A CANCER-TREATING RECOMBINANT PROTEIN THAT IMPROVES THE ACTIVITY OF CYTOTOXIC KILLER CELLS TARGETING CANCER CELLS AND USE THEREOF

• US 20190309036A1
• Filed: 08/27/2018
• Published: 10/10/2019
• Est. Priority Date: 04/10/2018
• Status: Active Grant

• Current Assignee: Gwangju Institute of Science and Technology
• Original Assignee: Gwangju Institute of Science and Technology
• Inventors: JUN, Chang-Duk, JEON, Bu Nam, KIM, hey-ran
• Granted Patent: US 11,440,941 B2
• Time in Patent Office: Days
• CPC Class Codes: A61K 2239/31 characterized by the route of administration
• CPC Class Codes: A61K 2239/38 characterised by the dose, timing or administration schedule
• CPC Class Codes: A61K 2239/49 Breast
• CPC Class Codes: A61K 38/17 from animals; from humans enzyme inhibitors A61K38/005
• CPC Class Codes: A61K 39/4611 T-cells, e.g. tumor infiltrating lymphocytes [TIL], lymphokine-activated killer cells [LAK] or regulatory T cells [Treg]
• CPC Class Codes: A61K 39/4644 Cancer antigens
• CPC Class Codes: A61K 39/464466 Adhesion molecules, e.g. NRCAM, EpCAM or cadherins
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: C07K 14/435 from animals; from humans
• CPC Class Codes: C07K 14/4716 Muscle proteins, e.g. myosin, actin
• CPC Class Codes: C07K 2319/10 containing a tag for extracellular membrane crossing, e.g. TAT or VP22
• CPC Class Codes: C12N 5/0638 Cytotoxic T lymphocytes [CTL] or lymphokine activated killer cells [LAK]

Abstract

A cancer killer cell in which a therapeutic recombinant protein or recombinant protein which improves cytotoxic activity of the cancer killer cell is loaded. In addition, a pharmaceutical composition including the recombinant protein or a recombinant protein-loaded cancer killer cell is disclosed. Further, disclosed is a method for preparing a recombinant protein-loaded cancer killer cell.

Claims

1. A recombinant protein for treating cancer consisting of an amino acid sequence of SEQ ID NO:

1, which is loaded in a cancer killer cell to improve cytotoxic activity of the cancer killer cell.

2. The recombinant protein of claim 1, wherein the recombinant protein is further fused with a protein transduction domain (PTD).

3. The recombinant protein of claim 2, wherein the fusion of PTD is by a peptide linker or direct fusion.

4. The recombinant protein of claim 1, capable of stabilizing actin polymerized in an immunological synapse by stimulation of a T-cell antigen receptor.

5. The recombinant protein of claim 1, wherein cytokine expression is increased when the recombinant protein is loaded in the cancer killer cell compared to a control group in which the recombinant protein is not loaded in the cancer killer cell.

6. The recombinant protein of claim 5, wherein the cytokine is at least one of mIL-2, mIFNγ

, and mGZMB.

7. A recombinant protein-loaded cancer killer cell, wherein the cancer killer cell comprises the recombinant protein of claim 1.

8. The recombinant protein-loaded cancer killer cell of claim 7, wherein the loading of the recombinant protein in the cancer killer cell is performed by:

i) transduction of a polynucleotide encoding the recombinant protein consisting of the amino acid sequence of SEQ ID NO;

1 by retrovirus.

9. A pharmaceutical composition for treating cancer, comprising:

i) a recombinant protein for treating cancer consisting of an amino acid sequence of SEQ ID NO;

1, which is loaded in a cancer killer cell to improve cytotoxic activity of the cancer killer cell;

ii) a recombinant protein, wherein a PTD is further fused into the i) protein;

or

iii) a cancer killer cell in which the recombinant protein of claim 7 is loaded.

10. The pharmaceutical composition for treating cancer of claim 9,

wherein the cancer is lung cancer, gastric cancer, colon cancer, breast cancer, bone cancer, pancreatic cancer, skin cancer, head cancer, head and neck cancer, melanoma, uterine cancer, ovarian cancer, colon cancer, small intestine cancer, rectal cancer, anal cancer, fallopian tube cancer, endometrial cancer, cervical cancer, vaginal cancer, vulva cancer, Hodgkin'"'"'s disease, esophageal cancer, lymph node cancer, bladder cancer, gall bladder cancer, endocrine cancer, prostate cancer, adrenal gland cancer, soft tissue sarcoma, urethral cancer, penile cancer, chronic or acute leukemia, lymphocytic lymphoma, renal cancer, ureteral cancer, renal pelvic cancer, central nervous system (CNS) tumor, spinal cord tumor, brainstem glioma, or pituitary adenoma.

11. A method for treating cancer, comprising administering the composition of claim 9 in a pharmaceutically effective amount to a subject suspected of having a cancer excluding humans.

14. The recombinant protein-loaded cancer killer cell of claim 7, wherein the loading of the recombinant protein in the cancer killer cell is performed by:

i) transduction of a polynucleotide encoding the recombinant protein consisting of the amino acid sequence of SEQ ID NO;

1 further fused with a protein transduction domain (PTD) by retrovirus.

15. The recombinant protein-loaded cancer killer cell of claim 7, wherein the loading of the recombinant protein in the cancer killer cell is performed by:

i) introduction in the form of the recombinant protein consisting of sequence of SEQ ID NO;

1 further fused with a protein transduction domain (PTD).

12. A method for producing a recombinant protein-loaded cancer killer cell, comprising:

(a) fusing a PTD into an amino acid of SEQ ID NO;

1; and

(b) introducing the fusion protein of (a) into a cancer killer cell.

13. A method for producing a recombinant protein-loaded cancer killer cell, comprising:

transducing into a cancer killer cell using retrovirus comprising a polynucleotide encoding an amino acid sequence of SEQ ID NO;

1.