# US 2023/0055475 A1 · Methods and compositions for making antibodies and antibody derivatives with reduced core fucosylation

> Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt

- Number: US 2023/0055475 A1
- Document ID: US20230055475A1
- Status: Published application
- Kind: A1
- Application number: 17/698,920
- Filed: 2022-03-18
- Published: 2023-02-23
- Earliest claimed: 2008-05-02
- Assignees: Seagen Inc.
- Applicants: Seagen Inc.
- Inventors: Stephen C. ALLEY; Scott C. JEFFREY; Django SUSSMAN; Dennis R. BENJAMIN; Brian TOKI; Patrick J. BURKE
- Web page: https://patentgrep.com/patent/US20230055475A1

## CPC

- C12N 5/0037: {Serum-free medium, which may still contain naturally-sourced components}
- C07H 13/04: having the esterifying carboxyl radicals attached to acyclic carbon atoms
- C07K 16/00: Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies
- C07K 2317/72: Increased effector function due to an Fc-modification
- C07K 2317/732: Antibody-dependent cellular cytotoxicity [ADCC]
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- C07H: SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS (derivatives of aldonic or saccharic acids C07C, C07D; aldonic acids, saccharic acids C07C59/105, C07C59/285; cyanohydrins C07C255/16; glycals C07D; compounds of unknown constitution C07G; polysaccharides, derivatives thereof C08B; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification C12N15/00; sugar industry C13)
- 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)

## Abstract

The invention provides methods and compositions for preparing antibodies and antibody derivatives with reduced core fucosylation.

## Claims

### Claim at position 1

1-95 (canceled)

### Claim 96

A method for inhibiting the growth of cells expressing CD70 antigen, comprising administering to the cells a population of humanized anti-CD70 antibodies in an amount sufficient to inhibit cell growth, wherein the antibodies each comprise:
(i) a humanized heavy chain comprising the three CDRs from SEQ ID NO:1 and a variable region framework sequence of human germline VH1-2 or VH1-18 and exon JH-6, provided that any of positions H46, H67, H68, H69, H70, H71, H80, H81, H82, H82A and H91 (Kabat numbering) can be occupied by the amino acid occupying the corresponding position from SEQ ID NO:1,
(ii) a humanized light chain comprising the three CDRs from SEQ ID NO:2 and a variable region framework sequence of human germline VK exon B3 and JK exon Jk−1, provided that any of positions L25 and L33 can be occupied by the amino acid occupying the corresponding position from SEQ ID NO:2, and
(iii) an Fc domain, wherein at least 50% of the antibodies in the population of antibodies lack core fucosylation.

### Claim 97 (depends on 96)

The method of claim 96, wherein at least 70% of the antibodies in the population of antibodies lack core fucosylation.

### Claim 98 (depends on 96)

The method of claim 96, wherein position H46 of the antibodies is occupied by the amino acid occupying the corresponding position from SEQ ID NO:1.

### Claim 99 (depends on 96)

The method of claim 96, wherein at least one of positions H46, H67, H68, H69, H70, H71, H80, H81, H82, H82A and H91 (Kabat numbering) in the humanized heavy chain variable region of the antibodies is occupied by the residue occupying the corresponding position in SEQ ID NO:1.

### Claim 100 (depends on 96)

The method of claim 96, wherein at least one of positions L25 and L33 in the humanized light chain variable region of the antibodies is occupied by the residue occupying the corresponding position in SEQ ID NO:2.

### Claim 101 (depends on 96)

The method of claim 96, wherein the antibodies comprise a human IgG1 constant domain, wherein the humanized heavy chain comprises the amino acid sequence corresponding to positions 20-467 of SEQ ID NO:3 or positions 20-467 of SEQ ID NO:4.

### Claim 102 (depends on 96)

The method of claim 96, wherein the antibodies each comprise an antibody effector domain.

### Claim 103 (depends on 102)

The method of claim 102, wherein the antibody effector domain mediates ADCP.

### Claim 104 (depends on 96)

The method of claim 96, wherein the antibodies are conjugated to therapeutic agents.

### Claim 105 (depends on 104)

The method of claim 104, wherein the therapeutic agents are chemotherapeutic agents or immunomodulatory agents.

### Claim 106

A method for inhibiting the growth of cells expressing CD70 antigen, comprising administering to the cells a population of humanized anti-CD70 antibodies in an amount sufficient to inhibit cell growth, wherein the antibodies each comprise:
(i) a humanized heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, or amino acids 20-137 of SEQ ID NO:9,
(ii) a humanized light chain variable region comprising the amino acid sequence of SEQ ID NO:10, and
(iii) an Fc domain, wherein at least 50% of the antibodies in the population of antibodies lack core fucosylation.

### Claim 107 (depends on 106)

The method of claim 106, wherein at least 70% of the antibodies in the population of antibodies lack core fucosylation.

### Claim 108 (depends on 106)

The method of claim 106, wherein the humanized heavy chain variable region of the antibodies comprise the sequence set forth in SEQ ID NO:7 or SEQ ID NO:8, and the humanized light chain variable region of the antibodies comprise the sequence set forth in SEQ ID NO:10.

### Claim 109 (depends on 106)

The method of claim 106, wherein the humanized heavy chain variable region of the antibodies comprise the sequence set forth in SEQ ID NO:7 and the humanized light chain variable region of the antibodies comprise the sequence set forth in SEQ ID NO:10.

### Claim 110 (depends on 109)

The method of claim 109, wherein at least 70% of the antibodies in the population of antibodies lack core fucosylation.

### Claim 111 (depends on 106)

The method of claim 106, wherein the antibodies are conjugated to therapeutic agents.

### Claim 112 (depends on 111)

The method of claim 111, wherein the therapeutic agents are chemotherapeutic agents or immunomodulatory agents.

### Claim 113

A method for inhibiting the growth of cells expressing CD70 antigen, comprising administering to the cells a population of humanized anti-CD70 antibodies in an amount sufficient to inhibit cell growth, wherein at least 50% of the antibodies in the population of antibodies lack core fucosylation, and wherein the antibodies are h1F6.

### Claim 114 (depends on 113)

The method of claim 113, wherein at least 70% of the antibodies in the population of antibodies lack core fucosylation.

### Claim 115 (depends on 113)

The method of claim 113, wherein the antibodies are conjugated to therapeutic agents.