Methods for induction of antigen-specific immune tolerance

• US 9,901,646 B2
• Filed: 08/09/2016
• Issued: 02/27/2018
• Est. Priority Date: 08/10/2010
• Status: Active Grant

• Current Assignee: Ecole polytechnique fãdãrale de lausanne epfl (Schweizerische Eidgenossenschaft)
• Original Assignee: Ecole polytechnique fãdãrale de lausanne epfl (Schweizerische Eidgenossenschaft)
• Inventors: Hubbell, Jeffrey A., Kontos, Stephan, Dane, Karen Y.
• Primary Examiner(s): Gebreyesus, Kagnew H
• Application Number:
  US15/232,724
• Publication Number:
  US 20160354454A1
• Time in Patent Office: 567 Days
• CPC Class Codes: A61K 2039/6031 Proteins
• CPC Class Codes: A61K 2039/605 MHC molecules or ligands thereof
• CPC Class Codes: A61K 2039/6056 Antibodies
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: A61K 39/0008 Antigens related to auto-immune diseases; Preparations to induce self-tolerance
• CPC Class Codes: A61K 39/001 Preparations to induce tolerance to non-self, e.g. prior to transplantation
• CPC Class Codes: A61K 47/64 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 Class Codes: A61K 47/6811 the drug being a protein or peptide, e.g. transferrin or bleomycin
• CPC Class Codes: A61K 47/6815 Enzymes
• CPC Class Codes: A61K 47/6843 the antibody targeting a material from animals or humans
• CPC Class Codes: A61K 47/6849 the antibody targeting a receptor, a cell surface antigen or a cell surface determinant
• CPC Class Codes: A61K 9/1075 Microemulsions or submicron emulsions; Preconcentrates or solids thereof; Micelles, e.g. made of phospholipids or block copolymers A61K9/0026 takes precedence
• CPC Class Codes: A61K 9/513 Organic macromolecular compounds; Dendrimers
• CPC Class Codes: C07K 14/62 Insulins
• CPC Class Codes: C07K 14/70539 MHC-molecules, e.g. HLA-molecules
• CPC Class Codes: C07K 16/18 against material from animals or humans
• CPC Class Codes: C07K 16/28 against receptors, cell surface antigens or cell surface determinants
• CPC Class Codes: C07K 2317/56 variable (Fv) region, i.e. VH and/or VL
• CPC Class Codes: C07K 2317/622 Single chain antibody (scFv)
• CPC Class Codes: C07K 2317/92 Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value
• CPC Class Codes: C07K 2319/00 : Fusion polypeptide
• CPC Class Codes: C07K 2319/21 : containing a His-tag
• CPC Class Codes: C07K 2319/22 : containing a Strep-tag
• CPC Class Codes: C07K 2319/23 : containing a GST-tag
• CPC Class Codes: C07K 2319/30 : Non-immunoglobulin-derived peptide or protein having an immunoglobulin constant or Fc region, or a fragment thereof, attached thereto
• CPC Class Codes: C07K 2319/33 : fusions for targeting to specific cell types, e.g. tissue specific targeting, targeting of a bacterial subspecies
• CPC Class Codes: C07K 2319/41 : containing a Myc-tag
• CPC Class Codes: C07K 2319/43 : containing a FLAG-tag
• CPC Class Codes: C07K 2319/74 : containing a fusion for binding to a cell surface receptor
• CPC Class Codes: C07K 7/08 : having 12 to 20 amino acids gastrins C07K14/595; somatostatins C07K14/655; melanotropins C07K14/68
• CPC Class Codes: C12N 9/82 : Asparaginase (3.5.1.1)
• CPC Class Codes: C12N 9/96 : Stabilising an enzyme by forming an adduct or a composition; Forming enzyme conjugates
• CPC Class Codes: C12Y 305/01001 : Asparaginase (3.5.1.1)

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.

Claims

1. A method for inducing antigen-specific tolerance in a subject, comprising:

administering a tolerogenic composition to a subject in an amount effective to induce antigen-specific tolerance in the subject, wherein the tolerogenic composition comprises;

an immunogenic component comprising an antigen recognizable by the immune system of a subject, the immune system of the subject responding to the antigen with an unwanted immune response, wherein the immunogenic component is proinsulin or an immunogenic portion thereof; and

an erythrocyte-binding moiety coupled to the immunogenic component, wherein the erythrocyte-binding moiety non-covalently and specifically binds glycophorin A on erythrocytes and does not specifically bind to other blood components, wherein said erythrocyte-binding moiety comprises a peptide, an antibody, or antibody fragment; and

wherein upon administration of the tolerogenic composition, the tolerogenic composition binds specifically to erythrocytes via the cell binding component, and

wherein the cell binding component has a dissociation constant of between about 10 μ

M and 0.1 nM as determined by equilibrium binding measurements between the cell binding component and erythrocytes, and upon presentation of the immunogenic component to the immune system, antigen-specific tolerance is induced.

2. The method of claim 1, wherein said administration is intravenous, wherein the immunogenic component is fused or chemically conjugated with the erythrocyte-binding moiety, wherein the erythrocyte-binding moiety comprises an antibody fragment, and wherein the erythrocyte-binding moiety is derived from a 10F7 clone.

3. The method of claim 2, wherein the antibody fragment is affinity matured.

4. The method of claim 1, wherein the erythrocyte-binding moiety is fused to the N- or C-terminus of the immunogenic component via a linker.

5. The method of claim 1, wherein the erythrocyte-binding moiety is chemically conjugated to the immunogenic component.

6. The method of claim 1, wherein the immunogenic component comprises a fragment of proinsulin.

7. The method of claim 1, wherein the tolerogenic composition comprises a 10F7-derived antibody fragment- proinsulin fusion protein.

8. The method of claim 1, wherein the erythrocyte-binding moiety comprises a peptide ligand, wherein the antigen comprises a fragment of proinsulin, wherein multiple copies of the erythrocyte-binding moiety are directly fused to the fragment of proinsulin at a plurality of sites on the fragment of proinsulin, and wherein the erythrocyte-binding moiety is capable of binding human glycophorin A.

9. A method for inducing antigen-specific tolerance in a subject, comprising:

administering a tolerogenic composition to a subject in an amount effective to induce antigen-specific tolerance in the subject, wherein the tolerogenic composition comprises;

an immunogenic component comprising an antigen recognizable by the immune system of a subject, the immune system of the subject responding to the antigen with an unwanted immune response, wherein the immunogenic component is a proinsulin or an immunogenic portion thereof comprising an epitope from proinsulin; and

an erythrocyte-binding moiety coupled to the immunogenic component, wherein the erythrocyte-binding moiety non-covalently and specifically binds glycophorin A on erythrocytes and does not specifically bind to other blood components, wherein said erythrocyte-binding moiety comprises an antibody fragment; and

,

wherein upon administration of the tolerogenic composition, the tolerogenic composition binds specifically to erythrocytes via the erythrocyte-binding moiety and the immunogenic component is presented to the immune system, thereby inducing antigen-specific tolerance.

10. The method of claim 9, wherein the immunogenic component comprises a fragment of proinsulin.

11. The method of claim 9, wherein the immunogenic component comprises a mimotope of proinsulin.

12. The method of claim 9, wherein said administration is intravenous.

13. The method of claim 9, wherein the antibody fragment has a dissociation constant of between about 10 μ

M and 0.1 nM as determined by equilibrium binding measurements between the antibody fragment and erythrocytes, and wherein the antibody fragment is capable of binding human glycophorin A.

14. A method for inducing antigen-specific tolerance in a subject, comprising:

administering a tolerogenic composition to a subject, the tolerogenic composition comprising an immunogenic component coupled to an erythrocyte-binding moiety;

wherein the immunogenic component comprises an antigen recognizable by the immune system of a subject, the immune system of the subject responding to the antigen with an unwanted immune response,

wherein the immunogenic component is associated with development of Type I diabetes,

wherein the erythrocyte-binding moiety is configured to non-covalently, specifically bind glycophorin A on erythrocytes and not specifically bind to other blood components,

wherein said erythrocyte-binding moiety comprises an antibody fragment, or a single chain variable fragment (scFv) of antibody; and

wherein upon administration of the tolerogenic composition, the tolerogenic composition binds specifically to erythrocytes via the erythrocyte-binding moiety and the immunogenic component is presented to the immune system, thereby inducing antigen-specific tolerance.

15. The method of claim 14, wherein said administration is intravenous, wherein the immunogenic component is chemically conjugated with the erythrocyte-binding moiety, wherein the erythrocyte-binding moiety has a dissociation constant of between about 10 μ

M and 0.1 nM as determined by equilibrium binding measurements between the erythrocyte-binding moiety and erythrocytes.

16. The method of claim 14, wherein the erythrocyte-binding moiety is derived from a 10F7 clone.

17. The method of claim 14, wherein the erythrocyte-binding moiety is an antibody, antibody fragment, or single chain variable fragment derived from an antibody, wherein the antibody, antibody fragment, or the single chain variable fragment is derived from a hybridoma that produces an antibody against a human erythrocyte.