Erythrocyte-binding therapeutics

• US 9,850,296 B2
• Filed: 08/15/2014
• Issued: 12/26/2017
• 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, Stphane, Dane, Karen Y.
• Primary Examiner(s): Gebreyesus, Kagnew H
• Application Number:
  US14/460,768
• Publication Number:
  US 20140356384A1
• Time in Patent Office: 1,229 Days
• CPC Class Codes: A61K 2039/505 comprising antibodies
• CPC Class Codes: A61K 2039/6031 Proteins
• CPC Class Codes: A61K 2039/6056 Antibodies
• CPC Class Codes: A61K 2039/627 characterised by the linker
• 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 47/60 the organic macromolecular compound being a polyoxyalkylene oligomer, polymer or dendrimer, e.g. PEG, PPG, PEO or polyglycerol
• CPC Class Codes: A61K 47/62 the modifying agent being a protein, peptide or polyamino acid
• 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 47/646 the entire peptide or protein drug conjugate elicits an immune response, e.g. conjugate vaccines
• CPC Class Codes: A61K 47/6811 the drug being a protein or peptide, e.g. transferrin or bleomycin
• CPC Class Codes: A61K 47/6849 the antibody targeting a receptor, a cell surface antigen or a cell surface determinant
• CPC Class Codes: A61K 47/6901 Conjugates being cells, cell fragments, viruses, ghosts, red blood cells or viral vectors
• 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 16/2896 against molecules with a "CD"-designation, not provided for elsewhere
• 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 2319/33 : 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: 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 pharmaceutically acceptable composition comprising an erythrocyte-binding moiety, a linker, and a tolerogenic antigen , which are recombinantly fused or chemically conjugated, wherein:

said erythrocyte-binding moiety is an antibody fragment having the ability to noncovalently and specifically bind to glycophorin A on a human erythrocyte in situ in blood,

said linker is a peptide, a covalent bond a nucleic acid or a particle; and

said tolerogenic antigen is a self antigen to which patients develop an unwanted immune response, wherein the self antigen comprises insulin, proinsulin, preproinsulin, or an antigenic fragment of insulin, proinsulin or preproinsulin.

2. The pharmaceutically acceptable composition of claim 1 wherein said linker is a covalent bond.

3. The pharmaceutically acceptable composition of claim 1 wherein administration of said composition results in reductions in diabetogenic T cells in a spleen or liver of a subject receiving the composition.

4. The pharmaceutically acceptable composition of claim 1 characterized by having the ability to induce apoptotic- or exhausted-fate proliferation and/or deletion of CD4+ and/or CD8+ T-cells specific for the antigen, or the ability to induce regulatory cell phenotypes.

5. The pharmaceutically acceptable composition of claim 1 wherein the tolerogenic antigen is proinsulin or an antigenic fragment thereof.

6. The pharmaceutically acceptable composition of claim 1 wherein the tolerogenic antigen comprises a mimotope of a protein.

7. The pharmaceutically acceptable composition of claim 1 wherein said linker is a branched polymer that increases the affinity of erythrocyte binding by avidity effects.

8. The pharmaceutically acceptable composition of claim 7 wherein said branched polymer is conjugated to more than one erythrocyte-binding moiety.

9. A method of treating an unwanted immune response comprising administering to a patient in need thereof a therapeutically effective amount of a pharmaceutically acceptable composition, the composition comprising an erythrocyte-binding moiety, a linker, and a tolerogenic antigen , which are recombinantly fused or chemically conjugated, wherein:

said erythrocyte-binding moiety is an antibody fragment having the ability to noncovalently specifically bind to glycophorin A on a human erythrocyte in situ in blood,

said linker is a peptide, a covalent bond, a nucleic acid or a particle;

said tolerogenic antigen is a self antigen to which patients develop an unwanted immune response, wherein the self antigen comprises insulin, proinsulin, preproinsulin, or an antigenic fragment of insulin, proinsulin or preproinsulin.

10. The method of claim 9 characterized by having the ability to induce apoptotic- or exhausted-fate proliferation and/or deletion of CD4+ and/or CD8+ T-cells specific for the antigen, or the ability to induce regulatory cell phenotypes.

11. The method of claim 9 wherein said composition is administered to a patient prior to an unwanted immune response to the antigen, wherein said treatment prevents or minimizes the unwanted immune response.

12. The method of claim 9 wherein said composition is administered to a patient having an unwanted immune response to the antigen, wherein said treatment reverses or minimizes the unwanted immune response.

13. The method of claim 9 where the unwanted immune response is type 1 diabetes mellitus.

14. The method of claim 9 wherein the tolerogenic antigen is proinsulin or an antigenic fragment thereof.

15. A pharmaceutically acceptable composition comprising an erythrocyte-binding moiety, and a tolerogenic antigen, which are recombinantly fused or chemically conjugated, wherein:

said erythrocyte-binding moiety is an antibody fragment binding to human glycophorin A; and

said tolerogenic antigen is a self antigen which induces an unwanted immune response, wherein the self antigen comprises insulin, proinsulin, preproinsulin, glutamic acid decarboxylase-65 (GAD65), GAD-67, islet-specific glucose-6-phosphatase catalytic subunit-related protein or fragments thereof.

16. A pharmaceutically acceptable composition comprising:

an erythrocyte-binding moiety;

said erythrocyte-binding moiety is an antibody directed to human glycophorin A thereby allowing the composition to specifically bind erythrocytes; and

a tolerogenic antigen,

wherein said tolerogenic 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 erythrocyte-binding moiety and the tolerogenic antigen are chemically conjugated to one another.

17. A pharmaceutically acceptable composition, comprising:

an erythrocyte-binding moiety,

said erythrocyte-binding moiety is an antibody fragment having the ability to noncovalently and specifically bind to glycophorin A on a human erythrocyte in situ in blood;

a tolerogenic antigen,

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

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

wherein the self-antigen is an islet antigen recognized by diabetogenic T cells, wherein the erythrocyte-binding moiety and tolerogenic antigen are recombinantly fused or chemically conjugated.

18. The pharmaceutically acceptable composition of claim 17, wherein the islet antigen recognized by diabetogenic T cells is selected from the group consisting of chromogranin A, insulin, proinsulin, preproinsulin, an antigenic fragment thereof, or a mimotope thereof.

19. The pharmaceutically acceptable composition of claim 17, wherein the erythrocyte-binding moiety and tolerogenic antigen are recombinantly fused.

20. The pharmaceutically acceptable composition of claim 17, wherein the tolerogenic antigen comprises an immunogenic mimotope of chromogranin A.

21. The pharmaceutically acceptable composition of claim 20, wherein the immunogenic mimotope of chromogranin A comprises the sequence of SEQ ID NO:

77.