Erythrocyte-binding therapeutics

Patent metadata

Field	Value
Publication number	US-12060414-B2
Application number	US-202017011321-A
Country	US
Kind code	B2
Filing date	Sep 3, 2020
Priority date	Aug 10, 2010
Publication date	Aug 13, 2024
Grant date	Aug 13, 2024

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.

First claim

The invention claimed is:

1. A method for inducing antigen-specific tolerance to an antigen in a subject, the method comprising:
administering a composition to a subject in an amount effective to induce antigen-specific tolerance, the composition comprising:
an antigen to which tolerance is desired;
wherein the antigen is selected from a group consisting of an immunogenic fragment of desmoglein 3, an immunogenic fragment of desmoglein 1, and an immunogenic fragment of desmoglein 4;

an erythrocyte-binding moiety,
wherein the erythrocyte-binding moiety comprises an antibody fragment,
wherein the antigen to which tolerance is desired is recombinantly fused or chemically conjugated to the erythrocyte-binding moiety at its antigen-binding site,
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 presents said antigen to an immune system of a human,
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 when the human is exposed to the antigen alone; and

wherein the erythrocyte-binding moiety specifically binds to a biomolecule selected from the group consisting of Band 3 (CD233), glycophorin A, glycophorin B (CD235b), glycophorin C (CD235c), glycophorin D (CD235d).

2. The method of claim 1 , wherein said administration is intravenous, wherein the antibody fragment is directed against glycophorin A, wherein the administration of the composition ameliorates myasthenia gravis or pemphigus vulgaris, and wherein the composition reduces a 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 method of claim 1 , wherein the erythrocyte-binding moiety is affinity matured, wherein the erythrocyte-binding moiety does not specifically bind to other blood components, wherein the other blood components comprise blood proteins, albumin, fibronectin, platelets, and white blood cells, wherein the binding of the erythrocyte-binding moiety to the erythrocyte cell surface does not alter cell morphology, wherein there is no cytoplasmic translocation of the composition upon binding of the erythrocyte-binding moiety to the erythrocyte cell surface, and wherein the binding of the erythrocyte-binding moiety to the erythrocyte cell surface does not cause the erythrocytes to become apoptotic.

4. The method of claim 1 , wherein the erythrocyte-binding moiety comprises an scFv, a divalent scFv, a diabody, a triabody, or a tetrabody.

5. The method of claim 1 , wherein the antibody fragment is affinity matured.

6. A method for inducing antigen-specific tolerance to an antigen in a subject, the method comprising:
administering a composition to a subject in an amount effective to induce antigen-specific tolerance, the composition comprising:
an antigen recombinantly fused or chemically conjugated with an erythrocyte-binding moiety;
said antigen being recognizable by an immune system of a subject, the immune system of the subject being able to respond to the antigen with an unwanted immune response wherein the antigen is selected from the group consisting of an immunogenic fragment of desmoglein 3, an immunogenic fragment of desmoglein 1, and an immunogenic fragment of desmoglein 4;
wherein said erythrocyte-binding moiety comprises a polypeptide;
wherein upon administration of the composition, said erythrocyte-binding moiety non-covalently and specifically binds a human erythrocyte in situ in blood and presents said antigen to the immune system of the subject;
wherein the erythrocyte-binding moiety does not specifically bind to other blood components, wherein the other blood components comprises blood proteins, albumin, fibronectin, platelets, and white blood cells;
wherein said composition elicits a tolerogenic response upon administration to said subject; and
wherein the erythrocyte-binding moiety comprises an antibody fragment, and wherein the antibody fragment has undergone affinity maturation.

7. The method of claim 6 , wherein the erythrocyte-binding moiety is chemically conjugated to the antigen.

8. The method of claim 6 , wherein the erythrocyte-binding moiety is fused to the antigen via recombinant DNA technology.

9. A method for inducing antigen-specific tolerance to an antigen in a subject, the method comprising:
administering a composition to a subject in an amount effective to induce antigen-specific tolerance, the composition comprising:
one or more antigens fused or chemically conjugated with an erythrocyte-binding moiety and comprises two or more antigens selected from the group consisting of an immunogenic fragment of desmoglein 3, an immunogenic fragment of desmoglein 1, and an immunogenic fragment of desmoglein 4;
wherein said one or more antigens is associated with an autoimmune disease;
wherein said one or more antigens being recognizable by an immune system of a subject, the immune system of the subject being able to respond to or previously having responded to the one or more antigens with an unwanted immune response,
said erythrocyte-binding moiety having the ability to non-covalently, specifically bind an erythrocyte surface in situ in blood and present said one or more antigens to the immune system of the subject,
wherein said erythrocyte-binding moiety comprises an antibody.

10. The method of claim 9 , wherein the erythrocyte-binding moiety has the ability to bind Band 3 (CD233), glycophorin-A, glycophorin B (CD235b), glycophorin C (CD235c), or glycophorin D (CD235d).

11. The method of Claim 9 , wherein said antigens are conjugated to a multimeric branched polymer.

12. The method of Claim 9 , wherein the erythrocyte-binding moiety is affinity-matured.

13. The method of claim 9 , wherein the erythrocyte-binding moiety is fused via a linker, to the N- or C-terminus of the one or more antigens.

14. The method of Claim 13 , wherein the linker is a peptide, a polymer, an aptamer, a nucleic acid, or a particle.

15. The method of claim 9 , wherein the erythrocyte-binding moiety binds to a protein generating a dissociation constant of between about 10 μM and 0.1 nM as determined by equilibrium binding measurements between the erythrocyte-binding moiety and erythrocytes.

Assignees

• Ecole Polytechnique Fed Lausanne Epfl

Inventors

• Hubbell Jeffrey A
• Kontos Stephan
• Dane Karen Y

Classifications

• A61K47/6901

  Conjugates being cells, cell fragments, viruses, ghosts, red blood cells or viral vectors · CPC title

• A61K47/646

  the entire peptide or protein drug conjugate elicits an immune response, e.g. conjugate vaccines · CPC title

• A61K47/6849

  the antibody targeting a receptor, a cell surface antigen or a cell surface determinant · CPC title

• A61K47/60

  the organic macromolecular compound being a polyoxyalkylene oligomer, polymer or dendrimer, e.g. PEG, PPG, PEO or polyglycerol · CPC title

• A61K47/64

  Drug-peptide, drug-protein or drug-polyamino acid conjugates, i.e. the modifying agent being a peptide, protein or polyamino acid which is covalently bonded or complexed to a therapeutically active agent (peptidic linkers A61K47/65) · CPC title