Compositions for inducing antigen-specific tolerance

• US 9,814,780 B2
• Filed: 08/09/2016
• Issued: 11/14/2017
• 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:
  US15/232,694
• Publication Number:
  US 20160354484A1
• Time in Patent Office: 462 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. An engineered composition for inducing antigen-specific tolerance in a subject, the engineered composition comprising:

an antigenic component comprising asparaginase coupled to a cell binding component;

the antigenic component comprising a polypeptide capable of eliciting an immune response from the immune system of the subject;

the cell binding component comprising an antibody fragment, the cell binding component being configured to specifically bind erythrocytes and not specifically bind to other blood components,

the cell binding component being configured to bind glycophorin A on the surface of human erythrocytes; and

wherein the cell binding component is an antibody fragment derived from a hybridoma,

wherein the antibody fragment is affinity matured,

wherein upon administration of the engineered composition, the cell binding component binds to erythrocytes,

wherein the cell binding component exhibits an erythrocyte equilibrium 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

wherein, upon administration of the engineered composition, the antigenic component is presented to the immune system and tolerance specific to the antigenic component is induced.

2. The composition of claim 1, wherein the cell binding component is derived from a 10F7 clone, wherein the antigenic portion is coupled to the cell binding component as a molecular fusion, and 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.

3. The composition of claim 1, wherein the cell binding component is derived from a 10F7 clone.

4. The composition of claim 1, wherein the cell binding component is fused to the N- or C-terminus of the antigenic component via a linker.

5. The composition of claim 1, wherein the cell binding component is chemically conjugated to the antigenic component.

6. An engineered composition for inducing antigen-specific tolerance in a subject, the engineered composition comprising:

an antigenic component coupled to a cell binding component;

the antigenic component comprising asparaginase or a polypeptide derived from asparaginase and capable of eliciting an immune response from the immune system of a subject;

the antigenic component being fused with the cell binding component;

the cell binding component comprising a polypeptide,

the cell binding component being configured to specifically bind erythrocytes and not specifically bind to other blood components,

the cell binding component being configured to bind to glycophorin A on the surface of erythrocytes with an equilibrium 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;

wherein the cell binding component is an antibody fragment derived from a hybridoma,

wherein the antibody fragment is affinity matured,

wherein the engineered composition is suitable for intravenous administration to the subject; and

wherein upon administration of the engineered composition, the cell binding component binds to erythrocytes, and the antigenic component is presented to the immune system and tolerance specific to the antigenic component is induced.

7. The composition of claim 6, wherein antigenic component comprises a fragment of asparaginase.

8. The composition of claim 6, wherein the antigenic portion is a mimotope of asparaginase.

9. The composition of claim 6, wherein cell binding component is fused to the N-terminus of the antigenic component.

10. The composition of claim 6, wherein cell binding component is fused to the antigenic component via a linker.

11. An engineered composition for inducing antigen-specific tolerance in a subject, the engineered composition comprising:

an antigenic component coupled to a cell binding component;

the antigenic component comprising a polypeptide that is capable of eliciting an immune response from the immune system of the subject, said antigenic component being chemically conjugated with the cell binding component;

the cell binding component comprising a polypeptide, the cell binding component being configured to specifically bind erythrocytes and not specifically bind to other blood components,

the cell binding component being configured to bind glycophorin A on the surface of erythrocytes,

wherein the cell binding component comprises an antibody fragment; and

wherein upon administration of the engineered composition, the cell binding component binds to erythrocytes, the antigenic component is presented to the immune system and tolerance specific to the antigenic component is induced.

12. The composition of claim 11, wherein the antigenic component comprises asparaginase.

13. The composition of claim 11, wherein the cell binding component is derived from a 10F7 clone.

14. An engineered composition for inducing antigen-specific tolerance in a subject, the engineered composition comprising:

an antigenic component coupled to a cell binding component;

the antigenic component comprising asparaginase or a polypeptide derived from asparaginase and capable of eliciting an immune response from the immune system of a subject;

the antigenic component being fused with the cell binding component;

the cell binding component comprising a polypeptide,

the cell binding component being configured to specifically bind erythrocytes and not specifically bind to other blood components,

the cell binding component comprises a sequence chosen from the group consisting of SEQ ID NO;

11, SEQ ID NO;

13, SEQ ID NO;

14, SEQ ID NO;

15, SEQ ID NO;

16, SEQ ID NO;

17, SEQ ID NO;

1, and conservative substitutions thereof,

the cell binding component being configured to bind to glycophorin A on the surface of erythrocytes with an equilibrium 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;

wherein the engineered composition is suitable for intravenous administration to the subject; and

wherein upon administration of the engineered composition, the cell binding component binds to erythrocytes, and the antigenic component is presented to the immune system and tolerance specific to the antigenic component is induced.

15. The composition of claim 14, wherein multiple copies of the cell binding component are directly fused to the antigen at a plurality of sites on the antigenic component.

16. The composition of claim 14, wherein antigenic component comprises a fragment of asparaginase.

17. The composition of claim 14, wherein the antigenic portion is a mimotope of asparaginase.

18. The composition of claim 14, wherein cell binding component is fused to the N-terminus of the antigenic component.

19. The composition of claim 14, wherein cell binding component is fused to the antigenic component via a linker.