# US 2016/0354484 A1 · Compositions for inducing antigen-specific tolerance > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 2016/0354484 A1 - Document ID: US20160354484A1 - Status: Published application - Kind: A1 - Application number: 15/232,694 - Filed: 2016-08-09 - Published: 2016-12-08 - Earliest claimed: 2010-08-10 - Assignees: Ecole Polytechnique Federale De Lausanne (Epfl) - Applicants: École Polytechnique Fédérale de Lausanne (EPFL) - Inventors: Jeffrey A. Hubbell; Stephan Kontos; Karen Y. Dane - Web page: https://patentgrep.com/patent/US20160354484A1 ## 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 (canceled) ### Claim 2 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 a peptide, antibody or 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 upon administration of the engineered composition, the cell binding component binds to erythrocytes, wherein the cell binding component exhibits an erythrocyte dissociation constant of between about 10 μM 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. ### Claim 3 (depends on 2) The composition of claim 2, 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. ### Claim 4 (depends on 2) The composition of claim 2, wherein the cell binding component comprises a peptide having 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. ### Claim 5 (depends on 2) The composition of claim 2, wherein the cell binding component comprises an antibody fragment. ### Claim 6 (depends on 5) The composition of claim 5, wherein the cell binding component is derived from a 10F7 clone. ### Claim 7 (depends on 5) The composition of claim 5, wherein the antibody fragment is affinity matured. ### Claim 8 (depends on 2) The composition of claim 2, wherein the erythrocyte-binding moiety comprises a peptide ligand, and wherein multiple copies of the erythrocyte-binding moiety are directly fused to the antigen at a plurality of sites on the antigenic component. ### Claim 9 (depends on 2) The composition of claim 2, wherein the cell binding component is fused to the N- or C-terminus of the antigenic component via a linker. ### Claim 10 (depends on 2) The composition of claim 2, wherein the cell binding component is chemically conjugated to the antigenic component. ### Claim 11 (depends on 2) The composition of claim 2, wherein the cell binding component is a an antibody fragment derived from a hybridoma, and wherein the antibody fragment is affinity matured. ### Claim 12 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 a dissociation constant of between about 10 μM 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. ### Claim 13 (depends on 12) The composition of claim 12, wherein the cell binding component comprises a peptide ligand, and wherein multiple copies of the cell binding component are directly fused to the antigen at a plurality of sites on the antigenic component. ### Claim 14 (depends on 12) The composition of claim 12, cell binding component comprises the 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. ### Claim 15 (depends on 12) The composition of claim 12, wherein antigenic component comprises a fragment of asparaginase. ### Claim 16 (depends on 12) The composition of claim 12, wherein the antigenic portion is a mimotope of asparaginase. ### Claim 17 (depends on 12) The composition of claim 12, wherein cell binding component is fused to the C-terminus of the antigenic component. ### Claim 18 (depends on 12) The composition of claim 12, wherein cell binding component is fused the antigenic component via a linker. ### Claim 19 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; 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. ### Claim 20 (depends on 19) The composition of claim 19, wherein the antigenic component comprises asparaginase. ### Claim 21 (depends on 19) The composition of claim 19, wherein the composition reduces, fails to induce or prevents inflammatory responses in antigen-specific T cells, when the subject is exposed to the antigen alone.