Compositions for generating immune tolerance by targeting erythrocytes

• US 9,878,048 B2
• Filed: 08/09/2016
• Issued: 01/30/2018
• 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:
  US15/232,714
• Publication Number:
  US 20160354453A1
• Time in Patent Office: 539 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 47/6803 Drugs conjugated to an antibody or immunoglobulin, e.g. cisplatin-antibody conjugates
• 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 composition for immunomodulation to achieve antigen-specific tolerance, the composition comprising:

an antigen that results in an unwanted immune response upon administration of the antigen to a subject and recognition of the antigen by an immune cell;

wherein said antigen comprises Factor VIII;

an erythrocyte-binding moiety,

wherein said erythrocyte-binding moiety comprises an antibody fragment

wherein the antigen is fused with the erythrocyte-binding moiety;

wherein said erythrocyte-binding moiety does not specifically bind to other blood components other than glycophorin-A;

wherein said composition is suitable for intravenous delivery to the subject;

wherein said erythrocyte-binding moiety has the ability to non-covalently bind an erythrocyte in situ in blood; and

wherein said binding results in the antigen being presented to the immune system and results in antigen-specific tolerance.

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

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

4. The composition of claim 3, wherein the erythrocyte-binding moiety is capable of binding said glycophorin A with an affinity generating 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.

5. The composition of claim 1, wherein the erythrocyte-binding moiety comprises an antibody fragment derived from a hybridoma that produces an antibody against an erythrocyte.

6. The composition of claim 5, wherein the erythrocyte-binding moiety is an affinity matured antibody fragment.

7. The composition of claim 5, wherein the erythrocyte-binding moiety comprises an antibody fragment that is affinity matured and derived from a 10F7 clone.

8. A composition for immunomodulation to achieve antigen-specific tolerance, the composition comprising:

an antigen selected from the group consisting of Factor VIII and fragments thereof, the antigen being chemically conjugated with at least one erythrocyte-binding moiety, an immune system of the subject being able to respond to or previously having responded to the antigen with an unwanted immune response,

said erythrocyte-binding moiety having the ability to non-covalently bind an erythrocyte surface in situ in blood and present said antigen to the immune system of the subject in a tolerogenic manner,

wherein said erythrocyte-binding moiety has the ability to bind to an erythrocyte surface molecule selected from the group consisting of Band 3 (CD233), glycophorin-A, glycophorin B (CD235b), glycophorin C (CD235c), and glycophorin D (CD235d),

wherein said erythrocyte-binding moiety comprises a protein or peptide ligand,

wherein said erythrocyte-binding moiety does not specifically bind to other blood components, and

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.

9. The composition of claim 8, wherein the erythrocyte-binding moiety comprises a peptide ligand, wherein multiple copies of the erythrocyte-binding moiety are conjugated to the antigen at a plurality of sites on the antigen.

10. The composition of claim 8, wherein the peptide ligand is selected from the group consisting of SEQ ID NO:

11, SEQ ID NO;

12, SEQ ID NO;

13, SEQ ID NO;

14, SEQ ID NO;

15, SEQ ID NO;

16, and SEQ ID NO;

17.

11. The composition of claim 8, wherein the protein comprises an antibody fragment, wherein the antibody fragment is derived from a 10F7 clone, and wherein the antibody fragment is capable of binding human glycophorin A.

12. The composition of claim 11, wherein the antibody fragment has undergone affinity maturation.

13. A composition for immunomodulation to achieve antigen-specific tolerance, the composition comprising:

an antigen fused or chemically conjugated with an erythrocyte-binding moiety;

said antigen comprising a therapeutic protein or peptide;

said antigen 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 antigen 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 antigen to the immune system of the subject,

wherein said erythrocyte-binding moiety has the ability to bind glycophorin A with an affinity generating 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, and

wherein said erythrocyte-binding moiety comprises a peptide, an antibody, or an antibody fragment.

14. The composition of claim 13, wherein the therapeutic protein or peptide is Factor VIII, wherein the antigen is fused with the erythrocyte-binding moiety, and wherein the erythrocyte-binding moiety is derived from a 10F7 clone.

15. The composition of claim 13, wherein the erythrocyte-binding moiety comprises a peptide ligand and wherein multiple copies of the erythrocyte-binding moiety are conjugated to the antigen at a plurality of sites on the antigen.

16. The composition of claim 13, wherein the erythrocyte-binding moiety is fused to the antigen at the N- or C-terminus of the antigen.

17. The composition of claim 13, wherein the erythrocyte-binding moiety comprises an antibody, an antibody fragment, or a single chain variable fragment (scFv).

18. The composition of claim 17, wherein the antibody, antibody fragment, or scFv has undergone affinity maturation.

19. The composition of claim 17, wherein the antibody, antibody fragment, or scFv is derived from a hybridoma that produces an antibody against an erythrocyte.

20. The composition of claim 17, wherein the antibody, antibody fragment, or scFv is derived from a 10F7 clone.

21. The composition of claim 13, wherein the erythrocyte-binding moiety comprises a peptide ligand selected from the group consisting of SEQ ID NO:

11, SEQ ID NO;

12, SEQ ID NO;

13, SEQ ID NO;

14, SEQ ID NO;

15, SEQ ID NO;

16, and SEQ ID NO;

17.