[go: up one dir, main page]
More Web Proxy on the site http://driver.im/

WO2024091962A1 - Compositions comprenant des enveloppes modifiées pour activer des précurseurs d'anticorps largement neutralisants de site de liaison à cd4 - Google Patents

Compositions comprenant des enveloppes modifiées pour activer des précurseurs d'anticorps largement neutralisants de site de liaison à cd4 Download PDF

Info

Publication number
WO2024091962A1
WO2024091962A1 PCT/US2023/077669 US2023077669W WO2024091962A1 WO 2024091962 A1 WO2024091962 A1 WO 2024091962A1 US 2023077669 W US2023077669 W US 2023077669W WO 2024091962 A1 WO2024091962 A1 WO 2024091962A1
Authority
WO
WIPO (PCT)
Prior art keywords
envelope
hiv
recombinant
composition
nucleic acid
Prior art date
Application number
PCT/US2023/077669
Other languages
English (en)
Inventor
James COUNTS
Barton F. Haynes
Kevin O. SAUNDERS
Original Assignee
Duke University
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Duke University filed Critical Duke University
Publication of WO2024091962A1 publication Critical patent/WO2024091962A1/fr

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/005—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from viruses
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/12—Viral antigens
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • A61P31/14—Antivirals for RNA viruses
    • A61P31/18—Antivirals for RNA viruses for HIV
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2740/00—Reverse transcribing RNA viruses
    • C12N2740/00011—Details
    • C12N2740/10011—Retroviridae
    • C12N2740/16011—Human Immunodeficiency Virus, HIV
    • C12N2740/16111—Human Immunodeficiency Virus, HIV concerning HIV env
    • C12N2740/16122—New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2740/00—Reverse transcribing RNA viruses
    • C12N2740/00011—Details
    • C12N2740/10011—Retroviridae
    • C12N2740/16011—Human Immunodeficiency Virus, HIV
    • C12N2740/16111—Human Immunodeficiency Virus, HIV concerning HIV env
    • C12N2740/16134—Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein

Definitions

  • the present invention relates in general, to a composition suitable for use in inducing anti-HIV-1 antibodies, and, in particular, to immunogenic compositions comprising envelope proteins and nucleic acids to induce cross-reactive neutralizing antibodies and increase their breadth of coverage.
  • the invention also relates to methods of inducing such broadly neutralizing anti-HIV-1 antibodies using such compositions.
  • the invention provides compositions and method for induction of immune response, for example cross-reactive (broadly) neutralizing Ab (bNAb) induction.
  • bNAb cross-reactive neutralizing Ab
  • the invention provides a CH505 envelope immunogen comprising an optimized D loop, V5 loop, and/or CD4 binding loop.
  • the invention 1 ActiveUS 201518670 Attorney Docket No.: 1234300.00424WO1 (DU7956PCT) provides CH505 T/F envelopes comprising an optimized D loop. In certain aspects the invention provides CH505 T/F envelopes comprising an optimized V5 loop. [0007] In certain aspects, the invention provides a recombinant protein or nucleic acid encoding a recombinant protein as described in Table 1. In certain aspects, the invention provides a selection of HIV-1 envelopes for use as prime and boost immunogens in methods to induce HIV-1 neutralizing antibodies. In certain aspects, the invention provides a selection of HIV-1 envelopes for use as boost immunogens in methods to induce HIV-1 neutralizing antibodies.
  • the invention provides one for more HIV-1 envelopes for use as a prime to induce HIV-1 neutralizing antibodies.
  • the invention provides a recombinant HIV-1 envelope sequence or nucleic acid encoding a recombinant protein HIV-1 envelope sequence comprising a mutation of one or more of amino acid residue R273, S274, N276, N279, I277, T278, K282, I284, R456, or M475.
  • the mutation is N276G.
  • the mutation is I277P.
  • the mutation is T278A.
  • the mutation is T278R.
  • the mutation is K282R.
  • the mutation is R456Q. In certain embodiments, the mutation is R273S. In certain embodiments, the mutation is S274Q. In certain embodiments, the mutation is S274V. In certain embodiments, the mutation is N276K. In certain embodiments, the mutation is N279H. In certain embodiments, the mutation is N279L. In certain embodiments, the mutation is N279R. In certain embodiments, the mutation is I284W. In certain embodiments, the mutation is M475L. In certain aspects, a recombinant HIV-1 envelope comprises any combination of mutations described herein. In certain aspects, a recombinant HIV-1 envelope comprises mutations N279H, K282R and R456Q.
  • a recombinant HIV-1 envelope comprises mutations N279H and K282R. In certain aspects, a recombinant HIV-1 envelope comprises mutations N279H and R456Q. In certain aspects, a recombinant HIV-1 envelope comprises mutations K282R and R456Q. In certain aspects, a recombinant HIV-1 envelope comprises mutations N279H, K282R, R456Q, and N276G. The amino acid numbering position is with respect to HXB2 envelope sequence. [0009] In certain embodiments, the compositions contemplate nucleic acid, as DNA and/or RNA, or proteins immunogens either alone or in any combination.
  • the methods contemplate genetic, as DNA and/or RNA, immunization either alone or in combination with envelope protein(s).
  • the nucleic acid encoding an envelope is operably linked to a promoter inserted in an expression vector.
  • the compositions comprise a suitable carrier.
  • the compositions comprise a suitable adjuvant.
  • the induced immune response includes induction of antibodies, including but not limited to autologous and/or cross-reactive (broadly) neutralizing antibodies against HIV-1 envelope.
  • the invention provides a nucleic acid sequence encoding any of the polypeptides of the invention, wherein the nucleic acid is operably linked to a promoter.
  • the invention provides a nucleic acid consisting essentially of a nucleic acid sequence encoding any of the polypeptides of the invention, wherein the nucleic acid is operably linked to a promoter.
  • the invention provides an expression vector comprising any of the nucleic acid sequences of the invention, wherein the nucleic acid is operably linked to a promoter.
  • the invention provides an expression vector comprising a nucleic acid sequence encoding any of the polypeptides of the invention, wherein the nucleic acid is operably linked to a promoter.
  • the invention provides an expression vector consisting essentially a nucleic acid sequence encoding any of the polypeptides of the invention, wherein the nucleic acid is operably linked to a promoter.
  • the nucleic acids are codon optimized for expression in a mammalian cell, in vivo or in vitro.
  • the invention provides nucleic acids comprising any one of the nucleic acid sequences of invention.
  • the invention provides nucleic acids consisting essentially of any one of the nucleic acid sequences of invention.
  • the invention provides nucleic acids consisting of any one of the nucleic acid sequences of invention.
  • the nucleic acid of the invention is operably linked to a promoter and is inserted in an expression vector.
  • the invention provides an immunogenic composition comprising the expression vector.
  • the invention provides a composition comprising at least one of the nucleic acid sequences of the invention.
  • the invention provides a composition comprising any one of the nucleic acid sequences of invention.
  • the invention provides a composition comprising at least one nucleic acid sequence encoding any one of the polypeptides of the invention.
  • the invention provides a composition comprising at least one nucleic acid encoding an HIV-1 envelope of the invention.
  • the compositions and methods employ an HIV-1 envelope as polypeptide instead of a nucleic acid sequence encoding the HIV-1 envelope.
  • the compositions and methods employ an HIV-1 envelope as polypeptide, a nucleic acid sequence encoding the HIV-1 envelope, or a combination thereof.
  • the polypeptides are recombinantly produced.
  • the envelope used in the compositions and methods of the invention can be a gp160, gp150, gp145, gp140, gp120, gp41, or N-terminal deletion variants thereof as described herein, cleavage resistant variants thereof as described herein, or codon optimized sequences thereof.
  • the composition comprises envelopes as trimers.
  • envelope proteins are multimerized, for example trimers are attached to a particle such that multiple copies of the trimer are attached and the multimerized envelope is prepared and formulated for immunization in a human.
  • the compositions comprise envelopes, including but not limited to trimers as particulate, high- density array on liposomes or other particles, for example but not limited to nanoparticles.
  • the trimers are in a well ordered, near native like or closed conformation.
  • the trimer compositions comprise a homogenous mix of native like trimers.
  • the trimer compositions comprise at least 65%, 70%, 75%, 80%, 85%, 90%, 95% native like trimers.
  • the polypeptide contemplated by the invention can be a polypeptide comprising any one of the polypeptides described herein.
  • the polypeptide contemplated by the invention can be a polypeptide consisting essentially of any one of the polypeptides described herein.
  • the polypeptide contemplated by the invention can be a polypeptide consisting of any one of the polypeptides described herein.
  • the polypeptide is recombinantly produced.
  • the polypeptides and nucleic acids of the invention are suitable for use as an immunogen, for example to be administered in a human subject.
  • the envelope is any of the forms of HIV-1 envelope.
  • the envelope is a gp120, gp140, gp145 (i.e. with a transmembrane), gp150 envelope.
  • gp140 is designed to form a stable trimer.
  • envelope protomers form a trimer which is not a SOSIP timer.
  • the trimer is a SOSIP based trimer wherein each protomer comprises additional modifications.
  • envelope trimers are recombinantly produced.
  • envelope trimers are purified from cellular recombinant 4 ActiveUS 201518670 Attorney Docket No.: 1234300.00424WO1 (DU7956PCT) fractions by antibody binding and reconstituted in lipid comprising formulations. See for example WO2015/127108 titled “Trimeric HIV-1 envelopes and uses thereof” which content is herein incorporated by reference in its entirety.
  • the envelopes of the invention are engineered and comprise non-naturally occurring modifications.
  • the envelope is in a liposome.
  • the envelope comprises a transmembrane domain with a cytoplasmic tail embedded in a liposome.
  • the nucleic acid comprises a nucleic acid sequence which encodes a gp120, gp140, gp145, gp150, or gp160.
  • the vectors are any suitable vector.
  • Non-limiting examples include, VSV, replicating rAdenovirus type 4, MVA, Chimp adenovirus vectors, pox vectors, and the like.
  • the nucleic acids are administered in NanoTaxi block polymer nanospheres.
  • the composition and methods comprise an adjuvant.
  • Non-limiting examples include, AS01 B, AS01 E, gla/SE, alum, Poly I poly C (poly IC), polyIC/long chain (LC) TLR agonists, TLR7/8 and 9 agonists, or a combination of TLR7/8 and TLR9 agonists (see Moody et al. (2014) J. Virol. March 2014 vol. 88 no. 63329-3339), or any other adjuvant.
  • Non-limiting examples of TLR7/8 agonist include TLR7/8 ligands, Gardiquimod, Imiquimod and R848 (resiquimod).
  • a non-limiting embodiment of a combination of TLR7/8 and TLR9 agonist comprises R848 and oCpG in STS (see Moody et al. (2014) J. Virol. March 2014 vol. 88 no. 63329-3339).
  • the adjuvant is an LNP. See e.g., without limitation Shirai et al.
  • LNPs used as adjuvants for protein compositions are composed of an ionizable lipid, cholesterol, lipid conjugated with polyethylene glycol, and a helper lipid.
  • Non-limiting embodiment include LNPs without polyethylene glycol.
  • the invention provides a cell comprising a nucleic acid encoding any one of the envelopes of the invention suitable for recombinant expression.
  • the invention provides a clonally derived population of cells encoding any one of the envelopes of the invention suitable for recombinant expression. In certain aspects, the invention provides a stable pool of cells encoding any one of the envelopes of the invention suitable for recombinant expression. 5 ActiveUS 201518670 Attorney Docket No.: 1234300.00424WO1 (DU7956PCT) [0022] In certain aspects, the invention provides a recombinant HIV-1 envelope polypeptide listed in Table 1 and/or Figs. 1 or 18. In certain embodiments, the polypeptide is a non- naturally occurring protomer designed to form an envelope trimer. The invention also provides nucleic acids encoding these recombinant polypeptides.
  • Non-limiting examples of amino acids and nucleic acids of such protomers are shown in Fig. 1.
  • the invention provides a recombinant trimer comprising three identical protomers of an envelope from Table 1 and/or Figs. 1 or 18.
  • the invention provides an immunogenic composition comprising the recombinant trimer and a carrier, wherein the trimer comprises three identical protomers of an HIV-1 envelope listed in Table 1 and/or Figs. 1 or 18.
  • the invention provides an immunogenic composition comprising a nucleic acid encoding these recombinant HIV-1 envelope and a carrier.
  • the invention provides nucleic acids encoding HIV-1 envelopes for immunization wherein the nucleic acid encodes a gp120 envelope, gp120D8 envelope, a gp140 envelope (gp140C, gp140CF, gp140CFI) as soluble or stabilized protomer of a SOSIP trimer, a gp145 envelope, a gp150 envelope, or a transmembrane bound envelope.
  • the invention provides a selection of HIV-1 envelopes for immunization wherein the HIV-1 envelope is a gp120 envelope or a gp120D8 variant.
  • a composition for immunization comprises protomers that form stabilized SOSIP trimers.
  • the compositions for use in immunization further comprise an adjuvant.
  • the compositions comprise a nucleic acid
  • the nucleic acid is operably linked to a promoter, and could be inserted in an expression vector.
  • the invention provides a composition for a prime boost immunization regimen comprising any of envelopes described herein, or any combination thereof wherein the envelope is a prime or boost immunogen.
  • the composition for a prime boost immunization regimen comprises one or more envelopes described herein, wherein the polypeptide is a non-naturally occurring protomer designed to form an envelope trimer, wherein the envelope is a prime or boost immunogen.
  • the invention provides a composition for a prime boost immunization regimen comprising one or more envelopes of the invention.
  • the invention provides a composition for a prime immunization comprising one or more envelopes of the invention.
  • the invention provides methods of inducing an immune response in a subject comprising administering a composition comprising a polypeptide and/or any suitable form of a nucleic acid(s) encoding an HIV-1 envelope(s) in an amount sufficient to induce an immune response.
  • the nucleic acid encodes a gp120 envelope, gp120D8 envelope, a gp140 envelope (gp140C, gp140CF, gp140CFI) as soluble or stabilized protomer of a SOSIP trimer, a gp145 envelope, a gp150 envelope, or a transmembrane bound envelope.
  • the polypeptide is gp120 envelope, gp120D8 envelope, a gp140 envelope (gp140C, gp140CF, gp140CFI) as soluble or stabilized protomer of a SOSIP trimer, a gp145 envelope, a gp150 envelope, or a transmembrane bound envelope.
  • the methods comprise administering an adjuvant.
  • the methods comprise administering an agent which modulates host immune tolerance.
  • the administered polypeptide is multimerized in a liposome or nanoparticle.
  • the methods comprise administering one or more additional HIV-1 immunogens to induce a T cell response.
  • the invention provides a recombinant HIV-1 envelope sequence or nucleic acid encoding a recombinant protein HIV-1 envelope sequence comprising a mutation of amino acid residue R273, S274, N276, N279, I277, T278, K282, I284, R456, or M475 (including, but not limited to, N276G, I277P, T278A, T278R, K282R, R456Q, R273S, S274Q, S274V, N276K, N279H, N279L, N279R, I284W, or M475L).
  • a recombinant HIV-1 envelope or nucleic acid encoding a recombinant protein HIV-1 envelope sequence comprises mutations N279H, K282R and R456Q. In certain aspects, a recombinant HIV-1 envelope or nucleic acid encoding a recombinant protein HIV-1 envelope sequence comprises mutations N279H and K282R. In certain aspects, a recombinant HIV-1 envelope or nucleic acid encoding a recombinant protein HIV-1 envelope sequence comprises mutations N279H and R456Q.
  • a recombinant HIV-1 envelope or nucleic acid encoding a recombinant protein HIV-1 envelope sequence comprises mutations K282R and R456Q. In certain aspects, a recombinant HIV-1 envelope or nucleic acid encoding a recombinant protein HIV-1 envelope sequence comprises mutations N279H, K282R, R456Q, and N276G. The amino acid numbering position is with respect to HXB2 envelope sequence. Non-limited examples of envelopes are listed in Table 1 and Figs. 1 or 18. In some embodiments, the amino acid sequence of one monomer comprised in the trimer is shown in Figs. 1 or 18. In some embodiments, the trimer is immunogenic.
  • the 7 ActiveUS 201518670 Attorney Docket No.: 1234300.00424WO1 (DU7956PCT) trimer binds to any one of the antibodies PGT145, PGT151, CH235UCA, CH235, 1-18, 2G12, or any combination thereof. In some embodiments the trimer does not bind to antibody 19B and/or 17B.
  • the invention provides a pharmaceutical composition comprising any one of the recombinant trimers of the invention. In certain embodiments the compositions comprising trimers are immunogenic. The percent trimer in such immunogenic compositions could vary.
  • the composition comprises 70%, 71%, 72%, 73%, 74%,75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% stabilized trimer.
  • the recombinant protein nanoparticle comprises the envelope and ferritin.
  • the inventive designs comprise modifications, including without limitation linkers between the envelope and ferritin designed to optimize ferritin nanoparticle assembly.
  • the invention provides a composition comprising any one of the inventive envelopes or nucleic acid sequences encoding the same.
  • the invention provides compositions comprising a nanoparticle which comprises any one of the envelopes of the invention.
  • the nanoparticle is a ferritin self-assembling nanoparticle.
  • the invention provides a method of inducing an immune response in a subject comprising administering an immunogenic composition comprising any one of the envelopes of the invention.
  • the composition is administered as a prime and/or a boost.
  • the composition comprises nanoparticles.
  • methods of the invention further comprise administering an adjuvant.
  • the invention provides a composition comprising a plurality of nanoparticles comprising a plurality of the envelopes/trimers of the invention.
  • the envelopes/trimers of the invention are multimeric when comprised in a nanoparticle.
  • the nanoparticle size is suitable for delivery.
  • the nanoparticles are ferritin-based nanoparticles.
  • the invention provides nucleic acids comprising sequences encoding polypeptides or proteins of the invention.
  • the nucleic acids are DNAs.
  • the invention provides expression vectors comprising the nucleic acids of the invention.
  • the invention provides a recombinant HIV-1 envelope polypeptide according to Table 1 and/or Figs. 1 or 18.
  • the polypeptide is a non- 8 ActiveUS 201518670 Attorney Docket No.: 1234300.00424WO1 (DU7956PCT) naturally occurring protomer.
  • the polypeptide is designed to form an envelope trimer.
  • the envelope is based on CH505 T/F envelope and comprises optimized sequence for binding to CH235 lineage members, including without limitation CH235 UCA.
  • the envelope comprises a mutation at position R273, S274, N276, N279, I277, T278, K282, I284, R456 or M475, or a combination thereof (including, but not limited to, N276G, I277P, T278A, T278R, K282R, R456Q, R273S, S274Q, S274V, N276K, N279H, N279L, N279R, I284W, or M475L).
  • a recombinant HIV-1 envelope comprises mutations N279H, K282R and R456Q.
  • a recombinant HIV-1 envelope comprises mutations N279H and K282R. In certain aspects, a recombinant HIV-1 envelope comprises mutations N279H and R456Q. In certain aspects, a recombinant HIV-1 envelope comprises mutations K282R and R456Q. In certain aspects, a recombinant HIV-1 envelope comprises mutations N279H, K282R, R456Q, and N276G.
  • the amino acid numbering position is with respect to HXB2 envelope sequence.
  • the envelope polypeptide is designed to multimerize.
  • the envelope sequence comprises a self-assembling protein. In certain embodiments, the self-assembling protein is a ferritin.
  • the self- assembling protein is added via a sortase A reaction.
  • the modifications described herein can be incorporated into any CH505 based envelope.
  • the invention provides a recombinant trimer comprising three identical protomers of an envelope from Table 1 and/or Fig. 1.
  • the invention provides an immunogenic composition comprising the recombinant trimer and a carrier, wherein the trimer comprises three identical protomers of an HIV-1 envelope listed in Table 1 and/or Fig. 1.
  • the invention provides an immunogenic composition comprising a nucleic acid encoding the recombinant HIV-1 envelope and a carrier.
  • the envelopes are or are designed as trimers, and/or nanoparticles.
  • the immunogenic composition further comprises an adjuvant.
  • the nucleic acid encoding one or more envelope selected from Fig. 1 or 18 any combination thereof is operably linked to a promoter.
  • the nucleic acid is inserted in an expression vector.
  • the invention provides a method of inducing an immune response in a subject comprising administering a composition comprising any suitable form of a nucleic acid(s) encoding one or more envelope selected from Table 1 or Figs. 1 or 18 or any combination thereof in an amount sufficient to induce an immune response.
  • the composition administered comprises a nucleic acid encoding a gp120 envelope, gp120D8 envelope, a gp140 envelope (gp140C, gp140CF, gp140CFI) as soluble or stabilized protomer of a SOSIP trimer, a gp145 envelope, a gp150 envelope, a transmembrane bound envelope, a gp160 envelope or an envelope designed to multimerize.
  • the composition administered comprises a polypeptide, wherein the polypeptide is gp120 envelope, gp120D8 envelope, a gp140 envelope (gp140C, gp140CF, gp140CFI) as soluble or stabilized protomer of a SOSIP trimer, a gp145 envelope, a gp150 envelope, a transmembrane bound envelope, or an envelope designed to multimerize.
  • the composition administered further comprises an adjuvant.
  • the method further comprises administering an agent which modulates host immune tolerance.
  • the polypeptide administered is multimerized in a liposome or nanoparticle.
  • the method further comprising administering one or more additional HIV-1 immunogens to induce a T cell response.
  • the invention provides a composition comprises a nanoparticle and a carrier, wherein the nanoparticle comprises an envelope, wherein the envelope is selected from Table 1 and/or Figs. 1 or 18 or any combination thereof.
  • the compositions comprises two, three, four or more different immunogens.
  • the immunogens target different CH235 lineage members.
  • the immunogens target the CH235 lineage UCA.
  • the different immunogens are selected from the various envelope designs described herein.
  • the nanoparticle of the composition is ferritin self-assembling nanoparticle.
  • the invention provides a composition comprising a nanoparticle and a carrier, wherein the nanoparticle comprises a nucleic acid encoding the recombinant HIV-1 envelope polypeptide from Table 1 and/or Figs. 1 or 18.
  • the nanoparticle of the composition is a ferritin self- assembling nanoparticle.
  • the nanoparticle of the composition comprises multimers of trimers.
  • the nanoparticle of the composition comprises 1-8 trimers.
  • the invention provides a method of inducing an immune response in a subject comprising administering an immunogenic composition comprising any one of the recombinant envelopes or compositions described herein.
  • the methods comprise administering two, three, four or more different immunogens.
  • the different immunogens target different CH235 lineage members.
  • the different immunogens target the CH235 lineage UCA or CH235 intermediate antibodies.
  • the different immunogens are selected from the envelope designs described herein—Tables 1 and/ or Figs. 1 or 18.
  • the subject is infected with HIV (e.g., HIV-1).
  • HIV e.g., HIV-1.
  • the subject is an HIV-uninfected individual.
  • the subject is an HIV-infected individual.
  • the administration to the HIV- infected individual induces broadly neutralizing antibodies.
  • the broadly neutralizing antibodies of the HIV-infected individual mediates viral (e.g., HIV-1) clearance from blood and tissues.
  • the composition is administered as a single prime or as repetitive immunization prime. In preferred embodiments, the repetitive immunization is administered 3 or 4 times.
  • the composition is administered as a single boost or as a repetitive series of boosts.
  • the repetitive series of boosts is administered 3 or 4 times.
  • the composition is a first composition administered as a prime.
  • the composition is a second composition administered as one or more boosts.
  • the method comprises administering the first composition as a prime and administering the second composition as one or more boosts.
  • the first composition and the second composition are different.
  • the invention provides a nucleic acid encoding any of the recombinant envelopes described herein.
  • the invention provides a composition comprising the nucleic acid and a carrier.
  • the invention provides a method of inducing an immune response in a subject comprising administering an immunogenic composition comprising the nucleic acid encoding any of the recombinant envelopes described herein.
  • the immunogenic composition further comprises a carrier.
  • the invention provides an immunogenic composition or composition, wherein the composition comprises at least two different HIV-1 envelope 11 ActiveUS 201518670 Attorney Docket No.: 1234300.00424WO1 (DU7956PCT) polypeptides or nucleic acids encoding a recombinant HIV-1 envelope polypeptide, or a combination thereof.
  • the invention provides an immunogenic composition comprising a first immunogen and a second immunogen, wherein the first immunogen is a recombinant HIV-1 envelope polypeptide from Table 1 and/or Figs. 1 or 18, or encoded by a nucleic acid encoding said recombinant HIV-1 envelope polypeptide, and wherein the second immunogen is a different recombinant HIV-1 envelope polypeptide from Table 1 and/or Figs. 1 or 18 or a nucleic acid encoding said different recombinant HIV-1 envelope polypeptide.
  • the invention provides a method of inducing an immune response in a subject comprising administering the immunogenic composition in an amount sufficient to induce an immune response.
  • the method further comprising administering an agent which modulates host immune tolerance.
  • at least one of the first immunogen and the second immunogen is a recombinant HIV-1 envelope polypeptide.
  • at least one of the first immunogen and the second immunogen is a recombinant trimer comprising three identical protomers of the recombinant HIV-1 envelope polypeptide.
  • the first immunogen and the second immunogen are a recombinant HIV-1 envelope polypeptide.
  • at least one of the first immunogen and the second immunogen is a nucleic acid. In certain embodiments, the first immunogen and the second immunogen are a nucleic acid.
  • the HIV-1 envelopes are in the form of a recombinant HIV-1 envelope polypeptides or nucleic acid, or a combination thereof.
  • one or more of the HIV-1 envelopes is a recombinant trimer comprising three identical protomers of the recombinant HIV-1 envelope polypeptide.
  • the composition comprises a carrier.
  • the composition further comprises an adjuvant.
  • Fig. 1 shows non-limiting embodiments of amino acid sequences of envelopes of the invention.
  • Signal peptide is underlined – the signal peptide is MPMGSLQPLATLYLLGMLVASVLA (SEQ ID NO: 1).
  • Avi-tag is boxed – Avi-tag is 12 ActiveUS 201518670 Attorney Docket No.: 1234300.00424WO1 (DU7956PCT) GGGGSGLNDIFEAQKIEWHE (SEQ ID NO: 2).
  • Fig. 1 discloses SEQ ID NOS 10-24, respectively, in order of appearance.
  • Fig. 2 shows study aims.
  • Fig. 3 shows mutagenized library construction.
  • Fig. 3 discloses SEQ ID NOS 25-28, respectively, in order of appearance.
  • Fig. 4 shows successive enrichment of SHM7 binding-enhanced mutants.
  • Fig. 5 shows that sequential selection of envelope variants enriched primarily D- Loop mutants.
  • Fig. 6 shows affinity of CH505 mutants (Bio-Layer Interferometry).
  • Fig. 7 shows that most mutations enhanced binding to CH235 lineage members.
  • Fig. 8 shows additional study aims.
  • Fig. 9 shows successive enrichment of UCA binding-enhanced mutants.
  • Fig. 10 shows that sequential selection of envelope variants again enriched primarily D-loop mutants.
  • Fig. 11 shows evolution of CH235.UCA-sorted mutants.
  • Fig. 12 shows methodology of creating site-scanning saturation library.
  • Fig.13 shows enrichments of CH235.SHM7 binding-enhanced mutants.
  • Fig. 14 shows enrichments of CH235.UCA binding-enhanced mutants.
  • Fig. 15 shows mutation enrichment data.
  • Fig. 16 shows evaluation of CH235.SHM7-sorted mutants.
  • Fig. 17A-B show evaluation of CH235.UCA-sorted mutants.
  • Fig. 18 shows non-limiting embodiments of nucleic acid sequences of envelopes of the invention. Signal peptide is underlined. Avi-tag is boxed.
  • Fig.13 shows enrichments of CH235.SHM7 binding-enhanced mutants.
  • Fig. 14 shows enrichments of CH235.UCA binding-enhanced mutants.
  • Fig. 15 shows mutation enrichment data.
  • Fig. 16 shows evaluation of CH235.SHM7-sorted mutants.
  • Fig. 17A-B show evaluation
  • Fig. 19A-C show that SHM7 heterologous reactivity requires subset of mutations.
  • Fig. 19B-C show that SHM7-selected double/triple mutants of CH505 place pressure on critical SHM7 mutations.
  • DETAILED DESCRIPTION OF THE INVENTION [0089] The development of a safe, highly efficacious prophylactic HIV-1 vaccine is of paramount importance for the control and prevention of HIV-1 infection.
  • a major goal of HIV-1 vaccine development is the induction of broadly neutralizing antibodies (bnAbs) 13 ActiveUS 201518670 Attorney Docket No.: 1234300.00424WO1 (DU7956PCT) (Immunol. Rev. 254: 225-244, 2013).
  • BnAbs are protective in rhesus macaques against SHIV challenge, but as yet, are not reproducibly induced by current vaccines.
  • the HIV vaccine development field has used single or prime boost heterologous Envs as immunogens, but to date has not found a regimen to induce high levels of bnAbs.
  • HIV-1 Envelopes Described herein are nucleic acid and amino acid sequences of HIV-1 envelopes. The sequences for use as immunogens are in any suitable form. In certain embodiments, the described HIV-1 envelope sequences are gp160s.
  • the described HIV-1 envelope sequences are gp120s.
  • Other sequences for example but not limited to stable SOSIP trimer designs, gp145s, gp140s, both cleaved and uncleaved, gp140 Envs with the deletion of the cleavage (C) site, fusion (F) and immunodominant (I) region in gp41-- QDPHG ⁇ DV ⁇ JS ⁇ &), (gp140CFI), gp140 Envs with the deletion of only the cleavage (C) site and fusion (F) domain -- QDPHG ⁇ DV ⁇ JS ⁇ &) (gp140CF), gp140 Envs with the deletion of only the cleavage (C)—QDPHG ⁇ JS ⁇ & ⁇ (gp140C) (See e.g.
  • An HIV-1 envelope has various structurally defined fragments/forms: gp160; gp140-- -including cleaved gp140 and uncleaved gp140 (gp140C), gp140CF, or gp140CFI; gp120 and gp41.
  • fragments/forms are defined not necessarily by their crystal structure, but by their design and bounds within the full length of the gp160 envelope. While the specific consecutive amino acid sequences of envelopes from different strains are different, the bounds and design of these forms are well known and characterized in the art. [0095] For example, it is well known in the art that during its transport to the cell surface, the gp160 polypeptide is processed and proteolytically cleaved to gp120 and gp41 proteins.
  • Envelope gp140 forms are designed by introducing a stop codon within the gp41 sequence. See Chakrabarti et al. at Figure 1.
  • Envelope gp140C refers to a gp140 HIV-1 envelope design with a functional deletion of the cleavage (C) site, so that the gp140 envelope is not cleaved at the furin cleavage site.
  • RRVVEREKR SEQ ID NO: 4
  • ERVVEREKE SEQ ID NO: 5
  • SEKS SEKS
  • Envelope gp140CF refers to a gp140 HIV-1 envelope design with a deletion of the cleavage (C) site and fusion (F) region.
  • Envelope gp140CFI refers to a gp140 HIV-1 envelope design with a deletion of the cleavage (C) site, fusion (F) and immunodominant (I) region in gp41. See Chakrabarti et al. Journal of Virology vol. 76, pp. 5357-5368 (2002) see for example Figure 1, and Second paragraph in the Introduction on p. 5357; Binley et al. Journal of Virology vol. 76, pp. 2606-2616 (2002) for example at Abstract; Gao et al.
  • the envelope design in accordance with the present invention involves deletion of residues (e.g., 5-11, 5, 6, 7, 8, 9, 10, or 11 amino acids) at the N- terminus.
  • residues e.g., 5-11, 5, 6, 7, 8, 9, 10, or 11 amino acids
  • amino acid residues ranging from 4 residues or even fewer to 14 residues or even more are deleted. These residues are between the maturation (signal peptide, usually ending with CX, X can be any amino acid) and "VPVXXXX".
  • CH505 T/F Env 8 amino acids (italicized and underlined in the below sequence) were deleted: 15 ActiveUS 201518670 Attorney Docket No.: 1234300.00424WO1 (DU7956PCT) MRVMGIQRNYPQWWIWSMLGFWMLMICNGMWVTVYYGVPVWKEAKTTLFCASDA KAYEKEVHNVWATHACVPTDPNPQE...(rest of envelope sequence is indicated as (SEQ ID NO: 7).
  • the delta N-design described for CH505 T/F envelope can be used to make delta N-designs of other CH505 envelopes.
  • the invention relates generally to an immunogen, gp160, gp120 or gp140, without an N-terminal Herpes Simplex gD tag substituted for amino acids of the N-terminus of gp120, with an HIV leader sequence (or other leader sequence), and without the original about 4 to about 25, for example 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25 amino acids of the N-terminus of the envelope (e.g. gp120). See US Patent 10,040,826, e.g. at pages 10-12, the contents of which is hereby incorporated by reference in its entirety.
  • N-terminal amino acids of envelopes results in proteins, for example gpl20s, expressed in mammalian cells that are primarily monomeric, as opposed to dimeric, and, therefore, solves the production and scalability problem of commercial gp120 Env vaccine production.
  • the amino acid deletions at the N-terminus result in increased immunogenicity of the envelopes.
  • the invention provides composition and methods which CH505 Envs, as gp120s, gp140s cleaved and uncleaved, gp145s, gp150s and gp160s, stabilized and/or multimerized trimers, as proteins, DNAs, RNAs, or any combination thereof, administered as primes and boosts to elicit immune response.
  • CH505 Envs as proteins would be co-administered with nucleic acid vectors containing Envs to amplify antibody induction.
  • the compositions and methods include any immunogenic HIV-1 sequences to give the best coverage for T cell help and cytotoxic T cell induction.
  • the compositions and methods include mosaic and/or consensus HIV-1 genes to give the best coverage for T cell help and cytotoxic T cell induction.
  • the compositions and methods include mosaic group M and/or consensus genes to give the best coverage for T cell help and cytotoxic T cell induction.
  • the mosaic genes are any suitable gene from the HIV-1 genome.
  • the mosaic genes are Env genes, Gag genes, Pol genes, Nef genes, or any combination thereof. See e.g. US Patent No. 7951377.
  • the mosaic genes are bivalent mosaics. In some embodiments the mosaic genes are trivalent.
  • the mosaic genes are administered in a suitable vector with each immunization with Env gene inserts in a suitable vector and/or as a protein.
  • the mosaic genes for example as bivalent mosaic Gag group M consensus genes, are 16 ActiveUS 201518670 Attorney Docket No.: 1234300.00424WO1 (DU7956PCT) administered in a suitable vector, for example but not limited to HSV2, would be administered with each immunization with Env gene inserts in a suitable vector, for example but not limited to HSV-2.
  • nucleic acid sequences [0104] In certain aspects the invention provides compositions and methods of Env genetic immunization either alone or with Env proteins to recreate the swarms of evolved viruses that have led to bnAb induction. Nucleotide-based vaccines offer a flexible vector format to immunize against virtually any protein antigen. Currently, two types of genetic vaccination are available for testing—DNAs and mRNAs. [0105] In certain aspects the invention contemplates using immunogenic compositions wherein immunogens are delivered as DNA. See Graham BS, Enama ME, Nason MC, Gordon IJ, Peel SA, et al.
  • DNA Vaccine Delivered by a Needle-Free Injection Device Improves Potency of Priming for Antibody and CD8+ T-Cell Responses after rAd5 Boost in a Randomized Clinical Trial.
  • DNA can be delivered as naked DNA.
  • DNA is formulated for delivery by a gene gun.
  • DNA is administered by electroporation, or by a needle-free injection technologies, for example but not limited to Biojector® device.
  • the DNA is inserted in vectors.
  • the DNA is delivered using a suitable vector for expression in mammalian cells.
  • the nucleic acids encoding the envelopes are optimized for expression.
  • DNA is optimized, e.g. codon optimized, for expression.
  • the nucleic acids are optimized for expression in vectors and/or in mammalian cells. In non-limiting embodiments these are bacterially derived vectors, adenovirus based vectors, rAdenovirus (e.g. Barouch DH, et al. Nature Med. 16: 319-23, 2010), recombinant mycobacteria (e.g.
  • rBCG or M smegmatis (Yu, JS et al. Clinical Vaccine Immunol. 14: 886- 093,2007; ibid 13: 1204-11,2006), and recombinant vaccinia type of vectors (Santra S. Nature Med.16: 324-8, 2010), for example but not limited to ALVAC, replicating (Kibler KV et al., PLoS One 6: e25674, 2011 nov 9.) and non-replicating (Perreau M et al. J.
  • the invention contemplates using immunogenic compositions wherein immunogens are delivered as DNA or RNA in suitable formulations.
  • DNA or RNA is administered as nanoparticles consisting of low dose antigen-encoding DNA formulated with a block copolymer (amphiphilic block copolymer 704).
  • a block copolymer amphiphilic block copolymer 704
  • the invention provides nucleic acids comprising sequences encoding envelopes of the invention.
  • the nucleic acids are DNAs.
  • the invention provides expression vectors comprising the nucleic acids of the invention.
  • the nucleic acid encoding an envelope is operably linked to a promoter inserted an expression vector.
  • the compositions comprise a suitable carrier.
  • the compositions comprise a suitable adjuvant.
  • the invention provides an expression vector comprising any of the nucleic acid sequences of the invention, wherein the nucleic acid is operably linked to a promoter.
  • the invention provides an expression vector comprising a nucleic acid sequence encoding any of the polypeptides of the invention, wherein the nucleic acid is operably linked to a promoter.
  • the nucleic acids are codon optimized for expression in a mammalian cell, in vivo or in vitro.
  • the invention provides nucleic acids comprising any one of the nucleic acid sequences of invention.
  • the invention provides nucleic acids consisting essentially of any one of the nucleic acid sequences of invention.
  • the invention provides nucleic acids consisting of any one of the nucleic acid sequences of invention.
  • the nucleic acid of the invention is operably linked to a promoter and is inserted in an expression vector.
  • the invention provides an immunogenic composition comprising the expression vector.
  • the invention provides a composition comprising at least one of the nucleic acid sequences of the invention.
  • the invention provides a 18 ActiveUS 201518670 Attorney Docket No.: 1234300.00424WO1 (DU7956PCT) composition comprising any one of the nucleic acid sequences of invention.
  • the invention provides a composition comprising at least one nucleic acid sequence encoding any one of the polypeptides of the invention.
  • the nucleic acid is an RNA molecule.
  • the RNA molecule is transcribed from a DNA sequence described herein.
  • the RNA molecule is encoded by one of the inventive sequences.
  • the nucleotide sequence comprises an RNA sequence transcribed by a DNA sequence encoding the polypeptide sequence of the sequences of the invention, or a variant thereof or a fragment thereof.
  • the invention provides an RNA molecule encoding one or more of inventive antibodies.
  • the RNA may be plus-stranded. Accordingly, in some embodiments, the RNA molecule can be translated by cells without needing any intervening replication steps such as reverse transcription.
  • a RNA molecule of the invention may have a 5' cap (e.g. but not limited to a 7-methylguanosine, 7mG(5')ppp(5')NlmpNp, CleanCap® (e.g., the AG, GG, AU, 3’OMe AG, or 3’OMe GG CleanCap®), or ARCA).
  • This cap can enhance in vivo translation of the RNA.
  • the 5' nucleotide of an RNA molecule useful with the invention may have a 5' triphosphate group. In a capped RNA this may be linked to a 7-methylguanosine via a 5'-to-5' bridge.
  • a RNA molecule may have a 3' poly-A tail. It may also include a poly-A polymerase recognition sequence (e.g. AAUAAA) near its 3' end.
  • a RNA molecule useful with the invention may be single-stranded.
  • a RNA molecule useful with the invention may comprise synthetic RNA.
  • the recombinant nucleic acid sequence can be an optimized nucleic acid sequence. Such optimization can increase or alter the immunogenicity of the envelope. Optimization can also improve transcription and/or translation.
  • Optimization can include one or more of the following: low GC content leader sequence to increase transcription; mRNA stability and codon optimization; addition of a Kozak sequence (e.g., GCC ACC) for increased translation; addition of an immunoglobulin (Ig) leader sequence encoding a signal peptide; and eliminating to the extent possible cis-acting sequence motifs (i.e., internal TATA boxes).
  • a Kozak sequence e.g., GCC ACC
  • Ig immunoglobulin leader sequence encoding a signal peptide
  • Ig immunoglobulin leader sequence encoding a signal peptide
  • eliminating to the extent possible cis-acting sequence motifs i.e., internal TATA boxes.
  • Methods for in vitro transfection of mRNA and detection of envelope expression are known in the art.
  • Methods for expression and immunogenicity determination of nucleic acid encoded envelopes are known in the art.
  • the invention contemplates using immunogenic compositions wherein immunogens are delivered
  • recombinant proteins are produced in CHO cells.
  • the immunogenic envelopes can also be administered as a protein prime in combination with a variety of nucleic acid envelope boosts (e.g., HIV -1 Envs delivered as DNA expressed in viral or bacterial vectors).
  • nucleic acid envelope boosts e.g., HIV -1 Envs delivered as DNA expressed in viral or bacterial vectors.
  • a single dose of nucleic acid can range from a few nanograms (ng) to a few micrograms ⁇ J ⁇ or milligram of a single immunogenic nucleic acid.
  • Recombinant protein dose can range from a few ⁇ J ⁇ micrograms to a few hundred micrograms, or milligrams of a single immunogenic polypeptide.
  • Administration The compositions can be formulated in designs that incorporate appropriate carriers such as peptides for enhancing CD4+ T cell help, known as PADRE, GTH1, GTH2, or any combination thereof.
  • the compositions are delivered via intramuscular (IM), via subcutaneous, via intravenous, via nasal, via mucosal routes, or any other suitable route of immunization.
  • compositions can be formulated with appropriate carriers and adjuvants using techniques to yield compositions suitable for immunization.
  • the compositions can include an adjuvant, such as, for example but not limited to, alum, 3M052, poly IC, MF-59 or other squalene-based adjuvant, ASOIB, or other liposomal based adjuvant suitable for protein or nucleic acid immunization.
  • the adjuvant is GSK AS01E adjuvant containing MPL and QS21.
  • compositions and methods comprise any suitable agent or immune modulation which could modulate mechanisms of host immune tolerance and release of the induced antibodies.
  • modulation includes PD-1 blockade; T regulatory cell depletion; CD40L hyperstimulation; soluble antigen administration, wherein the soluble antigen is designed such that the soluble agent eliminates B cells targeting dominant epitopes, or a combination thereof.
  • an immunomodulatory agent is administered in at time and in an amount sufficient for transient modulation of the subject's immune response so as to induce an immune response which comprises broad 20 ActiveUS 201518670 Attorney Docket No.: 1234300.00424WO1 (DU7956PCT) neutralizing antibodies against HIV-1 envelope.
  • Non-limiting examples of such agents is any one of the agents described herein: e.g.
  • the modulation includes administering an anti-CTLA4 antibody.
  • Non-limiting examples are ipilimumab and tremelimumab.
  • the methods comprise administering a second immunomodulatory agent, wherein the second and first immunomodulatory agents are different.
  • a second immunomodulatory agent wherein the second and first immunomodulatory agents are different.
  • highly somatically mutated antibodies become autoreactive and/or less fit (Immunity 8: 751, 1998; PloS Comp. Biol. 6 e1000800, 2010; J. Thoret. Biol. 164:37, 1993); Polyreactive/autoreactive na ⁇ ve B cell receptors (unmutated common ancestors of clonal lineages) can lead to deletion of Ab precursors (Nature 373: 252, 1995; PNAS 107: 181, 2010; J. Immunol.
  • envelope peptides referenced in various examples and figures comprise a signal peptide/leader sequence.
  • HIV-1 envelope peptides is a secretory protein with a signal peptide or leader sequence that is removed during processing and recombinant expression (without removal of the signal peptide, the protein is not secreted). See for example Li et al. Control of expression, glycosylation, and secretion of HIV-1 gp120 by homologous and heterologous signal sequences. Virology 204(1):266-78 (1994) (“Li et al. 1994”), at first paragraph, and Li et al.
  • the leader sequence is the endogenous leader sequence. Most of the gp120 and gp160 amino acid 21 ActiveUS 201518670 Attorney Docket No.: 1234300.00424WO1 (DU7956PCT) sequences include the endogenous leader sequence. In other non-limiting examples, the leader sequence is human Tissue Plasminogen Activator (TPA) sequence, human CD5 leader sequence (e.g.
  • MPMGSLQPLATLYLLGMLVASVLA (SEQ ID NO: 1)). Most of the chimeric designs include CD5 leader sequence. A skilled artisan appreciates that when used as immunogens, and for example when recombinantly produced, the amino acid sequences of these proteins do not comprise the signal peptide/leader sequences.
  • HIV-1 envelope trimers and other envelope designs [0126] Stabilized HIV-1 Env trimer immunogens show enhanced antigenicity for broadly neutralizing antibodies and are not recognized by non-neutralizing antibodies. Envelope modifications and designs include, but are not limited to, trimers that are further multimerized, and/or used as particulate, high-density array in liposomes or other particles, for example but not limited to nanoparticles.
  • a stabilized chimeric SOSIP designs can be used to generate CH505 trimers. This design is applicable to diverse viruses from multiple clades. SOSIP designs can be applied to the envelopes disclosed herein including those in Figs. 1 and 18. [0128] Elicitation of neutralizing antibodies is one goal for antibody-based vaccines. Neutralizing antibodies target the native trimeric HIV-1 Env on the surface virions. The trimeric HIV-1 envelope protein consists of three protomers each containing a gp120 and gp41 heterodimer. Recent immunogen design efforts have generated soluble near-native mimics of the Env trimer that bind to neutralizing antibodies but not non-neutralizing antibodies.
  • the recapitulation of the native trimer could be a key component of vaccine induction of neutralizing antibodies.
  • Neutralizing Abs target the native trimeric HIV-1 Env on the surface of viruses (Poignard et al. J Virol. 2003 Jan;77(1):353-65; Parren et al. J Virol. 1998 Dec;72(12):10270-4.; Yang et al. J Virol. 2006 Nov;80(22):11404-8.).
  • the HIV-1 Env protein consists of three protomers of gp120 and gp41 heterodimers that are noncovalently linked together (Center et al. J Virol. 2002 Aug;76(15):7863-7.).
  • Soluble near-native trimers preferentially bind neutralizing antibodies as opposed to non-neutralizing antibodies (Sanders et al. PLoS Pathog. 2013 Sep; 9(9): e1003618).
  • engineered trimeric immunogens derived from multiple viruses from CH505.
  • 6R.SOSIP.664 chimeric disulfide stabilized (DS) 6R.SOSIP.664 (Kwon et al Nat Struct Mol Biol. 2015 Jul; 22(7): 522–531.)
  • chimeric 6R.SOSIP.664v4.1 DeTaeye et al. Cell. 2015 Dec 17;163(7):1702-15.
  • the 6R.SOSIP.664 is the basis for all of these designs and is made as a chimera of C.CH0505 and A.BG505.
  • the gp120 of C.CH505 was fused with the BG505 inner domain gp120 sequence within the alpha helix 5 (D5) to result in the chimeric protein.
  • the chimeric gp120 is disulfide linked to the A.BG505 gp41 as outlined by Sanders et al. (PLoS Pathog. 2013 Sep; 9(9): e1003618). These immunogens were designed as chimeric proteins that possess the BG505 gp41 connected to the CH505 gp120, since the BG505 strain is particularly adept at forming well-folded, closed trimers.
  • This envelope design retains the CH505 CD4 binding site that is targeted by the CH103 and CH235 broadly neutralizing antibody lineages that were isolated from CH505.
  • any other suitable envelope for example but not limited to CH505 envelopes as described in US Patent 10,004,800, incorporated herein by reference, can be designed to include one or more of the mutations described herein (e.g., mutation of one or more of amino acid residue R273, S274, N276, N279, I277, T278, K282, I284, R456, or M475 (HXB2 numbering), including, but not limited to, N276G, I277P, T278A, T278R, K282R, R456Q, R273S, S274Q, S274V, N276K, N279H, N279L, N279R, I284W, or M475L).
  • the mutations described herein e.g., mutation of one or more of amino acid residue R273, S274, N276, N279, I277, T278, K282, I284, R456, or M475 (HXB2 numbering)
  • the envelope sequences further comprise SOSIP designs.
  • Recombinant envelopes as trimers could be produced and purified by any suitable method.
  • purification methods see Ringe RP, Yasmeen A, Ozorowski G, Go EP, Pritchard LK, Guttman M, Ketas TA, Cottrell CA, Wilson IA, Sanders RW, Cupo A, Crispin M, Lee KK, Desaire H, Ward AB, Klasse PJ, Moore JP. 2015. Influences on the design and purification of soluble, recombinant native-like HIV-1 envelope glycoprotein trimers. J Virol 89:12189 -12210. doi:10.1128/JVI.01768-15.
  • Multimeric Envelopes [0133] Presentation of antigens as particulates reduces the B cell receptor affinity necessary for signal transduction and expansion (see Baptista et al. EMBO J. 2000 Feb 15; 19(4): 513– 520). Displaying multiple copies of the antigen on a particle provides an avidity effect that can overcome the low affinity between the antigen and B cell receptor.
  • the initial B cell receptor specific for pathogens can be low affinity, which precludes vaccines from being able to stimulate and expand B cells of interest. In particular, very few na ⁇ ve B cells from which HIV-1 broadly neutralizing antibodies arise can bind to soluble HIV-1 Envelope.
  • envelopes including but not limited to trimers as particulate, high-density array on liposomes or other particles, for example but not limited to nanoparticles. See e.g. He et al. 23 ActiveUS 201518670 Attorney Docket No.: 1234300.00424WO1 (DU7956PCT) Nature Communications 7, Article number: 12041 (2016), doi:10.1038/ncomms12041; Bamrungsap et al. Nanomedicine, 2012, 7 (8), 1253-1271. [0134] Multimeric nanoparticles that comprise and/or display HIV envelope protein or fragments on their surface can be used a vaccine immunogens.
  • the nucleic acid encoding an antigen is fused via a linker/spacer to a nucleic acids sequence encoding a protein which can self-assemble.
  • a fusion protein is made that can self-assemble into a multimeric complex—also referred to as a nanoparticle displaying multiple copies of the antigen.
  • the protein antigen could be conjugated to the self-assembling protein via an enzymatic reaction, thereby forming a nanoparticle displaying multiple copies of the antigen.
  • Non-limiting embodiments of enzymatic conjugation include without limitation sortase mediated conjugation.
  • linkers for use in any of the designs of the invention could be 2-50 amino acids long, e.g. 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45, or 50 amino acids long.
  • these linkers comprise glycine and serine amino acid in any suitable combination, and/or repeating units of combinations of glycine, serine and/or alanine.
  • Ferritin is a well-known protein that self-assembles into a hollow particle composed of repeating subunits. In some species ferritin nanoparticles are composed of 24 copies of a single subunit, whereas in other species it is composed of 12 copies each of two subunits.
  • an envelope design is created so the envelope is presented on particles, e.g. but not limited to nanoparticle.
  • the HIV-1 envelope trimer could be fused to ferritin.
  • Ferritin protein self assembles into a small nanoparticle with three-fold axis of symmetry. At these axes the envelope protein is fused. Therefore, the assembly of the three-fold axis also clusters three HIV-1 envelope protomers together to form an envelope trimer.
  • Each ferritin particle has 8 axes which equates to 8 trimers being displayed per particle. See e.g., Sliepen et al.
  • Any suitable ferritin sequence could be used.
  • ferritin sequences are disclosed in US Patent 10,961,283, incorporated herein by reference.
  • Ferritin nanoparticle linkers The ability to form HIV-1 envelope ferritin nanoparticles relies self-assembly of 24 ferritin subunits into a single ferritin nanoparticle. The addition of a ferritin subunit to the C-terminus of HIV-1 envelope may interfere with the ability of the ferritin subunit to fold properly and or associate with other ferritin subunits.
  • ferritin When expressed 24 ActiveUS 201518670 Attorney Docket No.: 1234300.00424WO1 (DU7956PCT) alone ferritin readily forms 24-subunit nanoparticles, however appending it to envelope only yields nanoparticles for certain envelopes. Since the ferritin nanoparticle forms in the absence of envelope, the envelope could be sterically hindering the association of ferritin subunits. Thus, we designed ferritin with elongated glycine-serine linkers to further distance the envelope from the ferritin subunit. To make sure that the glycine linker is attached to ferritin at the correct position, we created constructs that attach at second amino acid position or the fifth amino acid position.
  • constructs can be designed constructs with and without the leucine, serine, and lysine amino acids following the glycine-serine linker. The goal will be to find a linker length that is suitable for formation of envelope nanoparticles when ferritin is appended to most envelopes. Any suitable linker between the envelope and ferritin could be used, so long as the fusion protein is expressed and the trimer is formed.
  • the nanoparticle immunogens are composed of various forms of HIV-1 envelope protein, e.g.
  • envelope trimer without limitation envelope trimer, and self-assembling protein, e.g. without limitation ferritin protein.
  • ferritin protein Any suitable ferritin could be used in the immunogens of the invention.
  • the ferritin is derived from Helicobacter pylori.
  • the ferritin is insect ferritin.
  • each nanoparticle displays 24 copies of the envelope protein on its surface.
  • Another approach to multimerize expression constructs uses staphylococcus Sortase A transpeptidase ligation to conjugate inventive envelope trimers, for e.g. but not limited to cholesterol. The trimers can then be embedded into liposomes via the conjugated cholesterol.
  • a C-terminal LPXTG tag SEQ ID NO: 8
  • a N-terminal pentaglycine repeat tag SEQ ID NO: 9
  • Cholesterol is also synthesized with these two tags.
  • a C- terminal tag is LPXTGG, where X signifies any amino acid but most commonly Ala, Ser, Glu (SEQ ID NO: 46).
  • Sortase A is then used to covalently bond the tagged envelope to the cholesterol.
  • the sortase A-tagged trimer protein can also be used to conjugate the trimer to other peptides, proteins, or fluorescent labels.
  • the sortase A tagged trimers are conjugated to ferritin to form nanoparticles. Any suitable ferritin can be used.
  • the invention provides design of envelopes and trimer designs wherein the envelope comprises a linker which permits addition of a lipid, such as but not limited to cholesterol, via 25 ActiveUS 201518670 Attorney Docket No.: 1234300.00424WO1 (DU7956PCT) a Sortase A reaction. See e.g., Tsukiji, S. and Nagamune, T. (2009), Sortase-Mediated Ligation: A Gift from Gram-Positive Bacteria to Protein Engineering. ChemBioChem, 10: 787–798.
  • the lipid modified envelopes and trimers could be formulated as liposomes. Any suitable liposome composition is contemplated.
  • the lipid modified and multimerized envelopes and trimers could be formulated as liposomes. Any suitable liposome composition is contemplated.
  • Table 1 shows a summary of sequences described herein. 26 ActiveUS 201518670 Attorney Docket No.: 1234300.00424WO1 (DU7956PCT) [0146] The trimer could be incorporated in a nanoparticle, including without limitation any ferritin based nanoparticle. [0147] Throughout the application amino acid positions numbers refer to HXB2 numbering.
  • any of the immunogens herein may be encoded by a nucleic acid. It will be understood that non-identical nucleic acid sequences may encode the same amino acid sequence. As such these examples do not exclude nucleic acid sequences that encode immunogens with the same amino acid sequence but possess different nucleic acid sequences.
  • Exemplary Embodiments [0150] In certain aspects, the invention provides a recombinant HIV-1 envelope comprising all the consecutive amino acids between the signal peptide and avi-tag of SEQ ID NOs:10-24. In some embodiments, the recombinant HIV-1 envelope comprises all the consecutive amino acids between the signal peptide and avi-tag of SEQ ID NO: 14.
  • the recombinant HIV-1 envelope comprising all the consecutive amino acids between the signal peptide and avi-tag of SEQ ID NO: 14 further comprises mutation K282R. In some embodiments, the recombinant HIV-1 envelope comprising all the consecutive amino acids between the signal peptide and avi-tag of SEQ ID NO: 14 further comprises mutation R456Q. In some embodiments, the recombinant HIV-1 envelope comprising all the consecutive amino acids between the signal peptide and avi-tag of SEQ ID NO: 14 further comprises mutation K282R and R456Q.
  • the recombinant HIV-1 envelope 27 ActiveUS 201518670 Attorney Docket No.: 1234300.00424WO1 (DU7956PCT) comprises all the consecutive amino acids between the signal peptide and avi-tag of SEQ ID NO: 15.
  • the recombinant HIV-1 envelope comprising all the consecutive amino acids between the signal peptide and avi-tag of SEQ ID NO: 15 further comprises mutation R456Q.
  • the recombinant HIV-1 envelope comprises all the consecutive amino acids between the signal peptide and avi-tag of SEQ ID NO: 16.
  • the recombinant HIV-1 envelope comprises all the consecutive amino acids between the signal peptide and avi-tag of SEQ ID NO: 10.
  • any of the recombinant HIV-1 envelopes further comprises mutation N276G.
  • the recombinant HIV-1 envelope further comprises the amino acid sequence of the avi-tag (e.g., in certain aspects, the invention provides a recombinant HIV-1 envelope comprising all the consecutive amino acids after the signal peptide of SEQ ID NOs:10-24).
  • the envelope is a protomer comprised in a trimer. [0151] In certain aspects, the invention provides a nucleic acid encoding any one of the recombinant HIV-1 envelopes described herein.
  • the nucleic acid comprises the portion of SEQ ID NOs: 31-45 encoding all the consecutive amino acids between the signal peptide and avi-tag. In some embodiments, the nucleic acid sequence further comprises a portion encoding a signal peptide. In some embodiments, the nucleic acid sequence further comprises a portion encoding an avi-tag. [0152] In certain aspects, the invention provides a recombinant trimer comprising three identical protomers of an HIV-1 envelope comprising all the consecutive amino acids between the signal peptide and avi-tag of SEQ ID NOs:10-24. In some embodiments, the HIV-1 envelope comprises all the consecutive amino acids between the signal peptide and avi-tag of SEQ ID NO: 14.
  • the HIV-1 envelope comprising all the consecutive amino acids between the signal peptide and avi-tag of SEQ ID NO: 14 further comprises mutation K282R. In some embodiments, the HIV-1 envelope comprising all the consecutive amino acids between the signal peptide and avi-tag of SEQ ID NO: 14 further comprises mutation R456Q. In some embodiments, the HIV-1 envelope comprising all the consecutive amino acids between the signal peptide and avi-tag of SEQ ID NO: 14 further comprises mutation K282R and R456Q. In some embodiments, the HIV-1 envelope comprises all the consecutive amino acids between the signal peptide and avi-tag of SEQ ID NO: 15.
  • the HIV-1 envelope comprising all the consecutive amino acids between the signal peptide and avi-tag of SEQ ID NO: 15 further comprises mutation R456Q. In some embodiments, the HIV-1 envelope comprises all the consecutive amino acids between the signal peptide and avi-tag of SEQ ID NO: 16. In some embodiments, the 28 ActiveUS 201518670 Attorney Docket No.: 1234300.00424WO1 (DU7956PCT) HIV-1 envelope comprises all the consecutive amino acids between the signal peptide and avi-tag of SEQ ID NO: 10. In some embodiments, any of the HIV-1 envelopes further comprises mutation N276G. [0153] In certain aspects, the invention provides an immunogenic composition comprising the recombinant trimer described herein and a carrier.
  • the invention provides an immunogenic composition comprising the nucleic acid described herein and a carrier. In certain aspects, the invention provides an immunogenic composition comprising the recombinant HIV-1 envelopes described herein and a carrier. In some embodiments, the immunogenic composition further comprises an adjuvant. In some embodiments, the nucleic acid is operably linked to a promoter, and optionally the nucleic acid is inserted in an expression vector. [0154] In certain aspects, the invention provides a method of inducing an immune response in a subject comprising administering a composition comprising any suitable form of a nucleic acid(s) described herein or the envelope described herein in an amount sufficient to induce an immune response.
  • the nucleic acid encodes a protomer of a SOSIP trimer.
  • the envelope is a protomer of a SOSIP trimer.
  • the composition further comprises an adjuvant.
  • the method further comprises administering an agent which modulates host immune tolerance.
  • the envelope administered is multimerized in a liposome or nanoparticle.
  • the method further comprises administering one or more additional HIV-1 immunogens to induce a T cell response.
  • the invention provides a composition comprising a nanoparticle and a carrier, wherein the nanoparticle comprises any one of the envelopes described herein.
  • the nanoparticle is ferritin self-assembling nanoparticle.
  • the composition is an immunogenic composition.
  • the invention provides a composition comprising a nanoparticle and a carrier, wherein the nanoparticle comprises any one of the trimers described herein.
  • the nanoparticle is ferritin self-assembling nanoparticle.
  • the nanoparticle comprises multimers of trimers.
  • the composition is an immunogenic composition.
  • the invention provides a composition comprising a nanoparticle and a carrier, wherein the nanoparticle comprises any one of the nucleic acids described herein.
  • the nanoparticle is a lipid nanoparticle.
  • the composition is administered as a prime. In some embodiments, the composition is 29 ActiveUS 201518670 Attorney Docket No.: 1234300.00424WO1 (DU7956PCT) administered as a boost. In some embodiments, the composition is an immunogenic composition.
  • the invention provides a method of inducing an immune response in a subject comprising administering an immunogenic composition described herein in an amount sufficient to induce an immune response. [0159] In certain aspects, the invention provides a composition comprising the recombinant trimer described herein and a carrier. In certain aspects, the invention provides a composition comprising the nucleic acid described herein and a carrier.
  • the invention provides a composition comprising the envelope described herein and a carrier.
  • EXAMPLES [0160] Example 1 - Evolving CD4 Binding Site Immunogens Via Antibody Lineage Selection and Mammalian Cell Display [0161] Background [0162] Anti-HIV broadly neutralizing antibodies (bnAbs) typically develop through extensive co-evolution with virus during chronic infection. Sequential vaccination, targeting these potent humoral responses, assumes an optimized sequence of differing envelopes can similarly mature B cell receptors toward neutralization breadth and potency.
  • CH235.SHM7 library sorting yielded seven stable single mutants, four targeting the 276 glycan sequon, as well as N279H, K282R, and R456Q with evidence of improved kinetics (down to picomolar affinity for CH235.12) and binding for CH235/8ANC131 lineage members.
  • K282R lower stringency sorting
  • higher stringency pointed toward glycan sequon dominance In early, lower stringency (higher antibody concentration) sorting K282R dominated, while later, higher stringency pointed toward glycan sequon dominance.
  • Example 2 CH505.TF Immunogen Design
  • Fig. 2-11 disclose CH505.TF immunogen design.
  • the existing priming/early boosting immunogen(s) in nonhuman primates (NHPs) were first characterized (Fig. 2). Key residues and somatic hypermutations required for VH1-46 bnAbs neutralization breadth were defined. Then, boosting immunogens were designed to select critical mutations.
  • SHM7 can select for a higher affinity immunogen
  • a mutagenesis library was constructed (Fig. 3). Saturation scanning mutagenesis was performed by replacing each target residues with all other possible amino acids, as shown in Fig. 3.
  • Fig. 4 shows successive enrichment of SHM7 binding-enhanced mutants. A gate for SHM7 was set using fluorescence with TF. Then, the mutagenesis library mutants with higher TF binding were sorted. [0174] Fig. 5 shows the sequential selection of envelope variants enriched primarily D-loop mutants. SMH7 x1 showed highly diverse mutations (24 mutations >0.5%) and early emergence of K282R (near HCDR3).
  • Fig. 6 shows the affinity of CH505 mutants by using bio-layer interferometry. It shows improvements in association rates for mature and early intermediates. There were also some improvements in dissociation for VH1-46 antibodies.
  • Fig. 7 shows that most mutations enhanced binding to CH235 lineage members. [0177] In summary, removal of glycan-276 is antigenically dominant. This should be excluded in the future.
  • T278R provides negative counter-selection against immature antibodies.
  • N279H, K282R and R456Q provide modest improvements for CH235 lineage members (limited evidence for other VH1-46s). Combinations should be tested for synergism.
  • the mutants were tested whether they show an enhancement of CH235.UCA- targeting boosts, resulting in improvements in germline breadth. See Fig. 8.
  • Fig. 9 shows successive enrichment of UCA binding-enhanced mutants.
  • Fig. 10 shows that sequential selection of envelope variants again enriched primarily D-loop mutants.
  • Fig. 11 shows ELISA and Bio-Layer Interferometry evaluation of CH235.UCA-sorted mutants.
  • loop D mutations are highly deterministic of CH235 antibody binding; glycan 276 knockout is not the dominant species: S274Q, N279R, I284W; N279L and M475L clearly dominate lower affinity population; and a new “N2” Scanning Saturation Mutagenesis Library with two variable sites in defined subepitopes and invariant glycans was developed.
  • Figs. 12-17 disclose evolving CD4 binding site immunogens via antibody lineage selection and mammalian cell display.
  • Fig. 12 shows a methodology. A site-scanning saturation library was generated by using saturation mutagenesis.
  • Fig. 13 shows the enrichment data of CH235.SHM7, where fluorescence with TF was used to set gate for SHM7. Mutagenesis library mutants with higher TF binding were sorted. [0186] Fig.
  • FIG. 14 shows the enrichment data of CH235.UCA.
  • Fig. 15 shows the mutation enrichment data of CH235.SHM7 sorting and CH235.UCA sorting.
  • Fig. 16 shows the evaluation of CH235.SHM7-sorted mutants by using ELISA and Bio-Layer Interferometry.
  • Fig. 17A shows the evaluation of CH235.UCA-sorted mutants by using ELISA
  • Fig. 17B shows the kinetics of CH235.UCA-sorted mutants. CH235.12 showed a higher association rate and a lower dissociation rate than CH235UCA.
  • Example 3 [0192] This example describes animal studies with HIV-1 envelopes designed to prime and boost CD4 binding antibodies lineages.
  • the envelopes described in Table 1, expressed as recombinant proteins are analyzed in animal studies including mouse and NHP animal models.
  • the mouse animal model could be any model, including an animal model comprising a CH235 UCA transgene.
  • Any suitable adjuvant will be used.
  • the envelopes in Table 1 will be produced under cGMP conditions as a recombinant protein for use in Phase I clinical trial.
  • Example 4 [0197] Fig. 19A shows that SHM7 heterologous reactivity requires a subset of mutations.
  • the ELISA binding data demonstrate the role of different SHM7 mutations on heterologous envelope binding reactivity by single mutation reversion. SHM7 binding is presented as a comparison for each individual reversion.
  • Fig. 19B shows that SHM7-selected double/triple mutants of CH505 place pressure on critical SHM7 mutations.
  • the ELISA binding data demonstrate the role of different SHM7-selected mutations coupled in binding specific mutations from SHM7. SHM7 binding is presented as a comparison for each individual reversion and CH505.TF is presented as a comparison for each of the SHM7-selected mutant pairings.
  • PGT145 is an envelope trimer 33 ActiveUS 201518670 Attorney Docket No.: 1234300.00424WO1 (DU7956PCT) apex bnAb that is a positive control for envelope reactivity and conformational integrity, while nCoV-NTD is an unrelated SARS-CoV-2 viral protein used as a negative control to ascertain binding specificity. Scale is in Log10 area under the curve as measured via trapezoid rule. [0199] Fig. 19C shows the continuation of Fig. 19B, showing difference in log10 area under the curve binding for each SHM7 reversion, minus SHM7 log10area under the curve. This demonstrates that certain envelope mutant pairings may present selective pressure for specific mutations in SHM7, particularly those also deemed important for heterologous reactivity. 34 ActiveUS 201518670

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Virology (AREA)
  • General Health & Medical Sciences (AREA)
  • Organic Chemistry (AREA)
  • Medicinal Chemistry (AREA)
  • Animal Behavior & Ethology (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Veterinary Medicine (AREA)
  • Molecular Biology (AREA)
  • Public Health (AREA)
  • AIDS & HIV (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Communicable Diseases (AREA)
  • Oncology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Biochemistry (AREA)
  • Genetics & Genomics (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Biophysics (AREA)
  • Immunology (AREA)
  • Microbiology (AREA)
  • Mycology (AREA)
  • Epidemiology (AREA)
  • Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)

Abstract

Dans certains aspects, l'invention concerne des immunogènes du VIH-1, comprenant des enveloppes du VIH-1 ayant une boucle D optimisée, une boucle V5 et/ou une boucle de liaison à CD4 pour l'induction d'anticorps.
PCT/US2023/077669 2022-10-24 2023-10-24 Compositions comprenant des enveloppes modifiées pour activer des précurseurs d'anticorps largement neutralisants de site de liaison à cd4 WO2024091962A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US202263418920P 2022-10-24 2022-10-24
US63/418,920 2022-10-24

Publications (1)

Publication Number Publication Date
WO2024091962A1 true WO2024091962A1 (fr) 2024-05-02

Family

ID=90831922

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2023/077669 WO2024091962A1 (fr) 2022-10-24 2023-10-24 Compositions comprenant des enveloppes modifiées pour activer des précurseurs d'anticorps largement neutralisants de site de liaison à cd4

Country Status (1)

Country Link
WO (1) WO2024091962A1 (fr)

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20170233441A1 (en) * 2014-09-04 2017-08-17 The United States Of America, As Represented By The Secretary, Department Of Health And Human Serv Recombinant hiv-1 envelope proteins and their use
US10968255B2 (en) * 2016-03-01 2021-04-06 Duke University Compositions comprising HIV envelopes to induce CH235 lineage antibodies
US11161895B2 (en) * 2016-10-03 2021-11-02 Duke University Methods to identify immunogens by targeting improbable mutations
US11318197B2 (en) * 2016-03-03 2022-05-03 Duke University Compositions and methods for inducing HIV-1 antibodies

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20170233441A1 (en) * 2014-09-04 2017-08-17 The United States Of America, As Represented By The Secretary, Department Of Health And Human Serv Recombinant hiv-1 envelope proteins and their use
US10968255B2 (en) * 2016-03-01 2021-04-06 Duke University Compositions comprising HIV envelopes to induce CH235 lineage antibodies
US11318197B2 (en) * 2016-03-03 2022-05-03 Duke University Compositions and methods for inducing HIV-1 antibodies
US11161895B2 (en) * 2016-10-03 2021-11-02 Duke University Methods to identify immunogens by targeting improbable mutations

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
MU, Z. ET AL.: "mRNA-encoded HIV-1 Env trimer ferritin nanoparticles inducemonoclonal antibodies that neutralize heterologous HIV-1 isolates in mice", CELL REPORT, vol. 110514, no. 1, 15 March 2022; Vol. 38, pages 1 *

Similar Documents

Publication Publication Date Title
US20210009640A1 (en) Compositions comprising hiv envelopes to induce hiv-1 antibodies
US20210379178A1 (en) Compositions comprising hiv envelopes to induce hiv-1 antibodies
US11773144B2 (en) Mosaic HIV-1 envelopes to induce ADCC responses
US20220380412A1 (en) Compositions comprising v2 opt hiv envelopes
US10322141B2 (en) Compositions comprising CH848 envelopes and uses thereof
US20230382952A1 (en) Compositions comprising hiv envelopes to induce hiv-1 antibodies
US10232034B2 (en) Compositions comprising CH505 envelopes, and trimers
EP4415752A2 (fr) Compositions comprenant des enveloppes de vih opt v2
WO2024091962A1 (fr) Compositions comprenant des enveloppes modifiées pour activer des précurseurs d'anticorps largement neutralisants de site de liaison à cd4
WO2022006095A2 (fr) Enveloppes mosaïques du vih-1 pour induire des réponses d'adcc
WO2024091968A1 (fr) Compositions comprenant des arnm codant pour des peptides de la région externe proximale de la membrane du vih-1 (mper)
WO2024091976A1 (fr) Compositions comprenant des peptides de région externe proximale de membrane de vih-1 (mper) et des acides nucléiques codant pour des peptides de mper
WO2024092058A1 (fr) Compositions comprenant des enveloppes de vih-1 avec v1v2 modifié ou des arnm codant pour celles-ci pour une liaison d'anticorps de neutralisation de v3-glycane large
WO2024215947A1 (fr) Compositions comprenant des enveloppes de vih opt v2
US20240197854A1 (en) Compositions comprising hiv envelopes to induce hiv-1 antibodies
US20180036400A1 (en) Double engineered hiv-1 envelopes
WO2024092061A1 (fr) Compositions comprenant des enveloppes de vih-1 ayant v1v2 modifié pour liaison d'anticorps neutralisant le v3-glycane large
WO2023235825A1 (fr) Enveloppes ch505 destinées à venir en prise avec des anticorps de neutralisation de site de liaison cd4 matures
WO2022192262A1 (fr) Nanoparticules de glycopeptides de l'enveloppe du vih-1 et leurs utilisations
WO2017152144A1 (fr) Immunisation d'essaim par des enveloppes de ch505
CA2983259A1 (fr) Immunisation en essaim au moyen d'enveloppes de ch505

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 23883660

Country of ref document: EP

Kind code of ref document: A1