Erythrocyte-binding therapeutics

• US 10,392,437 B2
• Filed: 11/21/2016
• Issued: 08/27/2019
• 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:
  US15/357,999
• Publication Number:
  US 20170066825A1
• Time in Patent Office: 1,009 Days
• CPC Class Codes: A61K 2039/505 comprising antibodies
• CPC Class Codes: A61K 2039/57 characterised by the type of response, e.g. Th1, Th2
• CPC Class Codes: A61K 2039/577 tolerising response
• CPC Class Codes: A61K 2039/6031 Proteins
• 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 47/6849 the antibody targeting a receptor, a cell surface antigen or a cell surface determinant
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 37/02 Immunomodulators
• CPC Class Codes: A61P 37/04 Immunostimulants
• CPC Class Codes: A61P 37/06 Immunosuppressants, e.g. drugs for graft rejection
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: B82Y 5/00 Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery
• CPC Class Codes: C07K 14/4701 not used
• CPC Class Codes: C07K 14/77 Ovalbumin
• 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 16/30 from tumour cells
• CPC Class Codes: C07K 2317/34 : Identification of a linear epitope shorter than 20 amino acid residues or of a conformational epitope defined by amino acid residues
• CPC Class Codes: C07K 2317/622 : Single chain antibody (scFv)
• CPC Class Codes: C07K 2319/00 : Fusion polypeptide
• CPC Class Codes: C07K 2319/01 : containing a localisation/targetting motif
• 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 2319/33 : 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

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.

Claims

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 insulinoma-associated protein 2 (IA-2) and an immunogenic portion of said IA-2;

an erythrocyte-binding moiety,

wherein the erythrocyte-binding moiety comprises an antibody fragment that has the ability to non-covalently, specifically bind an exterior erythrocyte surface in situ in blood,

wherein the erythrocyte-binding moiety is affinity matured,

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, and

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.

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

3. The composition of claim 2 wherein the erythrocyte-binding moiety is derived from 10F7.

4. The composition of claim 1 wherein the erythrocyte-binding moiety is derived from 10F7 and the antigen to which tolerance is desired is an immunogenic portion of said IA-2.

5. The composition of claim 4 wherein the administration of the composition ameliorates Type I Diabetes.

6. A composition comprising an antigen recombinantly fused or chemically conjugated with an erythrocyte-binding moiety;

wherein the erythrocyte-binding moiety comprises an antibody fragment that specifically binds to human erythrocytes; and

wherein the antigen is a self-antigen to which a subject develops an unwanted immune response,

wherein the self-antigen is associated with type 1 diabetes, and

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.

7. The composition of claim 6, wherein the self-antigen is selected from the group consisting of glutamic acid decarboxylase 65 (GAD-65), GAD-67, insulinoma-associated protein 2 (IA-2), insulinoma-associated protein 2β

(IA-2b), islet-specific glucose-6-phosphatase catalytic subunit-related protein, a portion of any one of said GAD-65, said GAD-67, said IA-2, said IA-2b, and said islet-specific glucose-6-phosphatase catalytic subunit-related protein, and mimotopes thereof.

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

9. The composition of claim 6, wherein the erythrocyte-binding moiety comprises an antibody fragment, and wherein the erythrocyte-binding moiety is derived from a 10F7 clone.

10. A composition for use in inducing tolerance comprising:

an antigen to which tolerance is desired;

wherein the antigen is a self-antigen to which a subject develops an unwanted immune response,

and wherein the self-antigen is associated with type 1 diabetes;

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 or antibody fragment directed against glycophorin A,

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 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.

11. The composition of claim 10 wherein the self-antigen is selected from the group consisting of glutamic acid decarboxylase 65 (GAD-65), GAD-67, insulinoma-associated protein 2 (IA-2), insulinoma-associated protein 2β

(IA-2b), islet-specific glucose-6-phosphatase catalytic subunit-related protein, portions thereof, and mimotopes thereof.