First-generation NK CAR structure and method
Patent Information
- Authority / Receiving Office
- JP · JP
- Patent Type
- Applications
- Current Assignee / Owner
- IMMUNITYBIO INC
- Filing Date
- 2026-03-18
- Publication Date
- 2026-06-23
Smart Images
Figure 2026102818000001_ABST
Abstract
Claims
1. Recombinant nucleic acid comprising a T7 promoter sequence, a 5' untranslated (5'-UTR) sequence, a signal peptide sequence, a single-chain antibody fragment sequence, a hinge region sequence, a transmembrane domain sequence, and one or more intracellular domain sequences.
2. The recombinant nucleic acid according to claim 1, wherein the signal peptide sequence portion further comprises a sequence encoding CD64.
3. The recombinant nucleic acid according to claim 1 or 2, wherein the 5'-UTR sequence portion further comprises a Kozak sequence.
4. The recombinant nucleic acid according to any one of claims 1 to 3, wherein the single-chain antibody fragment sequence portion comprises a sequence encoding a single-chain variable fragment adapted to bind to a PDL1 antigen or other tumor antigen.
5. The recombinant nucleic acid according to any one of claims 1 to 4, wherein the hinge sequence portion provides a range of movement for the single-chain antibody fragment sequence portion.
6. The recombinant nucleic acid according to any one of claims 1 to 5, wherein the transmembrane domain sequence portion enables insertion of the recombinant nucleic acid into the membrane.
7. The recombinant nucleic acid according to any one of claims 1 to 6, wherein the intracellular domain sequence portion includes a co-stimulatory or signal transduction sequence portion.
8. The recombinant nucleic acid according to any one of claims 1 to 7, wherein the intracellular domain sequence portion comprises CD28 and / or CD3ζ.
9. The recombinant nucleic acid according to any one of claims 1 to 8, wherein the intracellular domain sequence portion comprises CD28 and / or FcεRIγ.
10. The recombinant nucleic acid according to any one of claims 1 to 9, wherein the intracellular domain sequence portion provides enhanced cytotoxic activity against tumor cells.
11. The recombinant nucleic acid according to any one of claims 1 to 10, further comprising a 3'-untranslated region (3'-UTR).
12. The recombinant nucleic acid according to any one of claims 1 to 11, wherein the 3'-UTR sequence portion provides RNA stability and translation initiation.
13. The recombinant nucleic acid according to any one of claims 1 to 12, further comprising a poly-A sequence portion.
14. The recombinant nucleic acid according to any one of claims 1 to 13, wherein the poly-A sequence portion comprises at least 150 adenine nucleotides.
15. The poly-A sequence portion provides RNA stability and translation initiation, according to any one of claims 1 to 14.
16. The vector is a recombinant nucleic acid according to any one of claims 1 to 15, which targets a tumor antigen.
17. The recombinant nucleic acid according to any one of claims 1 to 16, further comprising a sequence portion encoding CD16a.
18. The recombinant nucleic acid according to any one of claims 1 to 17, further comprising a sequence portion encoding ER-IL2.
19. The recombinant nucleic acid according to claim 1, having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with SEQ ID NO:
1.
20. The recombinant nucleic acid according to claim 1, having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with SEQ ID NO:
2.
21. The recombinant nucleic acid according to claim 1, having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with SEQ ID NO:
3.
22. The recombinant nucleic acid according to claim 1, having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with SEQ ID NO:
4.
23. Modified NK cells comprising one or more nucleic acids encoding a 5'-untranslated (5'-UTR) sequence portion, a signal peptide sequence portion, a single-chain antibody fragment sequence portion, a hinge region sequence portion, a transmembrane domain sequence portion, and one or more intracellular domain sequence portions, wherein the nucleic acid sequences are operably linked to one another as a single polynucleotide.
24. Modified NK cells according to claim 23, which specifically target tumor cells.
25. Modified NK cells according to claim 23 or 24, which specifically target PDL1 or other tumor antigens.
26. The modified NK cell according to any one of claims 23 to 25, wherein the signal peptide sequence portion further comprises a sequence encoding CD64.
27. The modified NK cell according to any one of claims 23 to 26, wherein the single-chain antibody fragment sequence portion comprises a sequence encoding a single-chain variable fragment adapted to bind to the PDL1 antigen or other tumor antigen.
28. The modified NK cell according to any one of claims 23 to 27, wherein the hinge sequence portion provides a range of movement for the single-chain antibody fragment sequence portion.
29. The modified NK cell according to any one of claims 23 to 28, wherein the transmembrane domain sequence portion enables insertion of the recombinant nucleic acid into the membrane.
30. The modified NK cell according to any one of claims 23 to 29, wherein the intracellular domain sequence portion includes a co-stimulatory or signal transduction sequence portion.
31. The modified NK cell according to any one of claims 23 to 30, wherein the intracellular domain sequence portion includes CD28 and / or CD3ζ.
32. The modified NK cell according to any one of claims 23 to 31, wherein the intracellular domain sequence portion comprises CD28 and / or FcεRIγ.
33. The modified NK cell according to any one of claims 23 to 32, wherein the intracellular domain sequence portion provides enhanced cytotoxic activity against tumor cells.
34. A modified NK cell according to any one of claims 23 to 33, further comprising a 3'-untranslated region (3'-UTR).
35. The modified NK cell according to any one of claims 23 to 34, wherein the 3'-UTR sequence portion provides RNA stability and translation initiation.
36. Modified NK cells according to any one of claims 23 to 35, further comprising a poly-A sequence portion.
37. The modified NK cell according to any one of claims 23 to 36, wherein the poly-A sequence portion comprises at least 150 adenine nucleotides.
38. The modified NK cell according to any one of claims 23 to 37, wherein the poly-A sequence portion provides RNA stability and translation initiation.
39. The vector is a modified NK cell according to any one of claims 23 to 38, which targets a tumor antigen.
40. A modified NK cell according to any one of claims 23 to 39, further comprising a sequence portion encoding CD16a.
41. A modified NK cell according to any one of claims 23 to 40, further comprising a sequence portion encoding ER-IL2.
42. A method for generating CAR-NK cells, comprising transfecting primary NK cells with recombinant nucleic acid as described in any one of claims 1 to 22.
43. A composition comprising modified NK cells according to any one of claims 1 to 22 and a pharmaceutically acceptable excipient.
44. A kit comprising NK cells according to any one of claims 23 to 43 and instructions for use.
45. A method for treating cancer or a tumor in a subject, comprising administering to the subject a therapeutically effective amount of modified NK cells according to any one of claims 23 to 42 or the composition according to claim 43, wherein the administration treats the cancer in the subject or reduces the size of the tumor.
46. A method for reducing cancer metastasis in a subject, comprising administering a therapeutically effective amount of modified NK cells according to any one of claims 23 to 42 or the composition according to claim 43 to the subject, thereby reducing cancer metastasis in the subject.
47. 1m ２ 1 x 10 ３ ~1 x 10 １０ The method according to claim 45 or 46, wherein the individual NK cells are administered to the subject.
48. The method according to any one of claims 45 to 47, wherein the NK cells are administered parenterally, intravenously, around the tumor, or by injection.
49. The method according to any one of claims 45 to 48, further comprising administering an additional therapeutic agent to the subject.
50. Recombinant cells comprising recombinant nucleic acid according to any one of claims 1 to 22.
51. The recombinant cell according to claim 50, which is a bacterial cell.
52. The recombinant cells according to claim 50, which are autologous NK cells.
53. The recombinant cells according to claim 52, wherein the NK cells are genetically modified.
54. A method for treating cancer in a patient in need of such treatment, comprising administering a therapeutically effective amount of recombinant NK cells according to any one of claims 23 to 41 to the patient, thereby treating the cancer.
55. The method according to claim 54, further comprising the step of administering at least one additional therapeutic substance selected from the group consisting of viral cancer vaccines, bacterial cancer vaccines, yeast cancer vaccines, N-803, antibodies, stem cell transplants, and tumor-targeting cytokines.
56. The aforementioned cancers include leukemia, acute lymphoblastic leukemia, acute myeloid leukemia, chronic leukemia, chronic myeloid (granular) leukemia, chronic lymphocytic leukemia, polycythemia vera, lymphoma, Hodgkin's disease, non-Hodgkin's disease, multiple myeloma, Waldenström macroglobulinemia, heavy chain disease, and, without limitation, sarcomas and carcinomas, such as fibrosarcoma, myxosarcoma, liposarcoma, chondrosarcoma, osteosarcoma, chordoma, angiosarcoma, endosarcoma, lymphangiosarcoma, lymphangioendothelioma, synoviomas, mesothelioma, Ewing's tumor, leiomyosarcoma, rhabdomyosarcoma, colon cancer, and pancreatic cancer. The method according to claim 54 or 55, selected from solid tumors including cancer, breast cancer, ovarian cancer, prostate cancer, squamous cell carcinoma, basal cell carcinoma, adenocarcinoma, sweat gland carcinoma, sebaceous gland carcinoma, papillary carcinoma, papillary adenocarcinoma, cystadenocarcinoma, medullary carcinoma, bronchogenic carcinoma, renal cell carcinoma, hepatocellular carcinoma, cholangiocarcinoma, choriocarcinoma, seminomas, embryonic carcinoma, Wilms' tumor, cervical cancer, testicular cancer, lung cancer, small cell lung cancer, bladder cancer, epithelial carcinoma, glioma, astrocytoma, medulloblastoma, craniopharyngioma, ependymoma, pineal glandoma, hemangioblastoma, acoustic neuroma, oligodendroglioma, meningioma, melanoma, neuroblastoma, and retinoblastoma.