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JP-7856643-B2 - Proteins containing a delta-like ligand 3 (DLL3) antigen-binding domain and their use

JP7856643B2JP 7856643 B2JP7856643 B2JP 7856643B2JP-7856643-B2

Inventors

  • ヤン，ダンリン
  • ズウォラック，アダム
  • ヴェンカタラマニ，サチャデヴィ
  • パワーズ，ゴードン ディー．
  • シンハ，サンジャヤ
  • ブローダー，スコット アール．
  • カートン，ジル エム．
  • ガネサン，ラジクマール
  • ヘルツォホ，ジェニファー
  • マクデビット，テレサ
  • ピチャ，クリステン エム．
  • スミス，リャン エム．

Assignees

  • ヤンセン バイオテツク，インコーポレーテツド

Dates

Publication Date
20260511
Application Date
20211021
Priority Date
20201022

Claims (20)

  1. An isolated protein comprising an antigen-binding region that binds to delta-like protein 3 (DLL3), wherein the antigen-binding region is a. Sequence numbers 15, 16, 17, 33, 34, and 35, respectively b. Sequence numbers 18, 19, 20, 36, 37, and 38, respectively c. Sequence numbers 21, 22, 23, 39, 37, and 40, respectively d. Sequence numbers 24, 25, 26, 41, 42, and 43, respectively e. Sequence numbers 18, 28, 29, 44, 45, and 46, respectively f. Sequence numbers 30, 31, 32, 47, 48, and 49, respectively g. Sequence IDs 50, 51, 17, 33, 34, and 35, respectively h. Sequence numbers 52, 51, 17, 33, 34, and 35, respectively i. Sequence numbers 53, 54, 20, 36, 37, and 38, respectively j. Sequence numbers 55, 56, 23, 39, 37, and 40, respectively k. Sequence numbers 57, 58, 26, 41, 42, and 43, respectively l. Sequence numbers 59, 60, 29, 44, 45, and 46, respectively, or m. Sequence numbers 61, 62, 32, 47, 48, and 49, respectively. An isolated protein containing the amino acid sequence of heavy chain complementarity-determining region 1 (HCDR1), HCDR2, HCDR3, light chain complementarity-determining region 1 (LCDR1), LCDR2, and LCDR3.
  2. The isolated protein according to claim 1, wherein the antigen-binding region comprises a heavy chain variable region (VH) containing an amino acid sequence at least 90% identical to SEQ ID NOs: 1, 3, 5, 7, 9, 11, or 13, and a light chain variable region (VL) containing an amino acid sequence at least 90% identical to SEQ ID NOs: 2, 4, 6, 8, 10, 12, or 14.
  3. The VH and VL of the antigen-binding region are a. Sequence ID 1 and Sequence ID 2, respectively b. Sequence ID 3 and Sequence ID 4, respectively c. Sequence ID 5 and Sequence ID 6, respectively d. Sequence ID 7 and Sequence ID 8, respectively e. Sequence ID 9 and Sequence ID 10, respectively f. Sequence ID 11 and Sequence ID 12, respectively, or g. Sequence ID 13 and Sequence ID 14, respectively. The isolated protein according to claim 2, comprising the amino acid sequence.
  4. The isolated protein according to any one of claims 1 to 3, wherein the antigen-binding region comprises an scFv having an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 63 or 64.
  5. An immunoconjugate comprising an isolated protein according to any one of claims 1 to 4, conjugated to a therapeutic agent or contrast agent.
  6. A multispecific antigen-binding construct comprising an isolated protein according to any one of claims 1 to 4.
  7. The multispecific antigen-binding construct according to claim 6, further comprising a second antigen-binding region for binding to an antigen on a lymphocyte.
  8. The multispecific antigen-binding construct according to claim 7, wherein the antigen on the lymphocyte is CD3, CD3 epsilon (CD3ε), CD8, KI2L4, NKG2E, NKG2D, NKG2F, BTNL3, CD186, BTNL8, PD-1, CD195, or NKG2C.
  9. The second antigen-binding region is bound to CD3ε and includes a second VH containing HCDR1, HCDR2, and HCDR3, and a second VL containing LCDR1, LCDR2, and LCDR3, and the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 of the second antigen-binding region are a. Sequence numbers 98, 99, 100, 106, 107, and 108, respectively, or b. Sequence numbers 95, 96, 97, 101, 102, and 104, respectively. A multispecific antigen-binding construct according to claim 8, comprising the amino acid sequence.
  10. The VH and VL of the second antigen-binding region are a. Sequence ID 84 and Sequence ID 85, respectively, or b. Sequence ID 77 and Sequence ID 80, respectively. The multispecific antigen-binding construct according to claim 9, comprising an amino acid sequence that is at least 90% identical to that of the other construct.
  11. The VH and VL of the second antigen-binding region are a. Sequence ID 84 and Sequence ID 85, respectively, or b. Sequence ID 77 and Sequence ID 80, respectively. A multispecific antigen-binding construct according to claim 10, comprising the amino acid sequence.
  12. The multispecific antigen-binding construct according to any one of claims 6 to 11, wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 of the antigen-binding region that binds to DLL3 each contain the amino acid sequences of SEQ ID NOs: 15, 16, 17, 33, 34, and 35, respectively.
  13. A fusion or conjugate comprising a half-life extension portion fused to an isolated protein according to any one of claims 1 to 4 or a multispecific antigen-binding construct according to any one of claims 7 to 12, wherein the half-life extension portion is immunoglobulin (Ig), a fragment of Ig, an Ig constant region, a fragment of the Ig constant region, an Fc region, transferrin, albumin, an albumin-binding domain, or polyethylene glycol.
  14. The aforementioned half-life extension portion is T350V, L351Y, F405A, Y407V, T366Y, T366W, F405W, T394W, T394S, Y407T, Y407A, T366S/L368A/Y407V, L351Y/F405A/Y407V, T366I/K392M/T394W, F405A/Y407V, T366L/K392M/T394W, L351Y/Y407A, T366A/K409F, The fusion or conjugate according to claim 13, comprising a fragment of the Ig constant region containing at least one mutation selected from the group consisting of L351Y/Y407A, T366V/K409F, T366A/K409F, T350V/L351Y/F405A/Y407V, T350V/T366L/K392L/T394W, and L234A/L235A/D265S, wherein the residue numbering follows the EU index.
  15. A bispecific antigen-binding construct, (1) A first antigen-binding region that binds to DLL3, wherein the first antigen-binding region comprises a first VH including HCDR1, HCDR2, and HCDR3, and a first VL including LCDR1, LCDR2, and LCDR3, and the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 are (a) Sequence numbers 15, 16, 17, 33, 34, and 35, respectively (b) Sequence numbers 18, 19, 20, 36, 37, and 38, respectively (c) Sequence IDs 21, 22, 23, 39, 37, and 40, respectively (d) Sequence numbers 24, 25, 26, 41, 42, and 43, respectively (e) Sequence numbers 18, 28, 29, 44, 45, and 46, respectively (f) Sequence numbers 30, 31, 32, 47, 48, and 49, respectively (g) Sequence IDs 50, 51, 17, 33, 34, and 35, respectively (h) Sequence numbers 52, 51, 17, 33, 34, and 35, respectively (i) Sequence numbers 53, 54, 20, 36, 37, and 38, respectively (j) Sequence IDs 55, 56, 23, 39, 37, and 40, respectively (k) Sequence IDs 57, 58, 26, 41, 42, and 43 respectively, (l) Sequence numbers 59, 60, 29, 44, 45, and 46 respectively, or (m) Sequence numbers 61, 62, 32, 47, 48, and 49 respectively (2) A first antigen-binding region comprising the amino acid sequence of (1) and a second antigen-binding region that binds to CD3ε, (a) A second VH containing HCDR1, HCDR2, and HCDR3 of the amino acid sequences of SEQ ID NOs. 95, 96, and 97, respectively, and a second VL containing LCDR1, LCDR2, and HCDR3 of the amino acids of the sequences of SEQ ID NOs. 101, 102, and 104, respectively, or (b) A second VH containing HCDR1, HCDR2, and HCDR3 of the amino acid sequences of SEQ ID NOs. 98, 99, and 100, respectively, and a second VL containing LCDR1, LCDR2, and HCDR3 of the amino acid sequences of SEQ ID NOs. 106, 107, and 108, respectively. A bispecific antigen-binding construct comprising a second antigen-binding region.
  16. a. The first VH and the first VL are i. Sequence ID 1 and Sequence ID 2, respectively ii. Sequence ID 3 and Sequence ID 4, respectively iii. Sequence ID 5 and Sequence ID 6, respectively iv. Sequence ID 7 and Sequence ID 8, respectively v. Sequence ID 9 and Sequence ID 10, respectively vi. Sequence ID 11 and Sequence ID 12, respectively, or vii. Sequence ID 13 and Sequence ID 14 It contains an amino acid sequence that is at least 90% identical to, b. The second VH and the second VL are i. Sequence ID 77 and Sequence ID 80, respectively, or ii. Sequence ID 84 and Sequence ID 85, respectively The bispecific antigen-binding construct according to claim 15, comprising an amino acid sequence that is at least 90% identical to that of the other construct.
  17. The bispecific antigen-binding construct according to claim 15 or 16, wherein HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 of the first antigen-binding region each contain the amino acid sequences of SEQ ID NOs. 15, 16, 17, 33, 34, and 35, respectively.
  18. The bispecific antigen-binding construct according to claim 17, wherein the first VH comprises an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 3, and the first VL comprises an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 4.
  19. The bispecific antigen-binding construct according to any one of claims 15 to 18, wherein the first antigen-binding region comprises a first scFv or first Fab including the first VH and the first VL, and the second antigen-binding region comprises a second Fab or second scFv including the second VH and the second VL.
  20. The bispecific antigen-binding construct according to claim 19, wherein the first antigen-binding region comprises the first scFv, and the second antigen-binding region comprises the second Fab.

Description

(Cross-reference of related applications) This application claims priority to U.S. Provisional Application No. 63/094,933 and U.S. Provisional Application No. 63/094,934, filed on 22 October 2020, and the disclosures of each of these are incorporated herein by reference in their entirety. (Reference to electronically submitted sequence listings) This application includes a sequence listing submitted electronically via EFS-Web in ASCII format, the file name being "sequence listing JBI6411", the creation date being October 7, 2021, and the size being 275 KB. The sequence listing submitted via EFS-Web is part of this specification and is incorporated herein by reference in its entirety. (Field of invention) This application relates to a protein comprising an antigen-binding domain that binds to Delta-like canonical Notch ligand 3 (DLL3), as well as related compositions and methods. Prostate cancer is the second most common cancer, the sixth leading cause of cancer death in men, and accounts for 14% (903,500) of all new cancer cases and 6% (258,400) of all cancer deaths in men worldwide. Metastatic prostate cancer is the second leading cause of cancer death in men in the United States. The progression from diagnosis to death of prostate cancer is best classified into a series of clinical stages based on the severity of the disease, hormonal status, and the presence or absence of detectable metastasis: localized disease, elevated prostate-specific antigen (PSA) levels without detectable metastasis after radiotherapy or surgery, and clinical metastasis in the uncastrated or castrated stage. While surgery, radiation therapy, or a combination of both can be curative for patients with localized disease, a significant proportion of these patients have recurrent disease, evidenced by elevated PSA levels, which can also lead to metastatic formation and progression to the terminal stage of the disease, particularly in high-risk groups. Androgen depletion therapy (ADT) is the standard treatment, and the generally predictable outcomes are a decrease in PSA levels, a stable period with no tumor growth, followed by an increase in PSA levels and regrowth as a castration-resistant disease. For many years, ADT has been the standard treatment for patients with metastatic prostate cancer. However, recent clinical data suggest that androgen depletion therapy may lead to the emergence of an androgen-independent tumor phenotype known as neuroendocrine prostate cancer (NEPC) through the process of cellular redifferentiation. Delta-like canonical Notch ligand 3 (DLL3) has been shown to be abundant in NEPC tumors at both the RNA and protein levels. Therefore, strategies designed to target DLL3 may have clinical utility in the NEPC/small cell lung cancer patient population. Small cell lung cancer (SLC) accounts for approximately 20% of all lung cancers. SLC often progresses rapidly and is difficult to remove surgically because lymph node or distant metastasis has already occurred by the time of diagnosis. This cancer shows a high early response rate to anticancer drugs. Therefore, chemotherapy is considered the first-line treatment option. However, the cancer quickly becomes resistant to chemotherapy, recurs, and the three-year survival rate falls below 5%. Therefore, new therapies are needed to treat cancers such as NEPC, small cell carcinoma, or small cell lung cancer. In normal cells, DLL3 regulates intracellular notch signaling. In cancer cells, DLL3 is expressed extracellularly; for example, in humans, it has eight extracellular domains containing 618 amino acids and six EGF-like repeats. Human DLL3 is highly homologous to that of cynomolgus monkeys and mice/rats, sharing 96% and 83% amino acid sequence identity, respectively, but has less than 40% identity with DLL1 and DLL4. DLL3 has low to undetectable expression in normal tissues, but is highly expressed on the cell surface of neuroendocrine tumors, including small cell lung cancer, prostate cancer, large cell carcinoma, and bladder cancer, and is a target for T-cell redirection for the treatment of neuroendocrine cancers. The foregoing will become clear from a more specific description of the exemplary embodiments, as shown in the attached drawings. A schematic diagram of the DLL3 extracellular domain, which includes the DSL domain and six EGF domains, is shown. The amino acid sequences shown represent residues 176-215 of the DSL domain (SEQ ID NO: 246), residues 216-249 of the EGF-1 domain (SEQ ID NO: 247), residues 274-310 of the EGF-2 domain (SEQ ID NO: 248), residues 312-351 of the EGF-3 domain (SEQ ID NO: 249), residues 353-389 of the EGF-4 domain (SEQ ID NO: 250), residues 391-427 of the EGF-5 domain (SEQ ID NO: 251), residues 429-465 of the EGF-6 domain (SEQ ID NO: 252), and residues 429-618 of the EGF-6 domain + C-terminal domain (SEQ ID NO: 263). Figure 2A shows the cell binding of a bispecific anti-DLL3×CD3 antibody to DLL3 + tumor cell lines. Figure 2B shows the ce