Erythrocyte-binding therapeutics

Patent metadata

Field	Value
Publication number	US-9518087-B2
Application number	US-201113206034-A
Country	US
Kind code	B2
Filing date	Aug 9, 2011
Priority date	Aug 10, 2010
Publication date	Dec 13, 2016
Grant date	Dec 13, 2016

Abstract

Peptides that specifically bind erythrocytes are described. These are provided as peptidic ligands having sequences that specifically bind, or as antibodies or fragments thereof that provide specific binding, to erythrocytes. The peptides may be prepared as molecular fusions with therapeutic agents, tolerizing antigens, or targeting peptides. Immunotolerance may be created by use of the fusions and choice of an antigen on a substance for which tolerance is desired. Fusions with targeting peptides direct the fusions to the target, for instance a tumor, where the erythrocyte-binding ligands reduce or entirely eliminate blood flow to the tumor by recruiting erythrocytes to the target.

First claim

The invention claimed is:

1. A composition for use in inducing tolerance comprising:
an antigen to which tolerance is desired;
wherein the antigen is selected from the group consisting of gliadin and a portion thereof;
an erythrocyte-binding moiety,
wherein the erythrocyte-binding moiety has the ability to non-covalently, specifically bind an exterior erythrocyte surface in situ in blood,
wherein the erythrocyte-binding moiety comprises an antibody fragment directed against glycophorin A,

wherein the antigen to which tolerance is desired is recombinantly fused to the erythrocyte-binding moiety,
wherein, upon administration to a human in which tolerance to the antigen is desired:
the composition binds to CD45 negative cells, but not to CD45 positive cells, and the composition reduces, fails to induce, or prevents inflammatory responses in antigen-specific T cells as compared to when the human is exposed to the antigen alone.

2. The composition of claim 1 wherein the composition reduces the number of resident lymph node and spleen cells expressing interferon-gamma (IFNγ), as compared to the number of resident lymph node and spleen cells expressing IFNγ when the human is exposed to the antigen alone.

3. The composition of claim 1 wherein the erythrocyte-binding moiety is fused, optionally via a linker, to the N- or C-terminus of the antigen.

4. The composition of claim 3 wherein the erythrocyte-binding moiety is fused, optionally via a linker, to the N-terminus of the antigen.

5. The composition of claim 4 wherein the erythrocyte-binding moiety is derived from 10F7.

6. The composition of claim 1 wherein the antigen to which tolerance is desired is gliadin.

7. The composition of claim 1 wherein the erythrocyte-binding moiety is derived from 10F7 and the antigen to which tolerance is desired is a fragment of gliadin.

8. The composition of claim 7 wherein the administration of the composition ameliorates celiac disease.

9. A composition for use in inducing tolerance comprising:
an antigen chemically conjugated with an erythrocyte-binding moiety having the ability to non-covalently, specifically bind an exterior erythrocyte surface in situ in blood, said tolerogenic composition being pharmaceutically acceptable, wherein said antigen is selected from the group consisting of gliadin and a portion thereof, wherein, upon administration to a human in which tolerance to the antigen is desired, the composition reduces, fails to induce, or prevents inflammatory responses in antigen-specific T cells, as compared to the antigen alone, and wherein the erythrocyte-binding moiety comprises a peptide ligand that is directed against glycophorin A has a dissociation constant of between about 10 μM and 0.1 nM as determined by equilibrium binding measurements between the peptide and erythrocytes.

10. The composition of claim 9 , wherein the peptide ligand comprises SEQ ID NO:11, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, or SEQ ID NO:17.

11. The composition of claim 9 , wherein the antigen is a portion of gliadin.

12. A composition for use in inducing tolerance comprising:
an antigen to which tolerance is desired;
wherein the antigen is selected from the group consisting of gliadin and portions thereof;

a cell binding moiety,
wherein the cell binding moiety is an erythrocyte-binding moiety having the ability to non-covalently, specifically bind an exterior erythrocyte surface in situ in blood,
wherein the erythrocyte-binding moiety is directed against glycophorin A,
wherein the erythrocyte-binding moiety has erythrocyte dissociation constant of between about 10 μM and 0.1 nM,

wherein the antigen to which tolerance is desired is joined to the erythrocyte-binding moiety,
wherein, upon administration to a human, the composition:
(i) binds to CD45 negative cells, but not to CD45 positive cells,
(ii) induces greater proliferation of antigen-specific CD8+ T cells, as compared to the proliferation of antigen-specific CD8+ T cells induced by the antigen alone, or
(iii) reduces the number of resident lymph node and spleen cells expressing interferon-gamma (IFNγ), as compared to the number of resident lymph node and spleen cells expressing IFNγ when the human is exposed to the antigen alone.

13. The composition of claim 12 , wherein the erythrocyte-binding moiety is fused to the antigen via recombinant DNA technology.

14. The composition of claim 12 , wherein the erythrocyte-binding moiety is fused, optionally via a linker, to the N- or C-terminus of the antigen.

15. The composition of claim 12 , wherein the erythrocyte-binding moiety is covalently bonded to the antigen.

16. The composition of claim 12 , wherein the erythrocyte-binding moiety wherein is derived from 10F7.

17. The composition of claim 16 , wherein the erythrocyte-binding moiety comprises an antibody fragment.

18. The composition of claim 12 , wherein the erythrocyte-binding moiety comprises a peptide ligand.

19. The composition of claim 18 , wherein the peptide ligand comprises SEQ ID NO:11, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, or SEQ ID NO:17.

20. A composition comprising an antigen recombinantly fused or chemically conjugated with an erythrocyte-binding moiety;
wherein the erythrocyte-binding moiety comprises an antibody, antibody fragment, or a single chain variable fragment (scFv) that binds to human glycophorin A; and
wherein the antigen is selected from the group consisting of gliadin, α-gliadin and γ-gliadin, and a portion thereof, and
wherein, upon administration to a human in which tolerance to the antigen is desired:
the composition binds to CD45 negative cells, but not to CD45 positive cells, and the composition reduces, fails to induce, or prevents inflammatory responses in antigen-specific T cells as compared to when the human is exposed to the antigen alone.

21. The composition of claim 20 , wherein the erythrocyte-binding moiety is fused, optionally via a linker to the N- or C-terminus of the antigen.

22. A method for treating Celiac disease in a subject, comprising administering to said subject a composition of claim 20 .

23. A method for treating Celiac disease in a subject, comprising administering to said subject a composition of claim 21 .

Assignees

• Hubbell Jeffrey A
• Kontos Stéphane
• Dane Karen Y
• Ecole Polytechnique Fed De Lausanne (Epfl)

Inventors

• Hubbell Jeffrey A
• Kontos Stéphane
• Dane Karen Y

Classifications

• A61P43/00

  Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00 · CPC title

• A61P37/02

  Immunomodulators · CPC title

• A61P37/04

  Immunostimulants · CPC title

• A61P37/06

  Immunosuppressants, e.g. drugs for graft rejection · CPC title

• A61P35/00

  Antineoplastic agents · CPC title