# US 2017/0106096 A1 · Homogenous antibody drug conjugates via enzymatic methods > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 2017/0106096 A1 - Document ID: US20170106096A1 - Status: Published application - Kind: A1 - Application number: 15/317,907 - Filed: 2015-06-11 - Published: 2017-04-20 - Earliest claimed: 2014-06-12 - Assignees: CSPC Dophen Corporation - Applicants: CSPC Dophen Corporation - Inventors: Sean HU; Lisha ALLEN - Web page: https://patentgrep.com/patent/US20170106096A1 ## CPC - A61K 31/255: of sulfoxy acids or sulfur analogues thereof - A61K 38/16: Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof {(enzyme inhibitors A61K38/005)} - A61K 38/164: {from bacteria} - A61K 38/45: Transferases (2) - A61K 47/6803: {Drugs conjugated to an antibody or immunoglobulin, e.g. cisplatin-antibody conjugates} - A61K 47/68031: {the drug being an auristatin} - A61K 47/68033: {the drug being a maytansine} - A61K 47/6855: {the tumour determinant being from breast cancer cell} - A61P 35/00: Antineoplastic agents - C07K 1/00: General methods for the preparation of peptides {, i.e. processes for the organic chemical preparation of peptides or proteins of any length} - C07K 14/195: from bacteria - C07K 16/00: Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies - C07K 16/2863: {against receptors for growth factors, growth regulators} - C07K 16/30: from tumour cells - C07K 16/32: against translation products of oncogenes - C12N 9/1044: {Protein-glutamine gamma-glutamyltransferase (2.3.2.13), i.e. transglutaminase or factor XIII} - C12Y 203/02013: Protein-glutamine gamma-glutamyltransferase (2.3.2.13), i.e. transglutaminase or factor XIII - C07K 2317/24: containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered - C07K 2317/40: characterized by post-translational modification - C07K 2317/41: Glycosylation, sialylation, or fucosylation - C07K 2317/524: CH2 domain - C07K 2317/76: Antagonist effect on antigen, e.g. neutralization or inhibition of binding - 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 - 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 The present application in one aspect provide Fc-containing polypeptide conjugates comprising an Fc-containing polypeptide conjugated to a conjugate moiety, wherein the Fc-containing polypeptide comprises an N-glycosylated Fc region comprising an acceptor glutamine residue flanked by an N-glycosylation site and wherein the conjugate moiety is conjugated to the Fc-containing polypeptide via the acceptor glutamine residue. Also provided are methods of making such Fc-containing polypeptide conjugates by using a wildtype or engineered transglutaminases. Further provided are engineered transglutaminases specifically designed for carrying out such reactions. ## Claims ### Claim 1 An antibody drug conjugate comprising an antibody conjugated to a conjugation moiety via an endogenous acceptor glutamine residue on the antibody, wherein the antibody drug conjugate is glycosylated in the Fc region. ### Claim 2 (depends on 1) The antibody drug conjugate of claim 1, wherein the antibody drug conjugate is N-glycosylated in the Fc region. ### Claim at position 3 3-5. (canceled) ### Claim 6 (depends on 1) The antibody drug conjugate of claim 1, wherein the conjugate moiety comprises an active moiety selected from the group consisting of: a moiety that improves the pharmacokinetic property of the antibody, a therapeutic moiety, and a diagnostic moiety. ### Claim at position 5 7-10. (canceled) ### Claim 11 (depends on 1) A composition comprising the antibody drug conjugate of claim 1, wherein at least about 50% of antibody drug conjugates in the composition is glycosylated in the Fc region. ### Claim 12 (depends on 11) The composition of claim 11, wherein at least 80% of the antibody drug conjugates in the composition has the antibody to conjugate moiety molar ratio of 1:1, or 1:2. ### Claim 13 A method of making an antibody drug conjugate comprising an antibody specifically conjugated to a conjugate moiety comprising: contacting an antibody composition with the conjugate moiety in the presence of a transglutaminase under a condition sufficient to generate the antibody drug conjugate, wherein at least about 50% of the antibody in the composition is glycosylated in the Fc-region, and wherein the conjugate moiety is conjugated to the endogenous acceptor glutamine residue on the antibody. ### Claim 14 (depends on 13) The method of claim 13, wherein the conjugate moiety comprises a small molecule handle and an active moiety, and wherein the contacting comprises: a) contacting the antibody composition with the small molecule handle in the presence of the transglutaminase under the condition sufficient to generate an intermediate conjugate comprising the antibody specifically conjugated to the small molecule handle, and b) contacting the intermediate conjugate with the active moiety thereby obtaining the antibody drug conjugate. ### Claim 15 (depends on 13) The method of claim 13, wherein the transglutaminase is a wildtype transglutaminase. ### Claim 16 (depends on 15) The method of claim 15, wherein the wildtype transglutaminase has the amino acid sequence of SEQ ID NO:16. ### Claim 17 (depends on 13) The method of claim 13, wherein the transglutaminase is an engineered transglutaminase. ### Claim 18 (depends on 17) The method of claim 17, wherein the engineered transglutaminase comprises an amino acid sequence having at least about 80% identity to SEQ ID NO:16. ### Claim 19 (depends on 13) The method of claim 13, wherein the transglutaminase has a purity of at least about 90%. ### Claim 20 (depends on 13) The method of claim 13, wherein the molar ratio of the transglutaminase and the antibody composition is about 10:1 to about 1:10. ### Claim 21 (depends on 13) The method of claim 13, wherein the transglutaminase is immobilized on a solid support. ### Claim 22 (depends on 13) The method of claim 13, wherein the antibody is immobilized on a solid support. ### Claim 23 (depends on 13) The method of claim 13, where the antibody is a human or humanized antibody. ### Claim 24 (depends on 13) The method of claim 13, wherein the conjugate moiety comprises an active moiety selected from the group consisting of: a moiety that improves the pharmacokinetic property of the antibody composition, a therapeutic moiety, and a diagnostic moiety. ### Claim 25 (depends on 24) The method of claim 24, wherein the active moiety is a toxin. ### Claim 26 An engineered transglutaminase capable of conjugating an Fc-containing polypeptide to a conjugate moiety, wherein the Fc-containing polypeptide comprises an N-glycosylated Fc region, wherein the N-glycosylated Fc region comprises an acceptor glutamine residue flanked by an N-glycosylation site, wherein upon reaction the conjugate moiety is conjugated to the Fc-containing polypeptide via the acceptor glutamine residue, and wherein the conjugation is at least about 10% more active than a wildtype transglutaminase under the same reaction conditions. ### Claim 27 (depends on 26) The engineered transglutaminase of claim 26, comprising an amino acid sequence having at least about 80% identity to SEQ ID NO:16, wherein the transglutaminase comprises a deletion selected from the group consisting of: D1-E4; P244-P247; and N282-L285. ### Claim at position 23 28-29. (canceled)