# US 2021/0330802 A1 · Method for producing antibody-drug conjugate > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 2021/0330802 A1 - Document ID: US20210330802A1 - Status: Published application - Kind: A1 - Application number: 17/284,291 - Filed: 2019-10-10 - Published: 2021-10-28 - Earliest claimed: 2018-10-10 - Assignees: Takeda Pharmaceutical Company Limited - Applicants: Takeda Pharmaceutical Company Limited - Inventors: Masato SUENAGA; Kosuke TAKENAKA; Keiji IWAMOTO - Web page: https://patentgrep.com/patent/US20210330802A1 ## CPC - C07K 16/00: Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies - A61K 38/07: Tetrapeptides - A61K 47/545: {Heterocyclic compounds (A61K47/558 takes precedence)} - A61K 47/65: Peptidic linkers, binders or spacers, e.g. peptidic enzyme-labile linkers - A61K 47/68031: {the drug being an auristatin} - A61K 47/6811: {the drug being a protein or peptide, e.g. transferrin or bleomycin} - A61K 47/6835: {the modifying agent being an antibody or an immunoglobulin bearing at least one antigen-binding site} - A61K 47/6855: {the tumour determinant being from breast cancer cell} - A61K 47/6889: {Conjugates wherein the antibody being the modifying agent and wherein the linker, binder or spacer confers particular properties to the conjugates, e.g. peptidic enzyme-labile linkers or acid-labile linkers, providing for an acid-labile immuno conjugate wherein the drug may be released from its antibody conjugated part in an acidic, e.g. tumoural or environment} - A61K 51/1093: {conjugates with carriers being antibodies} - A61P 35/00: Antineoplastic agents - A61P 35/02: specific for leukemia - B01J 19/0093: {Microreactors, e.g. miniaturised or microfabricated reactors (laboratory containers with capillary fluid transport in microfabricated channels or chambers B01L3/5027)} - C07K 2317/52: Constant or Fc region; Isotype - 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 - B01J: CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS ## Abstract The present invention provides a method for producing an antibody-drug conjugate (ADC) comprising an antibody and a drug linked to each other via a linker. The present invention provides a method for producing, for example, an antibody-drug conjugate (ADC) comprising an antibody and a drug linked to each other via a linker, or a pharmaceutical comprising the ADC, the method comprising mixing, using a microreactor, a solution comprising tricarboxyethyl phosphine (TCEP) and an IgG antibody under reduction reaction with TCEP, with a solution comprising a stoichiometrically excessive amount of an inhibitor of TCEP based on TCEP. ## Claims ### Claim 1 A method for producing an antibody-drug conjugate (ADC) comprising an antibody and a drug linked to each other via a linker, or a pharmaceutical comprising the ADC, the method comprising mixing, using a microreactor, a solution comprising a reducing agent for reducing a disulfide bond of the antibody and a partially reduced IgG antibody under reduction reaction with the reducing agent, with a solution comprising an inhibitor of the reducing agent and/or a reduction terminator. ### Claim 2 A method for producing an antibody-drug conjugate (ADC) comprising an antibody and a drug linked to each other via a linker, or a pharmaceutical comprising the ADC, the method comprising: (b) mixing, using a microreactor, a solution comprising tricarboxyethyl phosphine (TCEP) and a partially reduced IgG antibody under reduction reaction with TCEP, with a solution comprising a stoichiometrically excessive amount of an inhibitor of TCEP based on TCEP. ### Claim 3 (depends on 2) The method according to claim 2, further comprising: (a) mixing, using a microreactor, a solution comprising an IgG antibody with a solution comprising tricarboxyethyl phosphine (TCEP) to generate a partially reduced antibody. ### Claim 4 (depends on 2, 3) The method according to claim 2 or 3, further comprising: (c) reacting the partially reduced antibody with a linker having a functional group reactive with an SH group of the antibody to generate the antibody linked to the linker. ### Claim 5 A method for producing an antibody-drug conjugate (ADC) comprising an antibody and a drug linked to each other via a linker, or a pharmaceutical comprising the ADC, the method comprising: (a) mixing, using a microreactor, a solution comprising an IgG antibody with a solution comprising tricarboxyethyl phosphine (TCEP) to generate a partially reduced antibody; (b) mixing, using a microreactor, a solution comprising tricarboxyethyl phosphine (TCEP) and the partially reduced IgG antibody under reduction reaction with TCEP with a solution comprising a stoichiometrically excessive amount of an inhibitor of TCEP based on TCEP; and (c) reacting the partially reduced antibody with a linker having a functional group reactive with an SH group of the antibody to generate the antibody linked to the linker. ### Claim 6 (depends on 2, 3, 4, 5) The method according to any one of claims 2 to 5, wherein the solution comprising the inhibitor of TCEP further comprises the linker. ### Claim 7 (depends on 4, 5) The method according to claim 4 or 5, further comprising, before (c), mixing, using a microreactor, a solution obtained in (b) with a solution comprising the linker. ### Claim 8 (depends on 2, 3, 4, 5, 6, 7) The method according to any one of claims 2 to 7, wherein the inhibitor of TCEP is one or more inhibitors selected from the group consisting of 4-azidobenzoic acid and 2-azidoethyl-2-acetamide-2-deoxy-β-D-glucopyranoside. ### Claim 9 (depends on 2, 3, 4, 5, 6, 7, 8) The method according to any one of claims 2 to 8, wherein the partially reduced antibody is an antibody having four SH groups. ### Claim 10 (depends on 4, 5, 6, 7, 8, 9) The method according to any one of claims 4 to 9, wherein the functional group reactive with an SH group of the antibody is a maleimide group. ### Claim 11 (depends on 4, 5, 6, 7, 8, 9, 10) The method according to any one of claims 4 to 10, wherein the linker is a linker linked to a drug. ### Claim 12 (depends on 4, 5, 6, 7, 8, 9, 10, 11) The method according to any one of claims 4 to 11, wherein the linker is linked to a drug, and the functional group reactive with an SH group of the antibody is a maleimide group.