# US 8,785,594 B2 · Glycoproteins and glycosylated cells and a method for the preparation of the same > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 8,785,594 B2 - Document ID: US8785594B2 - Status: Granted - Kind: B2 - Application number: 12/733,672 - Filed: 2008-09-24 - Published: 2010-10-14 - Granted: 2014-07-22 - Earliest claimed: 2007-09-25 - Assignees: Glycom A/S - Applicants: Gyula Dekany; Károly Ágoston; István Bajza; Marie Bøjstrup; Lars Kröger - Inventors: Gyula Dekany; Károly Ágoston; István Bajza; Marie Bøjstrup; Lars Kröger - Web page: https://patentgrep.com/patent/US8785594B2 ## CPC - C07K 1/1077: {by covalent attachment of residues other than amino acids or peptide residues, e.g. sugars, polyols, fatty acids} - A61K 47/549: {Sugars, nucleosides, nucleotides or nucleic acids} - A61P 43/00: Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00 - C07K 14/00: Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof - 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 ## Abstract The present application discloses novel glycoproteins and a related glycosylcarbamoylation methodology suitable for the preparation of glycopeptides (in particular glycoproteins and glycosylated cells), as well as the use of such glycoproteins in medicine, e.g. as pharmaceuticals and diagnostics or in diagnostic kits. A method for the preparation of a carbohydrate-peptide conjugate includes reacting a cyclic carbamate with a peptide which has at least one primary amino group. ## Claims ### Claim 1 A method for the preparation of a carbohydrate-peptide conjugate, said method comprising: reacting a cyclic carbamate (1) with a peptide, wherein R3 and R4 are independently selected from the group consisting of hydroxyl, acetamido, and a carbohydrate moiety, R5 is selected from the group consisting of hydrogen, methyl, hydroxymethyl, acetamidomethyl, carboxyl, and X—(CH2)r—, wherein X is a carbohydrate moiety and r is an integer selected from 0, 1, 2 and 3, and said peptide includes at least one primary amino group, and said peptide is a cell surface or cell membrane bound protein, and said carbohydrate-peptide conjugate includes at least one moiety selected from the group consisting of General Formulae 1 and 2 wherein, in General Formula 1, R1 and R2 together with the intervening lysine moiety represent a cell surface or cell membrane bound protein; R3, R4 and R5 are as defined at cyclic carbamate (1), and pharmaceutically acceptable salts thereof, and wherein, in General Formula 2, R11 is an amino acid side chain; R12 together with —NH—CHR11—C(═O)— represents a cell surface or cell membrane bound protein, R3, R4 and R5 are as defined at cyclic carbamate (1), and pharmaceutically acceptable salts thereof. ### Claim 2 (depends on 1) The method according to claim 1, wherein the reaction takes place in a polar solvent. ### Claim 3 (depends on 1) The method according to claim 1, wherein the reaction takes place at a pH of in the range of 6.5-10.5. ### Claim 4 (depends on 1) The method according to claim 1, wherein the carbohydrate-peptide conjugate includes at least one moiety selected from the group consisting of General Formulae 1a, 1b and 1c, wherein, R1 and R2 together with the intervening lysine moiety represent a cell surface or cell membrane bound protein, R6 and R7 are as defined for R3 and R4 in the General Formula 1, R9 is as defined for R5 in the General Formula 1, and R8 is selected from the group consisting of hydroxyl, C1-6-alkoxy, C2-20-acyloxy, acetamido, and a carbohydrate moiety. ### Claim 5 (depends on 1) The method according to claim 1, wherein the carbohydrate-peptide conjugate includes at least one moiety selected from the group consisting of General Formulae 2a, 2b and 2c, wherein, R11 and R12 are as defined in General Formula 2 R6 and R7 are as defined for R3 and R4 in the General Formula 2, R9 is as defined for R5 in the General Formula 2, and R8 is selected from the group consisting of hydroxyl, C1-6-alkoxy, C2-20-acyloxy, acetamido, and a carbohydrate moiety. ### Claim 6 (depends on 1) A carbohydrate-peptide conjugate obtainable by the method according to claim 1. ### Claim 7 A carbohydrate-peptide conjugate, comprising: at least one moiety of General Formula 1: wherein, R1 and R2 together with the intervening lysine moiety represent a cell surface or cell membrane bound protein; R3 and R4 are independently selected from the group consisting of hydroxyl, acetamido, and a carbohydrate moiety, R5 is selected from the group consisting of hydrogen, methyl, hydroxymethyl, acetamidomethyl, carboxyl, and X—(CH2)r-, wherein X is a carbohydrate moiety and r is an integer selected from 0, 1, 2 and 3 and pharmaceutically acceptable salts thereof. ### Claim 8 (depends on 7) The carbohydrate-peptide conjugate according to claim 7, further comprising at least one moiety of any of General Formulae 1a, 1b and 1c, wherein, R6 and R7 are as defined for R3 and R4 in claim 7, R9 is as defined for R5 in claim 7, R8 is selected from the group consisting of hydroxyl, C1-6-alkoxy, C2-20-acyloxy, acetamido, and a carbohydrate moiety. ### Claim 9 A carbohydrate-peptide conjugate, comprising: at least one moiety of the General Formula 2: wherein R11 is an amino acid side chain; R12 together with —NH—CHR11—C(═O)— represents a cell surface or cell membrane bound protein, R3 and R4 are independently selected from the group consisting of hydroxyl, acetamido, and a carbohydrate moiety, R5 is selected from the group consisting of hydrogen, methyl, hydroxymethyl, acetamidomethyl, carboxyl, and X—(CH2)r-, wherein X is a carbohydrate moiety and r is an integer selected from 0, 1, 2 and 3, and pharmaceutically acceptable salts thereof. ### Claim 10 (depends on 9) The carbohydrate-peptide conjugate according to claim 9, further comprising at least one moiety of any of the General Formulae 2a, 2b and 2c, wherein, R11 and R12 are as defined in claim 9, R6 and R7 are as defined for R3 and R4 in claim 9, R9 is as defined for R5 in claim 9, and R8 is selected from the group consisting of hydroxyl, C1-6-alkoxy, C2-20-acyloxy, acetamido, and a carbohydrate moiety. ### Claim 11 (depends on 6) The carbohydrate-peptide conjugate according to claim 6, wherein a carbohydrate moiety represents a non-immunogenic carbohydrate. ### Claim 12 (depends on 6) The carbohydrate-peptide conjugate according to claim 6, wherein a carbohydrate moiety represents an immunogenic carbohydrate. ### Claim 13 (depends on 6) The carbohydrate-peptide conjugate according to claim 6, wherein the peptide is a cell-surface or cell-membrane bound protein. ### Claim 14 (depends on 6) The carbohydrate-peptide conjugate as defined in claim 6, the peptide moiety has a total number of amino acid units of at least 30. ### Claim 15 (depends on 6) A pharmaceutical agent, a diagnostic agent, or a diagnostic kit comprising a carbohydrate-peptide conjugate according to claim 6. ### Claim 16 A cyclic carbamate of an oligosaccharide selected from (4a), (4b) and (4c) wherein the cyclic carbamate excludes (4-O-β-D-glucopyranosyl-1,2-dideoxy-β-D-glucopyranoso)[1,2-d]oxazolidine-2-one and (4-O-β-D-galactopyranosyl-1,2-dideoxy-β-D-glucopyranoso)[1,2-d]oxazolidine-2-one; and R3, R4, R6 and R7 are independently selected from the group consisting of hydroxyl, acetamido, and a carbohydrate moiety; R5 and R9 are independently selected from the group consisting of hydrogen, methyl, hydroxymethyl, acetamidomethyl, carboxyl, and X—(CH2)r-, wherein X is a carbohydrate moiety and r is an integer selected from 0, 1, 2 and 3; and R8 and R10 are independently selected from the group consisting of hydroxyl, C1-6-alkoxy, C1-6-acyloxy, acetamido, and a carbohydrate moiety; and pharmaceutically acceptable salts thereof. ### Claim 17 A carbohydrate-peptide conjugate selected from the group consisting of General Formulae 1 and 2: wherein, in General Formula 1, R1 and R2 together with the intervening lysine moiety represent a cell surface or cell membrane bound protein; R3 and R4 are independently selected from the group consisting of hydroxyl, acetamido, and a carbohydrate moiety, R5 is selected from the group consisting of hydrogen, methyl, hydroxymethyl, acetamidomethyl, carboxyl, and X—(CH2)r—, wherein X is a carbohydrate moiety and r is an integer selected from 0, 1, 2 and 3, and pharmaceutically acceptable salts thereof, and General Formula 2: wherein, in General Formula 2, R11 is an amino acid side chain; R12 together with —NH—CHR11—C(═O)— represents a cell surface or cell membrane bound protein, R3 and R4 are independently selected from the group consisting of hydroxyl, acetamido, and a carbohydrate moiety, R5 is selected from the group consisting of hydrogen, methyl, hydroxymethyl, acetamidomethyl, carboxyl, and X—(CH2)r—, wherein X is a carbohydrate moiety and r is an integer selected from 0, 1, 2 and 3, and pharmaceutically acceptable salts thereof. ### Claim 18 (depends on 17) The carbohydrate-peptide conjugate according to claim 17, further comprising at least one moiety of any of the General Formulae 1a, 1b and 1c, wherein, R1 and R2 are as defined in claim 17, R6 and R7 are as defined for R3 and R4 in claim 17, R9 is as defined for R5 in claim 17, R8 is selected from the group consisting of hydroxyl, C1-6-alkoxy, C2-20-acyloxy, acetamido, and a carbohydrate moiety. ### Claim 19 (depends on 17) The carbohydrate-peptide conjugate according to claim 17, further comprising at least one moiety of any of the General Formulae 2a, 2b and 2c, wherein, R11 and R12 are as defined in claim 17, R6 and R7 are as defined for R3 and R4 in claim 17, R9 is as defined for R5 in claim 17, and R8 is selected from the group consisting of hydroxyl, C1-6-alkoxy, C2-20-acyloxy, acetamido, and a carbohydrate moiety.