Neutralising antibody against dengue for use in a method of prevention and/or treatment of zika infection

Patent metadata

Field	Value
Publication number	US-11111274-B2
Application number	US-201716308745-A
Country	US
Kind code	B2
Filing date	Jun 9, 2017
Priority date	Jun 10, 2016
Publication date	Sep 7, 2021
Grant date	Sep 7, 2021

Abstract

A flavivirus Envelope Dimer Epitope (EDE) for use in vaccinating an individual against one or more flaviviruses wherein the EDE is a stabilized recombinant flavivirus, optionally dengue virus and/or zika envelope glycoprotein E ectodomain (sE) dimer, wherein the dimer is: covalently stabilized with at least one disulphide inter-chain bond between the two sE monomers, and/or non-covalently stabilized by substituting at least one amino acid residue in the amino acid sequence of at least one sE monomer with at least one bulky side chain amino acid, at the dimer interface or in domain 1 (D1)/domain 3 (D3) linker of each monomer, covalently stabilized with at least one sulfhydryl-reactive crosslinker between the two sE monomers, and/or covalently stabilised by being formed as a single polypeptide chain, optionally with a linker region, optionally a Glycine Serine rich linker region, separating the sE sequences, and/or covalently stabilized by linking the two sE monomers through modified sugars; and/or, wherein the dimer is a homodimer or heterodimer of native and/or mutant envelope polypeptides, from any one or two of DENV-1, DENV-2, DENV-3, DENV-4, Zika or other flavivirus; and wherein the one or more flaviviruses is selected from zika virus; zika virus and dengue virus; zika virus and other flaviviruses; flaviviruses other than dengue. The EDE may be a homodimer or heterodimer of native and/or mutant envelope polypeptides, from any one or two of DENV-1, DENV-2, DENV-3, DENV-4 and Zika. An isolated neutralizing antibody or antigen binding fragment thereof directed against the EDE as defined in any one of claims 1 to 29, optionally wherein said antibody or fragment thereof binds the five polypeptide segments of the dengue virus glycoprotein E ectodomain (sE) consisting of the residues 67-74, residues 97-106, residues 307-314, residues 148-159 and residues 243-251, or corresponding residues of the flavivirus or Zika virus glycoprotein E ectodomain, or consisting of Zika PF13 residues 67-77, residues 97-106, residues 313-315, residues 243-253, residue K373 or corresponding residues of the flavivirus glycoprotein E ectodomain, optionally wherein binding is unaffected by presence or absence of dengue N153 (Zika N154) glycan or corresponding residue, for use in a method for prevention and/or treatment of infection by one or more flaviviruses, wherein the one or more flaviviruses is selected from zika virus; zika virus and dengue virus; zika virus and other flaviviruses; flaviviruses other than dengue.

First claim

The invention claimed is:

1. A method for increasing an immune response against one or more flaviviruses in a subject, wherein the one or more flaviviruses is not a dengue virus, the method comprising administering a flavivirus Envelope Dimer Epitope (EDE) to the subject,
wherein the EDE is a stabilized recombinant flavivirus-envelope glycoprotein E ectodomain (sE) dimer, wherein the dimer is:
covalently stabilized with at least one disulphide inter-chain bond between the two sE monomers, and/or
non-covalently stabilized by substituting at least one amino acid residue in the amino acid sequence of at least one sE monomer with at least one bulky side chain amino acid, at the dimer interface or in domain I (DI)/domain III (DIII) linker of each monomer,
covalently stabilized with at least one sulfhydryl-reactive crosslinker between the two sE monomers, and/or
covalently stabilised by being formed as a single polypeptide chain, optionally with a linker region, optionally a Glycine Serine rich linker region, separating the sE sequences, and/or
covalently stabilized by linking the two sE monomers through modified sugars; and/or,
wherein the dimer is a homodimer or heterodimer of native and/or mutant envelope polypeptides, from any one or two of DENV-1, DENV-2, DENV-3, DENV-4, Zika or other flavivirus.

2. The method of claim 1 wherein the dimer is a homodimer or heterodimer of native and/or mutant envelope polypeptides, from any one or two of DENV-1, DENV-2, DENV-3, DENV-4 and Zika.

3. The method of claim 1 , wherein the subject is a human subject, wherein the human subject is a pregnant female.

4. The method of claim 1 , wherein the recombinant sE monomer is selected from the group consisting of
SEQ ID No: 1; SEQ ID No: 2; SEQ ID No: 3; SEQ ID No: 4; SEQ ID No: 5; SEQ ID No: 6; SEQ ID No: 7; SEQ ID No: 8; SEQ ID No: 9;
or a mutant sE thereof having at least one mutation selected among mutations #1 to #18 as set out in.

5. The method of claim 1 , wherein the dimer is: (a) glycosylated at the position corresponding to position 67 of DENV-2 and optionally, at the position corresponding to position 153 of DENV-2 or N154 of Zika PF13 of each sE monomer; or (b) covalently stabilized with at least one, two or three disulphide inter-chain bonds between the two sE monomers.

6. The method of claim 5 , wherein the dimer is:
(a) a homodimer of mutants sE having each the mutation:
(i) #2 corresponding to A259C of DENV-2/A264C of Zika PF13; or mutation #1 corresponding to S255C of DENV-2/S260C of Zika PF13, wherein the residues corresponding to 259C/264C or 255C/260C are linked together through a disulphide inter-chain bond; or
(ii) the mutations #4 corresponding to F108C and T315C of DENV-2/F108C and T321C of Zika PF13, or a homodimer of mutants sE having each the mutations #3 corresponding to L107C of DENV-2 and Zika PF13 and A313C of DENV-2/A319C of Zika PF13, wherein the residues corresponding to 108C and 315C/321C or the residues corresponding to 107C and 313C/319C are linked together through a disulphide inter-chain bond

(b) a heterodimer of a mutant sE having the mutations:
(i) #2 corresponding to A259C of DENV-2/A264C of Zika PF13 and a mutant sE having the mutation #1 corresponding to S255C of DENV-2/S260C of Zika PF13, and wherein the residues corresponding to 259C/264C and 255C/260C are linked together through a disulphide inter-chain bond; or
(ii) #4 corresponding to F108C of DENV-2 and Zika PF13 and mutation #3 corresponding to A313C of DENV-2/A319C of Zika PF13 and a mutant sE having the mutations #3 corresponding to L107C of DENV-2 and Zika PF13 and mutations #4 corresponding to T315C of DENV-2/321C of Zika PF13, and wherein the residues corresponding to 108C and 313C/319C are linked respectively to the residues corresponding to 315C/321C and 107C through a disulphide inter-chain bond between the two sE monomers.

7. The method of claim 5 , wherein the dimer is selected from the group consisting of:
(a) a homodimer of mutants sE having each the mutations #2 corresponding to A259C of DENV-2/A264C of Zika PF13, and/or #4 corresponding to F108C of DENV-2 and Zika PF13 and T315C of DENV-2/T321C of Zika PF13,
(b) a homodimer of mutants sE having each the mutations #1 corresponding to S255C of DENV-2/S260C of Zika PF13, and/or #4 corresponding to F108C of DENV-2 and Zika PF13 and T315C of DENV-2/T321C of Zika PF13,
(c) a homodimer of mutants sE having each the mutations #2 corresponding to A259C of DENV-2/A264C of Zika PF13, and/or #3 corresponding to L107C of DENV-2 and Zika PF13 and A313C of DENV-2/T319C of Zika PF13, and
(d) a homodimer of mutants sE having each the mutations #1 corresponding to S255C of DENV-2/S260C of Zika PF13, and/or #3 corresponding to L107C of DENV-2 and Zika PF13 and A313C of DENV-2/T319C of Zika PF13,
wherein the residues corresponding to 259C/264C, 255C/260C, 108C, 315C/321C, 107C and 313C/319C are linked respectively to the residues 259C/264C, 255C/260C, 315C/321C, 108C, 313C/319C and 107C through disulphide inter-chain bonds.

8. The method of claim 5 , wherein the dimer is a heterodimer of:
a mutant sE having the mutations #2 corresponding to A259C of DENV-2/A264C of Zika PF13, #4 corresponding to F108C of DENV-2 and Zika PF13 and T315C of DENV-2/T321C of Zika PF13 and
a mutant sE having the mutations #1 corresponding to S255C of DENV-2/S260C of Zika PF13, #4 corresponding to F108C of DENV-2 and Zika PF13 and T315C of DENV-2/T321C of Zika PF13, wherein the residues 259C/264C, 108C and 315C/321C are linked respectively to the residues 255C/260C, 315C/321C and 108C through disulphide inter-chain bonds.

9. The method of claim 5 , wherein the dimer is a heterodimer of a mutant sE having the mutations S255C/260C, L107C and A313C/319C and a mutant sE having the mutations A259C/264C, L107C and A313C/319C, and wherein the residues 255C/260C, 107C and 313C/319C are linked respectively to the residues 259C/264C, 313C/319C and 107C through disulphide inter-chain bonds.

10. The method of claim 1 , wherein said sulfhydryl-reactive crosslinker is selected from the group consisting of a maleimide, a haloacetyl, a pyridyl disulfide, a vinyl sulfone, an alkyl halide or an aziridine compound, an acryloyl derivative, an arylating agent, or a thiol-disulfide exchange reagent, and optionally, wherein the maleimide sulfhydryl-reactive crosslinker is BMOE, BMB, BMH, TMEA, BM(PEG) 2 , BM(PEG) 3 , BMDB, or DTME.

11. The method of claim 10 , wherein the dimer is: (a) a homodimer of mutant sE having each the mutation corresponding to T262C of DENV-2 or T265C of DENV-2, and the residues corresponding to 262C or 265C are linked together through a sulfhydryl-reactive crosslinker; or (b) a heterodimer of a mutant sE having the mutation corresponding to T/S262C of DENV-2 and a mutant sE having the mutation corresponding to T/A265C of DENV-2, and the residues corresponding to 262C and 265C are linked together through a sulfhydryl-reactive crosslinker.

12. The method of claim 1 , wherein the dimer is a homodimer or a heterodimer of
a mutant sE wherein at least one amino acid residue selected from the group consisting of the amino acid residues corresponding to amino acid residues 1-9, 25-30, 238-282, 96-111 and 311-318 of DENV-2 sE is mutated to cysteine and
a mutant sE wherein at least one amino acid residue selected from the group consisting of the amino acid residues corresponding to amino acid residues 1-9, 25-30, 238-282, 96-111 and 311-318 of DENV-2 sE is mutated to cysteine, and wherein the mutated cystein residues are linked together through a sulfhydryl-reactive crosslinker.

13. The method of claim 1 , wherein one of the recombinant sE

Assignees

• Imp Innovations Ltd
• Pasteur Institut
• Imperial College Innovations Ltd

Inventors

• Rey Félix
• Spaeth Giovanna Barba
• Vaney Marie-Christine
• Rouvinski Alexander
• Screaton Gavin
• Mongkolsapaya Juthathip

Classifications

• C07K16/116

  Togaviridae (F); Matonaviridae (F); Flaviviridae (F) · CPC title

• Y02A50/30

  Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change · CPC title

• C12N2770/24122

  New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes · CPC title

• A61P31/12

  Antivirals · CPC title

• C07K14/005 Primary

  from viruses · CPC title