# US 10,117,952 B2 · Antibody-drug conjugate

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- Number: US 10,117,952 B2
- Document ID: US10117952B2
- Status: Granted
- Kind: B2
- Application number: 14/435,114
- Filed: 2013-10-10
- Published: 2015-10-22
- Granted: 2018-11-06
- Earliest claimed: 2012-10-11
- Assignees: Daiichi Sankyo Company, Limited
- Applicants: Daiichi Sankyo Company, Limited
- Inventors: Takeshi Masuda; Hiroyuki Naito; Takashi Nakada; Masao Yoshida; Shinji Ashida; Hideki Miyazaki; Yuji Kasuya; Koji Morita; Yuki Abe; Yusuke Ogitani
- Web page: https://patentgrep.com/patent/US10117952B2

## CPC

- A61K 47/6803: {Drugs conjugated to an antibody or immunoglobulin, e.g. cisplatin-antibody conjugates}
- A61K 31/4745: condensed with ring systems having nitrogen as a ring hetero atom, e.g. phenantrolines (yohimbine derivatives, vinblastine A61K31/475; ergoline derivatives A61K31/48)
- A61K 31/48: Ergoline derivatives, e.g. lysergic acid, ergotamine
- C07K 16/2803: {against the immunoglobulin superfamily}
- C07K 16/2827: {against B7 molecules, e.g. CD80, CD86}
- C07K 16/2851: {against the lectin superfamily, e.g. CD23, CD72}
- C07K 16/2875: {against the NGF/TNF superfamily, e.g. CD70, CD95L, CD153, CD154 (against NGF C07K16/22, against TNF C07K16/241)}
- C07K 16/2878: {against the NGF-receptor/TNF-receptor superfamily, e.g. CD27, CD30, CD40, CD95}
- C07K 16/30: from tumour cells
- C07K 2317/30: characterized by aspects of specificity or valency
- C07K 2317/77: Internalization into the cell
- 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)

## Abstract

As an antitumor drug which is excellent in terms of antitumor effect and safety, there is provided an antibody-drug conjugate in which an antitumor compound represented by the following formula is conjugated to an antibody via a linker having a structure represented by the following formula: -L1-L2-LP-NH—(CH2)n1-La-Lb-Lc- wherein the antibody is connected to the terminal of L1, and the antitumor compound is connected to the terminal of Lc with the nitrogen atom of the amino group at position 1 as a connecting position.

## Claims

### Claim 1

An antibody-drug conjugate wherein an antitumor compound represented by the following formula:
is conjugated to an antibody via a linker having a structure represented by the following formula:
-L1-L2-LP-NH—(CH2)n1-La-Lb-Lc-
wherein the antibody is connected to the terminal of L1, the antitumor compound is connected to the terminal of Lc with the nitrogen atom of the amino group at position 1 of the antitumor compound as the connecting position,
wherein
L1 represents -(Succinimid-3-yl-N)—(CH2)n2-C(═O)— wherein n2 represents an integer of 2 to 8,
L2 represents —NH—(CH2—CH2—O)n5-CH2—CH2—C(═O) or a single bond, wherein n5 represents an integer of 1 to 6,
LP represents a tetrapeptide residue of GGFG,
—NH—(CH2)n1-La-Lb-Lc- is —NH—(CH2)3—C(═O)—, —NH—CH2—O—CH2—C(═O)—, or —NH—(CH2)2—O—CH2—C(═O)—,
-(Succinimid-3-yl-N)— has a structure represented by the following formula:
which is connected to the antibody at position 3 thereof and is connected to a methylene group in the linker structure containing this structure on the nitrogen atom at position 1.

### Claim 2 (depends on 1)

The antibody-drug conjugate according to claim 1, wherein n2 is an integer of 2 to 5, and L2 is a single bond.

### Claim 3 (depends on 1)

The antibody-drug conjugate according to claim 1, wherein n2 is an integer of 2 to 5, L2 is —NH—(CH2CH2O)n5-CH2—CH2—C(═O)—, and n5 is 2 or 4.

### Claim 4 (depends on 1)

The antibody-drug conjugate according to claim 1, wherein the structure of the linker is one structure selected from the following group:
-(Succinimid-3-yl-N)—CH2CH2—C(═O)-GGFG-NH—CH2CH2CH2—C(═O)—(NH-DX)
-(Succinimid-3-yl-N)—CH2CH2CH2CH2CH2—C(═O)-GGFG-NH—CH2CH2CH2—C(═O)—(NH-DX)
-(Succinimid-3-yl-N)—CH2CH2CH2CH2CH2—C(═O)-GGFG-NH—CH2—O—CH2—C(═O)—(NH-DX)
-(Succinimid-3-yl-N)—CH2CH2CH2CH2CH2—C(═O)-GGFG-NH—CH2CH2—O—CH2—C(═O)—(NH-DX)
-(Succinimid-3-yl-N)—CH2CH2—C(═O)—NH—CH2CH2O—CH2CH2O—CH2CH2—C(═O)-GGFG-NH—CH2CH2CH2—C(═O)—(NH-DX)
-(Succinimid-3-yl-N)—CH2CH2—C(═O)—NH—CH2CH2O—CH2CH2O—CH2CH2O—CH2CH2O—CH2CH2—C(═O)-GGFG-NH—CH2CH2CH2—C(═O)—(NH-DX)
wherein —(NH-DX) represents a group represented by the following formula:
wherein the nitrogen atom of the amino group at position 1 is the connecting position, and -GGFG- represents a peptide residue of -Gly-Gly-Phe-Gly-.

### Claim 5 (depends on 1)

The antibody-drug conjugate according to claim 1, wherein the structure of the linker is one structure selected from the following group:
-(Succinimid-3-yl-N)—CH2CH2CH2CH2CH2—C(═O)-GGFG-NH—CH2—O—CH2—C(═O)—(NH-DX)
-(Succinimid-3-yl-N)—CH2CH2CH2CH2CH2—C(═O)-GGFG-NH—CH2CH2—O—CH2—C(═O)—(NH-DX)
-(Succinimid-3-yl-N)—CH2CH2—C(═O)—NH—CH2CH2O—CH2CH2O—CH2CH2—C(═O)-GGFG-NH—CH2CH2CH2—C(═O)—(NH-DX)
wherein —(NH-DX) represents a group represented by the following formula:
wherein the nitrogen atom of the amino group at position 1 is the connecting position.

### Claim 6 (depends on 1)

The antibody-drug conjugate according to claim 1, wherein an average number of units of the selected one drug-linker structure conjugated per antibody is in a range of from 2 to 8.

### Claim 7 (depends on 1)

The antibody-drug conjugate according to claim 1, wherein an average number of units of the selected one drug-linker structure conjugated per antibody is in a range of from 3 to 8.

### Claim 8 (depends on 1)

The antibody-drug conjugate according to claim 1, wherein the antibody is an antibody having one or more of a property of recognizing a target cell, a property of binding to a target cell, a property of internalizing in a target cell, and a property of damaging a target cell.

### Claim 9 (depends on 1)

The antibody-drug conjugate according to claim 1, wherein a cell which is targeted by the antibody-drug conjugate is a tumor cell.

### Claim 10 (depends on 1)

The antibody-drug conjugate according to claim 1, wherein the antibody is an anti-A33 antibody, an anti-B7-H3 antibody, an anti-CanAg antibody, an anti-CD20 antibody, an anti-CD22 antibody, an anti-CD30 antibody, an anti-CD33 antibody, an anti-CD56 antibody, an anti-CD70 antibody, an anti-CEA antibody, an anti-Cripto antibody, an anti-EphA2 antibody, an anti-G250 antibody, an anti-MUC1 antibody, an anti-GPNMB antibody, an anti-integrin antibody, an anti-PSMA antibody, an anti-tenascin-C antibody, an anti-SLC44A4 antibody, or an anti-mesothelin antibody.

### Claim 11 (depends on 1)

The antibody-drug conjugate according to claim 1, wherein the antibody is an anti-B7-H3 antibody, an anti-CD30 antibody, an anti-CD33 antibody, or an anti-CD70 antibody.

### Claim 12 (depends on 1)

The antibody-drug conjugate according to claim 1, wherein the antibody is an anti-B7-H3 antibody.

### Claim 13 (depends on 1)

A drug containing the antibody-drug conjugate according to claim 1, or a salt thereof.

### Claim 14 (depends on 1)

An antitumor drug and/or anticancer drug containing the antibody-drug conjugate according to claim 1, or a salt thereof.

### Claim 15 (depends on 14)

The antitumor drug and/or anticancer drug according to claim 14, which is applied to lung cancer, kidney cancer, urothelial cancer, colorectal cancer, prostate cancer, glioblastoma multiforme, ovarian cancer, pancreatic cancer, breast cancer, melanoma, liver cancer, bladder cancer, stomach cancer, or esophageal cancer.

### Claim 16 (depends on 1)

A pharmaceutical composition containing the antibody-drug conjugate according to claim 1, or a salt thereof as an active component, and a pharmaceutically acceptable formulation component.

### Claim 17 (depends on 1)

A method for treating tumor and/or cancer comprising administering the antibody-drug conjugate according to claim 1, or a salt thereof.

### Claim 18

A drug-linker intermediate compound represented by the following formula:
Q—(CH2)nQ-C(═O)-L2a-LP-NH—(CH2)n1-La-Lb-Lc-(NH-DX)
wherein Q represents (maleimid-N-yl)-
nQ represents an integer of 2 to 8,
L2a represents —NH—(CH2—CH2—O)n5-CH2—CH2—C(═O)— or a single bond,
wherein n5 represents an integer of 1 to 6,
LP represents a tetrapeptide of GGFG,
—NH—(CH2)n1-La-Lb-Lc- is —NH—(CH2)3—C(═O)—, —NH—CH2—O—CH2—C(═O)—, or —NH—(CH2)2—O—CH2—C(═O)—,
(maleimid-N-yl)- is a group represented by the following formula:
wherein the nitrogen atom is a connecting position,
and
—(NH-DX) is a group represented by the following formula:
wherein the nitrogen atom of the amino group at position 1 is a connecting position.

### Claim 19 (depends on 18)

The drug-linker intermediate compound according to claim 18, wherein nQ is an integer of 2 to 6.

### Claim 20 (depends on 18)

The drug-linker intermediate compound according to claim 18, wherein
nQ is an integer of 2 to 5, and
L2a is a single bond.

### Claim 21 (depends on 18)

The drug-linker intermediate compound according to claim 18, wherein
nQ is an integer of 2 to 5,
L2a is —NH—(CH2—CH2—O)n5-CH2—CH2—C(═O)—, and
n5 is an integer of 2 to 4.

### Claim 22 (depends on 21)

The drug-linker intermediate compound according to claim 21, wherein n5 is an integer of 2 or 4.

### Claim 23

A compound of the following:
(maleimid-N-yl)-CH2CH2—C(═O)-GGFG-NH—CH2CH2CH2—C(═O)—(NH-DX)
(maleimid-N-yl)-CH2CH2CH2—C(═O)-GGFG-NH—CH2CH2CH2—C(═O)—(NH-DX)
(maleimid-N-yl)-CH2CH2CH2CH2—C(═O)-GGFG-NH—CH2CH2CH2—C(═O)—(NH-DX)
(maleimid-N-yl)-CH2CH2CH2CH2CH2—C(═O)-GGFG-NH—CH2CH2CH2—C(═O)—(NH-DX)
(maleimid-N-yl)-CH2CH2—C(═O)-GGFG-NH—CH2—O—CH2—C(═O)—(NH-DX)
(maleimid-N-yl)-CH2CH2CH2—C(═O)-GGFG-NH—CH2—O—CH2—C(═O)—(NH-DX)
(maleimid-N-yl)-CH2CH2CH2CH2—C(═O)-GGFG-NH—CH2—O—CH2—C(═O)—(NH-DX)
(maleimid-N-yl)-CH2CH2CH2CH2CH2—C(═O)-GGFG-NH—CH2—O—CH2—C(═O)—(NH-DX)
(maleimid-N-yl)-CH2CH2—C(═O)-GGFG-NH—CH2CH2—O—CH2—C(═O)—(NH-DX)
(maleimid-N-yl)-CH2CH2CH2—C(═O)-GGFG-NH—CH2CH2—O—CH2—C(═O)—(NH-DX)
(maleimid-N-yl)-CH2CH2CH2CH2—C(═O)-GGFG-NH—CH2CH2—O—CH2—C(═O)—(NH-DX)
(maleimid-N-yl)-CH2CH2CH2CH2CH2—C(═O)-GGFG-NH—CH2CH2—O—CH2—C(═O)—(NH-DX)
(maleimid-N-yl)-CH2CH2—C(═O)—NH—CH2CH2O—CH2CH2O—CH2CH2—C(═O)-GGFG-NH—CH2CH2CH2—C(═O)—(NH-DX) (maleimid-N-yl)-CH2CH2—C(═O)—NH—CH2CH2O—CH2CH2O—CH2CH2O—CH2CH2—C(═O)-GGFG-NH—CH2CH2CH2—C(═O)—(NH-DX), or
(maleimid-N-yl)-CH2CH2—C(═O)—NH—CH2CH2O—CH2CH2O—CH2CH2O—CH2CH2O—CH2CH2—C(═O)-GGFG-NH—CH2CH2CH2—C(═O)—(NH-DX)
wherein (maleimid-N-yl)- is a group represented by the following formula:
wherein the nitrogen atom is a connecting position, and
—(NH-DX) is a group represented by the following formula:
wherein the nitrogen atom of the amino group at position 1 is a connecting position.

### Claim 24

A compound of the following:
(maleimid-N-yl)-CH2CH2—C(═O)—NH—CH2CH2O—CH2CH2O—CH2CH2—C(═O)-GGFG-NH—CH2CH2CH2—C(═O)—(NH-DX),
(maleimid-N-yl)-CH2CH2CH2CH2CH2—C(═O)-GGFG-NH—CH2—O—CH2—C(═O)—(NH-DX) or
(maleimid-N-yl)-CH2CH2CH2CH2CH2—C(═O)-GGFG-NH—CH2CH2—O—CH2—C(═O)—(NH-DX),
wherein (maleimid-N-yl)- is a group represented by the following formula:
wherein the nitrogen atom is a connecting position, and
—(NH-DX) is a group represented by the following formula:
wherein the nitrogen atom of the amino group at position 1 is a connecting position.

### Claim 25

A linker represented by the following formula:
-L2-LP-NH—(CH2)n1-La-Lb-Lc-
for obtaining an antibody-drug conjugate in which a drug is conjugated to an antibody via the linker,
wherein L1 is a connecting position for the antibody, Lc is a connecting position for an antitumor compound,
wherein
L1 represents -(Succinimid-3-yl-N)—(CH2)n2-C(═O)—,
wherein n2 represents an integer of 2 to 8,
L2 represents —NH—(CH2—CH2—O)n5-CH2—CH2—C(═O) or a single bond, wherein n5 represents an integer of 1 to 6,
LP represents a tetrapeptide residue of GGFG,
—NH—(CH2)n1-La-Lb-Lc- is —NH—(CH2)3—C(═O)—, —NH—CH2—O—CH2—C(═O)—, or —NH—(CH2)2—O—CH2—C(═O)—,
-(Succinimid-3-yl-N)— has a structure represented by the following formula:
which is connected to the antibody at position 3 thereof and is connected to a methylene group in the linker structure containing this structure on the nitrogen atom at position 1.

### Claim 26 (depends on 5)

The antibody-drug conjugate according to claim 5, wherein the structure of the linker is:
-(Succinimid-3-yl-N)—CH2CH2CH2CH2CH2—C(═O)-GGFG-NH—CH2CH2—O—CH2—C(═O)—(NH-DX).

### Claim 27 (depends on 5)

The antibody-drug conjugate according to claim 5, wherein the structure of the linker is:
-(Succinimid-3-yl-N)—CH2CH2—C(═O)—NH—CH2CH2—O—CH2CH2—O—CH2CH2—C(═O)-GGFG-NH—CH2CH2CH2—C(═O)—(NH-DX).

### Claim 28 (depends on 24)

The compound according to claim 24, wherein the structure of the compound is:
(maleimid-N-yl)-CH2CH2—C(═O)—NH—CH2CH2O—CH2CH2O—CH2CH2—C(═O)-GGFG-NH—CH2CH2CH2—C(═O)—(NH-DX).

### Claim 29 (depends on 24)

The compound according to claim 24, wherein the structure of the compound is:
(maleimid-N-yl)-CH2CH2CH2CH2CH2—C(═O)-GGFG-NH—CH2—O—CH2—C(═O)—(NH-DX).

### Claim 30 (depends on 24)

The compound according to claim 24, wherein the structure of the compound is:
(maleimid-N-yl)-CH2CH2CH2CH2CH2—C(═O)-GGFG-NH—CH2CH2—O—CH2—C(═O)—(NH-DX).

### Claim 31 (depends on 25)

The linker according to claim 25, which is selected from the following group, provided that the left terminal is a connecting position with the antibody and the right terminal is a connecting position with the antitumor compound:
-(Succinimid-3-yl-N)—CH2CH2—C(═O)-GGFG-NH—CH2CH2CH2—C(═O)—
-(Succinimid-3-yl-N)—CH2CH2CH2—C(═O)-GGFG-NH—CH2CH2CH2—C(═O)—
-(Succinimid-3-yl-N)—CH2CH2CH2CH2—C(═O)-GGFG-NH—CH2CH2CH2—C(═O)—
-(Succinimid-3-yl-N)—CH2CH2CH2CH2CH2—C(═O)-GGFG-NH—CH2CH2CH2—C(═O)—
-(Succinimid-3-yl-N)—CH2CH2—C(═O)-GGFG-NH—CH2—O—CH2—C(═O)—
-(Succinimid-3-yl-N)—CH2CH2CH2—C(═O)-GGFG-NH—CH2—O—CH2—C(═O)—
-(Succinimid-3-yl-N)—CH2CH2CH2CH2—C(═O)-GGFG-NH—CH2—O—CH2—C(═O)—
-(Succinimid-3-yl-N)—CH2CH2CH2CH2CH2—C(═O)-GGFG-NH—CH2—O—CH2—C(═O)—
-(Succinimid-3-yl-N)—CH2CH2—C(═O)-GGFG-NH—CH2CH2—O—CH2—C(═O)—
-(Succinimid-3-yl-N)—CH2CH2CH2—C(═O)-GGFG-NH—CH2CH2—O—CH2—C(═O)—
-(Succinimid-3-yl-N)—CH2CH2CH2CH2—C(═O)-GGFG-NH—CH2CH2—O—CH2—C(═O)—
-(Succinimid-3-yl-N)—CH2CH2CH2CH2CH2—C(═O)-GGFG-NH—CH2CH2—O—CH2—C(═O)—
-(Succinimid-3-yl-N)—CH2CH2—C(═O)—NH—CH2CH2O—CH2CH2O—CH2CH2—C(═O)-GGFG-NH—CH2CH2CH2—C(═O)—
-(Succinimid-3-yl-N)—CH2CH2—C(═O)—NH—CH2CH2O—CH2CH2O—CH2CH2O—CH2CH2—C(═O)-GGFG-NH—CH2CH2CH2—C(═O)—,and
-(Succinimid-3-yl-N)—CH2CH2—C(═O)—NH—CH2CH2O—CH2CH2O—CH2CH2O—CH2CH2O—CH2CH2—C(═O)-GGFG-NH—CH2CH2CH2—C(═O)—.

### Claim 32 (depends on 25)

The linker according to claim 25, which is selected from the following group, provided that the left terminal is a connecting position with the antibody and the right terminal is a connecting position with the antitumor compound:
-(Succinimid-3-yl-N)—CH2CH2CH2CH2CH2—C(═O)-GGFG-NH—CH2—O—CH2—C(═O)—
-(Succinimid-3-yl-N)—CH2CH2CH2CH2CH2—C(═O)-GGFG-NH—CH2CH2—O—CH2—C(═O)—, and
-(Succinimid-3-yl-N)—CH2CH2—C(═O)—NH—CH2CH2O—CH2CH2O—CH2CH2—C(═O)-GGFG-NH—CH2CH2CH2—C(═O)—.

### Claim 33 (depends on 25)

The linker according to claim 25, which is selected from the following group, provided that the left terminal is a connecting position with the antibody and the right terminal is a connecting position with the antitumor compound:
-(Succinimid-3-yl-N)—CH2CH2CH2CH2CH2—C(═O)-GGFG-NH—CH2—O—CH2—C(═O)—.

### Claim 34 (depends on 25)

The linker according to claim 25, which is selected from the following group, provided that the left terminal is a connecting position with the antibody and the right terminal is a connecting position with the antitumor compound:
-(Succinimid-3-yl-N)—CH2CH2CH2CH2CH2—C(═O)-GGFG-NH—CH2CH2—O—CH2—C(═O)—.

### Claim 35 (depends on 25)

The linker according to claim 25, which is selected from the following group, provided that the left terminal is a connecting position with the antibody and the right terminal is a connecting position with the antitumor compound:
-(Succinimid-3-yl-N)—CH2CH2—C(═O)—NH—CH2CH2O—CH2CH2O—CH2CH2—C(═O)-GGFG-NH—CH2CH2CH2—C(═O)—.