Methods for reducing immune responses

• US 9,901,645 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 Fedrale De Lausanne
• Inventors: Hubbell, Jeffrey A., Kontos, Stephan, Dane, Karen Y.
• Primary Examiner(s): Gebreyesus, Kagnew H
• Application Number:
  US15/232,713
• Publication Number:
  US 20160361431A1
• 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 reducing an immune response of a subject to a foreign antigen, comprising:

administering a fusion molecule to a subject for reducing an immune response and inducing tolerogenesis, the fusion molecule comprising;

a foreign antigen comprising an immunogenic gliadin fragment, wherein exposure of the subject to the immunogenic gliadin fragment elicits an immune response by the subject and

a cell-binding moiety comprising an antibody fragment fused to the N- or C-terminus of the foreign antigen; and

wherein said administration is intravenous,

wherein an amount effective to reduce an immune response to the foreign antigen is administered,

wherein, upon administration the cell binding moiety non-covalently and specifically binds to glycophorin A on the surface of erythrocytes in blood of the subject with a dissociation constant of between about 10 μ

M and 0.1 nM as determined by equilibrium binding measurements between the cell-binding moiety and erythrocytes and does not specifically bind to other blood components, and

wherein the binding of the fusion molecule results in the presentation of the antigen to the immune system of the subject, which downregulates the degree to which the immune system responds to the foreign antigen, thereby reducing the immune response to the foreign antigen.

2. The method of claim 1, wherein the N- or C-terminus of the foreign antigen is fused with the cell-binding moiety via a linker.

3. The method of claim 1, wherein the antibody fragment is derived from a 10F7 clone.

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

5. A method for reducing an immune response of a subject to a foreign antigen, comprising:

administering a molecule for reducing an immune response and inducing tolerogenesis to a subject, the molecule comprising;

an immunogenic component comprising an immunogenic mimotope of gliadin wherein exposure of the subject to the immunogenic mimotope is capable of eliciting an immune response by the subject; and

a cell binding moiety coupled to the immunogenic component comprising an antibody fragment;

wherein the molecule is administered intravenously to the subject in an amount effective to reduce the immune response,

wherein, upon administration the cell binding moiety non-covalently and specifically binds to glycophorin A on the surface of erythrocytes in blood of the subject and does not specifically bind to other blood components, and

wherein the binding of the molecule results in the presentation of the immunogenic component to the immune system of the subject, which downregulates the degree to which the immune system responds to the immunogenic component, thereby reducing the immune response to the immunogenic component.

6. The method of claim 5, wherein the immunogenic component is fused with the cell-binding moiety.

7. The method of claim 6, wherein the N- or C-terminus of the immunogenic component is fused with the cell-binding moiety via a linker.

8. The method of claim 5, wherein the cell-binding moiety exhibits a dissociation constant of between about 10 μ

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

9. The method of claim 5, wherein the immune response is due to the exposure of the subject to one or more cereals, and wherein the antibody fragment is affinity matured.

10. A method for reducing an immune response of a subject to a foreign antigen, comprising:

administering a fusion molecule for reducing an immune response to a subject, the fusion molecule comprising an immunogenic component coupled to a cell binding moiety,

wherein the immunogenic component comprises an antigen foreign to a subject and to which the subject has developed or is likely to develop an unwanted immune response,

wherein the foreign antigen is a food antigen selected from the group consisting of high molecular weight glutenin, low molecular weight glutenin, gliadin, hordein, secalin, and avenin,

wherein the cell binding moiety comprises an antibody fragment that is capable of binding human glycophorin A; and

wherein the fusion molecule is administered intravenously to the subject,

wherein, upon administration the cell binding moiety non-covalently and specifically binds to the surface of erythrocytes in blood of the subject and does not specifically bind to other blood components, and

wherein the binding of the fusion molecule results in the presentation of the foreign antigen to the immune system of the subject, which downregulates the degree to which the immune system responds to the foreign antigen, thereby reducing the immune response to the foreign antigen.

11. The method of claim 10, wherein the cell binding moiety comprises an antibody fragment that is affinity matured and capable of binding human glycophorin A.

12. The method of claim 10, wherein the N- or C-terminus of the immunogenic component is fused with the cell-binding moiety via a linker.

13. The method of claim 10, wherein the cell-binding moiety exhibits a dissociation constant of between about 10 μ

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

14. A method for treating Celiac disease in a subject, comprising:

administering a composition to a subject, the composition comprising an antigen recombinantly fused or chemically conjugated with an erythrocyte-binding moiety;

wherein the antigen is an immunogenic gliadin fragment,

wherein the immunogenic gliadin fragment elicits an immune response, and

wherein the erythrocyte-binding moiety comprises an antibody fragment that binds to human glycophorin A.

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

16. The method of claim 14, wherein the composition:

reduces, fails to induce or prevents inflammatory responses in antigen-specific T cells when the subject is exposed to the antigenic component alone;

comprises the erythrocyte binding moiety which has a dissociation constant of between 10 μ

M and 0.1 nM as determined by equilibrium binding measurements between the erythrocyte-binding moiety and human glycophorin A on erythrocytes;

or

upon administration to a patient, induces the proliferation of CD8+ T cells having an apoptotic- or exhausted-fate phenotype.