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US-12617865-B2 - Activated antibodies targeting PSMA and effector cell antigens

US12617865B2US 12617865 B2US12617865 B2US 12617865B2US-12617865-B2

Abstract

Provided herein are multispecific antibodies that selectively bind to PSMA and effector cell antigens such as CD3, pharmaceutical compositions thereof, as well as nucleic acids, and methods for making and discovering the same.

Inventors

  • David Campbell
  • Thomas R. DIRAIMONDO

Assignees

  • Janux Therapeutics, Inc.

Dates

Publication Date
20260505
Application Date
20221116

Claims (20)

  1. 1 . A method of treating prostate cancer in a subject, the method comprising administering to the subject a therapeutically effective amount of a polypeptide or polypeptide complex according to Formula I: A 2 -A 1 -L 1 -P 1 —H 1 (Formula I) wherein: A 1 comprises a first antigen recognizing molecule that binds to an effector cell antigen, wherein A 1 comprises an anti-CD3 binding molecule comprising complementarity determining regions (CDRs): HC-CDR1, HC-CDR2, and HC-CDR3, wherein the HC-CDR1, the HC-CDR2, and the HC-CDR3 of A 1 comprise: HC-CDR1: SEQ ID NO: 1, HC-CDR2: SEQ ID NO: 2, and HC-CDR3: SEQ ID NO: 3; and A 1 comprises CDRs: LC-CDR1, LC-CDR2, and LC-CDR3, wherein the LC-CDR1, the LC-CDR2, and the LC-CDR3 of A 1 comprise LC-CDR1: SEQ ID NO: 4, LC-CDR2: SEQ ID NO: 5, and LC-CDR3: SEQ ID NO: 6; P 1 comprises a peptide that binds to A 1 , wherein Pi comprises an amino acid sequence according to U 1 -U 2 —C—U 4 —P—U 6 -U 7 —U 8 —U 9 -U 10 —U 11 —U 12 —C—U 14 and U 1 is selected from D, Y, F, I, N, V, H, L, A, T, S, and P; U 2 is selected from D, Y, L, F, I, N, A, V, H, T, and S; U 4 is selected from G and W; U 6 is selected from E, D, V, and P; U 7 is selected from W, L, F, V, G, M, I, and Y; U 8 is selected from E, D, P, and Q; U 9 is selected from E, D, Y, V, F, W, P, L, and Q; U 10 is selected from S, D, Y, T, I, F, V, N, A, P, L, and H; U 11 is selected from I, Y, F, V, L, T, N, S, D, A, and H; U 12 is selected from F, D, Y, L, I, V, A, N, T, P, S, G, and H; and U 14 is selected from D, Y, N, F, I, P, V, A, T, H, L, M, and S; L 1 comprises a linking moiety that connects A 1 to P 1 and is a substrate for a tumor specific protease; H 1 comprises a half-life extending molecule; and A 2 comprises a second antigen recognizing molecule that binds to prostate-specific membrane antigen (PSMA).
  2. 2 . The method of claim 1 , wherein the prostate cancer is metastatic castration-resistant prostate cancer (mCRPC).
  3. 3 . The method of claim 1 , wherein the administering comprises administering on a weekly basis.
  4. 4 . The method of claim 1 , wherein the administering comprises administering intravenously, intramuscularly, intralesionally, topically, subcutaneously, or orally.
  5. 5 . The method of claim 1 , wherein the administering comprises administering by continuous infusion or bolus injection.
  6. 6 . The method of claim 1 , wherein the administering comprises administering on a weekly basis through continuous intravenous infusion.
  7. 7 . The method of claim 1 , wherein A 2 comprises complementarity determining regions (CDRs): HC-CDR1, HC-CDR2, and HC-CDR3, wherein the HC-CDR1, the HC-CDR2, and the HC-CDR3 comprise: HC-CDR1: SEQ ID NO: 8, HC-CDR2: SEQ ID NO: 9, and HC-CDR3: SEQ ID NO: 10; and A 2 comprises CDRs: LC-CDR1, LC-CDR2, and LC-CDR3, wherein the LC-CDR1, the LC-CDR2, and the LC-CDR3 of A 2 comprise LC-CDR1: SEQ ID NO: 11, LC-CDR2: SEQ ID NO: 12, and LC-CDR3: SEQ ID NO: 13.
  8. 8 . The method of claim 1 , wherein the effector cell antigen comprises cluster of differentiation 3 (CD3).
  9. 9 . The method of claim 1 , wherein A 1 comprises an antibody format selected from single chain variable fragment and a Fab or Fab′ fragment.
  10. 10 . The method of claim 1 , wherein A 2 comprises an antibody format selected from single chain variable fragment, a single domain antibody, and a Fab or Fab′ fragment.
  11. 11 . The method of claim 1 , wherein A 1 comprises an antibody format of a single chain variable fragment (scFv), and A 2 comprises an antibody format of a Fab or Fab′.
  12. 12 . The method of claim 1 , wherein Pi becomes unbound from A 1 when L 1 is cleaved by the tumor specific protease thereby exposing A 1 to the effector cell antigen.
  13. 13 . The method of claim 1 , wherein the tumor specific protease is selected from the group consisting of a matrix metalloprotease (MMP), serine protease, cysteine protease, threonine protease, and aspartic protease.
  14. 14 . The method of claim 13 , wherein the matrix metalloprotease comprises MMP2, MMP7, MMP9, MMP13, or MMP14.
  15. 15 . The method of claim 13 , wherein the serine protease comprises matriptase (MTSP1), urokinase, or hepsin.
  16. 16 . The method of claim 1 , wherein L 1 comprises a urokinase cleavable amino acid sequence, a matriptase cleavable amino acid sequence, a matrix metalloprotease cleavable amino acid sequence, or a legumain cleavable amino acid sequence.
  17. 17 . The method of claim 1 , wherein L 1 comprises an amino acid sequence according to SEQ ID NO: 23.
  18. 18 . The method of claim 1 , wherein L 1 comprises an amino acid sequence according to any one of SEQ ID NOs: 20-49.
  19. 19 . The method of claim 1 , wherein L 1 comprises an amino acid sequence of Linker 25 (ISSGLLSGRSDAG) (SEQ ID NO: 45), Linker 26 (AAGLLAPPGGLSGRSDAG) (SEQ ID NO: 46), Linker 27 (SPLGLSGRSDAG) (SEQ ID NO: 47), or Linker 28 (LSGRSDAGSPLGLAG) (SEQ ID NO: 48), or an amino acid sequence that has 1, 2, or 3 amino acid substitutions, additions, or deletions relative to the amino acid sequences of Linker 25, Linker 26, Linker 27, or Linker 28.
  20. 20 . The method of claim 1 , wherein H 1 comprises serum albumin.

Description

CROSS-REFERENCE The present application is a continuation of U.S. patent application Ser. No. 17/544,539, filed Dec. 7, 2021, now U.S. Pat. No. 11,555,078, which claims the benefit of U.S. Provisional Application No. 63/187,699, filed May 12, 2021, and U.S. Provisional Application No. 63/123,329, filed Dec. 9, 2020, each of which is incorporated herein by reference in its entirety. SEQUENCE LISTING The instant application contains a Sequence Listing which has been submitted electronically in XML format and is hereby incorporated by reference in its entirety. Said XML copy, created on Nov. 11, 2022, is named 52426-730_301SL.xml and is 1,248,426 bytes in size. SUMMARY Disclosed herein, in certain embodiments, are isolated polypeptides or polypeptide complexes according to Formula I: A2-A1-L1-P1—H1  (Formula I) wherein: A1 comprises a first antigen recognizing molecule that binds to an effector cell antigen; P1 comprises a peptide that binds to A1; L1 comprises a linking moiety that connects A1 to P1 and is a substrate for a tumor specific protease; H1 comprises a half-life extending molecule; and A2 comprises a second antigen recognizing molecule that binds to prostate-specific membrane antigen (PSMA). In some embodiments, the first antigen recognizing molecule comprises an antibody or antibody fragment. In some embodiments, first antigen recognizing molecule comprises an antibody or antibody fragment that is human or humanized. In some embodiments, L1 is bound to a N-terminus of the first antigen recognizing molecule. In some embodiments, A2 is bound to a C-terminus of the first antigen recognizing molecule. In some embodiments, L1 is bound to a C-terminus of the first antigen recognizing molecule. In some embodiments, A2 is bound to a N-terminus of the first antigen recognizing molecule. In some embodiments, the antibody or antibody fragment comprises a single chain variable fragment, a single domain antibody, or a Fab fragment. In some embodiments, A1 is the single chain variable fragment (scFv). In some embodiments, the scFv comprises a scFv heavy chain polypeptide and a scFv light chain polypeptide. In some embodiments, A1 is the single domain antibody, In some embodiments, the antibody or antibody fragment comprises a single chain variable fragment (scFv), a heavy chain variable domain (VH domain), a light chain variable domain (VL domain), or a variable domain (VHH) of a camelid derived single domain antibody. In some embodiments, A1 comprises an anti-CD3e single chain variable fragment. In some embodiments, A1 comprises an anti-CD3e single chain variable fragment that has a KD binding of 1 μM or less to CD3 on CD3 expressing cells. In some embodiments, the effector cell antigen comprises CD3. In some embodiments, A1 comprises a variable light chain and variable heavy chain each of which is capable of specifically binding to human CD3. In some embodiments, A1 comprises complementary determining regions (CDRs) selected from the group consisting of muromonab-CD3 (OKT3), otelixizumab (TRX4), teplizumab (MGA031), visilizumab (Nuvion), SP34, X35, VIT3, BMA030 (BW264/56), CLB-T3/3, CRIS7, YTH12.5, F111-409, CLB-T3.4.2, TR-66, WT32, SPv-T3b, 11D8, XIII-141, XIII-46, XIII-87, 12F6, T3/RW2-8C8, T3/RW2-4B6, OKT3D, M-T301, SMC2, F101.01, UCHT-1, WT-31, 15865, 15865v12, 15865v16, and 15865v19. In some embodiments, the polypeptide or polypeptide complex of Formula I binds to an effector cell when L1 is cleaved by the tumor specific protease. In some embodiments, the polypeptide or polypeptide complex of Formula I binds to an effector cell when L1 is cleaved by the tumor specific protease and A1binds to the effector cell. In some embodiments, the effector cell is a T cell. In some embodiments, A1 binds to a polypeptide that is part of a TCR-CD3 complex on the effector cell. In some embodiments, the polypeptide that is part of the TCR-CD3 complex is human CD3E. In some embodiments, the effector cell antigen comprises CD3, wherein the scFv comprises complementarity determining regions (CDRs): HC-CDR1, HC-CDR2, and HC-CDR3, wherein the HC-CDR1, the HC-CDR2, and the HC-CDR3 of the scFv comprise: HC-CDR1: SEQ ID NO: 1, HC-CDR2: SEQ ID NO: 2, and HC-CDR3: SEQ ID NO: 3; and the scFv comprises CDRs: LC-CDR1, LC-CDR2, and LC-CDR3, wherein the LC-CDR1, the LC-CDR2, and the LC-CDR3 of the scFv comprises: LC-CDR1: SEQ ID NO: 4, LC-CDR2: SEQ ID NO:5, and LC-CDR3: SEQ ID NO: 6. In some embodiments, the effector cell antigen comprises CD3, and the scFv comprises an amino acid sequence according to SEQ ID NO: 7. In some embodiments, second antigen recognizing molecule comprises an antibody or antibody fragment. In some embodiments, the antibody or antibody fragment thereof comprises a single chain variable fragment, a single domain antibody, or a Fab. In some embodiments, the antibody or antibody fragment thereof comprises a single chain variable fragment (scFv), a heavy chain variable domain (VH domain), a light chain variable d