# US 9,517,257 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 9,517,257 B2 - Document ID: US9517257B2 - Status: Granted - Kind: B2 - Application number: 13/397,202 - Filed: 2012-02-15 - Published: 2012-07-12 - 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/US9517257B2 ## CPC - A61K 9/1075: {Microemulsions or submicron emulsions; Preconcentrates or solids thereof; Micelles, e.g. made of phospholipids or block copolymers (A61K9/0026 takes precedence)} - A61K 9/513: {Organic macromolecular compounds; Dendrimers} - 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 47/64: 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) - A61K 47/6811: {the drug being a protein or peptide, e.g. transferrin or bleomycin} - A61K 47/6815: {Enzymes} - A61K 47/6843: {the antibody targeting a material from animals or humans} - A61K 47/6849: {the antibody targeting a receptor, a cell surface antigen or a cell surface determinant} - C07K 7/08: having 12 to 20 amino acids (gastrins C07K14/595; somatostatins C07K14/655; melanotropins C07K14/68) - C07K 14/62: Insulins - C07K 14/70539: {MHC-molecules, e.g. HLA-molecules} - C07K 16/18: against material from animals or humans - C07K 16/28: against receptors, cell surface antigens or cell surface determinants - C12N 9/82: Asparaginase {(3.5.1.1)} - C12N 9/96: Stabilising an enzyme by forming an adduct or a composition; Forming enzyme conjugates - C12Y 305/01001: Asparaginase (3.5.1.1) - 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 2039/6031: {Proteins} - A61K 2039/605: {MHC molecules or ligands thereof} - A61K 2039/6056: {Antibodies} - C07K 2317/56: variable (Fv) region, i.e. VH and/or VL - C07K 2317/622: Single chain antibody (scFv) - C07K 2317/92: Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value - C07K 2319/00: Fusion polypeptide - C07K 2319/21: containing a His-tag - C07K 2319/22: containing a Strep-tag - C07K 2319/23: containing a GST-tag - C07K 2319/30: Non-immunoglobulin-derived peptide or protein having an immunoglobulin constant or Fc region, or a fragment thereof, attached thereto - 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 - 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) - 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) - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) - C12Y: ENZYMES ## 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 ### Claim 1 A pharmaceutically acceptable composition for inducing tolerance of an antigenic therapeutic protein comprising: a fusion molecule having an erythrocyte-binding moiety and a portion of the antigenic therapeutic protein, wherein said erythrocyte-binding moiety has the ability to non-covalently, specifically bind an exterior erythrocyte surface in situ in blood and wherein said antigenic therapeutic protein portion is a portion of human Factor VIII. ### Claim 2 (depends on 1) The composition of claim 1 wherein the erythrocyte-binding moiety specifically binds to a biomolecule chosen 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). ### Claim 3 (depends on 1) The composition of claim 1 wherein the erythrocyte-binding moiety is chosen from the group consisting of a peptide ligand, an antibody, an antibody fragment, and a single chain variable fragment (ScFv). ### Claim 4 (depends on 1) The composition of claim 1 wherein the erythrocyte-binding moiety comprises an antibody, an antibody fragment or an ScFv derived from a hybridoma that produces an antibody against an erythrocyte, with the hybridoma being chosen from the group consisting of BRIC 4, BRIC 5, BRIC 6, BRIC 10, BRIC 14, BRIC 18, BRIC 39, BRIC 66, BRIC 68, BRIC 69, BRIC 87, BRIC 108, BRIC 110, BRIC 111, BRIC 125, BRIC 126, BRIC 128, BRIC 145, BRIC 155, BRIC 157, BRIC 163, BRIC 170, BRIC 198, BRIC 203, BRIC 216, BRIC 220, BRIC 221, BRIC 222, BRIC 229, BRIC 230, BRIC 231, BRIC 235, BRIC 256, BRAC 17, BRAC 18, BGRL 1, BGRL 2, BGRL 11, BGRL 100, BRAD 3, BIRMA D6, BIRMA D10, BIRMA K3, BIRMA K3, BIRMA 84B; 6A7; COE; and KZ1. ### Claim 5 (depends on 1) A method of inducing immunotolerance, the method comprising administering the composition of claim 1 to a patient. ### Claim 6 (depends on 5) The method of claim 5 wherein the erythrocyte-binding moiety specifically binds to a biomolecule chosen 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). ### Claim 7 (depends on 5) The method of claim 5 wherein the erythrocyte-binding moiety is chosen from the group consisting of a peptide ligand, an antibody, an antibody fragment, and a single chain variable fragment (ScFv). ### Claim 8 (depends on 5) The method of claim 5 wherein the erythrocyte-binding moiety comprises an antibody, an antibody fragment or an ScFv derived from a hybridoma that produces an antibody against an erythrocyte, with the hybridoma being chosen from the group consisting of BRIC 4, BRIC 5, BRIC 6, BRIC 10, BRIC 14, BRIC 18, BRIC 39, BRIC 66, BRIC 68, BRIC 69, BRIC 87, BRIC 108, BRIC 110, BRIC 111, BRIC 125, BRIC 126, BRIC 128, BRIC 145, BRIC 155, BRIC 157, BRIC 163, BRIC 170, BRIC 198, BRIC 203, BRIC 216, BRIC 220, BRIC 221, BRIC 222, BRIC 229, BRIC 230, BRIC 231, BRIC 235, BRIC 256, BRAC 17, BRAC 18, BGRL 1, BGRL 2, BGRL 11, BGRL 100, BRAD 3, BIRMA D6, BIRMA D10, BIRMA K3, BIRMA K3, BIRMA 84B; 6A7; COE; and KZ1. ### Claim 9 (depends on 1) The composition of claim 1 wherein said erythrocyte-binding moiety comprises an antibody, antibody fragment or scFv derived from a 10F7 clone. ### Claim 10 A pharmaceutically acceptable composition for inducing tolerance to a therapeutic protein in a subject having a deficiency in production of a functional native protein, comprising: a fusion molecule comprising an erythrocyte-binding moiety and a portion of the antigenic therapeutic protein, wherein said erythrocyte-binding moiety has the ability to non-covalently, bind an exterior surface of erythrocytes in situ in blood and does not specifically bind to other blood components, wherein said therapeutic protein is Factor VIII, and wherein the antigenic portion consists of a portion of human Factor VIII. ### Claim 11 (depends on 10) The composition of claim 10, wherein the erythrocyte-binding moiety is chosen from the group consisting of a peptide ligand, an antibody, an antibody fragment, and a single chain variable fragment (ScFv). ### Claim 12 (depends on 10) The composition of claim 10, wherein the erythrocyte-binding moiety comprises an antibody, an antibody fragment or an ScFv derived from a hybridoma that produces an antibody against an erythrocyte, with the hybridoma being chosen from the group consisting of BRIC 4, BRIC 5, BRIC 6, BRIC 10, BRIC 14, BRIC 18, BRIC 39, BRIC 66, BRIC 68, BRIC 69, BRIC 87, BRIC 108, BRIC 110, BRIC 111, BRIC 125, BRIC 126, BRIC 128, BRIC 145, BRIC 155, BRIC 157, BRIC 163, BRIC 170, BRIC 198, BRIC 203, BRIC 216, BRIC 220, BRIC 221, BRIC 222, BRIC 229, BRIC 230, BRIC 231, BRIC 235, BRIC 256, BRAC 17, BRAC 18, BGRL 1, BGRL 2, BGRL 11, BGRL 100, BRAD 3, BIRMA D6, BIRMA D10, BIRMA K3, BIRMA K3, BIRMA 84B; 6A7; COE; and KZ1. ### Claim 13 (depends on 10) The composition of claim 10, wherein said erythrocyte-binding moiety comprises an single chain variable fragment (ScFv) derived from a 10F7 clone. ### Claim 14 A pharmaceutically acceptable composition for inducing tolerance to a therapeutic protein in a subject having a deficiency in production of a functional native protein, comprising: a fusion molecule comprising an erythrocyte-binding moiety and an antigenic portion of the therapeutic protein, wherein said erythrocyte-binding moiety has the ability to non-covalently, bind an exterior surface of erythrocytes in situ in blood and does not specifically bind to other blood components, wherein the erythrocyte-binding moiety is a single-chain variable fragment (scFv) derived from 10F7 that targets glycophorin A on erythrocytes, wherein said therapeutic protein is Factor VIII, and wherein the antigenic portion of the Factor VIII comprises a portion of human Factor VIII. ### Claim 15 (depends on 14) The composition of claim 14, wherein the erythrocyte-binding moiety is fused to the antigenic portion of the therapeutic protein. ### Claim 16 (depends on 14) The composition of claim 14, wherein the erythrocyte-binding moiety is fused to the N- or C-terminus of the antigenic portion of the therapeutic protein.