Compositions for generation of immune tolerance to specific antigens

• US 10,265,416 B2
• Filed: 02/23/2018
• Issued: 04/23/2019
• Est. Priority Date: 08/10/2010
• Status: Active Grant

• Current Assignee: Ecole polytechnique fãdãrale de lausanne epfl (Schweizerische Eidgenossenschaft)
• Original Assignee: Ecole Polytechnique Federal de Lausanne
• Inventors: Hubbell, Jeffrey A., Kontos, Stephan, Dane, Karen Y.
• Primary Examiner(s): Gebreysus, Kagnew H
• Application Number:
  US15/904,248
• Publication Number:
  US 20180185507A1
• Time in Patent Office: 424 Days
• Field of Search: None
• 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 to which tolerance is desired;

wherein the antigen is selected from the group consisting of proinsulin and immunogenic fragments thereof;

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 presents said antigen to the immune system of a human;

wherein the erythrocyte-binding moiety comprises an antibody fragment,

wherein the antigen to which tolerance is desired is recombinantly fused or chemically conjugated to the erythrocyte-binding moiety, and

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.

2. The composition of claim 1, wherein the antibody fragment is directed against glycophorin A, wherein the erythrocyte-binding moiety is derived from a 10F7 clone, wherein the administration of the composition ameliorates 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.

3. The composition of claim 1, wherein the erythrocyte-binding moiety specifically binds to a biomolecule selected from the group consisting of Band 3 (CD233), aquaporin-1, Glut-1, Kidd antigen, RhAg/Rh50 (CD241), Rh (CD240), Rh30CE (CD240CE), Rh30D (CD240D), Kx, glycophorin A, glycophorin B (CD235b), glycophorin C (CD235c), glycophorin D (CD235d), Kell (CD238), Duffy/DARCi (CD234), CR1 (CD35), DAF (CD55), Globoside, CD44, ICAM-4 (CD242), Lu/B-CAM (CD239), XG1/XG2 (CD99), EMMPRIN/neurothelin (CD147), JMH, Glycosyltransferase, Cartwright, Dombrock, C4A/CAB, Scianna, MER2, stomatin, BA-1 (CD24), GPIV (CD36), CD108, CD139, and H antigen (CD173).

4. The composition of claim 1, wherein the erythrocyte-binding moiety is affinity matured, wherein the erythrocyte-binding moiety does not specifically bind to other blood components, wherein the other blood components comprises blood proteins, albumin, fibronectin, platelets, and white blood cells, wherein the binding of the erythrocyte-binding moiety to the erythrocyte cell surface does not alter cell morphology, wherein there is no cytoplasmic translocation of the composition upon binding of the erythrocyte-binding moiety to the erythrocyte cell surface, and wherein the binding of the erythrocyte-binding moiety to the erythrocyte cell surface does not cause the erythrocytes to become apoptotic.

5. The composition of claim 1, wherein the erythrocyte-binding moiety comprises an scFv, a divalent scFv, a diabody, a triabody, or a tetrabody.

6. The composition of claim 1, wherein the erythrocyte-binding moiety is derived from a 10F7 clone.

7. The composition of claim 1, wherein the antibody fragment is affinity matured.

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

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

said antigen being recognizable by an immune system of a subject, the immune system of the subject being able to respond to the antigen with an unwanted immune response;

wherein said erythrocyte-binding moiety comprises a polypeptide;

wherein upon administration of the composition, said erythrocyte-binding moiety non-covalently and specifically binds an erythrocyte in situ in blood and presents said antigen to the immune system of the subject;

wherein the erythrocyte-binding moiety does not specifically bind to other blood components, wherein the other blood components comprises blood proteins, albumin, fibronectin, platelets, and white blood cells;

wherein said antigen is associated with Type 1 diabetes; and

wherein said composition elicits a tolerogenic response upon administration to said subject.

9. The composition of claim 8, wherein the antigen is selected from the group consisting of proinsulin, immunogenic fragments thereof, and immunogenic mimotopes thereof.

10. The composition of claim 8, wherein the erythrocyte-binding moiety comprises an antibody fragment, wherein the antibody fragment is derived from a 10F7 clone, and wherein the antibody fragment has undergone affinity maturation.

11. The composition of claim 8, wherein the erythrocyte-binding moiety is chemically conjugated to the antigen.

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

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

one or more antigens fused or chemically conjugated with an erythrocyte-binding moiety;

wherein said one or more antigens is selected from the group consisting of proinsulin and immunogenic fragments thereof, and immunogenic mimotopes thereof;

wherein said one or more antigens 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 one or more antigens 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 one or more antigens to the immune system of the subject,

wherein said erythrocyte-binding moiety comprises at least an antibody or an antibody fragment.

14. The composition of claim 13, wherein the erythrocyte-binding moiety has the ability to bind Band 3 (CD233), glycophorin-A, glycophorin B (CD235b), glycophorin C (CD235c), or glycophorin D (CD235d) 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.

15. The composition of claim 13, comprising two or more antigens selected from the group consisting of proinsulin, immunogenic fragments thereof, and immunogenic mimotopes thereof.

16. The composition of claim 15, wherein said antigens are conjugated to a multimeric branched polymer.

18. The composition of claim 13, wherein the erythrocyte-binding moiety is fused via a linker, to the N- or C-terminus of the one or more antigens.

19. The composition of claim 18, wherein the linker is a peptide, a polymer, an aptamer, a nucleic acid, or a particle.

20. The composition of claim 19, wherein the polymer is synthetic, and wherein the polymer is branched.

17. The composition of 16, wherein the erythrocyte-binding moiety is derived from a 10F7 clone, and wherein the erythrocyte-binding moiety is affinity-matured.