Antibody targeted to tissue factor, preparation method therefor, and use thereof

Patent metadata

Field	Value
Publication number	US-10676537-B2
Application number	US-201716326881-A
Country	US
Kind code	B2
Filing date	Feb 20, 2017
Priority date	Aug 22, 2016
Publication date	Jun 9, 2020
Grant date	Jun 9, 2020

Abstract

The present invention provides a tissue factor (TF) monoclonal antibody and a preparation method therefor. The monoclonal antibody provided by the present invention can specifically bind with a TF antigen, has high affinity and low immunogenicity, and has the activity of resisting tumors and the like.

First claim

The invention claimed is:

1. An anti-human tissue factor monoclonal antibody or an antigen-binding fragment thereof, wherein said antibody has
a heavy chain variable region comprising HCDR1 as set forth in SEQ ID NO: 1, HCDR2 as set forth in SEQ ID NO: 2, and HCDR3 as set forth in SEQ ID NO: 3, and
a light chain variable region comprising LCDR1 as set forth in SEQ ID NO: 4, LCDR2 as set forth in SEQ ID NO: 5, and LCDR3 as set forth in SEQ ID NO: 6.

2. The antibody according to claim 1 , wherein the antibody is an animal-derived antibody, a chimeric antibody, or a humanized antibody.

3. The antibody according to claim 1 , wherein the heavy chain variable region sequence of the antibody is selected from the group consisting of SEQ ID NOS: 7, 9, 10, 11, 12, and 13; and/or
the light chain variable region sequence of the antibody is selected from the group consisting of SEQ ID NOS: 8, 14, 15, 16, and 17.

4. A method for treating cancer or a thrombotic disease, comprising administering a therapeutically effective amount of an anti-human tissue factor monoclonal antibody selected from the group consisting of TF-mAb-SC1, TF-mAb-Ch, TF-mAb-H29, TF-mAb-H30, TF-mAb-H31, TF-mAb-H32, TF-mAb-H33, TF-mAb-H34, TF-mAb-H35, TF-mAb-H36, TF-mAb-H37, TF-mAb-H38, TF-mAb-H39, TF-mAb-H40, TF-mAb-H41, TF-mAb-H42, TF-mAb-H43, TF-mAb-H44, TF-mAb-H45, TF-mAb-H46, TF-mAb-H47, and TF-mAb-H48 to a subject in need thereof.

5. The method according to claim 4 , wherein the method is for treating a thrombotic disease.

6. The method according to claim 4 , wherein the method is for treating cancer, and wherein the cancer is a cancer with high TF expression.

7. An isolated polynucleotide, which comprises a nucleic acid molecule encoding the heavy chain variable region of an anti-human tissue factor monoclonal antibody, and a nucleic acid molecule encoding the light chain variable region of said monoclonal antibody, wherein:
the heavy chain variable region of the antibody comprises HCDR1 as set forth in SEQ ID NO: 1, HCDR2 as set forth in SEQ ID NO: 2, and HCDR3 as set forth in SEQ ID NO: 3, and
the light chain variable region of the antibody comprises LCDR1 as set forth in SEQ ID NO: 4, LCDR2 as set forth in SEQ ID NO: 5, and LCDR3 as set forth in SEQ ID NO: 6.

8. A vector, comprising the isolated polynucleotide according to claim 7 .

9. A genetically engineered host cell, comprising the vector according to claim 8 .

10. An immune cell, wherein the immune cell expresses the antibody according to claim 1 .

11. A pharmaceutical composition, comprising the antibody according to claim 1 and a pharmaceutically acceptable carrier.

12. A method for determining the presence or absence of a TF protein in a sample in vitro, the method comprises:
(1) contacting the sample with the antibody according to claim 1 in vitro; and
(2) determining whether an antigen-antibody complex is formed, wherein the formation of the complex indicates the presence of a TF protein in the sample.

13. A method for preparing an anti-human tissue factor monoclonal antibody, comprising:
(a) culturing the host cell according to claim 9 under conditions suitable for expressing an anti-human tissue factor monoclonal antibody; and
(b) separating the antibody from the culture.

14. The antibody according to claim 3 , wherein the antibody is selected from the group consisting of: TF-mAb-SC1, TF-mAb-Ch, TF-mAb-H29, TF-mAb-H30, TF-mAb-H31, TF-mAb-H32, TF-mAb-H33, TF-mAb-H34, TF-mAb-H35, TF-mAb-H36, TF-mAb-H37, TF-mAb-H38, TF-mAb-H39, TF-mAb-H40, TF-mAb-H41, TF-mAb-H42, TF-mAb-H43, TF-mAb-H44, TF-mAb-H45, TF-mAb-H46, TF-mAb-H47, and TF-mAb-H48.

15. The antibody according to claim 1 , wherein the antibody has an EC50 of 0.01-0.03 nM for the affinity to human TF protein.

16. The isolated polynucleotide according to claim 7 , wherein:
the heavy chain variable region sequence of the antibody is SEQ ID NO: 7, 9, 10, 11, 12, or 13; and/or
the light chain variable region sequence of the antibody is SEQ ID NO: 8, 14, 15, 16, or 17.

17. The isolated polynucleotide according to claim 7 , wherein the antibody is selected from the group consisting of: TF-mAb-SC1, TF-mAb-Ch, TF-mAb-H29, TF-mAb-H30, TF-mAb-H31, TF-mAb-H32, TF-mAb-H33, TF-mAb-H34, TF-mAb-H35, TF-mAb-H36, TF-mAb-H37, TF-mAb-H38, TF-mAb-H39, TF-mAb-H40, TF-mAb-H41, TF-mAb-H42, TF-mAb-H43, TF-mAb-H44, TF-mAb-H45, TF-mAb-H46, TF-mAb-H47, and TF-mAb-H48.

Assignees

• Univ Fudan
• Shanghai Miracogen Inc

Inventors

• Yu Ke
• Zhang Xuesai
• Lin Qing
• Li Qingrou

Classifications

• A61K47/6803

  Drugs conjugated to an antibody or immunoglobulin, e.g. cisplatin-antibody conjugates · CPC title

• A61P35/00

  Antineoplastic agents · CPC title

• C07K2317/77

  Internalization into the cell · CPC title

• A61P7/02

  Antithrombotic agents; Anticoagulants; Platelet aggregation inhibitors · CPC title

• C07K16/36 Primary

  against blood coagulation factors · CPC title