# US 9,518,087 B2 · Erythrocyte-binding therapeutics > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 9,518,087 B2 - Document ID: US9518087B2 - Status: Granted - Kind: B2 - Application number: 13/206,034 - Filed: 2011-08-09 - Published: 2012-02-16 - Granted: 2016-12-13 - Earliest claimed: 2010-08-10 - Assignees: Ecole Polytechnique Federale De Lausanne (Epfl) - Applicants: Jeffrey A. Hubbell; Stéphane Kontos; Karen Y. Dane - Inventors: Jeffrey A. Hubbell; Stéphane Kontos; Karen Y. Dane - Web page: https://patentgrep.com/patent/US9518087B2 ## CPC - C07K 16/28: against receptors, cell surface antigens or cell surface determinants - A61K 39/0008: {Antigens related to auto-immune diseases; Preparations to induce self-tolerance} - A61K 39/001: {Preparations to induce tolerance to non-self, e.g. prior to transplantation} - A61K 47/6849: {the antibody targeting a receptor, a cell surface antigen or a cell surface determinant} - A61P 35/00: Antineoplastic agents - A61P 37/02: Immunomodulators - A61P 37/04: Immunostimulants - A61P 37/06: Immunosuppressants, e.g. drugs for graft rejection - A61P 43/00: Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00 - B82Y 5/00: Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery - C07K 7/08: having 12 to 20 amino acids (gastrins C07K14/595; somatostatins C07K14/655; melanotropins C07K14/68) - C07K 14/4701: {not used} - C07K 14/77: Ovalbumin - C07K 16/18: against material from animals or humans - C07K 16/30: from tumour cells - 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) - A61K 2039/505: {comprising antibodies} - A61K 2039/6031: {Proteins} - A61K 2039/6056: {Antibodies} - C07K 2317/34: Identification of a linear epitope shorter than 20 amino acid residues or of a conformational epitope defined by amino acid residues - C07K 2317/622: Single chain antibody (scFv) - C07K 2319/00: Fusion polypeptide - C07K 2319/01: containing a localisation/targetting motif - C07K 2319/21: containing a His-tag - C07K 2319/22: containing a Strep-tag - C07K 2319/23: containing a GST-tag - C07K 2319/33: fusions for targeting to specific cell types, e.g. tissue specific targeting, targeting of a bacterial subspecies - C07K 2319/41: containing a Myc-tag - C07K 2319/43: containing a FLAG-tag - C07K 2319/74: containing a fusion for binding to a cell surface receptor - C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00) - A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D) - A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS - B82Y: SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES ## 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 ### Claim 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 gliadin and a portion 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 comprises an antibody fragment directed against glycophorin A, 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. ### Claim 2 (depends on 1) The composition of claim 1 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. ### Claim 3 (depends on 1) The composition of claim 1 wherein the erythrocyte-binding moiety is fused, optionally via a linker, to the N- or C-terminus of the antigen. ### Claim 4 (depends on 3) The composition of claim 3 wherein the erythrocyte-binding moiety is fused, optionally via a linker, to the N-terminus of the antigen. ### Claim 5 (depends on 4) The composition of claim 4 wherein the erythrocyte-binding moiety is derived from 10F7. ### Claim 6 (depends on 1) The composition of claim 1 wherein the antigen to which tolerance is desired is gliadin. ### Claim 7 (depends on 1) The composition of claim 1 wherein the erythrocyte-binding moiety is derived from 10F7 and the antigen to which tolerance is desired is a fragment of gliadin. ### Claim 8 (depends on 7) The composition of claim 7 wherein the administration of the composition ameliorates celiac disease. ### Claim 9 A composition for use in inducing tolerance comprising: an antigen chemically conjugated with an erythrocyte-binding moiety having the ability to non-covalently, specifically bind an exterior erythrocyte surface in situ in blood, said tolerogenic composition being pharmaceutically acceptable, wherein said antigen is selected from the group consisting of gliadin and a portion thereof, 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 the antigen alone, and wherein the erythrocyte-binding moiety comprises a peptide ligand that is directed against glycophorin A has a dissociation constant of between about 10 μM and 0.1 nM as determined by equilibrium binding measurements between the peptide and erythrocytes. ### Claim 10 (depends on 9) The composition of claim 9, wherein the peptide ligand comprises SEQ ID NO:11, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, or SEQ ID NO:17. ### Claim 11 (depends on 9) The composition of claim 9, wherein the antigen is a portion of gliadin. ### Claim 12 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 gliadin and portions thereof; a cell binding moiety, wherein the cell binding moiety is an erythrocyte-binding moiety having the ability to non-covalently, specifically bind an exterior erythrocyte surface in situ in blood, wherein the erythrocyte-binding moiety is directed against glycophorin A, wherein the erythrocyte-binding moiety has erythrocyte dissociation constant of between about 10 μM and 0.1 nM, wherein the antigen to which tolerance is desired is joined to the erythrocyte-binding moiety, wherein, upon administration to a human, the composition: (i) binds to CD45 negative cells, but not to CD45 positive cells, (ii) induces greater proliferation of antigen-specific CD8+ T cells, as compared to the proliferation of antigen-specific CD8+ T cells induced by the antigen alone, or (iii) 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. ### Claim 13 (depends on 12) The composition of claim 12, wherein the erythrocyte-binding moiety is fused to the antigen via recombinant DNA technology. ### Claim 14 (depends on 12) The composition of claim 12, wherein the erythrocyte-binding moiety is fused, optionally via a linker, to the N- or C-terminus of the antigen. ### Claim 15 (depends on 12) The composition of claim 12, wherein the erythrocyte-binding moiety is covalently bonded to the antigen. ### Claim 16 (depends on 12) The composition of claim 12, wherein the erythrocyte-binding moiety wherein is derived from 10F7. ### Claim 17 (depends on 16) The composition of claim 16, wherein the erythrocyte-binding moiety comprises an antibody fragment. ### Claim 18 (depends on 12) The composition of claim 12, wherein the erythrocyte-binding moiety comprises a peptide ligand. ### Claim 19 (depends on 18) The composition of claim 18, wherein the peptide ligand comprises SEQ ID NO:11, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, or SEQ ID NO:17. ### Claim 20 A composition comprising an antigen recombinantly fused or chemically conjugated with an erythrocyte-binding moiety; wherein the erythrocyte-binding moiety comprises an antibody, antibody fragment, or a single chain variable fragment (scFv) that binds to human glycophorin A; and wherein the antigen is selected from the group consisting of gliadin, α-gliadin and γ-gliadin, and a portion thereof, 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. ### Claim 21 (depends on 20) The composition of claim 20, wherein the erythrocyte-binding moiety is fused, optionally via a linker to the N- or C-terminus of the antigen. ### Claim 22 (depends on 20) A method for treating Celiac disease in a subject, comprising administering to said subject a composition of claim 20. ### Claim 23 (depends on 21) A method for treating Celiac disease in a subject, comprising administering to said subject a composition of claim 21.