Nucleic acid construct and use thereof

{
  "publication_number": "US2026103507A1",
  "country": "US",
  "kind": "A1",
  "title": "Nucleic acid construct and use thereof",
  "abstract": "Provided herein are a nucleic acid construct encoding an anti-VEGF fusion protein, such as an expression vector, as well as recombinant virus such as a recombinant lentivirus and a recombinant adeno-associated virus prepared by the construct. Also provided herein are uses of the fusion protein, the nucleic acid construct, and the recombinant virus in the treatment of age-related macular degeneration (AMD), such as wet AMD.",
  "claims": [
    "1. A nucleic acid construct, wherein the nucleic acid construct comprises a first nucleotide sequence encoding a first VEGF-binding region and/or a second nucleotide sequence encoding a second VEGF-binding region, wherein the first VEGF-binding region and the second VEGF-binding region are each independently selected from an antigen-binding fragment derived from an anti-VEGF antibody and a VEGF-binding fragment derived from a VEGF receptor (VEGFR) which comprises domain 2 and domain 3 (D2/3) of the VEGFR, wherein the first nucleotide sequence and the second nucleotide sequence are connected by a nucleotide sequence encoding a linker. 2. The nucleic acid construct of claim 1, wherein: the first VEGF-binding region is the antigen-binding fragment derived from the anti-VEGF antibody, and the second VEGF-binding region is an antigen-binding fragment derived from another different anti-VEGF antibody; or the first VEGF-binding region is the VEGF-binding fragment derived from the VEGFR, and the second VEGF-binding region is the antigen-binding fragment derived from the anti-VEGF antibody. 3. The nucleic acid construct of claim 1, wherein the nucleic acid construct further comprises a third nucleotide sequence encoding a third VEGF-binding region, wherein the third VEGF-binding region is selected from the antigen-binding fragment derived from the anti-VEGF antibody and the VEGF-binding fragment derived from the VEGFR which comprises the domain 2 and domain 3 (D2/3) of the VEGFR, wherein the first nucleotide sequence, the second nucleotide sequence and the third nucleotide sequence are connected to each other by a nucleotide sequence encoding a linker. 4. The nucleic acid construct of claim 3, wherein: the first VEGF-binding region is the antigen-binding fragment derived from the anti-VEGF antibody, the second VEGF-binding region is the antigen-binding fragment derived from another different anti-VEGF antibody, and the third VEGF-binding region is the VEGF-binding fragment derived from the VEGFR. 5. The nucleic acid construct of claim 1, wherein the nucleic acid construct comprises, from the 5′ end to the 3′ end, the following structures: (1) D2/3-linker-scFv1; (2) scFv1-linker-scFv2; (3) scFv1-linker-D2/3; (4) scFv1-linker-scFv2-linker-D2/3; (5) scFv1-linker-D2/3-linker-scFv2; or (6) D2/3-linker-scFv1-linker-scFv2, wherein, the scFv1 is a nucleotide sequence encoding the scFv derived from the anti-VEGF antibody, the scFv2 is a nucleotide sequence encoding the scFv derived from another different anti-VEGF antibody, the D2/3 is a nucleotide sequence encoding the VEGF-binding fragment derived from the VEGFR which comprises the D2/3 of the VEGFR, the linker is a nucleotide sequence encoding a linker, wherein each of the linkers may be same or different. 6. The nucleic acid construct of claim 1, wherein the nucleic acid construct further comprises a nucleotide sequence encoding the Fc region of an antibody. 7. The nucleic acid construct of claim 6, wherein the nucleic acid construct comprises, from the 5′ end to the 3′ end, the following structures: (1) D2/3-linker-scFv1-Fc; (2) scFv1-linker-D2/3-Fc; (3) scFv1-linker-scFv2-Fc; (4) scFv1-linker-scFv2-linker-D2/3-Fc; (5) scFv1-linker-D2/3-linker-scFv2-Fc; or (6) D2/3-linker-scFv1-linker-scFv2-Fc, wherein, the scFv1 is a nucleotide sequence encoding the scFv derived from the anti-VEGF antibody, the scFv2 is a nucleotide sequence encoding the scFv derived from another different anti-VEGF antibody, the D2/3 is a nucleotide sequence encoding the VEGF-binding fragment derived from the VEGFR which comprises the D2/3 of the VEGFR, the linker is a nucleotide sequence encoding a linker, wherein each of the linkers may be same or different, the Fc is a nucleotide sequence encoding the Fc region of the antibody. 8. The nucleic acid construct of claim 1, wherein the anti-VEGF antibody is selected from Brolucizumab, Ranibizumab or Aflibercept. 9. The nucleic acid construct of claim 8, wherein the nucleic acid construct comprises, from the 5′ end to the 3′ end, the following structures: (1) D2/3-linker-RaniScFv; (2) RaniScFv-linker-D2/3; (3) BrolscFv-linker-RaniScFv-Fc; (4) BrolscFv-linker-RaniScFv; (5) RaniScFv-linker-BrolscFv; (6) RaniScFv-linker-BrolscFv-Fc; (7) D2/3-linker-RaniScFv-Fc; (8) RaniScFv-linker-D2/3-Fc; (9) D2/3-linker-BrolscFv-linker-RaniScFv-Fc; (10) D2/3-linker-BrolscFv-linker-RaniScFv; (11) BrolscFv-linker-D2/3-linker-RaniScFv; (12) BrolscFv-linker-D2/3-linker-RaniScFv-Fc; (13) RaniScFv-linker-BrolscFv-linker-D2/3; (14) RaniScFv-linker-BrolscFv-linker-D2/3-Fc; (15) BrolscFv-linker-D2/3; (16) BrolscFv-linker-D2/3-Fc; (17) D2/3-linker-BrolscFv; or (18) BrolscFv-linker-D2/3-Fc, wherein, the BrolscFv is a nucleotide sequence encoding the scFv derived from Brolucizumab, the RaniScFv is a nucleotide sequence encoding the scFv derived from Ranibizumab. the D2/3 is a nucleotide sequence encoding the VEGF-binding fragment derived from the VEGFR which comprises the D2/3 of the VEGFR, the linker is a nucleotide sequence encoding a linker, wherein each of the linkers may be same or different, the Fc is a nucleotide sequence encoding the Fc region of the antibody. 10. The nucleic acid construct of claim 1, wherein the VEGF-binding fragment derived from the VEGFR has the amino acid sequence as set forth in SEQ ID NO: 5. 11. The nucleic acid construct of claim 1, wherein the linker is non-cleavable linker. 12. The nucleic acid construct of claim 11, wherein the linker each independently is (G 1 S) n, (G 2 S) n, (G 3 S) n or (G 4 S) n, wherein n is from 1 to 10. 13. (canceled) 14. The nucleic acid construct of claim 1, wherein the nucleic acid construct encodes a fusion protein comprising the amino acid sequence selected from SEQ ID NO: 13, SEQ ID NO: 7, SEQ ID NO: 9, SEQ ID NO: 11, SEQ ID NO: 15, SEQ ID NO: 17, SEQ ID NO: 19 and SEQ ID NO: 21. 15. The nucleic acid construct of claim 1, wherein the nucleic acid construct comprises the nucleotide sequence selected from SEQ ID NO: 14, SEQ ID NO: 8, SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 16, SEQ ID NO: 18, SEQ ID NO: 20 and SEQ ID NO: 22. 16. The nucleic acid construct of claim 1, wherein the nucleic acid construct further comprises a promoter sequence for the expression of the nucleotide sequence, and the promoter is a CAGG promoter. 17. The nucleic acid construct of claim 1, wherein the nucleic acid construct is a lentiviral vector or an adeno-associated viral vector. 18. (canceled) 19. (canceled) 20. (canceled) 21. (canceled) 22. A viral particle comprising the nucleic acid construct of claim 1, wherein the viral particle is a lentiviral particle or an adeno-associated viral particle. 23. A cell comprising the nucleic acid construct according to claim 1. 24. (canceled) 25. (canceled) 26. A vector system comprising the nucleic acid construct of claim 1, and optionally one or more additional vectors. 27. (canceled) 28. (canceled) 29. (canceled) 30. A composition comprising the viral particle according to claim 22 and optionally a pharmaceutically acceptable carrier and/or excipient. 31. A system for intraocular delivery comprising: a) the composition according to claim 30; and b) a device for intraocular delivery. 32. (canceled) 33. A method for preventing, ameliorating or treating age-related macular degeneration in a subject, which comprises the step of administering into the eye of the subject the composition according to claim 30. 34. The method according to claim 33, wherein the age-related macular degeneration is wet macular degeneration. 35. The method according to claim 33, wherein the composition are administered to the intravitreal and/or subretinal cavity and/or suprachoroidal cavity of the subject. 36. The nucleic acid construct of claim 8, wherein the scFv derived from Brolucizumab has the amino acid sequence as set forth in SEQ ID NO: 1, the scFv derived from Ranibizumab has the amino acid sequence as set forth in SEQ ID NO: 3, and/or the scFv derived from Aflibercept has the amino acid sequence as set forth in SEQ ID NO: 47."
  ],
  "cpc": [
    "C07K 16/22",
    "A61K 35/76",
    "A61P 27/02",
    "C07K 2317/52",
    "C07K 2317/622",
    "C12N 15/11",
    "C12N 15/8645",
    "C12N 15/867",
    "C12N 2740/15043",
    "C12N 2750/14143",
    "C12N 2830/00"
  ],
  "filing_date": "2023-09-28",
  "publication_date": "2026-04-16",
  "priority_date": "2022-09-29",
  "application_number": "US-202319115450-A"
}