Granted Patent B2

US 12,428,450 · App. 17/241,585 · Granted Sep 30, 2025

Inventors: Kei Yamada (Kawasaki, JP); Yoshihiko Matsuda (Kawasaki, JP); Kazutoshi Takahashi (Kawasaki, JP); Natsuki Shikida (Kawasaki, JP); Kazutaka Shimbo (Kawasaki, JP); Hayato Nagano (Kawasaki, JP)

Assignee: Ajinomoto Co., Inc.

C07K14/31C07K16/32C07K2317/24C07K2319/00

Abstract

Claims (79)

1. A compound having an affinity substance to an antibody, a cleavable portion, and a reactive group, or a salt thereof, the compound being represented by the following Formula (I):

A-L-B—R  (I),

wherein

A is the affinity substance to an antibody,

L is a cleavable linker that is a divalent group comprising the cleavable portion,

B is (a) a divalent group comprising a bioorthogonal functional group or (b) a divalent group comprising no bioorthogonal functional group, wherein the bioorthogonal functional group has a chemical structure selected from the group consisting of the following:

wherein

R 1f , one or a plurality of R 1g s, and one or a plurality of R 1h s are the same as or different from each other, and are each an atom or a group selected from the group consisting of the substituent (i) to (vii) or an electron-withdrawing group, and

● is a bond,

wherein the substituent (i) to (vii) are:

(i) a halogen atom;

(ii) a monovalent hydrocarbon group;

(iii) an aralkyl;

(iv) a monovalent heterocyclic group;

(v) R c —O—, R c —C(═O)—, R c —O—C(═O)—, or R c —C(═O)—O—, wherein R c represents a hydrogen atom or a monovalent hydrocarbon group;

(vi) NR d R e —, NR d R e —C(═O)—, NR d R e —C(═O)—O—, or R d —C(═O)—NR e —, wherein R d and R e are the same as or different from each other, and each represent a hydrogen atom or a monovalent hydrocarbon group; or

(vii) a nitro group, a sulfate group, a sulfonate group, a cyano group, or a carboxyl group, and

R is the reactive group to the antibody,

wherein the affinity substance to an antibody is a polypeptide comprising a glutamine residue (Q) at an N-terminal terminal and is selected from the group consisting of the following (1) to (4):

wherein Q is a glutamine residue at an N-terminal, E is a glutamic acid residue, T is a threonine residue, the solid line is a bond, the spacer portion consists of 1 to 50 amino acid residues, and the antibody-affinity polypeptide portion consists of 15 to 70 amino acid residues, wherein the antibody-affinity polypeptide portion is selected from the group consisting of protein A, protein G, protein L, protein Z, and a fragment thereof having affinity to an antibody, and a variant thereof having affinity to an antibody.

2. The compound or salt thereof according to claim 1 , wherein the glutamine residue is pyroglutamylated.

3. The compound or salt thereof according to claim 1 , wherein the antibody-affinity polypeptide portion comprises an amino acid sequence with any one amino acid residue substituted with a lysine residue in the amino acid sequence of FNMQCQRRFYEALHDPNLNEEQRNAXIXSIRDDC (SEQ ID NO: 5) and having at least 90% identity to the amino acid sequence of SEQ ID NO: 5, and

wherein two Xs are each independently one amino acid residue selected from the group consisting of an arginine residue, a glutamic acid residue, and a glutamine residue.

4. The compound or salt thereof according to claim 1 , wherein the antibody-affinity polypeptide portion comprises an amino acid sequence with any one amino acid residue substituted with a lysine residue in the amino acid sequence of FNMQCQRRFYEALHDPNLNEEQRNARIRSIRDDC (SEQ ID NO: 6) and having at least 90% identity to the amino acid sequence of SEQ ID NO: 6.

5. The compound or salt thereof according to claim 1 , wherein the affinity substance to an antibody comprises an amino acid sequence selected from the group consisting of the following:

(1) 

(SEQ ID NO: 7) 

QETNPTENLYFQQKNMQCQRRFYEALHDPNLNEEQRNARIRSIRDDC;

(2)

(SEQ ID NO: 8)

QTADNQKNMQCQRRFYEALHDPNLNEEQRNARIRSIRDDCSQSANLLAE

AQQLNDAQAPQA;

(3) 

(SEQ ID NO: 9)

QETKNMQCQRRFYEALHDPNLNEEQRNARIRSIRDDC;

(4) 

(SEQ ID NO: 10)

QETFNKQCQRRFYEALHDPNLNEEQRNARIRSIRDDC; 

(5) 

(SEQ ID NO: 22) 

QETENMQCQRRFYEALHDPNLNKEQRNARIRSIRDDC;

(6) 

(SEQ ID NO: 23) 

QETENMQCQRRFYEALHDPNLNEEQRNARIRSIKDDC;

(7) 

(SEQ ID NO: 51)

QETMQCQRRFYEALHDPNLNEEQRNARIRSIKDDC;

(8) 

(SEQ ID NO: 52)

QETQCQRRFYEALHDPNLNEEQRNARIRSIKDDC;

(9) 

(SEQ ID NO: 53) 

QETCQRRFYEALHDPNLNEEQRNARIRSIKDDC;

(10) 

(SEQ ID NO: 24) 

QETRGNCAYHKGQLVWCTYH; and

(11) 

(SEQ ID NO: 25)

QETRGNCAYHKGQIIWCTYH.

6. A reagent of regioselectively modifying an antibody, the reagent comprising a compound having an affinity substance to an antibody, a cleavable portion, and a reactive group, or a salt thereof, the compound being represented by the following Formula (I):

A-L-B—R  (I),

wherein

A is the affinity substance to an antibody,

L is a cleavable linker that is a divalent group comprising the cleavable portion,

B is (a) a divalent group comprising a bioorthogonal functional group or (b) a divalent group comprising no bioorthogonal functional group, wherein the bioorthogonal functional group corresponds to any one chemical structure selected from the group consisting of the following:

wherein

R 1f , one or a plurality of R 1g s, and one or a plurality of R 1h s are the same as or different from each other, and are each an atom or a group selected from the group consisting of the substituent (i) to (vii) or an electron-withdrawing group, and

● is a bond,

wherein the substituent (i) to (vii) are:

(i) a halogen atom;

(ii) a monovalent hydrocarbon group;

(iii) an aralkyl;

(iv) a monovalent heterocyclic group;

(v) R c —O—, R c —C(═O)—, R c —O—C(═O)—, or R c —C(═O)—O—, wherein R c represents a hydrogen atom or a monovalent hydrocarbon group;

(vi) NR d R e —, NR d R e —C(═O)—, NR d R e —C(═O)—O—, or R d —C(═O)—NR e —, wherein R d and R e are the same as or different from each other, and each represent a hydrogen atom or a monovalent hydrocarbon group; or

(vii) a nitro group, a sulfate group, a sulfonate group, a cyano group, or a carboxyl group, and

R is the reactive group to the antibody,

wherein the affinity substance to an antibody is a polypeptide comprising a glutamine residue (Q) at an N-terminal, and is selected from the group consisting of the following (1) to (4):

wherein Q is a glutamine residue at an N-terminal, E is a glutamic acid residue, T is a threonine residue, the solid line is a bond, the spacer portion consists of 1 to 50 amino acid residues, and the antibody-affinity polypeptide portion consists of 15 to 70 amino acid residues, wherein the antibody-affinity polypeptide portion is selected from the group consisting of protein A, protein G, protein L, protein Z, and a fragment thereof having affinity to an antibody, and a variant thereof having affinity to an antibody.

Assignments (1)

Priority Claims (1)

Continuity (2)

References Cited (48)