Anti-human MUC1 antibody fab fragment

• US 10,507,251 B2
• Filed: 05/17/2019
• Issued: 12/17/2019
• Est. Priority Date: 11/18/2016
• Status: Active Grant

• Current Assignee: Astellas Pharma Incorporated
• Original Assignee: Astellas Pharma Incorporated
• Inventors: Morinaka, Akifumi, Shirai, Hiroki, Hirayama, Kazunori, Hosogai, Naomi, Doihara, Hitoshi
• Primary Examiner(s): Puttlitz, Karl J
• Application Number:
  US16/414,895
• Publication Number:
  US 20190269804A1
• Time in Patent Office: 214 Days
• Field of Search: None
• CPC Class Codes: A61K 2039/505 comprising antibodies
• CPC Class Codes: A61K 39/395 Antibodies agglutinins A61K38/36 ; as drug carriers A61K47/50; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
• CPC Class Codes: A61K 47/54 the modifying agent being an organic compound
• CPC Class Codes: A61K 47/6851 the antibody targeting a determinant of a tumour cell
• CPC Class Codes: A61K 47/6855 the tumour determinant being from breast cancer cell
• CPC Class Codes: A61K 49/0021 the fluorescent group being a small organic molecule
• CPC Class Codes: A61K 49/0032 Methine dyes, e.g. cyanine dyes
• CPC Class Codes: A61K 49/0036 Porphyrins used in photodynamic therapy A61K41/0071 or A61K41/0076; used as targeting group or modifying agent for targeting a therapeutic compound A61K47/546
• CPC Class Codes: A61K 49/0058 Antibodies
• CPC Class Codes: A61K 49/16 Antibodies; Immunoglobulins; Fragments thereof
• CPC Class Codes: A61K 51/1051 the tumor cell being from breast, e.g. the antibody being herceptin
• CPC Class Codes: A61K 51/106 the tumor cell being from kidney or bladder
• CPC Class Codes: A61K 51/1093 conjugates with carriers being antibodies
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: C07K 16/18 against material from animals or humans
• CPC Class Codes: C07K 16/28 against receptors, cell surface antigens or cell surface determinants
• CPC Class Codes: C07K 16/30 from tumour cells
• CPC Class Codes: C07K 16/3092 against tumour-associated mucins
• CPC Class Codes: C07K 17/00 Carrier-bound or immobilised peptides carrier-bound or immobilised enzymes C12N11/00; Preparation thereof
• CPC Class Codes: C07K 2317/21 from primates, e.g. man
• CPC Class Codes: C07K 2317/24 : containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
• CPC Class Codes: C07K 2317/40 : characterized by post-translational modification
• CPC Class Codes: C07K 2317/55 : Fab or Fab'
• CPC Class Codes: C07K 2317/76 : Antagonist effect on antigen, e.g. neutralization or inhibition of binding
• CPC Class Codes: C07K 2317/92 : Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value
• CPC Class Codes: C12N 15/09 : Recombinant DNA-technology
• CPC Class Codes: C12N 15/85 : for animal cells
• CPC Class Codes: C12N 5/10 : Cells modified by introduction of foreign genetic material
• CPC Class Codes: C12P 21/02 : having a known sequence of two or more amino acids, e.g. glutathione
• CPC Class Codes: G01N 2333/4725 : Mucins, e.g. human intestinal mucin
• CPC Class Codes: G01N 33/57407 : Specifically defined cancers
• CPC Class Codes: G01N 33/57415 : of breast

Abstract

The problem to be solved is to provide an anti-human MUC1 antibody Fab fragment that is expected to be useful in the diagnosis and/or treatment of a cancer, particularly, the diagnosis and/or treatment of breast cancer or bladder cancer, and a diagnosis approach and/or a treatment approach using a conjugate comprising the Fab fragment. The solution is an anti-human MUC1 antibody Fab fragment comprising a heavy chain fragment comprising a heavy chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 8 or 10, and a light chain comprising a light chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 12, and a conjugate comprising the Fab fragment.

Claims

1. A conjugate comprising one or more labeling moiety and an anti-human MUC1 antibody Fab fragment, wherein the labeling moiety is (i) a fluorescent dye and a linker or (ii) a fluorescent dye, and wherein the anti-human MUC1 antibody Fab fragment is selected from the group consisting of the following (a) and (b):

(a) an anti-human MUC I antibody Fab fragment comprising a heavy chain fragment comprising a heavy chain variable region consisting of the amino acid sequence represented by SEQ ID NO;

8 or SEQ ID NO;

10 and a light chain comprising a light chain variable region consisting of the amino acid sequence represented by SEQ ID NO;

12; and

(b) an anti-human MUC1 antibody Fab fragment comprising a heavy chain fragment comprising a heavy chain variable region derived from a heavy chain variable region consisting of the amino acid sequence represented by SEQ ID NO;

8 or SEQ ID NO;

10 by the modification of glutamine at amino acid position 1 of SEQ ID NO;

8 or SEQ ID NO;

10 into pyroglutamic acid, and a light chain comprising a light chain variable region consisting of the amino acid sequence represented by SEQ ID NO;

12.

2. The conjugate according to claim 1 wherein the anti-human MUC1 antibody Fab fragment is selected from the group consisting of the following (a) and (b):

(a) an anti-human MUC1 antibody Fab fragment comprising a heavy chain fragment consisting of the amino acid sequence represented by SEQ ID NO;

2 or SEQ ID NO;

4 and a light chain consisting of the amino acid sequence represented by SEQ ID NO;

6; and

(b) an anti-human MUC1 antibody Fab fragment comprising a heavy chain fragment derived from a heavy chain fragment consisting of the amino acid sequence represented by SEQ ID NO;

2 or SEQ ID NO;

4 by the modification of glutamine at amino acid position 1 of SEQ ID NO;

2 or SEQ ID NO;

4 into pyroglutamic acid, and a light chain consisting of the amino acid sequence represented by SEQ ID NO;

6.

3. The conjugate according to claim 1 wherein the anti-human MUC1 antibody Fab fragment is selected from the group consisting of the following (a) and (b):

(a) an anti-human MUC1 antibody Fab fragment comprising a heavy chain fragment comprising a heavy chain variable region consisting of the amino acid sequence represented by SEQ ID NO;

10 and a light chain comprising a light chain variable region consisting of the amino acid sequence represented by SEQ ID NO;

12; and

(b) an anti-human MUC1 antibody Fab fragment comprising a heavy chain fragment comprising a heavy chain variable region derived from a heavy chain variable region consisting of the amino acid sequence represented by SEQ ID NO;

10 by the modification of glutamine at amino acid position 1 of SEQ ID NO;

10 into pyroglutamic acid, and a light chain comprising a light chain variable region consisting of the amino acid sequence represented by SEQ ID NO;

12.

4. The conjugate according to claim 3 wherein the anti-human MUC1 antibody Fab fragment is selected from the group consisting of the following (a) and (b):

(a) an anti-human MUC1 antibody Fab fragment comprising a heavy chain fragment consisting of the amino acid sequence represented by SEQ ID NO;

4 and a light chain consisting of the amino acid sequence represented by SEQ ID NO;

6; and

(b) an anti-human MUC1 antibody Fab fragment comprising a heavy chain fragment derived from a heavy chain fragment consisting of the amino acid sequence represented by SEQ ID NO;

4 by the modification of glutamine at amino acid position 1 of SEQ ID NO;

4 into pyroglutamic acid, and a light chain consisting of the amino acid sequence represented by SEQ ID NO;

6.

5. The conjugate according to claim 4 wherein the anti-human MUC1 antibody Fab fragment is an anti-human MUC1 antibody Fab fragment comprising a heavy chain fragment consisting of the amino acid sequence represented by SEQ ID NO:

4 and a light chain consisting of the amino acid sequence represented by SEQ ID NO;

6.

6. The conjugate according to claim 4 wherein the anti-human MUC1 antibody Fab fragment is an anti-human MUC1 antibody Fab fragment comprising a heavy chain fragment derived from a heavy chain fragment consisting of the amino acid sequence represented by SEQ ID NO:

4 by the modification of glutamine at amino acid position 1 of SEQ ID NO;

4 into pyroglutamic acid, and a light chain consisting of the amino acid sequence represented by SEQ ID NO;

6.

7. The conjugate according to claim 1, wherein the fluorescent dye is a fluorescent dye selected from the group consisting of the following formula (B) and the following formula (C):

8. The conjugate according to claim 7, wherein the wavy line represents binding to the anti-human MUC1 antibody Fab fragment, and the anti-human MUC1 antibody Fab fragment is bound via an amino group thereof to the carbon atom of a labeling moiety terminal C(═

O) group.

9. The conjugate according to claim 8, wherein the labeling moiety is a fluorescent dye represented by formula (B).

10. A conjugate selected from the group consisting of the following (a) to (c):

(a) the conjugate according to claim 9 wherein the anti-human MUC1 antibody Fab fragment is an anti-human MUC1 antibody Fab fragment comprising a heavy chain fragment consisting of the amino acid sequence represented by SEQ ID NO;

4 and a light chain consisting of the amino acid sequence represented by SEQ ID NO;

6;

(b) the conjugate according to claim 9 wherein the anti-human MUC1 antibody Fab fragment is an anti-human MUC1 antibody Fab fragment comprising a heavy chain fragment derived from a heavy chain fragment consisting of the amino acid sequence represented by SEQ ID NO;

4 by the modification of glutamine at amino acid position 1 of SEQ ID NO;

4 into pyroglutamic acid, and a light chain consisting of the amino acid sequence represented by SEQ ID NO;

6; and

(c) a conjugate which is a mixture of (a) and (b).

11. The conjugate according to claim 8, wherein the labeling moiety is a fluorescent dye represented by formula (C).

12. A conjugate selected from the group consisting of the following (a) to (c):

(a) the conjugate according to claim 11 wherein the anti-human MUC1 antibody Fab fragment is an anti-human MUC1 antibody Fab fragment comprising a heavy chain fragment consisting of the amino acid sequence represented by SEQ ID NO;

4 and a light chain consisting of the amino acid sequence represented by SEQ ID NO;

6;

(b) the conjugate according to claim 11 wherein the anti-human MUC1 antibody Fab fragment is an anti-human MUC1 antibody Fab fragment comprising a heavy chain fragment derived from a heavy chain fragment consisting of the amino acid sequence represented by SEQ ID NO;

4 by the modification of glutamine at amino acid position 1 of SEQ ID NO;

4 into pyroglutamic acid, and a light chain consisting of the amino acid sequence represented by SEQ ID NO;

6; and

(c) a conjugate which is a mixture of (a) and (b).

13. A composition comprising the conjugate according to claim 1, and a pharmaceutically acceptable carrier.

14. A composition comprising the conjugate according to claim 10 and a pharmaceutically acceptable carrier.

15. A composition comprising the conjugate according to claim 12 and a pharmaceutically acceptable carrier.

16. A method for diagnosing breast cancer or bladder cancer, comprising preoperatively or intraoperatively administering the conjugate according to claim 1 to a subject in need thereof.

17. A method for treating breast cancer or bladder cancer, comprising administering a therapeutically effective amount of the conjugate according to claim 1 to a subject in need thereof.