IL-15 heterodimeric protein and uses thereof

Patent metadata

Field	Value
Publication number	US-10206980-B2
Application number	US-201415109920-A
Country	US
Kind code	B2
Filing date	Dec 25, 2014
Priority date	Jan 8, 2014
Publication date	Feb 19, 2019
Grant date	Feb 19, 2019

Abstract

Provided is an IL-15 heterodimeric protein, such as an IL-15/IL-15Rα heterodimeric protein. The IL-15 heterodimeric protein contains protein (I) and protein (II). Protein (I) is recombinantly produced by combining IL-15 or a variant thereof with a first Fc variant, and protein (II) is a second Fc variant, or recombinantly produced by combining IL-15Rα or a variant thereof with a second Fc variant. The first Fc variant and the second Fc variant are preferably linked via a “Knob-into-Hole” mode. Also provided are methods of using the IL-15 heterodimeric proteins to modulate an immune response. The provided IL-15 heterodimeric proteins have significant anti-tumor activity, better biological activity and prolonged in vivo half-life compared to the IL-15 molecule alone.

First claim

The invention claimed is:

1. An IL-15 heterodimeric protein, comprising:
protein (I) and protein (H);
wherein protein (I) is recombinantly produced and comprises the sequence of IL-15 and the sequence of a first Fc variant;
protein (H) is a second Fc variant, or is recombinantly produced and comprises the sequence of IL-15Rα or a variant thereof and the sequence of a second Fc variant, wherein the IL-15Rα variant is an extracellular domain of IL-15Rα or a functional fragment thereof, the functional fragment being a C-terminal truncated form of the extracellular domain of IL-15Rα having the activity of IL-15Rα; and
protein (I) and protein (H) form a stable heterodimeric protein by an interaction between the first Fc variant and the second Fc variant.

2. The IL-15 heterodimeric protein according to claim 1 , wherein the sequence of IL-15 comprises SEQ ID NO: 1.

3. The IL-15 heterodimeric protein according to claim 1 , wherein protein (II) is a second Fc variant.

4. The IL-15 heterodimeric protein according to claim 1 , wherein protein (II) is recombinantly produced by combining IL-15Rα or a variant thereof with a second Fc variant.

5. The IL-15 heterodimeric protein according to claim 1 , wherein the sequence of the IL-15Rα variant is selected from the group consisting of SEQ ID NOs: 2-7.

6. The IL-15 heterodimeric protein according to claim 1 , wherein:
the first Fc variant is selected from the group consisting of a Knob modified Fc and a Hole modified Fc; and the second Fc variant is selected from the group consisting of a Hole modified Fc and a Knob modified Fc;
when the first Fc variant is a Knob modified Fc, the second Fc variant is a Hole modified Fc; and
when the second Fc variant is a Knob modified Fc, the first Fc variant is a Hole modified Fc.

7. The IL-15 heterodimeric protein according to claim 1 , wherein the sequence of the first Fc variant is selected from the group consisting of SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28 and SEQ ID NO: 29; and the sequence of the second Fc variant is selected from the group consisting of SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28 and SEQ ID NO: 29.

8. The IL-15 heterodimeric protein according to claim 1 , wherein the sequence of protein (I) is selected from the group consisting of SEQ ID NOs: 14-17; and the sequence of protein (H) is selected from the group consisting of SEQ ID NOs: 18-25 and 34-37.

9. The IL-15 heterodimeric protein according to claim 1 , wherein the sequence of protein (I) is selected from the group consisting of SEQ ID NOs: 30-31; and the sequence of protein (H) is selected from the group consisting of SEQ ID NOs: 32-33.

10. A nucleic acid encoding the IL-15 heterodimeric protein according to claim 1 .

11. A DNA vector comprising the nucleic acid according to claim 10 .

12. A host cell comprising the DNA vector according to claim 11 .

13. A method for preparing the IL-15 heterodimeric protein according to claim 1 , comprising:
culturing a host cell comprising a DNA vector comprising a nucleic acid encoding the IL-15 heterodimeric protein according to claim 1 under a condition sufficient for expression of the IL-15 heterodimeric protein; and expressing and purifying the IL-15 heterodimeric protein.

14. A pharmaceutical composition comprising the IL-15 heterodimeric protein according to claim 1 , and a pharmaceutically acceptable excipient, diluent or carrier.

15. A method of treating an IL-15 mediated disease or disorder comprising administering to a subject in need of the treatment the pharmaceutical composition of claim 14 , wherein the IL-15 mediated disease or disorder is selected from the group consisting of cancer, infectious disease, and blood disease; wherein the cancer is selected from the group consisting of melanoma, colorectal cancer, lymphoma, renal cell carcinoma, and lung cancer; the infectious disease is selected from the group consisting of variola virus infection, HIV infection, and HBV infection; and the blood disease is acute myeloid leukemia.

16. The method according to claim 15 , further comprising administering a small molecule inhibitor or an antibody to the subject, wherein the small molecule inhibitor is selected from the group consisting of alkylating agents, and the antibody is a monoclonal antibody.

17. A targeting protein molecule comprising the IL-15 heterodimeric protein according to claim 1 .

18. An IL-15 heterodimeric protein selected from the group consisting of the following dimeric proteins c-q, wherein the dimeric proteins c-q each comprise respective protein (I) and protein (II):

Dimeric

Protein
Protein (I)
Protein (II)

c
IL-15-Fc-Knob (SEQ ID NO: 14)
Fc-Hole (SEQ ID NO: 27)

d
IL-15-Fc-Hole (SEQ ID NO: 15)
Fc-Knob (SEQ ID NO: 26)

e
Fc-Knob-IL-15 (SEQ ID NO: 16)
Fc-Hole (SEQ ID NO: 27)

f
Fc-Hole-IL-15 (SEQ ID NO: 17)
Fc-Knob (SEQ ID NO: 26)

g
IL-15-Fc-Knob (SEQ ID NO: 14)
IL-15Rα ECD-Fc-Hole (SEQ ID NO: 19)

h
IL-15-Fc-Hole (SEQ ID NO: 15)
IL-15Rα ECD-Fc-Knob (SEQ ID NO: 18)

i
Fc-Knob-IL-15 (SEQ ID NO: 16)
Fc-Hole-IL-15Rα ECD (SEQ ID NO: 21)

j
Fc-Hole-IL-15 (SEQ ID NO: 17)
Fc-Knob-IL-15Rα ECD (SEQ ID NO: 20)

k
IL-15-Fc-Knob (SEQ ID NO: 14)
IL-15Rα-sushi(77)-Fc-Hole (SEQ ID NO: 23)

l
Fc-Knob(M)-IL-15 (SEQ ID NO: 30)
Fc-Hole(M)-IL-15Rα-sushi(65) (SEQ ID NO: 32)

m
IL-15-Fc-Knob(M) (SEQ ID NO: 31)
IL-15Rα-sushi (65)-Fc-Hole(M) (SEQ ID NO: 33)

n
IL-15-Fc-Knob (SEQ ID NO: 14)
IL-15Rα-sushi(73)-Fc-Hole (SEQ ID NO: 34)

o
IL-15-Fc-Knob (SEQ ID NO: 14)
IL-15Rα-sushi(65)-Fc-Hole (SEQ ID NO: 35)

p
IL-15-Fc-Knob (SEQ ID NO: 14)
IL-15Rα-sushi(86)-Fc-Hole (SEQ ID NO: 36)

q
IL-15-Fc-Knob (SEQ ID NO: 14)
IL-15Rα-sushi(102)-Fc-Hole (SEQ ID NO: 37).

Assignees

• Shanghai hengrui pharmaceutical co ltd
• Jiangsu Hengrui Medicine Co

Inventors

• Qu Xiangdong
• Ye Xin
• Yuan Jijun
• Zhang Lei
• Chen Kan
• Zhang Lianshan
• Cao Guoqing
• Xu Zhibin
• Tao Weikang

Classifications

• A61P37/02

  Immunomodulators · CPC title

• A61P35/02

  specific for leukemia · CPC title

• A61P35/00

  Antineoplastic agents · CPC title

• A61P31/20

  for DNA viruses · CPC title

• A61P31/18

  for HIV · CPC title