano-Santoro et al., J. Immunol. Methods, Vol. 202, p. 163, 1996.

[0051] "Neutralizing antibodies" are antibodies that can neutralize the pathogen's ability to cause and / or maintain infection in a host and / or target cell in vitro. Neutralizing monoclonal human antibodies and antigen-binding fragments thereof are described herein, wherein the antibody recognizes an antigen from HIV, for example, a gp120 polypeptide. In certain embodiments, "neutralizing antibodies" can inhibit entry of HIV-1 viruses, for example, SF162 and / or JR-CSF (Kostrikis LG et al., / Virol., 1996, Vol. 70, No. 1, pp. 445-458) with a neutralization index >1.5 or >2.0. So-called "broadly neutralizing antibodies" means antibodies that neutralize more than one HIV-1 virus species (from different clades and different strains within clades) in a neutralization assay. Broadly neutralizing antibodies can neutralize at least 2, 3, 4, 5, 6, 7, 8, 9 or more different strains of HIV-1, which belong to the same or different clades. In certain embodiments, broadly neutralizing antibodies can neutralize multiple HIV-1 species belonging to at least 2, 3, 4, 5, or 6 different clades. In certain embodiments, the inhibitory concentration of the monoclonal antibody can be less than about 0.0001 μg / ml, less than about 0.001 μg / ml, less than about 0.01 μg / ml, less than about 0.1 μg / ml, less than about 0.5 μg / ml, less than about 1.0 μg / ml, less than about 5 μg / ml, less than about 10 μg / ml, less than about 25 μg / ml, less than about 50 μg / ml, or less than about 100 μg / ml to neutralize about 50% of the input virus in a neutralization assay.

[0052] HIV viruses are divided into specific groups M, N, O and P, with M being the "major" group and responsible for most HIV / AIDS worldwide. Based on their genetic sequences, the M group is further subdivided into subtypes (also called clades) with prevalence in different geographic locations.

[0053] M group "subtype" or "clade" is a subtype of HIV-1 group M defined by genetic sequence data. Examples of group M subtypes include subtype AK. Some subtypes in the known subtypes are more toxic or resistant to different drugs. There are also "circulating recombinant forms" or CRFs derived from recombination between viruses of different subtypes, each of which has a number. For example, CRF12_BF is a recombination between subtypes B and F. Subtype A is common in West Africa. Subtype B is the main form in Europe, America, Japan, Thailand and Australia. Subtype C is the main form in parts of South Africa, East Africa, India, Nepal and China. Subtype D is usually only found in East and Central Africa. Subtype E has never been identified as a non-recombinant, and only recombined with subtype A as CRF01_AE. Subtype F has existed in Central Africa, South America and Eastern Europe. Subtype G (as well as CRF02_AG) has been found in Africa and Central Europe. Subtype H is limited to Central Africa. Subtype I was originally used to describe the strain now considered to be CRF04_cpx, where cpx is used for a "complex" recombination of several subtypes. Subtype J is mainly found in North Africa, Central Africa and West Africa, while Caribbean subtype K is limited to the Democratic Republic of the Congo and Cameroon. These subtypes are sometimes further divided into sub-subtypes, such as A1 and A2 or F1 and F2. In 2015, strain CRF19 was found to be a recombinant of subtypes A, D and G, and its subtype D protease is closely related to the rapid development of AIDS in Cuba.

[0054] "HIV tropism" refers to the specificity of the HIV virus for a particular host cell, which is determined in part by the interaction of viral surface structures with receptors present on the surface of the host cell. The HIV tropism of a patient's virus is measured using the monogrambio assay (monogrambio.com). (See, e.g., Lee et al., AIDS Res Hum Retroviruses., 2013, Vol. 29, No. 6, pp. 979-984).

[0055] HIV can infect a variety of cells, such as CD4+ helper T cells and macrophages, which express CD4 molecules on their surfaces. HIV-1 enters macrophages and T helper cells not only through the interaction of virion envelope glycoproteins (e.g., gp120) with CD4 molecules on target cells, but also mediated by the interaction of its chemokine co-receptors. HIV-1 macrophage (M-tropism) strains or non-syncytial inducible strains (NSI) use β-chemokine receptor CCR5 to enter and therefore can replicate in macrophages and CD4+T cells. These strains are referred to as R5 viruses. The CCR5 co-receptor is used by almost all primary HIV-1 isolates, and has nothing to do with viral genetic subtypes. In primary CD4+T cells and macrophages, replicate tropism T cell isolates or syncytial inducible (SI) strains, and use α-chemokine receptor CSCR4 to enter. These strains are referred to as X4 viruses. Viruses that use only the CCR5 receptor are called R5, those that use only CXCR4 are called X4, and those that use both are called X4R5 or dual / mixed tropism. However, co-receptor use alone does not explain viral tropism, as not all R5 viruses are able to use CCR5 on macrophages for productive infection.

[0056] Also described herein are "non-neutralizing antibodies," which in certain embodiments are antibodies that bind to one or more viral strains but do not neutralize the virus. However, in terms of Fc-mediated killing, non-neutralizing antibodies can still eliminate cells that express viral antigens that bind to the antibody but are not neutralized by the antibody. Thus, in certain embodiments, the antibody can bind to the viral antigen and eliminate virally infected cells without neutralizing the virus.

[0057] The term "nucleic acid molecule" refers to a polymeric form of nucleotides, and includes both sense and antisense strands of RNA, cDNA, genomic DNA, and the synthetic forms and mixed polymers described above. In a specific embodiment, nucleotides refer to ribonucleotides, deoxyribonucleotides, or modified forms of either type of nucleotides and combinations thereof. The term also includes, but is not limited to, DNA in single-stranded and double-stranded forms. In addition, polynucleotides, such as cDNA or mRNA, may include one or both of naturally occurring and modified nucleotides linked together by naturally occurring and / or non-naturally occurring nucleotide bonds. As will be readily appreciated by those skilled in the art, nucleic acid molecules may be chemically or biochemically modified, or may contain non-natural or derived nucleotide bases. Such modifications include, for example, labeling, methylation, substitution of one or more of the naturally occurring nucleotides with analogs, internucleotide modifications such as uncharged bonds (e.g., methylphosphonates, phosphotriesters, phosphoramidates, carbamates, etc.), charged bonds (e.g., phosphorothioates, phosphorodithioates, etc.), side chain moieties (e.g., polypeptides), intercalators (e.g., acridine, psoralens, etc.), chelators, alkylates, and modified bonds (e.g., α-anomeric nucleic acids, etc.). The above terms are also intended to include any topological conformation, including single-stranded conformation, double-stranded conformation, partially double-stranded conformation, triple-stranded conformation, hairpin conformation, cyclic conformation, and padlock conformation. Unless otherwise indicated, reference to a nucleic acid sequence encompasses its complementary sequence. Therefore, reference to a nucleic acid molecule with a specific sequence is understood to encompass its complementary chain and its complementary sequence. The term also includes codon-optimized nucleic acids.

[0058] The term "operably linked" refers to two or more nucleic acid sequence elements that are generally physically linked and in a functional relationship with each other. For example, a promoter is operably linked to a coding sequence if it is capable of initiating or regulating the transcription or expression of the coding sequence, in which case the coding sequence is understood to be "under the control of the promoter."

[0059] As used herein, "substitution" means that one or more amino acids or nucleotides are replaced by different amino acids or nucleotides, respectively.

[0060] An "isolated" nucleic acid refers to a nucleic acid molecule that has been separated from a component of its natural environment. An isolated nucleic acid includes a nucleic acid molecule contained in cells that normally contain the nucleic acid molecule, but the nucleic acid molecule is present extrachromosomally or at a chromosomal location that is different from its natural chromosomal location.

[0061] "Isolated nucleic acid encoding an antibody or fragment thereof" refers to one or more nucleic acid molecules encoding antibody heavy and light chains (or fragments thereof), including such nucleic acid molecules in a single vector or separate vectors, as well as such nucleic acid molecules present at one or more locations in a host cell.

[0062] As used herein, the term "vector" refers to a nucleic acid molecule capable of propagating another nucleic acid to which it is linked. The term includes vectors that are self-replicating nucleic acid structures as well as vectors that integrate into the genome of a host cell into which the vector has been introduced. Certain vectors are capable of directing the expression of nucleic acids to which they are operatively linked. Such vectors are referred to as

[0063] As used herein, the term polynucleotide "variant" is a polynucleotide that typically differs from a polynucleotide specifically disclosed herein in one or more substitutions, deletions, additions and / or insertions. Such variants may be naturally occurring or may be generated synthetically, for example, by modifying one or more of the polynucleotide sequences described herein and evaluating one or more biological activities of the encoded polypeptide as described herein and / or using any of a variety of techniques well known in the art.

[0064] As used herein, the term polypeptide "variant" is a polypeptide that generally differs from the polypeptides specifically disclosed herein in one or more substitutions, deletions, additions and / or insertions. Such variants may be naturally occurring or may be generated synthetically, for example, by modifying one or more of the polypeptide sequences of the invention described above and evaluating one or more biological activities of the polypeptides as described herein and / or using any of a variety of techniques well known in the art.

[0065] The term "variant" may also refer to any naturally occurring or engineered molecule comprising one or more nucleotide or amino acid mutations. In one embodiment, the molecule is an antibody. For example, somatic variants may encompass all related naturally occurring antibodies that are part of or derived from the same B cell lineage. Engineered variants may encompass all single or combined mutations made to an antibody.

[0066] Modifications can be made in the structures of the polynucleotides and polypeptides of the present invention, and still obtain functional molecules encoding variants or derivative polypeptides with desired properties. When it is desired to change the amino acid sequence of a polypeptide to produce an equivalent or even improved variant or part of a polypeptide of the present invention, those skilled in the art will typically change one or more of the codons encoding the DNA sequence.

[0067] For example, certain amino acids can be substituted for other amino acids in a protein structure without significantly losing its ability to bind to other polypeptides (e.g., antigens) or cells. Since it is the binding ability and properties of a protein that define the biological functional activity of that protein, certain amino acid sequence substitutions can be made in a protein sequence, and certainly also in its underlying DNA coding sequence, and yet still obtain a protein with similar properties. It is therefore conceivable that various changes can be made in the polypeptide sequences of the disclosed antibodies and antigen-binding fragments thereof, or the corresponding DNA sequences encoding the polypeptides, without significantly losing their biological utility or activity.

[0068] In many cases, polypeptide variants will contain one or more conservative substitutions. A "conservative substitution" is one in which an amino acid is substituted for another amino acid having similar properties, such that one skilled in the art of peptide chemistry would expect that the secondary structure and hydrophilicity of the polypeptide would remain essentially unchanged.

[0069] When comparing polynucleotide and polypeptide sequences, the two sequences are considered "identical" if the sequence of nucleotides or amino acids in the two sequences is the same when aligned for maximum correspondence, as described below. Comparisons between two sequences are typically performed by comparing the sequences over a comparison window to identify and compare local regions of sequence similarity. As used herein, a "comparison window" refers to a segment of at least about 20 consecutive positions (usually 30 to about 75, 40 to about 50) in which, after the two sequences are optimally aligned, the sequence can be compared to a reference sequence having the same number of consecutive positions.

[0070] Sequence alignments for comparison can be performed using the Megalign program in the Lasergene bioinformatics software package (DNASTAR, Inc., Madison, WI) using default parameters. This program embodies several alignment schemes described in the following references: Dayhoff, MO, 1978, A model of evolutionary change in proteins-Matrices for detecting distant relationships, Dayhoff, MO (ed.), Atlas of Protein Sequence and Structure, National Biomedical Research Foundation, Washington DC, Vol. 5 Supplement 3, pp. 345-358; Hein J., 1990, Unified Approach to Alignment and Phylogenes, pp. 626-645, Methods in Enzymology, Vol. 183, Academic Press, Inc., San Diego, CA; Higgins, DG and Sharp, PM, 1989, CABIOS, Vol. 5, pp. 151-153; Myers, EW and Muller W., 1988, CABIOS, Vol. 4, pp. 11-17; Robinson, ED, 1971, Comb. Theor, Vol. 77, p. 105; Santou, N. Nes, M., 1987, Mol. Biol. Evol., Vol. 4, pp. 406-425; Sneath, PHA and Sokal, RR, 1973, Numerical Taxonomy-the Principles and Practice of Numerical Taxonomy, Freeman Press, San Francisco, CA; Wilbur, WJ and Lipman, DJ, 1983, Proc. Natl. Acad., Sci. USA, Vol. 80, pp. 726-730.

[0071] Alternatively, alignments of sequences for comparison may be performed by the local identity algorithm of Smith and Waterman (1981, Add. APL. Math, Vol. 2, p. 482), by the identity alignment algorithm of Needleman and Wunsch (1970, J. Mol. Biol., Vol. 48, p. 443), by the search for similarity method of Pearson and Lipman (1988, Proc. Natl. Acad. Sci. USA, Vol. 85, p. 2444), by computerized implementations of these algorithms (GAP, BESTFIT, BLAST, FASTA, and TFASTA in the Wisconsin Genetics Software Package, Genetics Computer Group (GCG), 575 Science Dr., Madison, WI), or by inspection.

[0072] An example of an algorithm suitable for determining sequence identity percentage and sequence similarity is BLAST and BLAST2.0 algorithms, which are described in Altschul et al., 1977, Nucl. Acids Res., Vol. 25, pp. 3389-3402 and Altschul et al., 1990, J. Mol. Biol., Vol. 215, pp. 403-410, respectively. BLAST and BLAST 2.0 can be used, for example, with the parameters described herein to determine the sequence identity percentage of polynucleotides and polypeptides described herein. Software for performing BLAST analysis is publicly available through the National Center for Biotechnology Information.

[0073] In an illustrative example, for nucleotide sequences, the cumulative score can be calculated using the parameters M (reward score for a pair of matching residues; always > 0) and N (penalty score for mismatching residues; always < 0). Extension of the word hits in each direction stops when: the cumulative alignment score drops by the amount X from its maximum achieved value; the cumulative score becomes zero or lower due to the accumulation of one or more negative-scoring residue alignments; or the end of either sequence is reached. The BLAST algorithm parameters W, T, and X determine the sensitivity and speed of the alignment. The BLASTN program (for nucleotide sequences) uses a word length (W) of 11 and an expectation (E) of 10 by default, and an alignment with the BLOSUM62 scoring matrix (see Henikoff and Henikoff, 1989, Proc. Natl. Acad. Sci. USA, Vol. 89, p. 10915), in which (B) is 50, the expectation (E) is 10, M=5, N=-4, and a comparison of both chains is performed.

[0074] For amino acid sequences, a scoring matrix can be used to calculate the cumulative score. Extension of the word hits in each direction stops when: the cumulative alignment score drops by the amount X from its maximum achieved value; the cumulative score goes to zero or lower due to the accumulation of one or more negative-scoring residue alignments; or the end of either sequence is reached. The BLAST algorithm parameters W, T, and X determine the sensitivity and speed of the alignment.

[0075] In one method, the "percentage of sequence identity" is determined by comparing two optimally aligned sequences over a comparison window of at least 20 positions, wherein the portion of the polynucleotide or polypeptide sequence in the comparison window may contain 20% or less, typically 5% to 15%, or 10% to 12% additions or deletions (i.e., gaps) compared to the reference sequence (excluding additions or deletions) used to align the two sequences. The percentage is calculated by determining the number of positions where the same nucleic acid base or amino acid residue occurs in the two sequences to generate the number of matched positions, dividing the number of matched positions by the total number of positions in the reference sequence (i.e., the window size) and multiplying the result by 100 to generate the percentage of sequence identity.

[0076] "Homology" refers to the percentage of residues in a polynucleotide or polypeptide sequence variant that are identical with the non-variant sequence, after aligning the sequences and introducing gaps, if necessary, to achieve the maximum percent homology.

[0077] "Binding affinity" can refer to the binding dissociation constant (Kd) or the apparent affinity (eg, EC50) value. BRIEF DESCRIPTION OF THE DRAWINGS

[0078] Figure 1 The number of screened subjects from the Zurich Primary HIV Infection Cohort Study whose genotype predicts sensitivity to GS-9722 (Ipavir monoclonal antibody) is shown. Plasma samples from 92 individuals before ART were analyzed in the GenoSure HIV envelope RNA assay. "None" indicates that all screened individuals did not select specific amino acids in the HIV envelope gene. Amino acid positions indicated for each category.

[0079] Figure 2 The number of screened clade B subjects from the Zurich Primary HIV Infection Cohort Study whose genotype predicted sensitivity to GS-9722 is shown. Pre-ART plasma samples from 59 clade B infected individuals were analyzed in the GenoSure HIV Envelope RNA assay. "None" indicates that all screened individuals did not have a selection for a specific amino acid in the HIV Envelope gene. Amino acid positions indicated for each category.

[0080] Figure 3Figure 2 shows the sensitivity of a population of viruses derived from pre-ART plasma samples from the Zurich Primary HIV Infection Cohort Study to GS-9722. Viruses from 29 samples with a positive predictive value of 80.7% or higher were analyzed in the HIV Entry Assay (Monogram Biosciences). Amino acid positions are indicated for each category.

[0081] Figure 4 The sensitivity of subcloned viruses to GS-9722 is shown in a population of viruses derived from pre-ART plasma samples from the Zurich Primary HIV Infection Cohort Study. Twenty individual viruses from four pre-ART plasma samples were analyzed in the HIV Entry Assay (Monogram Biosciences), where group virus sensitivity was predicted by genotyping and tested by phenotyping. Solid lines indicate IC50 for group viruses. DETAILED DESCRIPTION

[0082] 1. Introduction

[0083] The methods of the present invention are based in part on the unexpected discovery of a population of HIV-infected patients that respond to administration of an anti-HIV gp120 V3 glycan-directed antibody or antigen-binding fragment thereof, in the absence of co-administration of additional anti-HIV antibodies directed against other HIV antigens (e.g., gp41) or non-overlapping epitopes of the same HIV antigen (e.g., directed against gp120 in the CD4 binding site region or the V2 apex region). Such patients are infected with a species of HIV that has a gp120 protein that is bound by the V3 glycan-directed antibody or antigen-binding fragment thereof.

[0084] Typically, these methods entail identifying a human subject infected with HIV or a population of HIV expressing gp120 comprising the following amino acid residues: a glycosylated asparagine at a position corresponding to amino acid residue position 332 (N332 glycan), an aspartic acid at a position corresponding to amino acid residue position 325 (D325), and one or more amino acid residues selected from the following amino acid residues: a threonine at a position corresponding to amino acid residue position 63 (T63), a leucine at a position corresponding to amino acid residue position 179 (L179), a threonine at a position corresponding to amino acid residue position 320 (T320), and a histidine at a position corresponding to amino acid residue position 330 (H330), wherein the amino acid positions are referenced to SEQ ID NO: 4 (i.e., residues 1-511 of NCBI Reference Sequence No. NP_057856.1). In various embodiments, the glycan is an oligomannose.

[0085] 2. Identification of subjects who respond to treatment with anti-HIV gp120V3 glycan-directed antibodies or antigen-binding fragments thereof Those .

[0086] In some embodiments, the patient is identified by receiving a report of the HIV species that infects the patient, the report identifying the HIV gp120 amino acid residues present at designated amino acid positions of interest, e.g., positions 332 and 325, and one or more amino acid positions selected from the following amino acid positions: 63, 179, 320, and 330, wherein the amino acid positions are referenced to SEQ ID NO: 4. In some embodiments, the patient is identified by performing one or more assays (e.g., polynucleotide or polypeptide sequencing) to determine the amino acid sequence of gp120 or the amino acid residues present at designated amino acid positions of interest of the gp120 protein of the HIV species that infects the patient. The identification of the full-length or partial sequence of the gp120 protein obtained from the subject can be determined at the polynucleotide or polypeptide level. In some embodiments, the amino acids present at the gp120 residue positions of interest are determined at the polypeptide level.

[0087] In various embodiments, the methods entail identifying a subject infected with HIV or a population of HIV that expresses a gp120 comprising the following amino acid residues: N332 glycan, D325, and T63, wherein the amino acid positions refer to SEQ ID NO:4.

[0088] In various embodiments, the methods entail identifying a subject infected with HIV or a population of HIV that expresses a gp120 comprising the following amino acid residues: N332 glycan, D325, and L179, wherein the amino acid positions refer to SEQ ID NO:4.

[0089] In various embodiments, the methods entail identifying a subject infected with HIV or a population of HIV that expresses a gp120 comprising the following amino acid residues: N332 glycan, D325, and T320, wherein the amino acid positions refer to SEQ ID NO:4.

[0090] In various embodiments, the methods entail identifying a subject infected with HIV or a population of HIV that expresses a gp120 comprising the following amino acid residues: N332 glycan, D325, and H330, wherein the amino acid positions refer to SEQ ID NO:4.

[0091] In various embodiments, the methods entail identifying a subject infected with HIV or a population of HIV that expresses a gp120 comprising the following amino acid residues: N332 glycan, D325, T63, and L179, wherein the amino acid positions refer to SEQ ID NO:4.

[0092] In various embodiments, the methods entail identifying a subject infected with HIV or a population of HIV that expresses a gp120 comprising the following amino acid residues: N332 glycan, D325, T63, and T320, wherein the amino acid positions refer to SEQ ID NO:4.

[0093] In some embodiments, the subject is infected with an HIV clade B virus. In various embodiments, the methods entail identifying a subject infected with HIV or a population of HIV that expresses a gp120 comprising the following amino acid residues: N332 glycan, D325, T63, and H330, wherein the amino acid positions are referenced to SEQ ID NO: 4. In various embodiments, the methods entail identifying a subject infected with HIV or a population of HIV that expresses a gp120 comprising the following amino acid residues: N332 glycan, D325, T63, L179, T320, and H330, wherein the amino acid positions are referenced to SEQ ID NO: 4.

[0094] In various embodiments, the methods entail identifying a subject infected with HIV or a population of HIV that expresses a gp120 comprising the following amino acid residues: N332 glycan, D325, T320, and H330, wherein the amino acid positions refer to SEQ ID NO:4.

[0095] In various embodiments, these methods entail identifying a subject infected with HIV or a population of HIV that expresses a gp120 comprising the following amino acid residues: N332 glycan, D325, L179, T320, and H330, wherein the amino acid positions are referenced to SEQ ID NO: 4. In some embodiments, the subject is infected with HIV clade A and / or HIV clade C virus. In some embodiments, the subject is infected with HIV clade A, clade B, and / or HIV clade C virus.

[0096] In various embodiments, the methods entail identifying a subject infected with HIV or a population of HIV that expresses a gp120 comprising the following amino acid residues: N332 glycan, D325, T63, L179, and T320, wherein the amino acid positions refer to SEQ ID NO:4.

[0097] In various embodiments, the methods entail identifying a subject infected with HIV or a population of HIV that expresses a gp120 comprising the following amino acid residues: N332 glycan, D325, T63, L179, and H330, wherein the amino acid positions refer to SEQ ID NO:4.

[0098] In some embodiments, the subject is infected with HIV or a population of HIV that expresses gp120 that further comprises one or more of the following amino acid residues: a glycan at amino acid residue 301 (glycan 301); a lysine at amino acid residue 677 (K677); an amino acid residue other than tryptophan (Trp, W) (e.g., alanine (Ala, A); cysteine ​​(Cys, C); aspartic acid or aspartic acid (Asp, D); glutamic acid or glutamic acid (Glu, E ); phenylalanine (Phe, F); glycine (Gly, G); histidine (His, H); isoleucine (Ile, I); lysine (Lys, K); leucine (Leu, L); methionine (Met, M); asparagine (Asn, N); proline (Pro, P); glutamine (Gln, Q); arginine (Arg, R); serine (Ser, S); threonine (Thr, T); valine (Val, V) or tyrosine at position 17 (Tyr, Y) (not_W17); an amino acid residue other than arginine at position 747 (e.g., A, C, D, E, F, G, H, I, K, L, M, N, P, Q, S, T, V, W, or Y) (not_R747); insertion_321.01 (e.g., insertion of any amino acid between positions G321 and K322 (e.g., A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, or Y) (not_R747); , W or Y)); glutamic acid at position 429 (E429); glutamine at position 442 (Q442); arginine at position 335 (R335); isoleucine at position 165 (I165); serine at position 393 (S393); isoleucine at position 307 (I307); glycan at position 295 (295 glycan); and / or asparagine at position 300 (N300), wherein the amino acid positions are referenced to SEQ ID NO:4.

[0099] gp120

[0100] Envelope glycoprotein gp120 (or gp120) is a 120kDa glycoprotein, which is a part of the HIV outer layer. It exists as a viral membrane spike, consisting of three gp120 molecules linked together, and anchored on the membrane by gp41 protein. Gp120 is essential for viral infection because it promotes HIV to enter host cells through its interaction with cell surface receptors. These receptors include DC-SIGN, heparan sulfate proteoglycans and CD4 receptors. The CD4 that is bound to the helper T cell induces the beginning of the cascade conformational changes of gp120 and gp41, which causes the virus to fuse with the host cell membrane.

[0101] Gp120 is encoded by the HIV env gene. The env gene encodes a gene product of approximately 850 amino acids. The major env product is the protein gp160, which is cleaved in the endoplasmic reticulum by the cellular protease furin into gp120 (approximately 480 amino acids) and gp41 (approximately 345 amino acids).

[0102] The V3 glycan site on gp120 is formed in part by a portion of the CCR5 co-receptor site and in part by surrounding camouflaged glycans (the so-called "high mannose patch") (Sok et al., Immunity, 2016, Vol. 45, pp. 31-45). Broadly neutralizing antibodies (bnAbs) to the V3 glycan site are the most common of all antibodies found in HIV infection (Walker et al., PLoS Pathog., 2010, Vol. 6, p. e1001028, 2010; Landais et al., PLoS Pathog., 2016, Vol. 12, p. e1005369; Georgiev et al., Science, 2013, Vol. 340, pp. 751-756). The consensus sequence of the V3 region of gp120 (Milich et al., J Virol., Vol. 67, No. 9, pp. 5623-5634, 1993) is provided as follows:

[0103] CTRPNNNTRKSIHIGPGRAFYTTGEIIGDIRQAHC (SEQ ID NO: 1).

[0104] The amino acid sequence of an exemplary gp160 polypeptide of HIV clone WITO is provided below (the V3 hypervariable loop is bolded, and the N332 potential N-linked glycosylation site is bolded and underlined):

[0105]

[0106]

[0107] The amino acid sequence of an exemplary gp160 polypeptide of the HIV clone identified in NCBI Reference Sequence No. NP_057856.1 is provided below (the V3 hypervariable loop is bolded, and the N332 potential N-linked glycosylation site is bolded and underlined):

[0108]

[0109]

[0110] The amino acid sequence of an exemplary gp120 polypeptide of a HXB2B subtype HIV-1 isolate (GenBank Accession No. K0345; corresponding to residues 1-511 of NCBI Reference Sequence No. NP_057856.1-) is provided below (the V3 hypervariable loop is bold, and the N332 potential N-linked glycosylation site is bold and underlined; the signal peptide is underlined):

[0111]

[0112] The amino acid sequence of an exemplary gp120 polypeptide is provided below:

[0113]

[0114] The amino acid sequence of another exemplary gp120 polypeptide (see, bioafrica.net / proteomics / ENV-GP120prot.html) is provided below:

[0115] TEKLWVTVYY GVPVWKEATT TLFCASDAKA YDTEVHNVWA

[0116] THACVPTDPN PQEVVLVNVT ENFNMWKNDM VEQMHEDIIS

[0117] LWDQSLKPCV KLTPLCVSLK CTDLKNDTNT NSSSGRMIME

[0118] KGEIKNCSFN ISTSIRGKVQ KEYAFFYKLD IIPIDNDTTS YKLTSCNTSV

[0119] ITQACPKVSF EPIPIHYCAP AGFAILKCNN KTFNGTGPCT NVSTVQCTHG

[0120] IRPVVSTQLL LNGSLAEEEV VIRSVNFTDN AKTIIVQLNT SVEINCTRPN

[0121] NNTRKRIRIQ RGPGRAFVTI GKIGNMRQAH CNISRAKWNN

[0122] TLKQIASKLR EQFGNNKTII FKQSSGGDPE IVTHSFNCGG EFFYCNSTQL

[0123] FNSTWFNSTW STEGSNNTEG SDTITLPCRI KQIINMWQKV

[0124] GKAMYAPPIS GQIRCSSNIT GLLLTRDGGN SNNESEIFRPGGGDMRDNWR SELYKYKVVKIEPLGVAPTK AKRRVVQREK R(SEQ ID NO:6)

[0125] Genomic diversity between independent human immunodeficiency virus type 1 (HIV-1) isolates, between serial isolates from the same patient, and even to a lesser extent within a single patient isolate, is a well-known feature of HIV-1. Although this sequence heterogeneity is distributed throughout the genome, most of the heterogeneity is localized in the env gene. Comparison of predicted amino acid sequences from several different isolates has shown that sequence heterogeneity is clustered in five variable regions (designated V1 to V5) of the surface glycoprotein gp120. The V3 region, although only 35 amino acids long, exhibits considerable sequence variability. Interestingly, despite this variability, the V3 region includes the proteins that mediate binding to CD4 + Determinants of cell interactions. Increased variability in gp120 leads to higher levels of viral replication, indicating increased viral fitness in individuals infected with different HIV-1 variants. Variability in potential N-linked glycosylation sites (PNGS) also leads to increased viral fitness. PNGS allows long-chain carbohydrates to bind to the hypervariable region of gp120. Therefore, the number of PNGS in env can affect the fitness of the virus by providing more or less sensitivity to neutralizing antibodies.

[0126] Biological samples

[0127] The HIV gp120 amino acid residues of interest are determined by HIV present in or suspected of being present in a biological sample from a subject. The biological sample may be from a solid tissue or biological fluid from a subject known or suspected of containing HIV. In various embodiments, the biological sample comprises or is from blood, peripheral blood mononuclear cells (PBMC), serum, plasma, semen, or lymph nodes. In some embodiments, the biological sample comprises or is from bile, blood, plasma, serum, breast milk, feces, pus, saliva, sebum, semen, sweat, tears, urine, or vomit. In patients whose viral levels are suppressed, for example, by antiretroviral (ART) therapy, the biological sample comprises solid tissue or biological fluid of a subject known or suspected to contain an HIV reservoir, for example, solid tissue and / or biological fluid comprising latent HIV-infected CD4+T cells (including memory and non-memory effector CD4+T cells), hematopoietic progenitors of CD4+T cells, γδT cells (including memory and non-memory effector γδT cells), natural killer (NK) cells, bone marrow cells (including monocytes and macrophages), hematopoietic progenitors of bone marrow cells and follicular dendritic cells. Anatomical reservoirs that may carry latent HIV-infected cells include lymphoid tissue, brain and central nervous system, gastrointestinal tract and gut-associated lymphoid tissue (GALT), reproductive tract, lungs, and skin. Tissues and cells in which latent HIV-infected cells and HIV reservoirs are found are described, for example, in Kuo et al., Curr Opin HIV AIDS., 2018, Vol. 13, No. 2, pp. 137-142; Mzingwane et al., Rev Med Virol., March 2017, Vol. 27, No. 2, doi: 10.1002 / rmv, 1924, (PMID 28128885); Churchill et al., Nat Rev Microbiol., 2016, Vol. 14, No. 1, pp. 55-60; Barton et al., Trends Microbiol., 2016, Vol. 24, No. 5, pp. 345-355, so these documents are hereby incorporated by reference in their entirety for all purposes.

[0128] In some embodiments, multiple biological samples from a single patient are evaluated. For example, in some embodiments, two or more biological samples from two or more different tissues or two or more different anatomical reservoirs from a single patient are evaluated.

[0129] Infection Stage

[0130] In various embodiments, the human subject is an adult, teenager, or infant. The subject may be symptomatic (e.g., viremia) or asymptomatic (e.g., acute infection or ART inhibition). In some embodiments, the human subject is acutely infected or recently infected with HIV. In certain embodiments, the subject does not seroconvert. In some embodiments, the human subject is chronically infected with HIV. The subject may or may not be receiving an antiretroviral therapy (ART) regimen.

[0131] Patients can be divided into Fiebig stages I to VI, which is based on the continuous gain of positive HIV-1 clinical diagnostic assays (viral RNA measured by PCR, p24 and p31 viral antigens measured by enzyme-linked immunosorbent assay (ELISA). The p24 antigen is a viral core protein that appears transiently in the blood during the rising phase once HIV-1 RNA levels rise above 10,000 copies / mL and before detectable HIV antibodies are produced. In Fiebig stage I, only HIV-1 RNA can be detected in the blood during the rising viremia. Fiebig stage II begins about 7 days later, when the test result for detecting p24 antigen is positive. In Fiebig stage III, IgM anti-HIV-1 antibodies can be detected with a sufficiently sensitive enzyme immunoassay (EIA) (e.g., third-generation EIA) within about 5 days after the p24 antigen test result is positive. Stage III usually occurs 1 to 2 weeks after the onset of acute retroviral symptoms. Fiebig stage IV represents the development of uncertain Western blot test, and occurs about 3 days after EIA test shows positive result. Transformation into obviously positive Western blot test, i.e. Fiebig stage V, usually occurs after 7 days or about 1 month after initial infection. The Fiebig stage of HIV infection is described in, for example, Fiebig et al., AIDS, 2003, Vol. 17, No. 13, pp. 1871-1879; Cohen et al., J Infect Dis., 2010, Vol. 202 Supplement 2, pp. S270-277; and McMichael et al., Nature Reviews Immunology, 2010, Vol. 10, pp. 11–23, so these documents are hereby incorporated by reference in their entirety for all purposes. In some embodiments, the biological sample evaluated is from a human subject with HIV infection of Fiebig stage IV or earlier, such as Fiebig stage I, Fiebig stage II, Fiebig stage III or Fiebig stage IV. In some embodiments, the biological sample evaluated is from a human subject with HIV infection at Fiebig stage V or Fiebig stage VI.

[0132] In some embodiments, the methods further comprise the step of obtaining a biological sample from a subject. In some embodiments, the methods entail receiving a report of HIV gp120 amino acid residues present at specified positions of interest, such as 332 and 325, and one or more amino acid positions selected from 63, 179, 320, and 330, wherein the amino acid positions are referenced to SEQ ID NO:4.

[0133] Identification of gp120 amino acids of interest

[0134] The amino acid residues at the designated positions of interest (e.g., 332 and 325, and one or more amino acid positions selected from 63, 179, 320, and 330) of the HIV gp120 sequence of the subject can be determined at the polynucleotide or polypeptide level, wherein the amino acid position is referenced to SEQ ID NO: 4. At the polynucleotide level, methods known in the art can be used to sequence HIV RNA or proviral DNA isolated from one or more biological samples. In some embodiments, HIV RNA or proviral DNA isolated from two or more biological samples of the subject are sequenced. In some embodiments, the two or more biological samples are obtained from different tissue sources (e.g., blood, peripheral blood mononuclear cells, lymph nodes, and / or semen). In some embodiments, at different time points, for example, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, or 8 weeks apart, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, or 12 months apart, the two or more biological samples are obtained.

[0135] As appropriate, primers annealed and amplified with HIV env coding sequences, especially the V3 variable region of gp120, can be used. In some embodiments, nested primer sets can be used. In various embodiments, RNA is directly sequenced or reverse transcriptase polymerase chain reaction (RT-PCR) can be performed. In some embodiments, Sanger sequencing can be performed, for example, when sequencing to determine the amino acid residues in the V3 region, or when sequencing samples from patients in the early Fiebig stage of the disease (for example, before Fiebig stage III, such as Fiebig stage I or II). In various embodiments, single genome amplification (SGA) and sequencing are performed. The method for single genome amplification (SGA) and sequencing for plasma HIV virion RNA is described in, for example, Salazar-Gonzalez et al., 2008, J Virol, Vol. 82, pp. 3952-3970; and Keele et al., Proc Natl Acad Sci U SA., 2008, Vol. 105 No. 21, pp. 7552-7557. The application of SGA for determining amino acid sequence changes in HIV gp120 sequences and for the methods described herein is described in, for example, Bar et al., N Engl J Med., 2016, Vol. 375, No. 21, pp. 2037-2050; and Mendoza et al., Nature, 2018, Vol. 561, No. 7724, pp. 479-484. In various embodiments, high-throughput next generation sequencing (NGS), massively parallel or deep sequencing technology are used to sequence the gp120 (including at least the V3 variable region) of a population of HIV species in one or more biological samples from a single patient or subject. In such cases, multiple nucleic acid sequences encoding at least the V3 variable region of gp120 are sequenced and compared. In some embodiments, the full length of gp120 is sequenced. Exemplary platforms for performing NGS sequencing that can be used to determine the gp120 sequence of HIV species in one or more biological samples from a patient include Illumina (Solexa) (illumina.com), Ion torrent: Proton / PGM sequencing (thermofisher.com), SOLiD (thermofisher.com), and Single Molecule, Real-Time (SMRT) sequencing (Pacific Biosciences, pacb.com).Methods for isolating and sequencing HIV gp120 (including at least the V3 glycan region) from patients and that can be applied to the methods of the present invention are described, for example, in Shioda et al., J Virol., 1997, Vol. 71, No. 7, pp. 4871-4881; Colón et al., J Virol Antivir Res., 2015, Vol. 4, No. 3, pii: p. 143 (PMID: 27358904); Kafando et al., PLoS One., 2017, Vol. 12, No. 12, p. e0189999; Hebberecht et al., PLoS One., 2018, Vol. 13, No. 4, p. e0195679; Andrews et al., Sci. Rep., 2018, Vol. 8, No. 1, p. 5743; and Landais et al., Immunity., 2017, Vol. 47, No. 5, pp. 990-1003. Optionally, shorter sequence reads ("contigs") of nucleic acid sequences can be assembled into longer sequences that include at least the V3 variable region of gp120. Methods for assembling contiguous groups of HIV genome sequences that can be applied to the methods of the present invention are described in, for example, Huang et al., Bioinformation, 2018, Vol. 14, No. 8, pp. 449-454; Hiener et al., J Vis Exp., Oct. 16, 2018, Vol. 140, doi: 10.3791 / 58016; and Wymant et al., Virus Evol., May 18, 2018, Vol. 4, No. 1, vey007, doi: 10.1093 / ve / vey007.

[0136] In some embodiments, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% of the sequenced V3 variable regions of gp120 in an HIV population obtained from one or more biological samples of a single patient comprise an amino acid sequence comprising: a glycosylated asparagine (N332 glycan) at a position corresponding to amino acid residue position 332, an aspartic acid (D325) at a position corresponding to amino acid residue position 325, and one or more of a threonine (T63) at a position corresponding to amino acid residue position 63, a leucine (L179) at a position corresponding to amino acid residue position 179, a threonine (T320) at a position corresponding to amino acid residue position 320, and a histidine (H330) at a position corresponding to amino acid residue position 330, wherein the amino acid positions are referenced to SEQ ID NO: 4. As used herein, when the position of any given polymer component (e.g., amino acid, nucleotide, also collectively referred to as "residue") is designated by reference to the same or equivalent position in a selected amino acid or nucleic acid polymer (e.g., based on an optimal alignment or consensus sequence), rather than by the actual numerical position of the component in a given polymer, the numbering of a given amino acid polymer or nucleic acid polymer "corresponds to," "corresponds to," or "relative to" the numbering of a selected or reference amino acid polymer or nucleic acid polymer. In some embodiments, HIV gp120 variants are detected at a frequency level of about 1% of the viral population (e.g., 1% mutation or variation frequency). In some embodiments, HIV gp120 variants are detected at a frequency level of about 0.5% of the viral population. As a rule of thumb, reliable detection of variants at a 1% frequency would require an HIV RNA level of at least 1000 copies / mL. See, for example, Casadellà et al., Virus Research, Vol. 239, 2017, pp. 69-81; Noguera-Julian et al., J Infect Dis., 2017, Vol. 216 Suppl. 9, pp. S829-S833; and Lee et al., Sci Rep., 2020, Vol. 10, No. 1, p. 1634.

[0137] 3. Administration of anti-HIV gp120V3 glycan-directed antibodies or antigen-binding fragments thereof

[0138] In certain embodiments, these methods entail administering an anti-HIV antibody or antigen-binding fragment or antigen-binding molecule that targets the V3 glycan binding region of gp120.

[0139] HIV-1 is the main family of HIV and accounts for 95% of all infections worldwide. HIV-2 is mainly found in a few West African countries.

[0140] HIV viruses are divided into specific groups, M, N, O and P, with M being the "major" group and responsible for most HIV / AIDS worldwide. Based on their genetic sequences, the M group is further subdivided into subtypes (also called clades) with prevalence in different geographic locations.

[0141] M group "subtype" or "clade" is a subtype of HIV-1 group M defined by genetic sequence data. Examples of group M subtypes include subtype AK. Some subtypes in the known subtypes are more toxic or resistant to different drugs. There are also "circulating recombinant forms" or CRFs derived from recombination between viruses of different subtypes, each of which has a number. For example, CRF12_BF is a recombination between subtypes B and F. Subtype A is common in West Africa. Subtype B is the main form in Europe, America, Japan, Thailand and Australia. Subtype C is the main form in parts of South Africa, East Africa, India, Nepal and China. Subtype D is usually only found in East and Central Africa. Subtype E has never been identified as a non-recombinant, and only recombined with subtype A as CRF01_AE. Subtype F has existed in Central Africa, South America and Eastern Europe. Subtype G (as well as CRF02_AG) has been found in Africa and Central Europe. Subtype H is limited to Central Africa. Subtype I was originally used to describe the strain now considered to be CRF04_cpx, where cpx is used for a "complex" recombination of several subtypes. Subtype J is mainly found in North Africa, Central Africa and West Africa, while Caribbean subtype K is limited to the Democratic Republic of the Congo and Cameroon. These subtypes are sometimes further divided into sub-subtypes, such as A1 and A2 or F1 and F2. In 2015, strain CRF19 was found to be a recombinant of subtypes A, D and G, and its subtype D protease is closely related to the rapid development of AIDS in Cuba.

[0142] The present disclosure provides, inter alia, methods for administering human anti-HIV neutralizing antibodies (e.g., broadly neutralizing Abs) that target the V3 glycan region of the gp120 polypeptide on the surface of HIV-infected cells. Neutralizing antibodies directed against viral envelope proteins provide adaptive immune defense against HIV-1 exposure by blocking infection of susceptible cells. Broad neutralization indicates that the antibody can neutralize HIV-1 isolates from different clades. Thus, the anti-HIV gp120 V3 glycan-directed antibodies or antigen-binding fragments described herein have cross-clade binding activity.

[0143] Antibodies and antigen-binding fragments thereof directed to the V3 glycan region of HIV gp120

[0144] In certain embodiments of the methods described herein, an antibody or antigen binding fragment thereof, or an antigen binding molecule that binds to the HIV gp120 protein within the V3 glycan region, e.g., an epitope or region of the third variable loop (V3) of gp120 and / or the high mannose patch comprising the N332 oligomannose glycan, is administered to the subject. In certain embodiments, the administered antibody or antigen binding fragment thereof, or antigen binding molecule binds to HIV-1 antigens expressed on the surface of cells and eliminates or kills infected cells.

[0145] In certain embodiments, the antibody or antigen-binding fragment thereof or antigen-binding molecule administered is or is derived from a human neutralizing antibody (e.g., a monoclonal antibody) targeting HIV-1. "Neutralizing antibodies" are antibodies that can neutralize HIV's ability to cause and / or maintain infection in hosts and / or target cells in vitro. The present disclosure provides neutralizing monoclonal human antibodies, wherein the antibody recognizes antigens from HIV, such as gp120 polypeptides. In certain embodiments, "neutralizing antibodies" can inhibit the entry of HIV-1 viruses, such as SF162 and / or JR-CSF, wherein the neutralization index is >1.5 or >2.0 (Kostrikis LG et al., J. Virol., Vol. 70, No. 1, pp. 445-458, 1996).

[0146] In some embodiments, the antibody or its antigen-binding fragment or antigen-binding molecule administered is or is derived from a broadly neutralizing antibody (e.g., monoclonal antibody) targeting HIV-1. So-called "broadly neutralizing antibody" means an antibody that neutralizes more than one HIV-1 virus species (from different clades and different strains within clades) in a neutralization assay. Broadly neutralizing antibodies can neutralize at least 2, 3, 4, 5, 6, 7, 8, 9 or more different strains of HIV-1, which belong to the same or different clades. In a specific embodiment, broadly neutralizing antibodies can neutralize a variety of HIV-1 species belonging to at least 2, 3, 4, 5 or 6 different clades. In certain embodiments, the inhibitory concentration of the anti-HIV gp120 V3 glycan-directed antibody or antigen-binding fragment may be less than about 0.0001 μg / ml, less than about 0.001 μg / ml, less than about 0.01 μg / ml, less than about 0.1 μg / ml, less than about 0.5 μg / ml, less than about 1.0 μg / ml, less than about 5 μg / ml, less than about 10 μg / ml, less than about 25 μg / ml, less than about 50 μg / ml, or less than about 100 μg / ml to neutralize about 50% of the input virus in a neutralization assay.

[0147] Exemplary broadly neutralizing antibodies that bind to gp120 in the third variable loop (V3) and / or the high mannose patch comprising the N332 oligomannose glycan and that can be used in the methods described herein include, but are not limited to, GS-9722 (ipavirumab), GS-9721, PGT-121, PGT-121.66, PGT-121.414, PGT-122, PGT-123, PGT-124, PGT-125, PGT-126, PGT-128, PGT-130, PGT-133, PGT-134, PGT-135, PGT-136, PGT-137, PGT-138, PGT-139, PGT-240, PGT-241, PGT-242, PGT-243, PGT-244, PGT-245, PGT-246, PGT-247, PGT-248, PGT-249, PGT-250, PGT-251, PGT-252, PGT-253, PGT-254, PGT-255, PGT-256, PGT-257, PGT-258, PGT-259, PGT-261, PGT-262, PGT-263, PGT-264, PGT-265 35. PGT-136, PGT-137, PGT-138, PGT-139, 10-1074, 10-1074-J, VRC24, 2G12, BG18, 354BG8, 354BG18, 354BG42, 354BG33, 354BG129, 354BG188, 354BG411, 354BG426, DH270.1, DH270.6, PGDM12, VRC41.01, PGDM21, PCDN-33A, BF520.1 and VRC29.03. Additional broadly neutralizing antibodies that bind to gp120 in the third variable loop (V3) and / or the high mannose patch comprising the N332 oligomannose glycan and can be used in the methods described herein are described, for example, in WO 2012 / 030904; WO 2014 / 063059; WO 2016 / 149698; WO 2017 / 106346; WO 2018 / 075564, WO 2018 / 125813; WO 2018 / 237148, WO 2019 / 226829, WO 2020 / 023827, WO2020 / 056145, and Kerwin et al., J Pharm Sci, January 2020, Vol. 109, No. 1, pp. 233-246, all of which are hereby incorporated by reference in their entirety for all purposes.

[0148] Exemplary sequences of complementarity determining regions (CDRs) of antibodies or antigen-binding fragments targeting HIV gp120 V3 glycan region are provided in Tables A1 to A4. Exemplary sequences of VH and VL of antibodies or antigen-binding fragments targeting HIV gp120 V3 glycan region are provided in Table B.

[0149]

[0150]

[0151]

[0152]

[0153]

[0154]

[0155]

[0156]

[0157]

[0158]

[0159]

[0160]

[0161]

[0162]

[0163]

[0164]

[0165]

[0166]

[0167]

[0168]

[0169] In some embodiments, an anti-HIV gp120 V3 glycan-directed antibody or antigen-binding fragment thereof comprises a VH comprising a VH-CDR1, a VH-CDR2, and a VH-CDR3; and a VL comprising a VL-CDR1, a VL-CDR2, and a second VH-CDR3; wherein VH-CDR1, VH-CDR2, VH-CDR3, VL-CDR1, VL-CDR2, and VH-CDR3 comprise sequences as follows: SEQ ID NOs: 7, 8, 9, 10, 11, and 12; SEQ ID NOs: 7, 13, 9, 10, 11, and 12; SEQ ID NOs: 14, 15, 16, 17, 11, and 18; SEQ ID NOs: 14, 19, 20, 17, 11, and 18; SEQ ID NOs: 21, 22, 23, 24, 25, and 26; SEQ ID NOs: 2 SEQ ID NO:21, 22, 27, 24, 25 and 26; SEQ ID NO:28, 29, 30, 31, 32 and 33; SEQ ID NO:34, 35, 36, 37, 25 and 38; SEQ ID NO:39, 40, 41, 42, 43 and 44; SEQ ID NO:45, 46, 47, 48, 49 and 50; SEQ ID NO:45, 51, 52, 53, 49 and 54; SEQ ID NO:55, 56, 57, 58, 59 and 44; SEQ ID NO:61, 46, 63, 58, 49 and 44; SEQ ID NO:64, 65, 66, 67, 68 and 69; SEQ ID NO:70, 71, 72, 73, 74 and 75; SEQ ID NO:76, 77, 78, 79, 80 and 75; NO:81, 82, 83, 84, 85 and 75; SEQ ID NO:85, 86, 87, 88, 89 and 90; SEQ ID NO:85, 91, 92, 93, 94 and 90; SEQ ID NO:85, 96, 92, 93, 94 and 90; SEQ ID NO:85, 86, 87, 97, 98 and 90; SEQ ID NO:85, 99, 100, 101, 102 and 95; SEQ ID NO:85, 99, 100, 101, 102 and 103; SEQ ID NO:85, 99, 100, 104, 102 and 90; SEQ ID NO:85, 105, 92, 93, 94 and 90; SEQ ID NO:85, 99, 100, 101, 102 and 107; SEQ ID NO: 108, 109, 110, 111, 112 and 113; SEQ ID NO: 108, 114, 115, 111, 116 and 117;or SEQ ID NO: 108, 118, 119, 111, 120 and 121 (CDR according to Kabat). ;

[0170] In some embodiments, an anti-HIV gp120 V3 glycan-directed antibody or antigen-binding fragment thereof comprises a VH comprising a VH-CDR1, a VH-CDR2, and a VH-CDR3; and a VL comprising a VL-CDR1, a VL-CDR2, and a second VH-CDR3; wherein VH-CDR1, VH-CDR2, VH-CDR3, VL-CDR1, VL-CDR2, and VH-CDR3 comprise sequences as follows: SEQ ID NOs: 123, 124, 9, 10, 11, and 12; SEQ ID NOs: 125, 126, 16, 17, 11, and 18; SEQ ID NOs: 127, 128, 20, 17, 11, and 18; SEQ ID NOs: 129, 130, 23, 24, 25, and 26; SEQ ID NOs: 129, 130, 27, 24, 25, and 26; SEQ ID NO:131, 132, 30, 31, 32 and 33; SEQ ID NO:133, 134, 36, 37, 25 and 38; SEQ ID NO:135, 136, 41, 42, 43 and 44; SEQ ID NO:137, 138, 47, 48, 49 and 50; SEQ ID NO:137, 138, 52, 53, 49 and 54; SEQ ID NO:139, 56, 57, 58, 59 and 44; SEQ ID NO:141, 138, 63, 58, 49 and 44; SEQ ID NO:142, 143, 66, 67, 68 and 69; SEQ ID NO:144, 145, 72, 73, 74 and 75; SEQ ID NO:146, 147, 78, 79, 80 and 75; SEQ ID NO:146, 147, 83, 84, 85 and 75; SEQ ID NO:149, 150, 87, 88, 89 and 90; SEQ ID NO:151, 150, 87, 88, 89 and 90; SEQ ID NO:152, 153, 92, 93, 94 and 90; SEQ ID NO:151, 150, 87, 97, 98 and 90; SEQ ID NO:152, 153, 100, 101, 102 and 95; SEQ ID NO:152, 153, 100, 101, 102 and 103; SEQ ID NO:152, 153, 100, 104, 102 and 90; SEQ ID NO:152, 153, 100, 101, 102 and 107; NO: 154, 155, 110, 111, 116 and 117; SEQ ID NO: 156, 157, 115, 111, 116 and 117;or SEQ ID NO: 158, 159, 119, 111, 120 and 121 (CDR according to Chothia). ;

[0171] In some embodiments, an anti-HIV gp120 V3 glycan-directed antibody or antigen-binding fragment thereof comprises a VH comprising a VH-CDR1, a VH-CDR2, and a VH-CDR3; and a VL comprising a VL-CDR1, a VL-CDR2, and a second VH-CDR3; wherein VH-CDR1, VH-CDR2, VH-CDR3, VL-CDR1, VL-CDR2, and VH-CDR3 comprise sequences as follows: SEQ ID NOs: 160, 161, 162, 163, 164, and 12; SEQ ID NOs: 165, 166, 167, 168, 164, and 18; SEQ ID NOs: 169, 170, 171, 172, 173, 174, and 186; SEQ ID NO:172, 173, 174, 175, 164 and 26; SEQ ID NO:172, 173, 176, 175, 164 and 26; SEQ ID NO:177, 178, 179, 180, 164 and 38; SEQ ID NO:181, 182, 183, 184, 185 and 33; SEQ ID NO:186, 187, 188, 189, 190 and 44; SEQ ID NO:191, 192, 193, 194, 195 and 50; SEQ ID NO:191, 196, 197, 198, 195 and 54; SEQ ID NO:199, 200, 201, 202, 399 and 44; SEQ ID NO:203, 204, 205, 202, 195 and 44; SEQ ID NO:206, 207, 208, 209, 210 and 69; 211, 212, 213, 214, 215 and 75; SEQ ID NO:216, 217, 218, 219, 220 and 75; SEQ ID NO:221, 217, 83, 223, 224 and 75; SEQ ID NO:225, 226, 87, 227, 228 and 90; SEQ ID NO:229, 226, 87, 227, 228 and 90; SEQ ID NO:230, 231, 92, 232, 233 and 90; SEQ ID NO:230, 234, 92, 232, 233 and 90; SEQ ID NO:229, 226, 87, 235, 398 and 90; NO: 230, 236, 100, 232, 233 and 95; SEQ ID NO: 230, 236, 100, 232, 233 and 103; SEQ ID NO: 230, 236, 100, 237, 233 and 90; SEQ ID NO: 230, 238, 92, 232, 233 and 107;SEQ ID NO: 239, 240, 110, 241, 242 and 113; SEQ ID NO: 243, 244, 115, 241, 245 and 117; or SEQ ID NO: 243, 246, 119, 241, 247 and 121 (according to CDRs of IMGT). ;

[0172] In some embodiments, an anti-HIV gp120 V3 glycan-directed antibody or antigen-binding fragment thereof comprises a VH comprising a VH-CDR1, a VH-CDR2, and a VH-CDR3; and a VL comprising a VL-CDR1, a VL-CDR2, and a second VH-CDR3; wherein VH-CDR1, VH-CDR2, VH-CDR3, VL-CDR1, VL-CDR2, and VH-CDR3 comprise sequences as follows: SEQ ID NOs: 248, 249, 250, 251, 252, and 253; SEQ ID NOs: 248, 254, 250, 251, 252, and 253; SEQ ID NOs: 255, 256, 257, 258, 252, and 259; SEQ ID NOs: 260, 261, 262, 258, 252, and 259; SEQ ID NOs: 261, 262, 258, 252, and 259; SEQ ID NO:263, 264, 265, 266, 267 and 268; SEQ ID NO:263, 264, 397, 266, 267 and 268; SEQ ID NO:269, 270, 271, 272, 273 and 274; SEQ ID NO:275, 276, 277, 278, 279 and 280; SEQ ID NO:281, 282, 283, 284, 285 and 286; SEQ ID NO:287, 288, 289, 290, 291 and 286; SEQ ID NO:287, 292, 293, 294, 295 and 296; SEQ ID NO:297, 298, 299, 300, 301 and 286; SEQ ID NO: SEQ ID NO:302, 288, 303, 300, 295 and 286; SEQ ID NO:304, 305, 306, 307, 308 and 309; SEQ ID NO:310, 311, 312, 313, 314 and 315; SEQ ID NO:316, 316, 318, 319, 320 and 315; SEQ ID NO:321, 322, 323, 324, 325 and 315; 326, 327, 328, 329, 330 and 331; SEQ ID NO:332, 327, 328, 329, 330 and 331; SEQ ID NO:333, 334, 335, 336, 337 and 338; SEQ ID NOs: 333, 339, 335, 336, 337 and 338; SEQ ID NOs: 332, 327, 328, 340, 341 and 338; SEQ ID NOs: 342, 343, 344, 336, 345 and 346; SEQ ID NOs: 342, 343, 344, 336, 347 and 348; SEQ ID NOs: 342, 343, 344, 349, 350 and 338;SEQ ID NO:333, 351, 335, 336, 337 and 338; SEQ ID NO:342, 343, 344, 336, 347 and 352; SEQ ID NO:353, 354, 355, 356, 357 and 358; SEQ ID NO:359, 360, 361, 356, 362 and 363; or SEQ ID NO:359, 364, 365, 356, 366 and 358. (CDRs according to Honegger). ;

[0173] Exemplary embodiments of CDR sequences of anti-HIV gpl20 V3 glycan-directed antibodies or antigen-binding fragments thereof that can be used in the methods described herein are provided in Tables A1 to A4.

[0174] In some embodiments, the anti-HIV gp120 V3 glycan-directed antibody or antigen-binding fragment thereof comprises a VH and a VL comprising an amino acid sequence that is at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to an amino acid sequence selected from the group consisting of SEQ ID NOs: 400 and 401; SEQ ID NOs: 402 and 403; SEQ ID NOs: 402 and 404; SEQ ID NOs: 462 and 463; SEQ ID NOs: 464 and 465; SEQ ID NOs: 405 and 406; SEQ ID NOs: 466 and 467; SEQ ID NOs: 407 and 408; SEQ ID NOs: 409 and 410; SEQ ID NOs: 411 and 412; SEQ ID NOs: 413 and 414; SEQ ID NOs: 415 and 416; SEQ ID NOs: 417 and 418; SEQ ID NOs: 419 and 420; SEQ ID NOs: 421 and 422; SEQ ID NOs: 423 and 424; SEQ ID NOs: 425 and 426 SEQ ID NOs:413 and 414; SEQ ID NOs:415 and 416; SEQ ID NOs:417 and 418; SEQ ID NOs:419 and 420; SEQ ID NOs:421 and 422; SEQ ID NOs:423 and 424; SEQ ID NOs:425 and 426; SEQ ID NOs:427 and 428; SEQ ID NOs:429 and 430; SEQ ID NOs:431 and 432; SEQ ID NOs:433 and 434; SEQ ID NOs:435 and 436; SEQ ID NOs:437 and 438; SEQ ID NOs:439 and 440; SEQ ID NOs:441 and 442; SEQ ID NOs:443 and 444; SEQ ID NOs:445 and 446; SEQ ID NOs:447 and 448; SEQ ID NOs:449 and 450; SEQ ID NOs:451 and 452; SEQ ID NOs:453 and 454; SEQ ID NOs:455 and 456; SEQ ID NOs:457 and 458; SEQ ID NOs:459 and 460; SEQ ID NOs:461 and 462; SEQ ID NOs:463 and 464; SEQ ID NOs:465 and 466; SEQ ID NOs:467 and 468; SEQ ID NOs:469 and 470; SEQ ID NOs:471 and 472; SEQ ID NOs:473 and 474; SEQ ID NOs:475 and 476 SEQ ID NOs: 453 and 454; SEQ ID NOs: 455 and 456; SEQ ID NOs: 457 and 458; or SEQ ID NOs: 459 and 460. Exemplary embodiments of variable domain VH and VL sequences of anti-HIV gpl20 V3 glycan-directed antibodies or antigen-binding fragments thereof that can be used in the methods described herein are provided in Table B.

[0175] In some embodiments, the anti-HIV gp120 V3 glycan-directed antibodies comprise VH and VL amino acid sequences that are at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequences shown below, respectively, and comprise one or more of the following amino acids at the indicated positions (according to Kabat position numbering): SEQ ID NO: NO:400 or 402, which comprises one or more of the following amino acids: Ser-Ser-Val (SSV) or Thr-Gly-Val (TGV) at positions 82a-82c, Gln (Q) at position 39, Asn (N) at position 60, His (H) at position 68, Lys (K), any of His (H) or Thr (T) at position 105, Leu (L) at position 2, Ala (A) at position 32 and / or Ala (A) at position 95; and SEQ ID NO:401, 403 or 404, which comprises one or more of the following amino acids: Gly (G) at position 67, Tyr (Y), Phe (F) or Thr (T) at position 67a, Arg (R) at position 67b, Pro (P) at position 67c and / or Lys (K) at position 103. In some embodiments, the anti-HIV gp120 V3 glycan-directed antibody comprises VH and VL amino acid sequences that are at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence shown below, respectively, and comprise one or more of the following amino acids at the indicated positions (according to Kabat position numbering): SEQ ID NO: 400 or 402, which comprises any one of Thr-Gly-Val (TGV) at positions 82a-82c, Asn (N) at position 60, His (H) at position 68, Lys (K), His (H) and / or Thr (T) at position 105; and SEQ ID NO: 401 or 402, which comprises any one of Thr-Gly-Val (TGV) at positions 82a-82c, Asn (N) at position 60, His (H) at position 68, Lys (K), His (H) and / or Thr (T) at position 105; and NO:401, 403 or 404, which comprises one or more of the following amino acids: Gly (G) at position 67, Tyr (Y), Phe (F) or Thr (T) at position 67a, Arg (R) at position 67b, Pro (P) at position 67c.

[0176] Fc mutations that increase serum half-life

[0177] In some embodiments, the Fc region or Fc domain of the anti-HIV gp120 V3 glycan-directed antibody comprises an amino acid modification that promotes an increase in the serum half-life of the anti-binding molecule. Mutations that increase the half-life of antibodies have been described. In one embodiment, the Fc region or Fc domain of one or both of the CD3-targeted heavy chain and the HIV antigen-targeted heavy chain comprises a substitution of methionine to tyrosine at position 252 (EU numbering), a substitution of serine to threonine at position 254 (EU numbering), and a substitution of threonine to glutamate at position 256 (EU numbering). See, e.g., U.S. Patent 7,658,921. This type of mutant, named "YTE mutant", exhibits a four-fold increase in half-life relative to the wild-type version of the same antibody (Dall'Acqua et al., J Biol Chem, Vol. 281, pp. 23514-23524, 2006; Robbie et al., Antimicrob Agents Chemotherap., Vol. 57, No. 12, pp. 6147-6153, 2013. In certain embodiments, the Fc region or Fc domain of one or both of the CD3 targeting heavy chain and the HIV antigen targeting heavy chain comprises an IgG constant domain comprising one, two, three or more amino acid substitutions of amino acid residues at positions 251-257, 285-290, 308-314, 385-389 and 428-436 (EU numbering). Alternatively, M428L and N434S ("LS") substitutions can increase the multispecific antigen Pharmacokinetic half-life of binding molecules. In other embodiments, the Fc region or Fc domain of one or both of CD3 targeting heavy chain and HIV antigen targeting heavy chain comprises M428L and N434S substitutions (EU numbering). In other embodiments, the Fc region or Fc domain of one or both of CD3 targeting heavy chain and HIV antigen targeting heavy chain comprises T250Q and M428L (EU numbering) mutants. In other embodiments, the Fc region or Fc domain of one or both of CD3 targeting heavy chain and HIV antigen targeting heavy chain comprises H433K and N434F (EU numbering) mutants.

[0178] Fc mutants with enhanced effector activity

[0179] In some embodiments, the Fc region or Fc domain of the anti-HIV gp120 V3 glycan-directed antibody comprises post-translational and / or amino acid modifications that increase effector activity, such as improved FcγIIIa binding and increased antibody-dependent cellular cytotoxicity (ADCC). In some embodiments, the Fc region or Fc domain of the anti-HIV gp120 V3 glycan-directed antibody comprises a DE modification in the Fc region (i.e., S239D and I332E of EU numbering). In some embodiments, the Fc region or Fc domain of the anti-HIV gp120 V3 glycan-directed antibody comprises a DEL modification in the Fc region (i.e., S239D, I332E, and A330L of EU numbering). In some embodiments, the Fc region or Fc domain of the anti-HIV gp120 V3 glycan-directed antibody comprises a DEA modification in the Fc region (i.e., S239D, I332E, and G236A of EU numbering). In some embodiments, the Fc region or Fc domain of an anti-HIV gpl20 V3 glycan-directed antibody comprises a DEAL modification (i.e., S239D, I332E, G236A, and A330L by EU numbering) in the Fc region. See, e.g., U.S. Patents 7,317,091; 7,662,925; 8,039,592; 8,093,357; 8,093,359; 8,383,109; 8,388,955; 8,735,545; 8,858,937; 8,937,158; 9,040,041; 9,353,187; 10,184,000; and 10,584,176. Additional amino acid modifications that increase effector activity, such as improved FcγIIIa binding and increased antibody-dependent cellular cytotoxicity (ADCC), include but are not limited to (EU numbering) F243L / R292P / Y300L / V305I / P396L; S298A / E333A / K334A; or L234Y / L235Q / G236W / S239M / H268D / D270E / S298A on the first Fc domain and D270E / K326D / A330M / K334E on the second Fc domain. Amino acid mutations that increase C1q binding and complement-dependent cytotoxicity (CDC) include but are not limited to (EU numbering) S267E / H268F / S324T or K326W / E333S. Fc region mutations that enhance effector activity are reviewed in, for example, Wang et al., Protein Cell, 2018, Vol. 9, No. 1, pp. 63–73; and Saunders, Front Immunol., 2019, Vol. 10, p. 1296.

[0180] In other embodiments, the anti-HIV gp120 V3 glycan-directed antibody or antigen-binding fragment thereof has a modified glycosylation, which can be introduced, for example, post-translationally or by genetic engineering. In some embodiments, the anti-HIV gp120 V3 glycan-directed antibody or antigen-binding fragment thereof is non-fucosylated, for example, at a glycosylation site present in the antibody or antigen-binding fragment thereof. Most approved monoclonal antibodies have an IgG1 isotype, in which two N-linked bibranched complex-type oligosaccharides are bound to the Fc region. The Fc region exerts the effector function of ADCC through its interaction with leukocyte receptors of the FcγR family. Non-fucosylated monoclonal antibodies are monoclonal antibodies that are engineered so that the oligosaccharides in the Fc region of the antibody do not have any fucose units.

[0181] In some embodiments, the Fc region or Fc domain of the anti-HIV gp120 V3 glycan-directed antibody may comprise post-translational and / or amino acid modifications, as appropriate, for increasing serum half-life and enhancing effector activity.

[0182] 4. Combination therapy with two or more anti-HIV antibodies

[0183] In certain embodiments, the present disclosure provides a method for treating or preventing HIV infection in a human subject with HIV infection or at risk of HIV infection. The method includes administering to a human subject a therapeutically effective amount of an anti-HIV 120V3 polysaccharide-directed antibody or antigen-binding fragment as disclosed herein, or a pharmaceutical composition thereof, and a therapeutically effective amount of one or more (e.g., one, two, three, one or two, or one to three) additional therapeutic agents. In one embodiment, a method for treating HIV infection in a human subject with infection or at risk of infection is provided, the method comprising administering to a human subject a therapeutically effective amount of one or more antibodies disclosed herein, or a pharmaceutically acceptable salt thereof, and a therapeutically effective amount of one or more (e.g., one, two, three, one or two, or one to three) additional therapeutic agents.

[0184] Antibody combination therapy

[0185] In some embodiments, the anti-V3 glycan antibody or antigen binding fragment thereof is co-administered with a second anti-HIV antibody. In some embodiments, the anti-V3 glycan antibody or antigen binding fragment thereof is co-administered with a second anti-HIV antibody that binds to an epitope or region of gp120 selected from: (i) the second variable loop (V2) and / or the Env trimer apex; (ii) the CD4 binding site (CD4bs); (iii) the gp120 / gp41 interface; or (v) the gp120 silent face. The aforementioned epitopes or regions of gp120 bound by broadly neutralizing antibodies are described in, for example, McCoy, Retrovirology, 2018, Vol. 15, p. 70; Sok and Burton, Nat Immunol., 2018, Vol. 19, No. 11, pp. 1179-1188; Possas et al., Expert Opin Ther Pat., July 2018, Vol. 28, No. 7, pp. 551-560; and Stephenson and Barouch, Curr HIV / AIDS Rep, 2016, Vol. 13, pp. 31-37, all of which are hereby incorporated by reference in their entirety for all purposes.

[0186] In some embodiments, the combination therapy requires co-administration of an anti-V3 glycan antibody or antigen-binding fragment thereof and another anti-HIV broadly neutralizing antibody or bNAb (i.e., a neutralizing antibody that neutralizes multiple HIV-1 viral strains). Various bNAbs are known in the art and can be used as combination therapeutic agents. Additional exemplary bNAbs used include antibodies comprising VH and VL that bind to or compete with an epitope or region of gp120 selected from: (i) the second variable loop (V2) and / or the Env trimer vertex; (ii) the CD4 binding site (CD4bs); (iii) the gp120 / gp41 interface; or (v) the gp120 silent face. Exemplary bNAbs for anti-HIV antibody combination therapy include antibodies comprising VH and VL that bind to or compete with: 2F5, 4E10, M66.6, CAP206-CH12, 10E8, 10E8v4, 10E8-5R-100cF, DH511.11P, 7b2, and LN01 (all of which bind to the MPER of gp41); PG9, PG16, CH01-04 (all of which bind to V1V2-glycans), 2G12 (which binds to the outer domain glycans); b12, F105, VRC01, VRC07, VRC07-523, VRC03, VRC06, VRC06b01 VRC08, VRC0801, NIH45-46, GS-9723, GS-5423, 3BNC117, 3BNC60, VRC-PG04, PGV04; CH103, 44-VRC13.01, 1NC9, 12A12, N6, N6LS (VRC-HIVMAB091-00-AB), N49-P7, NC-Cow1, IOMA, CH235 and CH235.12, N49P6, N49P7, N49P11, N49P9 and N60P25 (all of which bind to the CD4 binding site).

[0187] In some embodiments, the combination therapy includes an antibody that binds to an epitope or region of gp120 in the second variable loop (V2) and / or the apex of the Env trimer and competes with or comprises the CDRs and / or VH and VL regions from an antibody selected from the group consisting of PG9, PG16, PGC14, PGG14, PGT-142, PGT-143, PGT-144, PGT-145, CHO1, CH59, PGDM1400, CAP256, CAP256-VRC26.08, CAP256-VRC26.09, CAP256-VRC26.25, PCT64-24E, and VRC38.01.

[0188] In some embodiments, the combination therapy includes an antibody that binds to a gp120 epitope or region in the CD4 binding site (CD4bs) and competes with or comprises CDRs and / or VH and VL regions from an antibody selected from the group consisting of b12, F105, VRC01, VRC07, VRC07-523, VRC03, VRC06, VRC06b01, VRC08, VRC0801, NIH45-46 , GS-9723, GS-5423, 3BNC117, 3BNC60, VRC-PG04, PGV04; CH103, 44-VRC13.01, 1NC9, 12A12, N6, N6LS (VRC-HIVMAB091-00-AB), N49-P7, NC-Cow1, IOMA, CH235 and CH235.12, N49P6, N49P7, N49P11, N49P9 and N60P25.

[0189] In some embodiments, the combination therapy includes an antibody that binds to an epitope or region of gp120 in the gp120 / gp41 interface and competes with or comprises CDRs and / or VH and VL regions from an antibody selected from the group consisting of PGT-151, CAP248-2B, 35022, 8ANC195, ACS202, VRC34, and VRC34.01.

[0190] In some embodiments, the combination therapy includes an antibody that binds to an epitope or region of the silent face of gp120 and competes with or comprises the second VH and VL regions from antibody VRC-PG05.

[0191] In some embodiments, the combination therapy includes an antibody that binds to an epitope or region of gp41 in the membrane proximal region (MPER) and competes with or includes a second VH region and a VL region from an antibody selected from the group consisting of 10E8, 10E8v4, 10E8-5R-100cF, 4E10, DH511.11P, 2F5, 7b2, and LN01. In some embodiments, the combination therapy includes an antibody that binds to an epitope or region of KLIC ("KLIC" is disclosed as SEQ ID NO: 468) (an invariable site of the transmembrane protein gp41) and competes with or includes a second VH region and a VL region from a clone 3 human monoclonal antibody (Cl3hmAb) (Protheragen). See, e.g., Vanini et al., AIDS, 1993, Vol. 7, No. 2, pp. 167-174.

[0192] In some embodiments, the combination therapy includes an antibody that binds to an epitope or region of the gp41 fusion peptide and competes with or comprises a second VH and VL region from an antibody selected from VRC34 and ACS202.

[0193] In some embodiments, the combination therapy includes a multispecific antibody, such as a bispecific or trispecific antibody that binds to an HIV antigen. Examples of HIV bispecific and trispecific antibodies include: MGD014, B12BiTe, BiIA-SG, TMB-bispecific, SAR-441236, VRC-01 / PGDM-1400 / 10E8v4, 10E8.4 / iMab, and 10E8v4 / PGT121-VRC01.

[0194] Prior to administration, bNAbs can be improved to have enhanced drug-like properties, reduced immunogenicity, enhanced ADCC, and suitable pharmacokinetic properties. Such antibodies are shown to bind to HIV envelope glycoproteins expressed on the surface of virions or infected cells, and mediate direct neutralization of the virus and potent NK, monocyte, and PBMC killing of these cells. This property allows antibodies to treat HIV infection by neutralizing the virus, and also kill and eliminate latent HIV-infected cells in infected individuals, potentially leading to sterilization therapy of HIV.

[0195] In various embodiments, all antibodies administered in a combination anti-HIV antibody therapy may have Fc and / or post-translational modifications that increase serum half-life and / or enhance effector activity, as described above.

[0196] In various embodiments, anti-HIV gp120 V3 glycan-directed antibodies or antigen-binding fragments, and optionally combined bNAbs, can be delivered in vivo, for example, expressed in vivo by administered mRNA or engineered B cells. Examples of bNAbs delivered in vivo include AAV8-VRC07; mRNA encoding anti-HIV antibody VRC01; and engineered B cells encoding 3BNC117 (Hartweger et al., J. Exp. Med., 2019, p. 1301).

[0197] 5. Combination therapy with other anti-HIV therapeutic agents

[0198] In certain embodiments, methods for treating or preventing HIV infection in a person who has an infection or is at risk of infection are provided, the method comprising administering to the person a therapeutically effective amount of an anti-HIV 120V3 glycan-directed antibody or antigen-binding fragment, as disclosed herein, and a therapeutically effective amount of one or more (e.g., one, two, three, one or two, or one to three) additional therapeutic agents. In some embodiments, methods for treating HIV infection in a person who has an infection or is at risk of infection are provided, the method comprising administering to the person a therapeutically effective amount of an anti-HIV 120V3 glycan-directed antibody or antigen-binding fragment, as disclosed herein, and a therapeutically effective amount of one or more (e.g., one, two, three, one or two, or one to three) additional therapeutic agents.

[0199] In one embodiment, pharmaceutical compositions are provided that comprise anti-HIV 120V3 glycan-directed antibodies or antigen-binding fragments, as disclosed herein, and one or more (e.g., one, two, three, one or two, or one to three) additional therapeutic agents, and a pharmaceutically acceptable carrier, diluent, or excipient.

[0200] In certain embodiments, methods for treating HIV infection are provided, comprising administering to a patient in need thereof a therapeutically effective amount of an anti-HIV gp120 V3 glycan-directed antibody or antigen-binding fragment thereof, as described herein, and a therapeutically effective amount of one or more additional therapeutic agents suitable for treating HIV infection.

[0201] In certain embodiments, an anti-HIV gp120 V3 glycan-directed antibody or antigen-binding fragment thereof is combined with one, two, three, four or more additional therapeutic agents. In certain embodiments, an anti-HIV gp120 V3 glycan-directed antibody or antigen-binding fragment thereof is combined with two additional therapeutic agents. In other embodiments, an anti-HIV gp120 V3 glycan-directed antibody or antigen-binding fragment thereof is combined with three additional therapeutic agents. In other embodiments, an anti-HIV gp120 V3 glycan-directed antibody or antigen-binding fragment thereof is combined with four additional therapeutic agents. The one, two, three, four or more additional therapeutic agents may be different therapeutic agents selected from the same class of therapeutic agents (e.g., one or more anti-HIV broadly neutralizing antibodies), and / or they may be selected from different classes of therapeutic agents.

[0202] Administration of HIV combination therapy

[0203] In certain embodiments, as described herein, an anti-HIV gp120 V3 glycan-directed antibody or antigen-binding fragment thereof is co-administered with one or more additional therapeutic agents. Co-administration of an anti-HIV gp120 V3 glycan-directed antibody or antigen-binding fragment disclosed herein with one or more additional therapeutic agents generally refers to the simultaneous or sequential administration of an anti-HIV gp120 V3 glycan-directed antibody or antigen-binding fragment disclosed herein with one or more additional therapeutic agents, such that a therapeutically effective amount of an anti-HIV gp120 V3 glycan-directed antibody or antigen-binding fragment disclosed herein and the one or more additional therapeutic agents are both present in the patient's body. When administered sequentially, the combination can be administered in two or more administrations.

[0204] Co-administration includes simultaneous administration as well as administration of a unit dose of an anti-HIV gp120 V3 glycan-directed antibody or antigen-binding fragment thereof, as described herein, before or after administration of a unit dose of one or more additional therapeutic agents. For example, as described herein, an anti-HIV 120 V3 glycan-directed antibody or antigen-binding fragment thereof can be administered within seconds, minutes, hours, or days of administration of the one or more additional therapeutic agents. In some embodiments, a unit dose of an anti-HIV gp120 V3 glycan-directed antibody or antigen-binding fragment disclosed herein is administered first, followed by administration of a unit dose of one or more additional therapeutic agents within seconds, minutes, hours, or days. Alternatively, a unit dose of one or more additional therapeutic agents is administered first, followed by administration of a unit dose of an anti-HIV gp120 V3 glycan-directed antibody or antigen-binding fragment disclosed herein within seconds, minutes, hours, or days. In other embodiments, a unit dose of an anti-HIV gp120 V3 glycan-directed antibody or antigen-binding fragment disclosed herein is administered first, and then a unit dose of one or more additional therapeutic agents is administered after a period of hours (e.g., 1 hour-12 hours, 1 hour-24 hours, 1 hour-36 hours, 1 hour-48 hours, 1 hour-60 hours, 1 hour-72 hours). In other embodiments, a unit dose of one or more additional therapeutic agents is administered first, and then a unit dose of an anti-HIV gp120 V3 glycan-directed antibody or antigen-binding fragment disclosed herein is administered after a period of hours (e.g., 1 hour-12 hours, 1 hour-24 hours, 1 hour-36 hours, 1 hour-48 hours, 1 hour-60 hours, 1 hour-72 hours).

[0205] In certain embodiments, the anti-HIV 120V3 glycan-directed antibodies or antigen-binding fragments disclosed herein are combined with one or more additional therapeutic agents in a single dosage form for simultaneous administration to a patient, e.g., as a solid, liquid or suspension dosage form for oral, intravenous, intramuscular or subcutaneous administration.

[0206] In certain embodiments, the anti-HIV gp120 V3 glycan-directed antibody or antigen-binding fragment is formulated as a liquid solution or suspension, which may optionally contain one or more other compounds useful for treating HIV. In certain embodiments, the liquid solution or suspension may contain another active ingredient for treating HIV, such as HIV protease inhibitors, non-nucleoside or non-nucleotide inhibitors of HIV reverse transcriptase, nucleoside or nucleotide inhibitors of HIV reverse transcriptase, HIV integrase inhibitors, HIV non-catalytic site (or allosteric) integrase inhibitors, pharmacokinetic enhancers, and combinations thereof.

[0207] In certain embodiments, such liquid solutions or suspensions are suitable for dosing or dosing intervals once daily, once weekly (i.e., QW), once every two weeks (i.e., once every other week, or once every two weeks or Q2W), once monthly (i.e., QM), or once every two months (i.e., once every other month, or once every two months or Q2M). In some embodiments, the anti-HIVgp120 V3 glycan-directed antibody or antigen-binding fragment is administered once daily, once weekly (i.e., QW), once every two weeks (i.e., once every other week, or once every two weeks or Q2W), once monthly (i.e., QM), once every two months (i.e., once every other month, or once every two months or Q2M), once every three months (i.e., Q3M), once every four months (i.e., Q4M).

[0208] HIV combination therapy

[0209] In the above embodiments, the additional therapeutic agent can be an anti-HIV agent. HIV protease inhibitors, non-nucleoside or non-nucleotide inhibitors of HIV reverse transcriptase, nucleoside or nucleotide inhibitors of HIV reverse transcriptase, HIV integrase inhibitors, HIV non-catalytic site (or allosteric) integrase inhibitors, HIV entry inhibitors, HIV maturation inhibitors, HIV capsid inhibitors, HIV Tat or Rev inhibitors, immunomodulators (e.g., immunostimulators), immunotherapeutics, immunomodulators, immunotherapeutics, antibody-drug conjugates, gene regulators, gene editors (such as CRISPR / Cas9, zinc finger nucleases, homing nucleases, synthetic nucleases, TALEN), cell therapies (such as chimeric antigen receptor T-cells, CAR-T and engineered T-cell receptors, TCR-T, autologous T-cell therapy), latency reversal agents, compounds targeting HIV capsid, immune-based therapies, phosphatidylinositol 3-kinase (PI3K) inhibitors, HIV antibodies, bispecific antibodies and "antibody-like" therapeutic proteins, HIV p17 matrix protein inhibitors, IL-13 antagonists, peptidyl prolyl cis-trans isomerase A modulators, protein disulfide isomerase inhibitors, complement C5a receptor antagonists, DNA methyltransferase inhibitors, fatty acid synthase inhibitors, HIV vif gene modulators, Vif dimerization antagonists, HIV-1 viral infection factor inhibitors, TAT protein inhibitors, HIV-1 Nef modulators (e.g., Nef inhibitors), Hck tyrosine kinase modulators, mixed lineage kinase-3 (MLK-3) inhibitors, HIV-1 splicing inhibitors, Rev protein inhibitors, integrin antagonists, nuclear protein inhibitors, splicing factor modulators, COMM domain-containing protein 1 modulators, HIV ribonuclease H inhibitors, defensin modulators, CDK-4 inhibitors, CDK-6 inhibitors, CDK-9 inhibitors, dendritic ICAM-3 grabbing non-integrin 1 inhibitors, HIV GAG protein inhibitors, HIV POL protein inhibitors, complementary factor H regulators, ubiquitin ligase inhibitors, deoxycytidine kinase inhibitors, cyclin-dependent kinase inhibitors, proprotein convertase PC9 stimulators, ATP-dependent RNA helicase DDX3X inhibitors, reverse transcriptase initiation complex inhibitors, G6PD and NADH oxidase inhibitors, mTOR complex 1 inhibitors, mTOR complex 2 inhibitors, P-glycoprotein regulators, TAT protein inhibitors, prolyl endopeptidase inhibitors, phospholipase A2 inhibitors, pharmacokinetic enhancers, HIV gene therapy, TNFα ligand inhibitors, IFN antagonists, HIV vaccines and combinations thereof.

[0210] In some embodiments, the additional therapeutic agent is selected from a combination drug for HIV, other drugs for treating HIV, HIV protease inhibitors, HIV reverse transcriptase inhibitors, HIV integrase inhibitors, HIV non-catalytic site (or allosteric) integrase inhibitors, HIV entry (fusion) inhibitors, HIV maturation inhibitors, latency reversal agents, HIV capsid inhibitors, HIV Tat or Rev inhibitors, immunomodulators (e.g., immunostimulators), immunotherapeutics, immune-based therapies, PI3K inhibitors, HIV antibodies and bispecific antibodies and "antibody-like" therapeutic proteins, and combinations thereof.

[0211] HIV combination drugs

[0212] In certain embodiments, the anti-HIV gp120 V3 glycan-directed antibodies or antigen-binding fragments described herein are combined with one, two, three, four or more additional anti-HIV therapeutic agents. Exemplary anti-HIV therapeutic agents that can be combined include, but are not limited to (efavirenz, tenofovir disoproxil fumarate, and emtricitabine); ( rilpivirine, tenofovir disoproxil fumarate, and emtricitabine); (elvitegravir, cobicistat, tenofovir disoproxil fumarate, and emtricitabine); (tenofovir disoproxil fumarate and emtricitabine; TDF+FTC); (tenofovir alafenamide and emtricitabine); (tenofovir alafenamide, emtricitabine, and rilpivirine); (tenofovir alafenamide, emtricitabine, cobicistat and elvitegravir); BIKTARVY (bicagvir + emtricitabine + tenofovir alafenamide), adefovir; adefovir dipivoxil; cobicistat; emtricitabine; tenofovir; tenofovir alafenamide and elvitegravir; tenofovir disoproxil; tenofovir disoproxil fumarate; tenofovir alafenamide; tenofovir alafenamide hemifumarate; (dolutegravir, abacavir, and lamivudine); dolutegravir, abacavir sulfate, and lamivudine; raltegravir; pegylated raltegravir; raltegravir and lamivudine; maraviroc; tenofovir + emtricitabine + maraviroc, enfuvirtide; ( lopinavir and ritonavir); (zidovudine and lamivudine; AZT+3TC); ( abacavir sulfate and lamivudine; ABC+3TC); (abacavir sulfate, zidovudine and lamivudine; ABC+AZT+3TC); atazanavir and cobicistat; atazanavir sulfate and cobicistat; atazanavir sulfate and ritonavir; darunavir; darunavir and cobicistat; dolutegravir and rilpivirine; dolutegravir and rilpivirine hydrochloride; dolutegravir, abacavir sulfate and lamivudine; lamivudine, nevirapine and zidovudine; raltegravir and lamivudine; doravirine, lamivudine and tenofovir disoproxil fumarate; doravirine, lamivudine and tenofovir disoproxil fumarate ; Dolutegravir + Lamivudine, Lamivudine + Abacavir + Zidovudine, Lamivudine + Abacavir, Lamivudine + Tenofovir Disoproxil Fumarate, Lamivudine + Zidovudine + Nevirapine, Lopinavir + Ritonavir, Lopinavir + Ritonavir + Abacavir + Lamivudine, Lopinavir + Ritonavir + Zidovudine + Lamivudine, Tenofovir + Lamivudine and Tenofovir Disoproxil Fumarate + Emtricitabine + Rilpivirine Hydrochloride, Lopinavir, Ritonavir, Zidovudine and Lamivudine; Cabotegravir + Rilpivirine; Erbi elsulfavirine; VM-1500; VM-1500A; rilpivirine; rilpivirine hydrochloride; atazanavir sulfate and cobicistat; atazanavir and cobicistat; darunavir and cobicistat; atazanavir; atazanavir sulfate; dolutegravir; elvitegravir; ritonavir; atazanavir sulfate and ritonavir; darunavir; lamivudine; α1-proteinase inhibitor; fosamprenavir; fosamprenavir calcium efavirenz; efavirenz, lamivudine, and emtricitabine; etravirine; nelfinavir; nelfinavir mesylate ; interferon; didanosine; stavudine; indinavir; indinavir sulfate; tenofovir and lamivudine; zidovudine; nevirapine; saquinavir; saquinavir mesylate; aldesleukin; zalcitabine; tipranavir; amprenavir; delavirdine; delavirdine mesylate; Radha-108 (receptol); lamivudine and tenofovir disoproxil fumarate; efavirenz, lamivudine, and tenofovir disoproxil fumarate; phosphinothricin; lamivudine, nevirapine, and zidovudine; abacavir; and abacavir sulfate.

[0213] Other HIV medicines

[0214]

[04] Examples of other drugs for treating HIV that can be combined with the agents of the present disclosure include aspernigin C, acetomorphan, alisporivir, BanLec, deferiprone, Gamimune, metenfarin, naltrexone, α1-proteinase inhibitors, REP 9. RPI-MN, VSSP, H1 viral, SB-728-T, 1,5-dicaffeoylquinic acid, rHIV7-shl-TAR-CCR5RZ, AAV-eCD4-Ig gene therapy, MazF gene therapy, BlockAide, bevirimab derivatives, ABX-464, AG-1105, APH-0812, bryostatin analogs, BIT-225, CYT-107, CS-TATI-1, fluoro-β-D-arabinose nucleic acid (FANA)-modified antisense oligonucleotides, FX-101, Grefsen, HGTV-43, HPH-116, HS-10234, hydroxychloroquine, IMB-10035, IMO-3100, IND-02, JL-18008, LADAVRU, MK-1376, MK-204 8. MK-4250, MK-8507, MK-8558, MK-8591 (islavir), NOV-205, OB-002H, ODE-Bn-TFV, M1-TFV , PA-1050040 (PA-040), PC-707, PGN-007, QF-036, S-648414, SCY-635, SB-9200, SCB-71 9, TR-452, TEV-90110, TEV-90112, TEV-90111, TEV-90113, RN-18, DIACC-1010, Fasnall, Immuglo, 2-CLIPS peptide, HRF-4467, thrombospondin analog, TBL-1004HI, VG-1177, xl-081, rfhSP-D, [ 18 F]-MC-225, URMC-099-C, RES-529 and VIR-576.

[0215] HIV protease inhibitors

[0216] In certain embodiments, the anti-HIV gp120 V3 glycan-directed antibodies or antigen-binding fragments described herein are combined with HIV protease inhibitors. Examples of HIV protease inhibitors include amprenavir, atazanavir, brenavir, darunavir, fosamprenavir, fosamprenavir calcium, indinavir, indinavir sulfate, lopinavir, nelfinavir, nelfinavir mesylate, ritonavir, saquinavir, saquinavir mesylate, tipranavir, AEBL-2, DG-17, GS-1156, TMB-657 (PPL-100), T-169, BL-008, MK-8122, TMB-607, GRL-02031, and TMC-310911.

[0217] HIV RNase H Inhibitors

[0218] In certain embodiments, the anti-HIV gp120 V3 glycan-directed antibodies or antigen-binding fragments described herein are combined with HIV RNase H inhibitors. Examples of HIV RNase H inhibitors that can be combined include NSC-727447.

[0219] HIV Nef inhibitors

[0220] In certain embodiments, the anti-HIV gpl20 V3 glycan-directed antibodies or antigen-binding fragments described herein are combined with HIV Nef inhibitors. Examples of HIV Nef inhibitors that can be combined include FP-1.

[0221] HIV reverse transcriptase inhibitors

[0222] In certain embodiments, the anti-HIV gp120 V3 glycan-directed antibodies or antigen-binding fragments described herein are combined with non-nucleoside or non-nucleotide inhibitors. Examples of non-nucleoside or non-nucleotide inhibitors of HIV reverse transcriptase include dapivirine, delavirdine, delavirdine mesylate, doravirine, efavirenz, etravirine, lentinan, nevirapine, rilpivirine, ACC-007, ACC-008, AIC-292, F-18, KM-023, PC-1005, VM-1500A-LAI, PF-3450074, elsulfavirine (slow-release oral, HIV infection), elsulfavirine (long-acting injectable nanosuspension, HIV infection) and elsulfavirine (VM-1500).

[0223] In certain embodiments, the anti-HIV gp120 V3 glycan-directed antibodies or antigen-binding fragments described herein are combined with HIV nucleoside or nucleotide inhibitors. Examples of nucleoside or nucleotide inhibitors of HIV reverse transcriptase include: adefovir, adefovir dipivoxil, azvudine, emtricitabine, tenofovir, tenofovir alafenamide, tenofovir alafenamide fumarate, tenofovir alafenamide hemifumarate, tenofovir disoproxil, tenofovir disoproxil fumarate, tenofovir octadecyloxyethyl ester (AGX-1009), tenofovir disoproxil fumarate, and VIDEX (didanosine, ddl), abacavir, abacavir sulfate, alovudine, aritibine, censavudine, didanosine, elvucitabine, festinavir, fosalvudine tidoxil, CMX-157, dapivirine, doravirine, etravirine, OCR-5753, tenofovir disoproxil orotate, fozivudine tidoxil, lamivudine, phosphinothricin, stavudine, zalcitabine, zidovudine, rovafovir etalafenamide (GS-9131), GS-9148, MK-8504, MK-8591, MK-858, VM-2500 and KP-1461.

[0224] HIV integrase inhibitors

[0225] In certain embodiments, the anti-HIV gp120 V3 glycan-directed antibodies or antigen-binding fragments described herein are combined with HIV integrase inhibitors. Examples of HIV integrase inhibitors include elvitegravir, elvitegravir (extended release microcapsules), curcumin, curcumin derivatives, chicoric acid, chicoric acid derivatives, 3,5-dicaffeoylquinic acid, 3,5-dicaffeoylquinic acid derivatives, aurintricarboxylic acid, aurintricarboxylic acid derivatives, caffeic acid phenethyl ester, caffeic acid phenethyl ester derivatives, tyrosine phosphorylation inhibitors, tyrosine phosphorylation inhibitor derivatives, quercetin, quercetin derivatives, raltegravir, pegylated raltegravir, dolutegravir, JTK-351, bicagvir, AVX-1556 7. Cabotegravir (long-acting injectable), diketoquinoline-4-1 derivatives, integrase-LEDGF inhibitors, ledgins, M-522, M-532, MK-0536, NSC-310217, NSC-371056, NSC-48240, NSC-642710, NSC-699171, NSC-699172, NSC-699173, NSC-699174, stilbene disulfonic acid, T-169, STP-0404, VM-3500 and cabotegravir.

[0226] In certain embodiments, the anti-HIV gp120 V3 glycan-directed antibodies or antigen-binding fragments described herein are combined with HIV non-catalytic site or allosteric integrase inhibitors (NCINIs). Examples of HIV non-catalytic site or allosteric integrase inhibitors (NCINIs) include CX-05045, CX-05168, and CX-14442.

[0227] HIV entry inhibitors

[0228] In certain embodiments, the anti-HIV gp120 V3 glycan-directed antibodies or antigen-binding fragments described herein are combined with HIV entry inhibitors. Examples of HIV entry (fusion) inhibitors include AAR-501, LBT-5001, Cinevir, CCR5 inhibitors, gp41 inhibitors, CD4 attachment inhibitors, gp120 inhibitors, gp160 inhibitors, and CXCR4 inhibitors.

[0229] In certain embodiments, the anti-HIV gp120 V3 glycan-directed antibodies or antigen-binding fragments described herein are combined with CCR5 inhibitors. Examples of CCR5 inhibitors include apravirox, vicovirox, maraviroc, maraviroc (long-acting nanoemulsion for injection), cinevirox, lelizumab (PRO-140), adaptavir (RAP-101), nifevirox (TD-0232), anti-GP120 / CD4 or CCR5 bispecific antibodies, B-07, MB-66, polypeptide C25P, TD-0680, seravirox, and vMIP (Haimipu).

[0230] In certain embodiments, the anti-HIV gp120 V3 glycan-directed antibodies or antigen-binding fragments described herein are combined with a CXCR4 inhibitor. Examples of CXCR4 inhibitors include plerixafor, ALT-1188, N15 peptide, and vMIP (Haimipu).

[0231] In certain embodiments, the anti-HIV gp120 V3 glycan-directed antibodies or antigen-binding fragments described herein are combined with a gp41 inhibitor. Examples of gp41 inhibitors include Abuvirtide, Enfuvirtide, Greifsom (gp41 / gp120 / gp160 inhibitor), BMS-986197, Enfuvirtide improved biosimilars, Enfuvirtide biosimilars, HIV-1 fusion inhibitors, (P26-Bapc), ITV-1, ITV-2, ITV-3, ITV-4, CPT-31, Cl3hmAb, PIE-12 trimer, and Sifuvirtide.

[0232] In certain embodiments, the anti-HIV gp120 V3 glycan-directed antibodies or antigen-binding fragments described herein are combined with a CD4 attachment inhibitor. Examples of CD4 attachment inhibitors include ibalizumab and CADA analogs

[0233] In certain embodiments, the anti-HIV gp120 V3 glycan-directed antibodies or antigen-binding fragments described herein are combined with a gp120 inhibitor. Examples of gp120 inhibitors include anti-HIV microbicides, Radha-108 (receptol) 3B3-PE38, BanLec, bentonite-based nanomedicines, fostemsavir tromethamine, IQP-0831, VVX-004, and BMS-663068.

[0234] In certain embodiments, the anti-HIV gp120 V3 glycan-directed antibodies or antigen-binding fragments described herein are combined with a gp160 inhibitor. Examples of gp160 inhibitors that can be combined include tetrandrine.

[0235] HIV maturation inhibitors

[0236] In certain embodiments, the anti-HIV gpl20 V3 glycan-directed antibodies or antigen-binding fragments described herein are combined with an HIV maturation inhibitor. Examples of HIV maturation inhibitors include BMS-955176, GSK-3640254, and GSK-2838232.

[0237] Delayed reversal agents

[0238] In certain embodiments, the anti-HIV gp120 V3 glycan-directed antibodies or antigen-binding fragments described herein are combined with HIV latency reversal agents. Examples of latency reversal agents that can be combined with the one or more multispecific antigen-binding molecules described herein include IL-15 receptor agonists (e.g., ALT-803; interleukin-15 / Fc fusion proteins (e.g., XmAb24306); recombinant interleukin-15 (e.g., AM0015, NIZ-985); PEGylated IL-15 (e.g., NKTR-255)); toll-like receptor (TLR) agonists (including TLR7 agonists, such as GS-9620 and TLR8 agonists, such as GS-9688), histone deacetylase (HDAC) inhibitors, proteasome inhibitors such as bortezomib, protein kinase C (PKC) activators, Smyd2 inhibitors, BET -Bromodomain 4 (BRD4) inhibitors, ionomycin, IAP antagonists (apoptosis protein inhibitors, such as APG-1387, LBW-242), SMAC mimetics (including TL32711, LCL161, GDC-0917, HGS1029, AT-406), Debio-1143, PMA, SAHA (vorinostat or suberoyl, anilide and hydroxamic acid), NIZ-985, IL-15 modulating antibodies (including IL-15, IL-15 fusion protein and IL-15 receptor agonists, such as ALT-803), JQ1, disulfiram, amphotericin B and ubiquitin inhibitors such as ragozola analogs, APH-0812 and GSK-343. Examples of HDAC inhibitors include romidepsin, vorinostat and panobinostat. Examples of PKC activators include indole lactams, prostratin, ingenol B, and DAG-lactone.

[0239] Toll-like receptor (TLR) agonists

[0240] In certain embodiments, the anti-HIV gp120 V3 glycan-directed antibodies or antigen-binding fragments described herein are combined with a toll-like receptor (TLR) agonist, e.g., a TLR1 agonist (NCBI gene ID: 7096), a TLR2 agonist (NCBI gene ID: 7097), a TLR3 agonist (NCBI gene ID: 7098), a TLR4 agonist (NCBI gene ID: 7099), a TLR5 agonist (NCBI gene ID: 7100), a TLR6 agonist (NCBI gene ID: 10333), a TLR7 agonist (NCBI gene ID: 51284), a TLR8 agonist (NCBI gene ID: 51311), a TLR9 agonist (NCBI gene ID: 54106), and / or a TLR10 agonist (NCBI gene ID: 81793).Exemplary TLR7 agonists that can be co-administered or combined with the one or more multispecific antigen-binding molecules described herein include, but are not limited to, AL-034, DSP-0509, GS-9620 (visamod), visamod analogs, LHC-165, TMX-101 (imiquimod), GSK-2245035, resiquimod, DSR-6434, DSP-3025, IMO-4200, MCT-465, MEDI-9197, 3M-051, SB-9922, 3M-052, Limtop, TMX-30X, TMX-202, RG-7863, RG-7854, RG-7795, and compounds disclosed in the following patents: US20100143301 (Gilead Sciences), US20110098248 (Gilead Sciences) and US20090047249 (Gilead Sciences), US20140045849 (Janssen), US20140073642 (Janssen), WO2014 / 056953 (Janssen), WO2014 / 076221 (Ja nssen), WO2014 / 128189(Janssen), US20140350031(Janssen), WO2014 / 023813(Janssen), US20080234251(Array Biopharma), US20080306050 (Array Biopharma), US20100029585 (Ventirx Pharma), US20110092485 (Ventirx Pharma), US20110118235 (Ventirx Pharma), US20120082658 (Ventirx Pharma), US20120219615 (Ventirx Pharma), US20140066432 (Ventirx Pharma), US20140088085 (Ventirx Pharma), US20140275167 (Novira Therapeutics) and US20130251673 (Novira Therapeutics). TLR7 / TLR8 agonists that can be co-administered are NKTR-262, telratolimod and BDB-001.Exemplary TLR8 agonists that can be co-administered or combined with one or more of the multispecific antigen-binding molecules described herein include, but are not limited to, E-6887, IMO-4200, IMO-8400, IMO-9200, MCT-465, MEDI-9197, motomod, resiquimod, GS-9688, VTX-1463, VTX-763, 3M-051, 3M-052, and compounds disclosed in the following patents: US20140045849 (Janssen), U S20140073642(Janssen)、WO2014 / 056953(Janssen)、WO2014 / 076221(Janssen)、WO2014 / 128189(Janssen)、US 20140350031(Janssen)、WO2014 / 023813(Janssen)、US20080234251(ArrayBiopharma)、US20080306050(Array Biopharma), US20100029585 (Ventirx Pharma), US20110092485 (Ventirx Pharma), US20110118235 (Ventirx Pharma), US20120082658 (Ventirx Pharma), US20120219615 (Ventirx Pharma), US20140066432 (Ventirx Pharma), US20140088085 (Ventirx Pharma), US20140275167 (Novira Therapeutics) and US20130251673 (Novira Therapeutics). Exemplary TLR9 agonists that can be co-administered include, but are not limited to, AST-008, cotomod, CMP-001, IMO-2055, IMO-2125, linimod, MGN-1601, BB-001, BB-006, IMO-3100, IMO-8400, IR-103, IMO-9200, atomomod, DIMS-9054, DV-1079, DV-1179, AZD-1419, letomomod (MGN-1703), CYT-003, CYT-003-QbG10, tilsotolimod, and PUL-042. Examples of TLR3 agonists include: lintamod, polyICLC,. Apoxxim, IPH-33, MCT-465, MCT-475 and ND-1.1. Examples of TLR4 agonists include: G-100 and GSK-1795091.

[0241] Histone deacetylase (HDAC) inhibitors

[0242] In certain embodiments, the anti-HIV gp120 V3 glycan-directed antibodies or antigen-binding fragments described herein are combined with an inhibitor of histone deacetylase, such as histone deacetylase 1, histone deacetylase 9 (HDAC9, HD7, HD7b, HD9, HDAC, HDAC7, HDAC7B, HDAC9B, HDAC9FL, HDRP, MITR; Gene ID: 9734). Examples of HDAC inhibitors include, but are not limited to, abexinostat, ACY-241, AR-42, BEBT-908, belinostat, CKD-581, CS-055 (HBI-8000), CT-101, CUDC-907 (feminostat), entinostat, givista, motistat, panobinostat, premotistat, quinostat (JNJ-26481585), resminostat, linostat, romidepsin, SHP-141, TMB-ADC, valproic acid (VAL-001), vorinostat, tinostamustine, resminostat, and entinostat.

[0243] Cyclin-dependent kinase (CDK) inhibitors or antagonists

[0244] In certain embodiments, the anti-HIV gp120 V3 glycan-directed antibodies or antigen-binding fragments described herein are combined with a cyclin-dependent kinase (CDK) inhibitor or antagonist, such as cyclin-dependent kinase 4 (CDK4; NCBI gene ID: 1019), cyclin-dependent kinase 6 (CDK6; NCBI gene ID: 1021), cyclin-dependent kinase 9 (CDK9; NCBI gene ID: 1025). In some embodiments, the CDK4 / CDK6 / CDK9 inhibitor or antagonist is selected from VS2-370.

[0245] Stimulator of interferon genes (STING) agonists

[0246] In some embodiments, the anti-HIV gp120 V3 glycan-directed antibodies or antigen-binding fragments described herein are combined with stimulator of interferon genes (STING). In some embodiments, the STING receptor agonist or activator is selected from ADU-S100 (MIW-815), SB-11285, MK-1454, SR-8291, AdVCA0848, GSK-532, SYN-STING, MSA--1, SR-8291, 5,6-dimethylxanthone-4-acetic acid (DMXAA), cyclic GAMP (cGAMP) and cyclic diAMP.

[0247] RIG-I agonists

[0248] In certain embodiments, the anti-HIV gp120 V3 glycan-directed antibodies or antigen-binding fragments described herein are combined with DExD / H-box helicase 58 (DDX58; also known as RIG-I, RIG1, RIGI, RLR-1, SGMRT2; NCBI Gene ID: 23586). In some embodiments, the agents described herein are combined with RIG-I modulators such as RGT-100 or NOD2 modulators such as SB-9200 (also known as GS 9992; inarigivir) and IR-103. An exemplary RIG-I agonist is KIN1148, described in Hemann et al., J Immunol, May 1, 2016, Vol. 196 Suppl. 1) 76.1. Additional RIG-I agonists are described in, for example, Elion et al., Cancer Res., 2018, Vol. 78, No. 21, pp. 6183-6195; and Liu et al., J Virol., 2016, Vol. 90, No. 20, pp. 9406-9419. RIG-I agonists are commercially available, for example, from Invivogen (invivogen.com).

[0249] LAG-3 and TIM-3 inhibitors

[0250] In certain embodiments, the anti-HIV gp120 V3 glycan-directed antibodies or antigen-binding fragments described herein are combined with anti-TIM-3 (also known as hepatitis A virus cellular receptor 2 antibody (HAVCR2; NCBI gene ID: 84868) such as TSR-022, LY-3321367, MBG-453, INCAGN-2390. In some embodiments, the anti-HIV gp120 V3 glycan-directed antibodies or antigen-binding fragments described herein are combined with anti-LAG-3 (lymphocyte activation) (NCBI gene ID: 3902) antibodies such as relalizumab (ONO-4482), LAG-525, MK-4280, REGN-3767, INCAGN2385.

[0251] Capsid inhibitors

[0252] In certain embodiments, the anti-HIV gp120 V3 glycan-directed antibodies or antigen-binding fragments described herein are combined with capsid inhibitors. Examples of capsid inhibitors that can be combined with the disclosed agents include capsid polymerization inhibitors or capsid interfering compounds, HIV nucleocapsid P7 (NCp7) inhibitors such as azodicarbonamide, HIV p24 capsid protein inhibitors, GS-6207, GS-CA1, AVI-621, AVI-101, AVI-201, AVI-301 and AVI-CAN1-15 series, PF-3450074, and compounds described in International Patent Publication WO2019 / 087016.

[0253] Immune-based therapies

[0254] In certain embodiments, the anti-HIV gp120 V3 glycan-directed antibodies or antigen-binding fragments described herein are combined with immune-based therapies. Examples of immune-based therapies include toll-like receptor (TLR) modulators, such as TLR1, TLR2, TLR3, TLR4, TLR5, TLR6, TLR7, TLR8, TLR9, TLR10, TLR11, TLR12, and TLR13; programmed cell death protein 1 (PD-1) modulators; programmed death ligand 1 (PD-L1) inhibitors; IL-15 inhibitors (e.g., IL-15 receptor agonists (e.g., ALT-803; interleukin-15 / Fc fusion proteins (e.g., XmAb24306); recombinant interleukin-15 (e.g., AM0015, NIZ-985); pegylated IL-15 (e.g., NKTR-255)); ermaVir; interleukin-7; hydroxychloroquine (hydroxychloroquine); interleukins (aldesleukin, IL-2); interferon alpha; interferon alpha-2b; interferon alpha-n3; pegylated interferon alpha; interferon gamma; hydroxyurea; mycophenolate mofetil (MPA) and its ester derivative mycophenolate mofetil (MMF); ribavirin; polymer polyethyleneimine (PEI); gepon; IL-12; WF-10; VGV-1; MOR-22; BMS-936559; CYT-107, nomofuron, pegylated interferon alpha-2a, pegylated interferon alpha-2b, RPI-MN, STING modulators, RIG-I modulators, NOD2 modulators, SB-9200 and IR-103.

[0255] In certain embodiments, the anti-HIV gp120 V3 glycan-directed antibodies or antigen-binding fragments described herein are combined with a TLR agonist. Examples of TLR agonists include, but are not limited to, visammod (GS-9620), letomod, tilsotolimod, lintamod, DSP-0509, AL-034, G-100, coutomod, AST-008, motomod, GSK-1795091, GSK-2245035, VTX-1463, GS-9688, LHC-165, BDB-001, RG-7854, telratolimod.

[0256] Immune checkpoint receptor protein modulators

[0257] In various embodiments, the anti-HIV gp120 V3 polysaccharide-directed antibodies or antigen-binding fragments described herein are combined with one or more blockers or inhibitors of inhibitory immune checkpoint proteins or receptors and / or with one or more stimulators, activators or agonists of one or more stimulatory immune checkpoint proteins or receptors. Blocking or inhibition of inhibitory immune checkpoints can positively regulate T cell or NK cell activation and prevent immune escape of infected cells. Activation or stimulation of stimulating immune checkpoints can enhance the effect of immune checkpoint inhibitors in infectious treatments. In various embodiments, immune checkpoint proteins or receptors regulate T cell responses (e.g., reviewed in Xu et al., J Exp Clin Cancer Res., 2018, Vol. 37, p. 110). In various embodiments, the immune checkpoint protein or receptor modulates NK cell responses (e.g., reviewed in Davis et al., Semin Immunol., 2017, Vol. 31, pp. 64-75, and Chiossone et al., Nat Rev Immunol., 2018, Vol. 18, No. 11, pp. 671-688).

[0258] Examples of immune checkpoint proteins or receptors that can be combined with the anti-HIV gp120 V3 glycan-directed antibodies or antigen-binding fragments described herein include, but are not limited to, CD27, CD70; CD40, CD40LG; CD47, CD48 (SLAMF2), transmembrane and immunoglobulin domain 2 (TMIGD2, CD28H), CD84 (LY9B, SLAMF5), CD96, CD160, MS4A1 (CD20), CD244 (SLAMF4); CD276 (B7H3); V-set domain-containing T cell activation inhibitor 1 (VTCN1, B7H4); V-set immunomodulatory receptor (VSIR, B7H5, VISTA); immunoglobulin superfamily member 11 (IGSF11, VSIG3); natural killer cytotoxicity receptor 3 ligand 1 (NCR3LG1, B7H6); HERV-H LTR-associated 2 (HHLA2, B7H7); inducible T cell co-stimulator (ICOS, CD278); inducible T cell co-stimulatory ligand (ICOSLG, B7H2); TNF receptor superfamily member 4 (TNFRSF4, OX40); TNF superfamily member 4 (TNFSF4, OX40L); TNFRSF8 (CD30), TNFSF8 (CD30L); TNFRSF10A (CD261, DR4, TRAILR1), TNFRSF9 (CD137), TNFSF9 (CD137L); TNFRSF10B (CD262, DR5, TRAILR2), TNFRSF10 (TRAIL); TNFRSF14 (HVEM, CD270), TNFSF14 (HVEML); CD272 (B and T lymphocyte-associated (BTLA)); TNFRSF17 (BCMA, CD269), TNFSF1 3B (BAFF); TNFRSF18 (GITR), TNFSF18 (GITRL); MHC class I polypeptide-related sequence A (MICA); MHC class I polypeptide-related sequence B (MICB); CD274 (CD274, PDL1, PD-L1); programmed cell death protein 1 (PDCD1, PD1, PD-1); cytotoxic T-lymphocyte-associated protein 4 (CTLA4, CD152); CD80 (B7-1), CD28; fibronectin cell adhesion molecule 2 (NECTIN2, CD112); CD226 (DNAM-1); poliovirus receptor (PVR) cell adhesion molecule (PVR, CD155); containing PVR-related immunoglobulin domain (PVRIG, CD112R); T cell immunoreceptor with Ig and ITIM domains (TIGIT); containing T cell immunoglobulin and mucin domain 4 (TIMD4; TIM4);Hepatitis A virus cell receptor 2 (HAVCR2, TIMD3, TIM3); galectin 9 (LGALS9); lymphocyte activation 3 (LAG3, CD223); signal transducer lymphocyte activation molecule family member 1 (SLAMF1, SLAM, CD150); lymphocyte antigen 9 (LY9, CD229, SLAMF3); SLAM family member 6 (SLAMF6, CD352); SLAM family member 7 (SLAMF7, CD319 ); UL16 binding protein 1 (ULBP1); UL16 binding protein 2 (ULBP2); UL16 binding protein 3 (ULBP3); retinoic acid early transcript 1E (RAET1E; ULBP4); retinoic acid early transcript 1G (RAET1G; ULBP5); retinoic acid early transcript 1L (RAET1L; ULBP6); lymphocyte activation 3 (CD223); killer cell immunoglobulin-like receptor, three Ig domains and long cytoplasmic tail 1 (KIR , CD158E1); killer cell lectin-like receptor C1 (KLRC1, NKG2A, CD159A); killer cell lectin-like receptor K1 (KLRK1, NKG2D, CD314); killer cell lectin-like receptor C2 (KLRC2, CD159c, NKG2C); killer cell lectin-like receptor C3 (KLRC3, NKG2E); killer cell lectin-like receptor C4 (KLRC4, NKG2F); killer cell immunoglobulin-like receptor, two Ig domains and long cytoplasmic tail 1 (KIR2DL1); killer cell immunoglobulin-like receptor, two Ig domains and long cytoplasmic tail 2 (KIR2DL2); killer cell immunoglobulin-like receptor, two Ig domains and long cytoplasmic tail 3 (KIR2DL3); killer cell immunoglobulin-like receptor, three Ig domains and long cytoplasmic tail 1 (KIR3DL1); killer cell lectin-like receptor D1 (KLRD1); and SLAM family member 7 (SLAMF7).

[0259] In various embodiments, the anti-HIV gpl20 V3 glycan-directed antibodies or antigen-binding fragments described herein are combined with one or more blockers or inhibitors of one or more T cell inhibitory immune checkpoint proteins or receptors. Exemplary T-cell inhibitory immune checkpoint proteins or receptors include, but are not limited to, CD274 (CD274, PDL1, PD-L1); programmed cell death protein 1 ligand 2 (PDCD1LG2, PD-L2, CD273); programmed cell death protein 1 (PDCD1, PD1, PD-1); cytotoxic T-lymphocyte-associated protein 4 (CTLA4, CD152); CD276 (B7H3); V-set domain-containing T cell activation inhibitor 1 (VTCN1, B7H4); V-set immunoregulatory receptor (VSIR, B7H5, VISTA); immunoglobulin superfamily member 11 (IGSF11, VSIG3); TNFRSF14 (HVEM, CD270), TNFSF14 (HVEML); CD272 (B and T lymphocyte-associated (BTLA)); PVR-containing related immunoglobulin domains (PVRIG, CD112R); T cell immunoreceptor with Ig and ITIM domains (TIGIT); lymphocyte activation 3 (LAG3, CD223); hepatitis A virus cellular receptor 2 (HAVCR2, TIMD3, TIM3); galectin 9 (LGALS9); killer cell immunoglobulin-like receptor, three Ig domains and long cytoplasmic tail 1 (KIR, CD158E1); killer cell immunoglobulin-like receptor, two Ig domains and long cytoplasmic tail 1 (KIR2DL1); killer cell immunoglobulin-like receptor, two Ig domains and long cytoplasmic tail 2 (KIR2DL2); killer cell immunoglobulin-like receptor, two Ig domains and long cytoplasmic tail 3 (KIR2DL3); and killer cell immunoglobulin-like receptor, three Ig domains and long cytoplasmic tail 1 (KIR3DL1). In various embodiments, the anti-HIV gpl20 V3 glycan-directed antibodies or antigen-binding fragments described herein are combined with one or more agonists or activators of one or more T cell stimulating immune checkpoint proteins or receptors.Exemplary T cell stimulatory immune checkpoint proteins or receptors include, but are not limited to, CD27, CD70; CD40, CD40LG; inducible T cell co-stimulator (ICOS, CD278); inducible T cell co-stimulatory ligand (ICOSLG, B7H2); TNF receptor superfamily member 4 (TNFRSF4, OX40); TNF superfamily member 4 (TNFSF4, OX40L); TNFRSF9 (CD137), TNFSF9 (CD137L); TNFRSF18 (GITR), TNFSF18 (GITRL); CD80 (B7-1), CD28; fibronectin cell adhesion molecule 2 (NECTIN2, CD112); CD226 (DNAM-1); CD244 (2B4, SLAMF4), poliovirus receptor (PVR) cell adhesion molecule (PVR, CD155). See, e.g., Xu et al., J Exp Clin Cancer Res., 2018, vol. 37, p. 110.

[0260] In various embodiments, the anti-HIV gpl20 V3 glycan-directed antibodies or antigen-binding fragments described herein are combined with one or more blockers or inhibitors of one or more NK cell inhibitory immune checkpoint proteins or receptors. Exemplary NK cell inhibitory immune checkpoint proteins or receptors include, but are not limited to, killer cell immunoglobulin-like receptor, three Ig domains and a long cytoplasmic tail 1 (KIR, CD158E1); killer cell immunoglobulin-like receptor, two Ig domains and a long cytoplasmic tail 1 (KIR2DL1); killer cell immunoglobulin-like receptor, two Ig domains and a long cytoplasmic tail 2 (KIR2DL2); killer cell immunoglobulin-like receptor, two Ig domains and a long cytoplasmic tail 3 (KIR2DL3); killer cell immunoglobulin-like receptor, three Ig domains and a long cytoplasmic tail 1 (KIR3DL1); killer cell lectin-like receptor C1 (KLRC1, NKG2A, CD159A); and killer cell lectin-like receptor D1 (KLRD1, CD94). In various embodiments, the anti-HIV gp120 V3 polysaccharide-directed antibodies or antigen-binding fragments described herein are combined with one or more agonists or activators of one or more NK cell-stimulating immune checkpoint proteins or receptors. Exemplary NK cell-stimulating immune checkpoint proteins or receptors include, but are not limited to, CD16, CD226 (DNAM-1); CD244 (2B4, SLAMF4); Killer cell lectin-like receptor K1 (KLRK1, NKG2D, CD314); SLAM family member 7 (SLAMF7). See, for example, Davis et al., Semin Immunol., 2017, Vol. 31, pp. 64-75; Fang et al., Semin Immunol., 2017, Vol. 31, pp. 37-54; and Chiossone et al., Nat Rev Immunol., 2018, Vol. 18, No. 11, pp. 671-688.

[0261] In some embodiments, the one or more immune checkpoint inhibitors include proteinaceous (e.g., antibodies or fragments thereof, or antibody mimetics) inhibitors of PD-L1 (CD274), PD-1 (PDCD1), or CTLA4. In some embodiments, the one or more immune checkpoint inhibitors include small organic molecule inhibitors of PD-L1 (CD274), PD-1 (PDCD1), or CTLA4.

[0262] Examples of CTLA4 inhibitors that can be co-administered include, but are not limited to, ipilimumab, tremelimumab, BMS-986218, AGEN1181, AGEN1884, BMS-986249, MK-1308, REGN-4659, ADU-1604, CS-1002, BCD-145, APL-509, JS-007, BA-3071, ONC-392, AGEN-2041, JHL-1155, KN-044, CG-0161, ATOR -1144, PBI-5D3H5, BPI-002, and the multispecific inhibitors FPT-155 (CTLA4 / PD-L1 / CD28), PF-06936308 (PD-1 / CTLA4), MGD-019 (PD-1 / CTLA4), KN-046 (PD-1 / CTLA4), MEDI-5752 (CTLA4 / PD-1), XmAb-20717 (PD-1 / CTLA4), and AK-104 (CTLA4 / PD-1).

[0263] Examples of inhibitors of PD-L1 (CD274) or PD-1 (PDCD1) that can be co-administered include, but are not limited to, pembrolizumab, nivolumab, simumab, pidilizumab, AMP-224, MEDI0680 (AMP-514), spartalizumab, atezolizumab, avelumab, durvalumab, BMS-936559, CK-301, PF-06801591, BGB-A317 (tislelizumab), GLS-010 (WBP-3055), AK-103 (HX-008), AK-105, CS-1003, HLX-10, MGA-012, BI-754091, A GEN-2034, JS-001 (toripalizumab), JNJ-63723283, genol monoclonal antibody (CBT-501), LZM-009, BCD-100, LY-3300054, SHR-1201, SHR-1210 (camelizumab), Sym-021, ABBV-181 (bugliizumab), PD1-PIK, BAT-1306, (MSB0010718C), CX-072, CBT-502, TSR-042 (dotalimumab), MSB-2311, JTX-4014, BGB-A333, SHR-1316, CS-1001 (WBP-3 155), KN-035, IBI-308 (sintilimab), HLX-20, KL-A167, STI-A1014, STI-A1015 (IMC-001), BCD-135, FAZ-053, TQB-2450, MDX1105-01, GS-4224, GS-4416, INCB086550, MAX10181, and multispecific inhibitors FPT-155 (CTLA4 / PD-L1 / CD28), PF-06936308 (PD-1 / CTLA4), MGD-013 (PD-1 / LAG-3), FS-118 (LAG-3 / PD -L1)MGD-019(PD-1 / CTLA4), KN-046(PD-1 / CTLA4), MEDI-5752(CTLA4 / PD-1), RO-7121661(PD-1 / TIM-3), XmAb-20717(PD-1 / CTLA4), AK-104(C TLA4 / PD-1), M7824 (PD-L1 / TGFβ-EC domain), CA-170 (PD-L1 / VISTA), CDX-527 (CD27 / PD-L1), LY-3415244 (TIM3 / PDL1) and INBRX-105 (4-1BB / PDL1).

[0264] In some embodiments, the small molecule inhibitor of CD274 or PDCD1 is selected from GS-4224, GS-4416, INCB086550, and MAX10181. In some embodiments, the small molecule inhibitor of CTLA4 comprises BPI-002.

[0265] In various embodiments, an antibody or antigen-binding fragment as described herein is combined with an anti-TIGIT antibody, such as BMS-986207, RG-6058, AGEN-1307.

[0266] TNF receptor superfamily (TNFRSF) member agonists or activators

[0267] In various embodiments, the anti-HIV gp120 V3 glycan-directed antibodies or antigen-binding fragments described herein are combined with an agonist of one or more TNF receptor superfamily (TNFRSF) members, e.g., TNFRSF1A (NCBI gene ID: 7132), TNFRSF1B (NCBI gene ID: 7133), TNFRSF4 (OX40, CD134; NCBI gene ID: 7293), TNFRSF5 (CD40; NCBI gene ID: 958), TNFRSF6 (FAS, NCBI gene ID: 355), TNFRSF7 (CD27, N NCBI gene ID: 939), TNFRSF8 (CD30, NCBI gene ID: 943), TNFRSF9 (4-1BB, CD137, NCBI gene ID: 3604), TNFRSF10A (CD261, DR4, TRAILR1, NCBI gene ID: 8797), TNFRSF10B (CD262, DR5, TRAILR2, NCBI gene ID: 8795), TNFRSF10C (CD263, TRAILR3, NCBI gene ID: 8794), TNFRSF10D (CD264, TRAILR4, NCBI gene ID: 8793), TNFRSF11A (CD265, RANK, NCBI gene ID: 8792), TNFRSF11B (NCBI gene ID: 4982), TNFRSF12A (CD266, NCB gene ID: 51330), TNFRSF13B (CD267, NCBI gene ID: 23495), TNFRSF13C (CD268, NCBI gene ID: 115650), TNF Agonists of one or more of RSF16 (NGFR, CD271, NCBI gene ID: 4804), TNFRSF17 (BCMA, CD269, NCBI gene ID: 608), TNFRSF18 (GITR, CD357, NCBI gene ID: 8784), TNFRSF19 (NCBI gene ID: 55504), TNFRSF21 (CD358, DR6, NCBI gene ID: 27242) and TNFRSF25 (DR3, NCBI gene ID: 8718).

[0268] Exemplary anti-TNFRSF4 (OX40) antibodies that may be co-administered include, but are not limited to, MEDI6469, MEDI6383, MEDI0562 (tavulizumab), MOXR0916, PF-04518600, RG-7888, GSK-3174998, INCAGN1949, BMS-986178, GBR-8383, ABBV-368, and antibodies described in the following patents: WO2016179517, WO2017096179, WO2017096182, WO2017096281, and WO2018089628.

[0269] Exemplary anti-TNFRSF5 (CD40) antibodies that may be co-administered include, but are not limited to, RG7876, SEA-CD40, APX-005M, and ABBV-428.

[0270] In some embodiments, the anti-TNFRSF7 (CD27) antibody varilutumab (CDX-1127) is co-administered.

[0271] Exemplary anti-TNFRSF9 (4-1BB, CD137) antibodies that may be co-administered include, but are not limited to, usurelumab, utolumab (PF-05082566), AGEN2373, and ADG-106.

[0272] Exemplary anti-TNFRSF18 (GITR) antibodies that can be co-administered include, but are not limited to, MEDI1873, FPA-154, INCAGN-1876, TRX-518, BMS-986156, MK-1248, GWN-323, and antibodies described in the following patents: WO2017096179, WO2017096276, WO2017096189, and WO2018089628. In some embodiments, antibodies or fragments thereof that co-target TNFRSF4 (OX40) and TNFRSF18 (GITR) are co-administered. Such antibodies are described, for example, in WO2017096179 and WO2018089628.

[0273] Interleukin receptor agonists

[0274] In certain embodiments, the anti-HIV gp120 V3 glycan-directed antibodies or antigen-binding fragments described herein are combined with an interleukin receptor agonist, such as an IL-2 antagonist, an IL-7 antagonist, an IL-15 antagonist, an IL-10 antagonist, an IL-12 antagonist; examples of IL-2 receptor antagonists are interleukins (aldesleukin, IL-2); PEGylated IL-2 (e.g., NKTR-214); modified variants of IL-2 (e.g., THOR-707), bempegaldesleukin, AIC-284, ALKS-4230, CUI-101, Neo-2 / 15; IL-15 receptor antagonists such as ALT-803, NKTR-255 and hetIL-15, interleukin 15 / Fc fusion protein, AM-0015, NIZ-985, SO-C101, IL-15Synthorin (pegylated IL-15), P-22339 and IL-15-PD-1 fusion protein N-809; examples of IL-7 include CYT-107.

[0275] Additional interleukin receptor agonists that can be combined with the anti-HIV gp120 V3 glycan-directed antibodies or antigen-binding fragments described herein include interferon alpha; interferon alpha-2b; interferon alpha-n3; pegylated interferon alpha; interferon gamma; Flt3 agonists such as CDX-301; gepon; nomofuron, pegylated interferon alpha-2a, pegylated interferon alpha-2b, RPI-MN.

[0276] Bispecific and trispecific natural killer (NK) cell engagers

[0277] In various embodiments, the anti-HIV gp120 V3 glycan-directed antibodies or antigen-binding fragments described herein are combined with bispecific NK cell engagers (BiKEs) or trispecific NK cell engagers (TriKEs) (e.g., without Fc) or bispecific antibodies (e.g., with Fc) to NK cell activating receptors such as CD16A, C-type lectin receptors (CD94 / NKG2C, NKG2D, NKG2E / H, and NKG2F), natural cytotoxicity receptors (NKp30, NKp44, and NKp46), killer cell C-type lectin-like receptors (NKp65, NKp80), Fc receptors FcγR (which mediate antibody-dependent cellular cytotoxicity), SLAM family receptors (e.g., 2B4, SLAM6, and SLAM7), killer cell immunoglobulin-like receptors (KIRs) (KIR-2DS and KIR-3DS), DNAM-1, and CD137 (4-1BB). Exemplary anti-CD16 bispecific antibodies, BiKEs or TriKEs that can be co-administered include AFM26 (BCMA / CD16A) and AFM-13 (CD16 / CD30). Depending on the circumstances, the anti-CD16 binding bispecific molecule may or may not have an Fc. BiKE and TriKE are described in, for example, Felices et al., Methods Mol Biol., 2016, Vol. 1441, pp. 333-346; Fang et al., Semin Immunol., 2017, Vol. 31, pp. 37-54. Examples of trispecific NK cell engagers (TRiKEs) include OXS-3550 and CD16-IL-15-B7H3 TriKe.

[0278] Phosphatidylinositol 3-kinase (PI3K) inhibitors

[0279] In certain embodiments, the anti-HIV gp120 V3 glycan-directed antibodies or antigen-binding fragments described herein are combined with a PI3K inhibitor. Examples of PI3K inhibitors include idelalisib, apellisib, buparlisib, CAI orotic acid, cupanisib, duvelisib, gedatolisib, neratinib, panulisib, perifosine, pictilisib, pilaralisib, praquintinib mesylate, regocerti, regocerti sodium, sonolisib, taselisib, AMG-319, AZD-8186, BAY-1082439, CLR-1401, CLR-457, CUDC-90 7. DS-7423, EN-3342, GSK-2126458, GSK-2269577, GSK-2636771, INCB-040093, LY-3023414, MLN-1117, PQR-309, R G-7666, RP-6530, RV-1729, SAR-245409, SAR-260301, SF-1126, TGR-1202, UCB-5857, VS-5584, XL-765 and ZSTK-474.

[0280] Alpha-4 / beta-7 antagonists

[0281] In certain embodiments, the anti-HIV gp120 V3 glycan-directed antibodies or antigen-binding fragments described herein are combined with an alpha-4 / beta-7 antagonist. Examples of integrin alpha-4 / beta-7 antagonists include PTG-100, TRK-170, alirocumab, edrelimumab, methyl caltastat, and vedolizumab.

[0282] Pharmacokinetic enhancers

[0283] In certain embodiments, the anti-HIV gp120 V3 glycan-directed antibodies or antigen-binding fragments described herein are combined with a pharmacokinetic enhancer. Examples of pharmacokinetic enhancers include cobicistat and ritonavir.

[0284] Additional therapeutic agents

[0285] Examples of additional therapeutic agents include compounds disclosed in WO 2004 / 096286 (Gilead Sciences); WO 2006 / 015261 (Gilead Sciences); WO 2006 / 110157 (Gilead Sciences); WO 2012 / 003497 (Gilead Sciences); WO 2012 / 003498 (Gilead Sciences); WO 2012 / 145728 (Gilead Sciences); WO 2013 / 006738 (Gilead Sciences); WO 2013 / 159064 (Gilead Sciences); WO 2014 / 100323 (Gilead Sciences), US 2013 / 0165489 (University of Pennsylvania), US 2014 / 0221378 (Japan); Tobacco), US2014 / 0221380 (Japan Tobacco); WO 2009 / 062285 (Boehringer Ingelheim); WO 2010 / 130034 (Boehringer Ingelheim); WO 2013 / 006792 (Pharma Resources), US20140221356 (Gilead Sciences), US20100143301 (GileadSciences) and WO 2013 / 091096 (Boehringer Ingelheim).

[0286] HIV combination therapy

[0287] In a specific embodiment, the anti-HIV 120V3 glycan-directed antibodies or antigen-binding fragments described herein are combined with one, two, three, four or more additional therapeutic agents selected from (efavirenz, tenofovir disoproxil fumarate, and emtricitabine); ( rilpivirine, tenofovir disoproxil fumarate, and emtricitabine); (elvitegravir, cobicistat, tenofovir disoproxil fumarate, and emtricitabine); (tenofovir disoproxil fumarate and emtricitabine; TDF+FTC); (tenofovir alafenamide and emtricitabine); (tenofovir alafenamide, emtricitabine, and rilpivirine); (tenofovir alafenamide, emtricitabine, cobicistat and elvitegravir); adefovir; adefovir disoproxil; cobicistat; emtricitabine; tenofovir; tenofovir disoproxil; tenofovir disoproxil fumarate; tenofovir alafenamide; tenofovir alafenamide hemifumarate; (dulutegravir, abacavir, and lamivudine); dolutegravir, abacavir sulfate, and lamivudine; raltegravir; raltegravir and lamivudine; maraviroc; enfuvirtide; ( lopinavir and ritonavir); (zidovudine and lamivudine; AZT+3TC); ( abacavir sulfate and lamivudine; ABC+3TC); (abacavir sulfate, zidovudine and lamivudine; ABC+AZT+3TC); rilpivirine; rilpivirine hydrochloride; atazanavir sulfate and cobicistat; atazanavir and cobicistat; darunavir and cobicistat; atazanavir; atazanavir sulfate; dolutegravir; elvitegravir; ritonavir; atazanavir sulfate and ritonavir; darunavir; lamivudine; α1-proteinase inhibitor; fosamprenavir; fosamprenavir calcium efavirenz; etravirine; nelfinavir; nelfinavir mesylate; interferon; didanosine ; stavudine; indinavir; indinavir sulfate; tenofovir and lamivudine; zidovudine; nevirapine; saquinavir; saquinavir mesylate; aldesleukin; zalcitabine; tipranavir; amprenavir; delavirdine; delavirdine mesylate; Radha-108 (receptol); lamivudine and tenofovir disoproxil fumarate; efavirenz, lamivudine, and tenofovir disoproxil fumarate; phosphinothricin; lamivudine, nevirapine, and zidovudine; abacavir; and abacavir sulfate.

[0288] It will be appreciated by those skilled in the art that the additional therapeutic agents listed above may be included in more than one of the categories listed above. The specific categories are not intended to limit the functionality of those compounds listed in those categories.

[0289] In one specific embodiment, the anti-HIV gp120 V3 glycan-directed antibodies or antigen-binding fragments described herein are combined with nucleoside or nucleotide inhibitors of HIV reverse transcriptase and non-nucleoside inhibitors of HIV reverse transcriptase. In another specific embodiment, the anti-HIV gp120 V3 glycan-directed antibodies or antigen-binding fragments described herein are combined with nucleoside or nucleotide inhibitors of HIV reverse transcriptase and HIV protease inhibitory compounds. In another embodiment, the anti-HIV gp120 V3 glycan-directed antibodies or antigen-binding fragments described herein are combined with nucleoside or nucleotide inhibitors of HIV reverse transcriptase, non-nucleoside inhibitors of HIV reverse transcriptase, and pharmacokinetic enhancers. In certain embodiments, the anti-HIV gp120 V3 glycan-directed antibodies or antigen-binding fragments described herein are combined with at least one nucleoside inhibitor of HIV reverse transcriptase, an integrase inhibitor, and a pharmacokinetic enhancer. In another embodiment, the anti-HIV gp120 V3 glycan-directed antibodies or antigen-binding fragments described herein are combined with two nucleoside or nucleotide inhibitors of HIV reverse transcriptase.

[0290] In a specific embodiment, the anti-HIV gp120 V3 glycan-directed antibodies or antigen-binding fragments described herein are combined with abacavir sulfate, tenofovir, tenofovir disoproxil, tenofovir disoproxil fumarate, tenofovir disoproxil hemifumarate, tenofovir alafenamide, or tenofovir alafenamide hemifumarate.

[0291] In a specific embodiment, an anti-HIV gp120 V3 glycan-directed antibody or antigen-binding fragment described herein is combined with tenofovir, tenofovir disoproxil, tenofovir disoproxil fumarate, tenofovir alafenamide, or tenofovir alafenamide hemifumarate.

[0292] In a specific embodiment, the anti-HIV gp120 V3 glycan-directed antibodies or antigen-binding fragments described herein are combined with a first additional therapeutic agent selected from abacavir sulfate, tenofovir, tenofovir disoproxil, tenofovir disoproxil fumarate, tenofovir alafenamide, and tenofovir alafenamide hemifumarate, and a second additional therapeutic agent selected from emtricitabine and lamivudine.

[0293] In a specific embodiment, the anti-HIV gp120 V3 glycan-directed antibodies or antigen-binding fragments described herein are combined with a first additional therapeutic agent and a second additional therapeutic agent, the first additional therapeutic agent being selected from the group consisting of tenofovir, tenofovir disoproxil, tenofovir disoproxil fumarate, tenofovir alafenamide, and tenofovir alafenamide hemifumarate, wherein the second additional therapeutic agent is emtricitabine.

[0294] In certain embodiments, the anti-HIV gp120 V3 glycan-directed antibodies or antigen-binding fragments described herein are combined with one or more additional therapeutic agents at a therapeutically effective dose, which is, for example, in the following range: 1 mg to 50 mg, 75 mg, 100 mg, 150 mg, 200 mg, 250 mg, 300 mg, 400 mg, 500 mg, 1000 mg or 1500 mg of anti-HIV gp120 V3 glycan-directed antibodies or antigen-binding fragments. In certain embodiments, the anti-HIV gp120 V3 glycan-directed antibodies or antigen-binding fragments described herein are combined with one or more additional therapeutic agents at a therapeutically effective dose, which is in the range of, for example, about 0.1 mg / kg to about 0.5 mg / kg, 1 mg / kg, 2 mg / kg, 3 mg / kg, 4 mg / kg, 5 mg / kg, 8 mg / kg, 10 mg / kg, 15 mg / kg, 20 mg / kg, 25 mg / kg, 30 mg / kg, 35 mg / kg, 40 mg / kg, 45 mg / kg or 50 mg / kg of the anti-HIV gp120 V3 glycan-directed antibodies or antigen-binding fragments. In certain embodiments, the anti-HIV gp120 V3 glycan-directed antibodies or antigen-binding fragments described herein are combined with one or more additional therapeutic agents at a therapeutically effective dose, which is, for example, in the following range: about 5 mg to about 10 mg, 20 mg, 25 mg, 50 mg, 100 mg, 125 mg, 150 mg, 250 mg, 300 mg, 500 mg, 1000 mg or 1500 mg of anti-HIV gp120 V3 glycan-directed antibodies or antigen-binding fragments.

[0295] In certain embodiments, the anti-HIV gp120 V3 glycan-directed antibodies or antigen-binding fragments described herein are combined with 5mg-30mg of tenofovir alafenamide fumarate, tenofovir alafenamide hemifumarate, or tenofovir alafenamide and 200mg of emtricitabine. In certain embodiments, the anti-HIV gp120 V3 glycan-directed antibodies or antigen-binding fragments described herein are combined with 5mg-10mg, 5mg-15mg, 5mg-20mg, 5mg-25mg, 25mg-30mg, 20mg-30mg, 15mg-30mg, or 10mg-30mg of tenofovir alafenamide fumarate, tenofovir alafenamide hemifumarate, or tenofovir alafenamide and 200mg of emtricitabine. In certain embodiments, the anti-HIV gp120 V3 glycan-directed antibodies or antigen-binding fragments described herein are combined with 10 mg of tenofovir alafenamide fumarate, tenofovir alafenamide hemifumarate, or tenofovir alafenamide and 200 mg of emtricitabine. In certain embodiments, the anti-HIV gp120 V3 glycan-directed antibodies or antigen-binding fragments described herein are combined with 25 mg of tenofovir alafenamide fumarate, tenofovir alafenamide hemifumarate, or tenofovir alafenamide and 200 mg of emtricitabine. In some embodiments, an anti-HIV gp120 V3 glycan-directed antibody or antigen-binding fragment described herein is combined with an agent provided herein at any dose of the anti-HIV gp120 V3 glycan-directed antibody or antigen-binding fragment (e.g., 1 mg to 500 mg of the anti-HIV gp120 V3 glycan-directed antibody or antigen-binding fragment, as described herein), as if each combination of doses were specifically and individually listed.

[0296] In certain embodiments, the anti-HIV gp120 V3 glycan-directed antibodies or antigen-binding fragments described herein are combined with 200 mg-400 mg of tenofovir disoproxil fumarate, tenofovir disoproxil hemifumarate, or tenofovir disoproxil and 200 mg of emtricitabine. In certain embodiments, the anti-HIV gp120 V3 glycan-directed antibodies or antigen-binding fragments described herein are combined with 200 mg-250 mg, 200 mg-300 mg, 200 mg-350 mg, 250 mg-350 mg, 250 mg-400 mg, 350 mg-400 mg, 300 mg-400 mg, or 250 mg-400 mg of tenofovir disoproxil fumarate, tenofovir disoproxil hemifumarate, or tenofovir disoproxil and 200 mg of emtricitabine. In certain embodiments, the anti-HIV gp120 V3 glycan directed antibodies or antigen-binding fragments described herein are combined with 300 mg of tenofovir disoproxil fumarate, tenofovir disoproxil hemifumarate, or tenofovir disoproxil fumarate and 200 mg of emtricitabine. Anti-HIV gp120 V3 glycan directed antibodies or antigen-binding fragments can be combined with the agents provided herein at any dose (e.g., 1 mg to 500 mg of anti-HIV gp120 V3 glycan directed antibodies or antigen-binding fragments), as if each combination of doses is specifically and individually listed.

[0297] Long-acting HIV inhibitors

[0298] In some embodiments, the anti-HIV gp120 V3 glycan directed antibodies or antigen-binding fragments described herein can be co-administered with long-acting HIV inhibitors. Examples of drugs being developed as long-acting HIV inhibitors include, but are not limited to, cabotegravir LA, rilpivirine LA, any integrase LA, VM-1500 LAI, maraviroc (LAI), tenofovir implant, MK-8591 implant, long-acting dulutegravir.

[0299] In one embodiment, the kit includes an anti-HIV 120V3 glycan-directed antibody or antigen-binding fragment described herein, and one or more (eg, one, two, three, one or two, or one to three) additional therapeutic agents.

[0300] HIV vaccine

[0301] In certain embodiments, the anti-HIV gp120 V3 glycan directed antibodies or antigen-binding fragments described herein are combined with HIV vaccines. Examples of HIV vaccines include peptide vaccines, recombinant subunit protein vaccines, live vector vaccines, DNA vaccines, HIV MAG DNA vaccines, CD4-derived peptide vaccines, vaccine combinations, adenovirus vector vaccines (e.g., Ad5, Ad26, or Ad35), monkey adenovirus (chimpanzee, gorilla, rhesus monkey, i.e., rhAd), adeno-associated virus vector vaccines, chimpanzee adenovirus vaccines (e.g., ChAdOX1, ChAd68, ChAd3, ChAd63, ChAd83, ChAd155, ChAd157, Pan5, Pan6, Pan7, Pan9), coxsackievirus-based vaccines, enterovirus-based vaccines, gorilla adenovirus vaccines, lentiviral vector-based vaccines, vaccines based on two or three parts of arenaviruses (e.g., LCMV, Pichinde virus), trimer-based HIV-1 vaccines , measles virus-based vaccines, flavivirus vector-based vaccines, tobacco mosaic virus vector-based vaccines, varicella zoster virus-based vaccines, human parainfluenza virus 3 (PIV3)-based vaccines, poxvirus-based vaccines (modified Ankara vaccinia virus (MVA), orthopoxvirus-derived NYVAC and fowlpox virus-derived ALVAC (canarypox virus) strains); fowlpox virus-based vaccines, rhabdovirus-based vaccines, such as vesicular stomatitis virus (VSV) and Maraba virus; recombinant human CMV (rhCMV)-based vaccines, alpha-virus-based vaccines, such as Semliki Forest virus, Venezuelan equine encephalitis virus and Sindbis virus (see, e.g., Lauer et al., Clin Vaccine Immunol., 2017, Vol. 24, No. 1, p. e00298-16). LNP-formulated mRNA-based therapeutic vaccines; and LNP-formulated self-replicating RNA / self-amplifying RNA vaccines.

[0302] Examples of HIV vaccines include, but are not limited to, anti-CD40.Env-gp140 vaccine, Ad4-EnvC150, BG505SOSIP.664 gp140 adjuvant vaccine, BG505 SOSIP.GT1.1 gp140 adjuvant vaccine, Chimigen HIV vaccine, ConMSOSIP.v7 gp140, rgp120 (AIDSVAX), ALVAC HIV (vCP1521) / AIDSVAX B / E (gp120) (RV144), monomeric gp120 HIV-1 subtype C vaccine, MPER-656 liposomal subunit vaccine, Remune, ITV-1, Contre Vir, Ad5-ENVA-48, DCVax-001 (CDX-2401), Vacc-4x, Vacc-C5, VAC-3S, multiclade DNA recombinant adenovirus-5 (rAd5), rAd5 gag-pol env A / B / C vaccines, Pennvax-G, Pennvax-GP, Pennvax-G / MVA-CMDR, HIV-TriMix-mRNA vaccine, HIV-LAMP-vax, Ad35, Ad35-GRIN, NAcGM3 / VSSPISA-51, polyICLC adjuvant vaccine, TatImmune, GTU-multiHIV(FIT-06), ChAdV63.HIVconsv, gp140[delta]V2.TV1+MF-59, rVSVIN HIV-1gag vaccine, SeV-EnvF, SeV-Gag vaccine, AT-20, DNK-4, ad35-Grin / ENV, TBC-M4, HIVAX, HIVAX-2, N123-VRC-34.01 inducible epitope-based HIV vaccine, NYVAC-HIV-PT1, NYVAC-HIV-PT4, DNA-HIV-PT123, rAAV1-PG9DP, GOVX-B11, G OVX-B21, GOVX-C55, TVI-HIV-1, Ad-4 (Ad4-env clade C + Ad4-mGag), Paxvax, EN41-UGR7C, EN41-FPA2, ENOB-HV-11, PreVaxTat, AE-H, MYM-V101, CombiHIVvac, ADVAX, MYM-V201, MVA-CMDR, MagaVax, DNA-Ad5 gag / pol / nef / nev (HVTN505), MVATG-17401, ETV-01, CDX-1401, DNA and Sev vector vaccines expressing SCaVII, rcAD26.MOS1.HIV-Env, Ad26.Mod.HIV vaccine.HIV+MVA mosaic vaccine+gp140, AGS-004, AVX-101, AVX-201, PEP-6409, SAV-001, ThV-01, TL-01, TUTI-16, VGX-3300, VIR-1111, IHV-001 and virus-like particle vaccines such as pseudovirion vaccines, CombiVICHvac, LFn-p24 B / C fusion vaccine, GTU-based DNA vaccine, HIV gag / pol / nef / env DNA vaccine, anti-TAT HIV vaccine, combined peptide vaccine, dendritic cell vaccine, gag-based DNA vaccine, GI-2010, gp41 HIV-1 vaccine, HIV vaccine (PIKA adjuvant), I i-key / MHC class II epitope hybrid peptide vaccine, ITV-2, ITV-3, ITV-4, LIPO-5, multiclade Env vaccine, MVA vaccine, Pennvax-GP, pp71-deficient HCMV vector HIV gag vaccine, recombinant peptide vaccine (HIV infection), NCI, rgp160 HIV vaccine, RNActive HIV vaccine, SCB-703, Tat Oyi vaccine, TBC-M4, therapeutic HIV vaccine, UBI HIV gp120, Vacc-4x+romidepsin, variant gp120 peptide vaccine, rAd5gag-pol env A / B / C vaccine; DNA.HTI and MVA.HTI, VRC-HIVDNA016-00-VP+VRC-HIVADV014-00-VP, INO-6145, JNJ-9220, gp145C.6980; based on eOD-GT8 60mer vaccine, PD-201401, ENV (A, B, C, A / E) / gag (C) DNA vaccine, gp120 (A, B, C, A / E) protein vaccine, PDPHV-201401, Ad4-EnvCN54, EnvSeq-1 Env HIV-1 vaccine (GLA-SE adjuvant), HIV p24gag primer boost plasmid DNA vaccine, HIV-1iglb12 neutralizing VRC-01 antibody stimulated anti-CD4 vaccine, MVA-BNHIV-1 vaccine regimen, UBI HIV gp120, mRNA-based preventive vaccine, VPI-211 and TBL-1203HI. .

[0303] Birth control (contraceptive pills) combination therapy

[0304] In certain embodiments, the anti-HIV gp120 V3 glycan directed antibodies or antigen binding fragments described herein are combined with birth control or contraceptive regimens. Therapeutic agents for birth control (contraceptives) include cyproterone acetate, desogestrel, dienogest, drospirenone, estradiol valerate, ethinyl estradiol, norethindrone, etonogestrel, levonorgestrel, levonorgestrel, linegestrel, medroxyprogesterone acetate, ethinyl estradiol methyl ether, mifepristone, misoprostol, nomegestrol acetate, norgestrel, norgestrel, ormeloxifene, segestersone acetate, ulipristal acetate, and any combination thereof.

[0305] Gene therapy and cell therapy

[0306] In certain embodiments, the anti-HIV gp120 V3 glycan-directed antibodies or antigen-binding fragments described herein are combined with gene or cell therapy regimens. Gene therapy and cell therapy include, but are not limited to, gene modifications that silence genes; gene methods that directly kill infected cells; infusion of immune cells designed to replace most of the patient's own immune system to enhance the immune response to infected cells, or activate the patient's own immune system to kill infected cells, or find and kill infected cells; genetic methods that modify cell activity to further change the endogenous immune response to infection. Examples of cell therapies include LB-1903, ENOB-HV-01, GOVX-B01, and SupT1 cell-based therapies. Examples of dendritic cell therapies include AGS-004. CCR5 gene editors include SB-728T. CCR5 gene inhibitors include Cal-1. In some embodiments, CD4-positive T cells expressing C34-CCR5 / C34-CXCR4 are co-administered with anti-HIV gp120 V3 glycan-directed antibodies or antigen-binding fragments. In some embodiments, an anti-HIV gpl20 V3 glycan-directed antibody or antigen-binding fragment is co-administered with AGT-103-transduced autologous T cell therapy or AAV-eCD4-Ig gene therapy.

[0307] Gene editing

[0308] In certain embodiments, the anti-HIV gp120 V3 glycan-directed antibodies or antigen-binding fragments described herein are combined with gene editors, such as HIV targeted gene editors. In various embodiments, the genome editing system can be selected from: CRISPR / Cas9 complexes, zinc finger nuclease complexes, TALEN complexes, homing endonuclease complexes, and meganuclease complexes. Exemplary HIV targeting CRISPR / Cas9 systems include, but are not limited to, EBT-101.

[0309] CAR-T cell therapy

[0310] In some embodiments, the anti-HIV gp120 V3 polysaccharide-directed antibodies or antigen-binding fragments described herein can be co-administered with an immune effector cell population engineered to express a chimeric antigen receptor (CAR), wherein the CAR comprises an HIV antigen binding domain. HIV antigens include HIV envelope proteins or portions thereof, gp120 or portions thereof, CD4 binding sites on gp120, CD4-induced binding sites on gp120, N polysaccharides on gp120, V2 of gp120, and membrane proximal regions on gp41. Immune effector cells are T cells or NK cells. In some embodiments, T cells are CD4+T cells, CD8+T cells, or combinations thereof. Cells may be autologous or allogeneic. Examples of HIV CAR-T include convertible CAR-T, VC-CAR-T, anti-CD4 CART cell therapy, and autologous hematopoietic stem cells genetically engineered to express CD4 CAR and C46 peptides.

[0311] TCR-T cell therapy

[0312] In certain embodiments, the anti-HIV gpl20 V3 glycan-directed antibodies or antigen-binding fragments described herein are combined with a population of TCR-T cells. The TCR-T cells are engineered to target HIV-derived peptides present on the surface of virus-infected cells.

[0313] 6. Test kit

[0314] Also provided are kits for performing diagnostic and therapeutic methods as described herein. In some embodiments, the kit includes primers for amplifying and sequencing at least the gp120 V3 glycan region of HIV species in biological samples. In some embodiments, the kit includes a set or a group of nested primers for amplifying and sequencing at least the gp120 V3 glycan region of HIV species in biological samples. In some embodiments, the kit includes a pair of primers or a set of nested primers for amplifying and sequencing the full-length gp120. In some embodiments, the kit includes sample preparation, nucleic acid quantification, amplification and / or sequencing reagents, such as nucleic acid separation reagents for separating RNA and / or DNA, protein denaturing solvents, buffers, dNTPs, reverse transcriptases, polymerases and / or detection labels. In some embodiments, the kit includes library preparation reagents, such as barcode reagents and / or target-specific primers. In some embodiments, the kit includes analysis guides and / or software, such as to facilitate the implementation of the diagnostic methods described herein. In some embodiments, the kit includes instructions for sequencing at least the gp120 V3 glycan region of an HIV species in a biological sample and detecting or identifying an HIV species expressing a gp120 comprising the following amino acid residues: a glycosylated asparagine at a position corresponding to amino acid residue position 332 (N332 glycan), an aspartic acid at a position corresponding to amino acid residue position 325 (D325), and one or more amino acid residues selected from the following amino acid residues: a threonine at a position corresponding to amino acid residue position 63 (T63), a leucine at a position corresponding to amino acid residue position 179 (L179), a threonine at a position corresponding to amino acid residue position 320 (T320), and a histidine at a position corresponding to amino acid residue position 330 (H330), wherein the amino acid positions are referenced to SEQ ID NO:4 (i.e., residues 1-511 of NCBI Reference Sequence No. NP_057856.1), as described herein.

[0315] In one embodiment, the kit includes one or more pharmaceutical packages, which include one or more containers (e.g., vials, ampoules, preloaded syringes) containing one or more ingredients of the pharmaceutical compositions described herein, such as antibodies or antigen-binding fragments thereof against the HIV gp120 V3 glycan region, or one or more polynucleotides encoding such antibodies or antigen-binding fragments, as provided herein. In some cases, the kit includes a pharmaceutical composition described herein. In some embodiments, the kit includes one or more containers, which include antibodies or antigen-binding fragments thereof against the HIV gp120 V3 glycan region, or one or more polynucleotides encoding such antibodies or antigen-binding fragments in aqueous or lyophilized form. Optionally associated with such containers may be a notice in a form prescribed by a governmental agency regulating the manufacture, use, or sale of drugs or biological products, which reflects the agency's approval of manufacture, use, or sale for human administration.

[0316] Example

[0317] The following examples are provided to illustrate but not to limit the claimed invention.

[0318] Example 1

[0319] HIV-infected individuals responsive to therapy with anti-HIV gp120V3 glycan-directed antibodies or antigen-binding fragments thereof Identification of infected patients

[0320] This example demonstrates the identification of Env genotypes associated with viral susceptibility to neutralization by PGT121 and its derivative GS-9722 (ipavirumab) for pre-screening of HIV-infected subjects for susceptibility to PGT121 / GS-9722.

[0321] The high level of sequence diversity in the HIV Env gene makes it attractive to prescreen subjects in clinical trials for broadly neutralizing antibodies (bNAbs) to increase the likelihood of high response rates. To identify Env genotypes that predict viral susceptibility to PGT121 and GS-9722, we examined the neutralization data and corresponding Env sequences for 206 clade B Envs against PGT121 and GS-9722.

[0322] GS-9722 is an engineered variant of PGT121 that retains the same neutralizing activity as PGT121, as demonstrated by the highly statistically significant correlation of neutralization IC50s of PGT121 and GS-9722 across 397 HIV strains tested with PGT121 and GS-9722 (r 2=0.9698, P<0.0001). Therefore, we combined the GS-9722 neutralization data obtained from 140 clade B Envs isolated from viremic subjects participating in Gilead-sponsored clinical trials with the publicly available PGT121 neutralization data (n=66) obtained from the Los Alamos HIV sequence database to increase statistical power.

[0323] Full-length Env amino acid sequences were aligned using ClustalW and manually adjusted upon visual inspection. To identify genotypes associated with susceptibility to neutralization by PGT121 / GS-9722, we compared the frequency of amino acids and potential N-linked glycosylation sites (PNGS) at each residue in PGT121 / GS-9722-susceptible viruses with that in PGT121 / GS-9722-resistant viruses by Fisher's exact test. The N-linked glycosylation motif is NXS / T, where X is any residue except proline. Neutralization sensitivity to PGT121 / GS-9722 was defined as IC50 < 1 μg / mL. For residues with statistically significant associations with sensitivity to PGT121 / GS-9722, positive predictive value (PPV; i.e., the probability that Env is sensitive to PGT121 / GS-9722 when the genotype is present) and sensitivity (i.e., the probability that the genotype is present when Env is sensitive to PGT121 / GS-9722) were calculated as follows:

[0324] Table 1

[0325] 2 for calculating PPV, NPV, sensitivity and specificity of genotypic determinants of PGT121 / GS-9722 susceptibility ×2 Table

[0326]

[0327]

[0328] The Mann-Whitney test was also applied to identify determinants of susceptibility independent of the 1 μg / mL cutoff value defining Env as "susceptible" versus "resistant."

[0329] Table 2 lists the residues that are statistically associated with PGT121 / GS-9722 susceptibility and / or have been previously reported to be associated with PGT121 susceptibility, in descending order of PPV. Of the residues previously reported to confer susceptibility to PGT121, 307I, 295PNGS, and 300PNGS were not statistically associated with susceptibility to PGT121 / GS-9722 in this clade B dataset. We identified a number of residues that have not been previously reported to be significantly associated with susceptibility to PGT121 / GS-9722.

[0330] Table 2

[0331] Individual genotypes associated with susceptibility to PGT121 / GS-9722 neutralization in clade B Env

[0332]

[0333] 1 Viral genotype, indicating the presence of specific amino acid residues translated from the HIV Env gene

[0334] *Residues reported in the literature to confer susceptibility to neutralization of PGT121 (Julg et al., Sci Transl Med., 2017, Vol. 9, p. 408).

[0335] Since the epitope consists of more than one residue, combinations of genotypic determinants that were statistically associated with PGT121 / GS-9722 susceptibility were evaluated to see if combining individual genotypic determinants would improve the PPV by preferentially enriching true positives over false positives. Sensitivity was also considered, as genotypes with low sensitivity would require screening of a larger number of subjects in order to enroll a sufficient number of subjects in the clinical trial.

[0336] The combined genotypes that provided the highest PPV and sensitivity are listed in Table 3 and are Figure 1 Several combined genotypes that included genotypes not previously reported to be associated with susceptibility to PGT121 / GS-9722 neutralization provided higher PPVs than could be obtained using only previously described genotypes. The highest PPV obtained was 98.4% (for viruses containing amino acid N332 glycan / D325 / H330 / T63 / T320 / L179), which was a 57% increase over the positive predictive value of 62.6% without genotype selection.

[0337] Table 3

[0338] Individual genotypes and combined genes associated with susceptibility to PGT121 / GS-9722 neutralization in clade B Env Cause type

[0339]

[0340] 1 Viral genotype, indicating the presence of specific amino acid residues translated from the HIV Env gene

[0341] 3 None, indicating that the 206 subtype B viruses did not select specific amino acids in the HIV envelope gene

[0342] *Indicates genotypes consisting of residues previously reported in the literature to be associated with susceptibility to PGT121. See, e.g., Julg et al., Sci Transl Med., 2017, Vol. 9, p. 408.

[0343] Using the neutralization data and the corresponding Env sequences of 66 clade A Envs and 258 clade C Envs, the combined genotypes of PGT121 / GS-9722 of clade B in Table 3 were used to determine the PPV, sensitivity and prevalence of clade A (Table 4) and clade C (Table 5). The clade A and clade C data sets are public data obtained from the Los Alamos HIV sequence database. The highest PPV of clade A obtained was 93.8% (for viruses containing amino acid N332 polysaccharide / D325 / H330 / T320 / L179), which increased by 88% compared to the positive predictive value of 50% when there was no genotype selection. The highest PPV of clade C obtained was 89.3% (for viruses containing amino acid N332 polysaccharide / D325 / H330 / T320 / L179), which increased by 53% compared to the positive predictive value of 58.5% when there was no genotype selection.

[0344] Table 4

[0345] Individual and combined genotypes associated with susceptibility to PGT121 neutralization in clade A Env

[0346]

[0347] 1 Viral genotype, indicating the presence of specific amino acid residues translated from the HIV Env gene 2 None, indicating that 66 clade A viruses did not select for a specific amino acid in the HIV envelope gene

[0348] Table 5

[0349] Individual and combined genotypes associated with susceptibility to PGT121 neutralization in clade C Env

[0350]

[0351] 1 Viral genotype, indicating the presence of specific amino acid residues translated from the HIV Env gene

[0352] 2 None, indicating that 258 clade C viruses did not select for a specific amino acid in the HIV envelope gene

[0353] For the evolutionary branch A, evolutionary branch B and evolutionary branch C virus sequences, the prevalence of individual amino acids (T63, L179, T320, D325, H330, N332, NotP333 and S / T334) used in the PGT121 / GS-9722 combined genotype was determined (Table 6). The prevalence of all amino acids in evolutionary branch B, evolutionary branch A except L179 (51.5%) and in evolutionary branch C except T63 (10.1%)) was higher than 60%.

[0354] Table 6

[0355] Prevalence of individual amino acids in clade A, clade B, and clade C viruses

[0356]

[0357] 1 Analysis based on 66 clade A, 206 clade B, and 258 clade C viruses from the PGT121 / GS-9722 dataset

[0358] 10-1074 is a broadly neutralizing antibody that targets the V3 glycan region of HIV gp120 and is associated with PGT121 / GS-9722. See, e.g., Mouquet et al., Proc Natl Acad Sci U SA. November 20, 2012, Vol. 109, No. 47, pp. 3268-3277, and Walker et al., Nature, September 22, 2011, Vol. 477, No. 7365, pp. 466-470. Using the neutralization data and the corresponding Env sequences of 315 clade B Envs (Table 7), the combined genotypes of PGT121 / GS-9722 in Table 3 were used to determine the PPV, sensitivity, and prevalence of 10-1074. The 315 clade B dataset included 143 clade B Envs isolated from viremic subjects participating in Gilead-sponsored clinical trials and 172 clade B Envs from public data obtained from the Los Alamos HIV sequence database. The highest PPV obtained was 100% (for viruses containing amino acid N332 glycan / D325 / H330 / T63 / T320 / L179), which was a 61% increase over the positive predictive value of 62.2% without genotype selection.

[0359] Table 7

[0360] Individual and combined genotypes associated with susceptibility to 10-1074 neutralization in clade B Env

[0361]

[0362] 1Viral genotype, indicating the presence of specific amino acid residues translated from the HIV Env gene 2 None, indicating that 315 clade B viruses did not select for a specific amino acid in the HIV envelope gene

[0363] Subsequently, the highest scoring genotype algorithm (Table 3) was applied to analyze plasma samples from HIV-infected individuals before ART from the Zurich Primary HIV Infection Cohort Study (ZPHI) to predict whether they would be sensitive to GS-9722 treatment. A total of 92 individual plasma samples were analyzed in an NGS assay for HIV envelope genes (GenoSure HIV envelope RNA assay, Monogram Biosciences, South SanFrancisco, CA). A subject was characterized as positive for a given genotype if the derived viral sequence contained the amino acids specified by the algorithm without sequence variability (sequence variability at the specified position was zero). Based on these criteria, 47 / 92, 37 / 92, 32 / 92, 27 / 92, 22 / 92, and 16 / 92 subjects were predicted to be sensitive to GS-9722 ( Figure 1 ), with corresponding positive predictive values ​​of 80.7%, 83.5%, 86.1%, 91.6%, 93.7%, and 98.4%, respectively (Table 3). For subjects infected with clade B (59 of 92 subjects), 35 / 59, 27 / 59, 22 / 59, 23 / 59, 18 / 59, and 12 / 59 were predicted to be susceptible to GS-9722 ( Figure 2 ), with the corresponding positive predictive values ​​being 80.7%, 83.5%, 86.1%, 91.6%, 93.7% and 98.4%, respectively (Table 3).

[0364] For pre-ART plasma samples of all subjects (n=92) and a subset of subjects infected with evolutionary branch B (n=59), 100% conservation (zero sequence variation at the specified position) of each amino acid (T63, L179, T320, D325, H330, N332, NotP333 and S / T334) in the combined genotype used for GS-9722 susceptibility prediction was determined (Table 8).

[0365] Table 8 100% conservation of individual amino acids among ZPHI subjects

[0366]

[0367] 1 Analysis based on 92 (all subjects) and 59 (clade B subjects) pre-ART plasma samples from ZPHI individuals

[0368] To confirm genotypic predictions for GS-9722 sensitivity, viral populations from pre-ART plasma samples from ZPHI were cloned and evaluated in a GS-9722 neutralization assay (PhenoSense HIV Entry Assay, Monogram Biosciences, South San Francisco, CA). Viruses were derived from 29 clade B samples with a positive predictive value of 80.7% or higher. Derived viruses were characterized as GS-9722 sensitive when the IC50 was 1 μg / ml or lower. Based on these criteria, 25 / 29, 20 / 22, 16 / 18, 18 / 20, 14 / 16, and 10 / 11 viruses were confirmed to be sensitive to GS-9722 ( Figure 2 ), with the corresponding positive predictive values ​​being 80.7%, 83.5%, 86.1%, 91.6%, 93.7% and 98.4%, respectively (Table 3).

[0369] To further confirm the genotypic prediction and phenotypic sensitivity to GS-9722, 20 individual viruses from four viral swarms from pre-ART plasma samples from ZPHI were subcloned and evaluated in a GS-9722 neutralization assay (PhenoSense HIV Entry Assay, Monogram Biosciences, South San Francisco, CA). All individual viruses were sensitive to GS-9722 with IC50 values ​​comparable to those of the swarm viruses ( Figure 4 ).

[0370] It should be understood that the embodiments and implementations described herein are for illustrative purposes only, and those skilled in the art will propose various modifications or changes based on these embodiments and implementations, and include them in the spirit and scope of this patent application and the scope of the appended claims. All publications, patents and patent applications cited herein are hereby incorporated by reference in their entirety for all purposes.

[0371] Preferred embodiment:

[0372] 1. A method for treating or preventing HIV in a human subject in need thereof, the method comprising:

[0373] a) identifying a human subject infected with HIV or a population of HIV expressing a gp120 comprising the following amino acid residues: N332 glycan, D325, and one or more amino acid residues selected from the group consisting of: T63, L179, T320, and H330, wherein the amino acid positions are referenced to SEQ ID NO: 4; and

[0374] b) administering to the subject an effective amount of an antibody or antigen-binding fragment thereof that competes with or comprises a VH region and a VL region that bind to an epitope of gp120 within the third variable loop (V3) and / or the high mannose patch comprising the N332 oligomannose glycan.

[0375] 2. The method according to item 1, comprising identifying a subject infected with HIV or a population of HIV expressing gp120 comprising the following amino acid residues:

[0376] i. N332 glycan, D325 and T63;

[0377] ii. N332 glycan, D325 and L179;

[0378] iii. N332 glycan, D325 and T320;

[0379] iv. N332 glycan, D325 and H330;

[0380] v.N332 glycan, D325, T63, and L179;

[0381] vi. N332 glycan, D325, T63 and T320;

[0382] vii. N332 glycan, D325, T63 and H330;

[0383] viii. N332 glycan, D325, L179 and T320;

[0384] ix. N332 glycan, D325, L179 and H330;

[0385] x.N332 glycan, D325, T320, and H330;

[0386] xi. N332 glycan, D325, T63, T320, and H330;

[0387] xii. N332 glycan, D325, T63, L179 and T320;

[0388] xiii. N332 glycan, D325, T63, L179 and H330;

[0389] xiv. N332 glycan, D325, L179, T320 and H330; or

[0390] xv. N332 glycan, D325, T63, L179, T320 and H330.

[0391] 3. The method according to item 1, comprising identifying an infected cell expressing a gene comprising the following amino acid sequence:

[0392] HIV or HIV population subjects with gp120 containing amino acid residues:

[0393] i. N332 glycan, D325 and T63;

[0394] ii. N332 glycan, D325 and L179;

[0395] iii. N332 glycan, D325 and T320; or

[0396] iv. N332 glycan, D325 and H330.

[0397] 4. A method according to any one of items 1 to 2, comprising identifying an infected cell expressing

[0398] A subject with HIV or a population of HIV whose gp120 contains the following amino acid residues:

[0399] i. N332 glycan, D325, T63 and L179;

[0400] ii. N332 glycan, D325, T63 and T320;

[0401] iii. N332 glycan, D325, T63 and H330;

[0402] iv. N332 glycan, D325, L179 and T320;

[0403] v. N332 glycan, D325, L179 and H330; or

[0404] vi. N332 glycan, D325, T320 and H330.

[0405] 5. A method according to any one of items 1 to 5, comprising identifying an infected cell expressing

[0406] A subject with HIV or a population of HIV whose gp120 contains the following amino acid residues:

[0407] i. N332 glycan, D325, L179, T320 and H330;

[0408] ii. N332 glycan, D325, T63, T320 and H330;

[0409] iii. N332 glycan, D325, T63, L179 and T320; or

[0410] iv. N332 glycan, D325, T63, L179 and H330.

[0411] 6. A method according to any one of items 1 to 5, comprising identifying an infected cell expressing

[0412] A subject with HIV or a population of HIV whose gp120 contains the following amino acid residues:

[0413] i. N332 glycan, D325, T63 and H330;

[0414] ii. N332 glycan, D325, T320 and H330;

[0415] iii. N332 glycan, D325, L179, T320 and H330; or

[0416] iv. N332 glycan, D325, T63, L179, T320 and H330.

[0417] 7. The method according to any one of items 1 to 6, wherein the subject is infected with HIV or a population of HIV that expresses gp120 further comprising one or more of the following amino acid residues:

[0418] i. Polysaccharide 301;

[0419] ii.K677;

[0420] iii.not_W17;

[0421] iv.not_R747;

[0422] v.insertion_321.01;

[0423] vi.E429;

[0424] vii.Q442;

[0425] viii.R335;

[0426] ix.I165;

[0427] x.S393;

[0428] xi.I307;

[0429] xii.295 polysaccharides; and / or

[0430] xiii.N300.

[0431] 8. A method according to any one of items 1 to 7, wherein at least 90%, for example, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the HIV species in the HIV population contain the amino acid residue.

[0432] 9. The method according to any one of items 1 to 8, wherein the antibody or antigen-binding fragment thereof administered competes with or comprises the VH region and VL region of an antibody selected from the group consisting of GS-9722, GS-9721, PGT-121, PGT-121.66, PGT-121.414, PGT-122, PGT-123, PGT-124, PGT-125, PGT-126, PGT-128, PGT-130, PGT-133, PGT-134, PGT-135, PGT- 136. PGT-137, PGT-138, PGT-139, GS-2872, 10-1074, 10-1074-J, VRC24, 2G12, BG18, 354BG8, 354BG18, 354BG42, 354BG33, 354BG129, 354BG188, 354BG411, 354BG426, DH270.1, DH270.6, PGDM12, VRC41.01, PGDM21, PCDN-33A, BF520.1 and VRC29.03.

[0433] 10. The method according to any one of items 1 to 9, wherein the antibody or antigen-binding fragment thereof competes with or comprises a VH region and a VL region from an antibody selected from the group consisting of:

[0434] Zone and VL zone: GS-9722, GS-9721, PGT-121, PGT-121.66, PGT-121.414, PGT-124, PGT-134, GS-2872, 10-1074, 10-1074-J, PGT-122 and PGT-123.

[0435] 11. The method according to any one of items 1 to 10, wherein the antibody comprises an Fc region,

[0436] The Fc region comprises the following amino acids at the indicated positions (EU index numbering):

[0437] i. tyrosine at position 252, threonine at position 254, and glutamic acid (YTE) at position 256; or

[0438] ii. Leucine at position 428 and serine (LS) at position 434.

[0439] 12. The method according to any one of items 1 to 11, wherein the antibody comprises an Fc region,

[0440] The Fc region comprises the following amino acids at the indicated positions (EU index numbering):

[0441] i. Aspartic acid at position 239 and glutamic acid at position 332 (DE);

[0442] ii. Aspartic acid at position 239, glutamic acid at position 332, and leucine at position 330 (DEL);

[0443] iii. Aspartic acid at position 239, glutamic acid at position 332, alanine (DEA) at position 236; or

[0444] iv. Aspartic acid at position 239, glutamic acid at position 332, alanine at position 236 and leucine at position 330 (DEAL).

[0445] 13. The method according to any one of items 1 to 10, comprising administering an antigen binding fragment.

[0446] 14. The method according to item 13, wherein the antigen binding fragment is selected from: scFv, Fab, Fab2, Fab', F(ab')2, Fv and bivalent antibody.

[0447] 15. The method according to any one of items 1 to 14, wherein the antibody is a multispecific antibody.

[0448] 16. The method according to any one of items 1 to 15, wherein the human subject is acutely infected with HIV.

[0449] 17. The method according to item 16, wherein the antibody is administered to a human subject having Fiebig stage IV or earlier HIV infection.

[0450] 18. The method according to item 16, wherein the antibody is administered to a human subject who has not seroconverted.

[0451] 19. A method according to any one of items 1 to 15, wherein the human subject has recently been infected with HIV.

[0452] 20. The method according to item 19, wherein the antibody is administered to a human subject with HIV infection at Fiebig stage V or Fiebig stage VI.

[0453] 21. The method according to any one of items 1 to 15, wherein the human subject is chronically infected with HIV.

[0454] 22. A method according to any one of items 1 to 21, wherein the human subject is infected with an HIV clade B virus.

[0455] 23. The method according to item 22, comprising identifying an infected cell expressing an amino acid comprising

[0456] Subjects with HIV or HIV populations containing gp120 containing amino acid residues:

[0457] i. N332 glycan, D325, T63 and H330;

[0458] ii. N332 glycan, D325, L179, T320 and H330; or

[0459] iii. N332 glycan, D325, T63, L179, T320 and H330.

[0460] 24. A method according to any one of items 1 to 23, wherein the human subject is infected with an HIV clade A virus.

[0461] 25. A method according to any one of items 1 to 24, wherein the human subject is infected with an HIV clade C virus.

[0462] 26. The method according to any one of items 1 to 25, further comprising administering to the subject one or more additional therapeutic agents for treating HIV infection.

[0463] 27. A method according to any one of items 1 to 26, wherein the subject is not receiving antiretroviral therapy (ART), or has discontinued ART prior to administration of the antibody.

[0464] 28. The method according to any one of items 1 to 26, wherein ART is discontinued after one or more administrations of the antibody or antigen-binding fragment thereof.

[0465] 29. A method according to any one of items 1 to 26, further comprising administering one or more antiretroviral therapy (ART) agents to the subject.

[0466] 30. The method according to any one of items 1 to 29, further comprising administering to the subject a second antibody or antigen-binding fragment thereof, wherein the second antibody or antigen-binding fragment thereof

[0467] The invention relates to a fragment that binds to an epitope or region of gp120 selected from the group consisting of:

[0468] i. the second variable loop (V2) and / or the Env trimer vertex;

[0469] ii.CD4 binding site (CD4bs);

[0470] iii. gp120 / gp41 interface; or

[0471] iv.gp120 Silent side.

[0472] 31. A method according to claim 30, wherein the second antibody or its antigen-binding fragment binds to an epitope or region of gp120 in the second variable loop (V2) and / or the apex of the Env trimer, and competes with or comprises the VH and VL regions of an antibody selected from the group consisting of: PG9, PG16, PGC14, PGG14, PGT-142, PGT-143, PGT-144, PGT-145, CH01, CH59, PGDM1400, CAP256, CAP256-VRC26.08, CAP256-VRC26.09, CAP256-VRC26.25, PCT64-24E and VRC38.01.

[0473] 32. The method according to item 30, wherein the second antibody or antigen-binding fragment thereof binds to an epitope or region of gp120 in the CD4 binding site (CD4bs) and competes with or comprises the VH region and VL region of an antibody selected from the group consisting of:

[0474] b12, F105, VRC01, VRC07, VRC07-523, VRC03, VRC06, VRC06b01 VRC08, VRC0801, NIH45-46, GS-9723, GS-5423, 3BNC117, 3BNC60, VRC-PG04, PGV04; CH103, 44-VRC13.01, 1NC9, 12A12, N6, N6LS (VRC-HIVMAB091-00-AB), N49-P7, NC-Cow1, IOMA, CH235 and CH235.12, N49P6, N49P7, N49P11, N49P9 and N60P25.

[0475] 33. The method of item 30, wherein the second antibody or antigen-binding fragment thereof binds to an epitope or region of gp120 in the gp120 / gp41 interface and competes with or comprises the VH and VL regions of an antibody selected from:

[0476] PGT-151, CAP248-2B, 35O22, 8ANC195, ACS202, VRC34, and VRC34.01.

[0477] 34. The method according to item 30, wherein the second antibody or antigen-binding fragment thereof binds to an epitope or region of the silent face of gp120 and competes with or comprises the VH and VL regions from antibody VRC-PG05.

[0478] 35. The method of item 30, wherein the second antibody or antigen-binding fragment thereof binds to an epitope or region of gp41 in the membrane proximal region (MPER) and competes with or comprises the VH region and VL region of an antibody selected from the group consisting of: 10E8,

[0479] 10E8v4, 10E8-5R-100cF, 4E10, DH511.11P, 2F5, 7b2, and LN01.

[0480] 36. The method according to item 30, wherein the second antibody or antigen-binding fragment thereof binds to an epitope or region of the gp41 fusion peptide and binds to a VH from an antibody selected from the group consisting of:

[0481] The VH and VL regions compete with or comprise the VH and VL regions: VRC34 and ACS202.

[0482] 37. The method according to any one of items 1 to 36, further comprising administering a TLR agonist to the subject.

[0483] 38. The method according to item 37, wherein the TLR agonist is a TLR2 agonist, a TLR3 agonist, a TLR7 agonist, a TLR8 agonist or a TLR9 agonist.

[0484] 39. The method according to item 38, wherein the TLR7 agonist is selected from: visammod,

[0485] Imiquimod and resiquimod.

[0486] 40. The method according to any one of items 1 to 39, comprising administering the antibody or antigen-binding fragment thereof multiple times at predetermined intervals, optionally together with a TLR agonist.

[0487] 41. A method according to any one of items 1 to 40, wherein after one or more administrations of the antibody or antigen-binding fragment thereof, in the absence of antiretroviral therapy (ART), the subject does not show symptoms of HIV or AIDS for at least 6 months, at least 1 year, at least 2 years, at least 3 years or longer.

[0488] 42. A method according to any one of items 1 to 41, wherein after one or more administrations of the antibody, in the absence of antiretroviral therapy (ART), the subject has a viral load copy / ml blood of less than 500, such as less than 400, less than 300, less than 200, less than 100, less than 50 for at least 6 months, at least 1 year, at least 2 years, at least 3 years or longer.

[0489] 43. A method for identifying a human subject infected with HIV or a population of HIV susceptible to an antibody or antigen binding fragment thereof, said antibody or antigen binding fragment thereof competing with or comprising a VH region and a VL region that bind to an epitope of gp120 within the third variable loop (V3) and / or a high mannose patch comprising an N332 oligomannose glycan, said method comprising identifying in a biological sample from said human subject HIV expressing gp120 comprising the following amino acid residues: the N332 glycan, D325, and one or more amino acid residues selected from the group consisting of: T63, L179, T320, and H330, wherein said amino acid positions are referenced to SEQ ID NO:4.

[0490] 44. A method according to item 43, wherein the subject is infected with a gene expressing a gene comprising the following amino acid sequence:

[0491] Acid residues of gp120 HIV or HIV groups:

[0492] i. N332 glycan, D325 and T63;

[0493] ii. N332 glycan, D325 and L179;

[0494] iii. N332 glycan, D325 and T320;

[0495] iv. N332 glycan, D325 and H330;

[0496] v.N332 glycan, D325, T63, and L179;

[0497] vi. N332 glycan, D325, T63 and T320;

[0498] vii. N332 glycan, D325, T63 and H330;

[0499] viii. N332 glycan, D325, L179 and T320;

[0500] ix. N332 glycan, D325, L179 and H330;

[0501] x.N332 glycan, D325, T320, and H330;

[0502] xi. N332 glycan, D325, T63, T320, and H330;

[0503] xii. N332 glycan, D325, T63, L179 and T320;

[0504] xiii. N332 glycan, D325, T63, L179 and H330;

[0505] xiv. N332 glycan, D325, L179, T320 and H330; or

[0506] xv. N332 glycan, D325, T63, L179, T320 and H330.

[0507] 45. A method according to any one of items 43 to 44, wherein the subject is infected with a virus expressing

[0508] HIV or HIV populations having gp120 comprising the following amino acid residues:

[0509] i. N332 glycan, D325 and T63;

[0510] ii. N332 glycan, D325 and L179;

[0511] iii. N332 glycan, D325 and T320; or

[0512] iv. N332 glycan, D325 and H330.

[0513] 46. ​​A method according to any one of items 43 to 45, wherein the subject is infected with a virus expressing

[0514] HIV or HIV populations having gp120 comprising the following amino acid residues:

[0515] i. N332 glycan, D325, T63 and L179;

[0516] ii. N332 glycan, D325, T63 and T320;

[0517] iii. N332 glycan, D325, T63 and H330;

[0518] iv. N332 glycan, D325, L179 and T320;

[0519] v. N332 glycan, D325, L179 and H330; or

[0520] vi. N332 glycan, D325, T320 and H330.

[0521] 47. The method according to any one of items 43 to 46, comprising identifying a subject infected with HIV or a population of HIV expressing gp120 comprising the following amino acid residues:

[0522] i. N332 glycan, D325, L179, T320 and H330;

[0523] ii. N332 glycan, D325, T63, T320 and H330;

[0524] iii. N332 glycan, D325, T63, L179 and T320; or

[0525] iv. N332 glycan, D325, T63, L179 and H330.

[0526] 48. The method according to any one of items 43 to 47, comprising identifying a subject infected with HIV or a population of HIV expressing gp120 comprising the following amino acid residues:

[0527] i. N332 glycan, D325, T63 and H330;

[0528] ii. N332 glycan, D325, T320 and H330;

[0529] iii. N332 glycan, D325, L179, T320 and H330; or

[0530] iv. N332 glycan, D325, T63, L179, T320 and H330.

[0531] 49. The method according to any one of items 43 to 48, comprising identifying a subject infected with HIV or a population of HIV expressing gp120 comprising the following amino acid residues:

[0532] i. Polysaccharide 301;

[0533] ii.K677;

[0534] iii.not_W17;

[0535] iv.not_R747;

[0536] v.insertion_321.01;

[0537] vi.E429;

[0538] vii.Q442;

[0539] viii.R335;

[0540] ix.I165;

[0541] x.S393;

[0542] xi.I307;

[0543] xii.295 polysaccharides; and / or

[0544] xiii.N300.

[0545] 50. A method according to any one of items 43 to 49, wherein at least 90%, for example, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the HIV species in the HIV population contain the amino acid residue.

[0546] 51. The method according to any one of items 43 to 50, wherein the antibody or antigen-binding fragment thereof competes with or comprises the VH region and VL region of an antibody selected from the group consisting of: GS-9722, GS-9721, PGT-121, PGT-121.66, PGT-121.414, PGT-122, PGT-123, PGT-124, PGT-125, PGT-126, PGT-128, PGT-130, PGT-133, PGT-134, PGT-135, PGT-136, PGT-137, PGT-138, PGT-139, 10-1074, 10-1074-J, VRC24, 2G12, BG18, 354BG8, 354BG18, 354BG42, 354BG33, 35 4BG129, 354BG188, 354BG411, 354BG426, DH270.1, DH270.6, PGDM12, VRC41.01, PGDM21, PCDN-33A, BF520.1 and VRC29.03.

[0547] 52. A method according to any one of items 43 to 51, wherein the antibody or antigen-binding fragment thereof competes with or comprises the VH region and VL region of an antibody selected from the following antibodies: GS-9722, GS-9721, PGT-121, PGT-121.66, PGT-121.414, PGT-124, PGT-134, GS-2872, 10-1074, 10-1074-J, PGT-122 and PGT-123.

[0548] 53. A method according to any one of items 43 to 52, wherein the human subject is acutely infected with HIV.

[0549] 54. The method according to item 53, wherein the antibody is administered to a human subject having Fiebig stage IV or earlier HIV infection.

[0550] 55. The method according to item 53, wherein the antibody is administered to a human subject who has not seroconverted.

[0551] 56. A method according to any one of items 43 to 52, wherein the human subject has recently been infected with HIV.

[0552] 57. The method according to item 56, wherein the antibody is administered to a human subject with HIV infection at Fiebig stage V or Fiebig stage VI.

[0553] 58. A method according to any one of items 43 to 52, wherein the human subject is chronically infected with HIV.

[0554] 59. A method according to any one of items 43 to 58, wherein the human subject is infected with an HIV clade B virus.

[0555] 60. The method according to item 59, comprising identifying a cell infected with an expression comprising the following amino acid

[0556] Subjects with HIV or HIV populations containing gp120 containing amino acid residues:

[0557] i. N332 glycan, D325, T63 and H330;

[0558] ii. N332 glycan, D325, L179, T320 and H330; or

[0559] iii. N332 glycan, D325, T63, L179, T320 and H330.

[0560] 61. A method according to any one of items 43 to 60, wherein the human subject is infected with an HIV clade A virus.

[0561] 62. A method according to any one of items 43 to 61, wherein the human subject is infected with an HIV clade C virus.

[0562] 63. The method according to any one of items 1 to 63, wherein the gp120 amino acid is identified in one or more gp120 polypeptide sequences expressed in HIV or a population of HIV isolated from the subject.

[0563] 64. The method according to any one of items 1 to 64, wherein the gp120 amino acid is identified in one or more gp120 polynucleotide sequences in HIV or a population of HIV isolated from the subject.

[0564] 65. The method according to item 64, comprising performing next generation sequencing (NGS) on the polynucleotide sequence encoding gp120 from the HIV population.

[0565] 66. The method according to item 65, wherein the gp120 variant is detected at a frequency level of about 1% of the viral population.

[0566] 67. A method according to any one of items 1 to 66, wherein the gp120 amino acid is identified in one or more biological samples from the subject, wherein the one or more biological samples are obtained from blood, peripheral blood mononuclear cells (PBMC), serum, plasma, semen or lymph nodes.

[0567] 68. A method according to any one of items 1 to 67, comprising identifying HIV RNA populations in a serum or plasma sample.

[0568] 69. The method according to any one of items 1 to 68, further comprising the step of obtaining one or more biological samples from the subject.

[0569] 70. The method according to item 69, wherein two or more biological samples are obtained from the subject.

[0570] 71. The method according to item 70, wherein the two or more biological samples are obtained from the same tissue or fluid at two or more different time points.

[0571] 72. The method according to item 70, wherein the two or more biological samples are obtained from different tissues or fluids or from different anatomical locations.

Claims

1. An antibody or antigen-binding fragment thereof comprising a VH region and a VL region, wherein the VH region and the VL region bind to an epitope of HIV gp120 within the third variable loop (V3) comprising the N332 oligomannosyl glycan, for use in a method of treating or preventing HIV in a human subject in need thereof, wherein the human subject is infected with HIV or a population of HIV that expresses HIV gp120 comprising the following amino acid residues: N332 glycan, D325, L179 and H330, wherein the amino acid positions are referenced to SEQ ID NO: 4, and wherein the antibody or antigen-binding fragment thereof comprises a VH region and a VL region, wherein the VH region and the VL region respectively comprise the amino acid sequences shown below: i. SEQ ID NO: 400 and 401; ii. SEQ ID NO: 402 and 404; or iii. SEQ ID NO:405 and 406.

2. The antibody or antigen-binding fragment thereof for use according to claim 1, wherein: a) the antibody comprises an Fc region comprising the following amino acids at the indicated positions, wherein the positions are numbered according to the EU index: i. tyrosine at position 252, threonine at position 254, and glutamic acid (YTE) at position 256; or ii. Leucine at position 428 and serine (LS) at position 434; and / or b) the antibody comprises an Fc region comprising the following amino acids at the specified positions, wherein the positions are numbered according to the EU index: i. Aspartic acid at position 239 and glutamic acid at position 332 (DE); ii. Aspartic acid at position 239, glutamic acid at position 332, and leucine at position 330 (DEL); iii. Aspartic acid at position 239, glutamic acid at position 332, alanine (DEA) at position 236; or iv. Aspartic acid at position 239, glutamic acid at position 332, alanine at position 236 and leucine at position 330 (DEAL).

3. The antibody or antigen-binding fragment thereof for use according to claim 1 or claim 2, wherein: a) the method comprises administering an antigen binding fragment, optionally wherein the antigen binding fragment is selected from the group consisting of: scFv, Fab, Fab2, Fab', F(ab')2, Fv and a diabody; b) the antibody is a multispecific antibody; and / or c) the human subject: i) acutely infected with HIV, optionally wherein: A) administering the antibody to a human subject having Fiebig stage IV or earlier HIV infection; or B) administering the antibody to a human subject who has not seroconverted; ii) recently infected with HIV, optionally wherein the antibody is administered to a human subject with HIV infection at Fiebig stage V or Fiebig stage VI; or iii) Chronic infection with HIV.

4. The antibody or antigen-binding fragment thereof for use according to any one of claims 1 to 3, wherein: a) the human subject is infected with an HIV clade B virus, optionally wherein the subject is infected with HIV or a population of HIV that expresses HIV gp120 comprising the following amino acid residues: i. N332 glycan, D325, L179, T320 and H330; or ii. N332 glycan, D325, T63, L179, T320 and H330; b) the human subject is infected with HIV clade A virus; c) the human subject is infected with an HIV clade C virus; and / or d) The method further comprises administering to the subject one or more additional therapeutic agents for treating HIV infection.

5. The antibody or antigen-binding fragment thereof for use according to any one of claims 1 to 4, wherein: a) the subject is not receiving antiretroviral therapy (ART) or has discontinued ART before administration of the antibody; b) ART is discontinued after one or more administrations of the antibody or antigen-binding fragment thereof; or c) The method further comprises administering to the subject one or more antiretroviral therapy (ART) agents.

6. The antibody or antigen-binding fragment thereof for use according to any one of claims 1 to 5, wherein: a) the method further comprises administering to the subject a TLR agonist, optionally wherein the TLR agonist is a TLR2 agonist, a TLR3 agonist, a TLR7 agonist, a TLR8 agonist or a TLR9 agonist, optionally wherein the TLR7 agonist is selected from the group consisting of: visammod, imiquimod and resiquimod; b) the method comprises administering the antibody or antigen-binding fragment thereof multiple times at predetermined intervals, optionally together with a TLR agonist; c) the subject does not exhibit symptoms of HIV or AIDS for at least 6 months, at least 1 year, at least 2 years, at least 3 years, or longer, in the absence of antiretroviral therapy (ART) following one or more administrations of the antibody or antigen-binding fragment thereof; and / or d) the subject has a viral load copy / ml blood of less than 500, e.g., less than 400, less than 300, less than 200, less than 100, less than 50, for at least 6 months, at least 1 year, at least 2 years, at least 3 years or longer after one or more administrations of the antibody, in the absence of antiretroviral therapy (ART).

7. An in vitro method for identifying a human subject infected with HIV or a population of HIV susceptible to an antibody or antigen-binding fragment thereof, the antibody or antigen-binding fragment thereof comprising a VH region and a VL region, the VH region and the VL region binding to an epitope of HIV gp120 within the third variable loop (V3) comprising an N332 oligomannosyl glycan, the method comprising identifying in a biological sample from the human subject HIV expressing HIV gp120 comprising the following amino acid residues as determined by polynucleotide or polypeptide sequencing: N332 glycan, D325, L179 and H330, wherein the amino acid positions are referenced to SEQ ID NO: 4, and wherein the antibody or antigen-binding fragment thereof comprises a VH region and a VL region, the VH region and the VL region respectively comprising the amino acid sequences shown below: i. SEQ ID NO: 400 and 401; ii. SEQ ID NO: 402 and 404; or iii. SEQ ID NO:405 and 406.

8. The method according to claim 7, wherein: a) the human subject is acutely infected with HIV, optionally wherein the antibody has been administered to a human subject: i) have Fiebig stage IV or earlier HIV infection; or ii) no seroconversion; b) the human subject has recently been infected with HIV, optionally wherein the antibody has been administered to a human subject with HIV infection at Fiebig stage V or Fiebig stage VI; or c) The human subject is chronically infected with HIV.

9. The method according to claim 7 or 8, wherein: a) the human subject is infected with an HIV clade B virus, optionally wherein the method comprises identifying a subject infected with HIV or a population of HIV that expresses HIV gp120 comprising the following amino acid residues: i. N332 glycan, D325, L179, T320 and H330; or ii. N332 glycan, D325, T63, L179, T320 and H330; b) the human subject is infected with an HIV clade A virus; and / or c) the human subject is infected with an HIV clade C virus.

10. The method according to any one of claims 7 to 9, wherein: a) identifying said HIV gp120 amino acid in one or more HIV gp120 polypeptide sequences expressed in HIV isolated from said subject or in a HIV population; b) identifying said HIV gp120 amino acid in one or more HIV gp120 polynucleotide sequences in HIV isolated from said subject or in a HIV population, optionally wherein said method comprises performing next generation sequencing (NGS) of polynucleotide sequences encoding HIV gp120 from a HIV population, optionally wherein HIV gp120 variants are detected at a frequency level of about 1% of said viral population; and / or c) identifying the HIV gp120 amino acid in one or more biological samples from the subject, wherein the one or more biological samples are obtained from blood, peripheral blood mononuclear cells (PBMC), serum, plasma, semen, or lymph nodes.

11. The method according to any one of claims 7 to 10: a) involves identifying HIV RNA populations in serum or plasma samples; and / or b) wherein one or more biological samples are obtained from the subject, optionally wherein two or more biological samples are obtained from the subject, optionally wherein the two or more biological samples are obtained from: i) from the same tissue or fluid at two or more different time points; or ii) Different tissues or fluids or different anatomical locations.

Images