Compositions and methods related to tumor-activating antibodies targeting PSMA and effector cell antigens

Abstract

The present invention provides CD3, its pharmaceutical composition, and polyspecific antibodies that selectively bind to PSMA and effector cell antigens such as nucleic acids, as well as methods for producing and discovering them. [Solution] The present invention provides an isolated polypeptide or polypeptide complex according to formula I:A2-A1-L1-P1-H1, 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 portion that connects A1 to P1 and is a substrate for a tumor-specific protease, H1 comprises a half-life extension molecule, and A2 comprises a second antigen-recognizing molecule that binds to prostate-specific membrane antigen (PSMA).

Description

[Technical Field]

[0001] Cross-references to related applications This application is U.S. Provisional Patent Application No. 63 / 123,329, filed on December 9, 2020. and benefits under U.S. Provisional Patent Application No. 63 / 187,699 filed on May 12, 2021 These applications assert that they do so, and these applications are incorporated herein by reference to the whole.

[0002] Sequence List This application was filed electronically in ASCII format and is incorporated herein by reference. It includes a sequence list. This is a copy of the ASCII created on November 30, 2021. The file is named 52426-730_601_SL.txt and has a size of 321,067 bytes. ru. [Overview of the project]

[0003] In certain embodiments of this specification, Formula I: A2-A1-L1-P1-H1 (Formula I) A polypeptide or polypeptide complex isolated by the formula, wherein A1 is Ef It contains a first antigen-recognition molecule that binds to the ecta ​​cell antigen, and P1 is a peptide that binds to A1. It includes L1, which connects A1 to P1 and is a substrate of tumor-specific proteases. The molecule contains a ligation portion, H1 contains a half-life extension molecule, and A2 contains prostate-specific membrane antigen (PSMA). An isolated polypeptide or polypeptide complex containing a second antigen recognition molecule that binds to ). The combination is disclosed. In some embodiments, the first antigen recognition molecule is an antibody or antibody- Includes a lagment. In some embodiments, the first antigen recognition molecule is human or humanized. It comprises an antibody or antibody fragment which is a first antibody. In some embodiments, L1 is a first antibody It is bound to the N-terminus of the original recognition molecule. In some embodiments, A2 is the first antigen recognition component It is bound to the C-terminus of the child. In some embodiments, L1 is the C-terminus of the first antigen recognition molecule. It is bound to the end. In some embodiments, A2 is bound to the N-terminus of the first antigen recognition molecule. In some embodiments, the antibody or antibody fragment is a single-chain variable fragment. The Fab fragment comprises a single-domain antibody or a Fab fragment. In some embodiments, A 1 is a single-chain variable fragment (scFv). In some embodiments, scFv is , including scFv heavy chain polypeptides and scFv light chain polypeptides. Several implementations In this configuration, A1 is a single-domain antibody; in some embodiments, an antibody or antibody filament The ligment consists of a single-chain variable fragment (scFv) of a single-domain antibody derived from the camelid family, and a heavy chain. Variable domain (VH domain), light chain variable domain (VL domain), or variable domain It includes (VHH). In some embodiments, A1 is an anti-CD3e single-chain variable fragment. It includes. In some embodiments, A1 is 1 μM relative to CD3 on CD3-expressing cells. The following K D It comprises an anti-CD3e single-chain variable fragment having a bond. In some embodiments, The effector cell antigen contains CD3. In some embodiments, A1 is, It includes variable light chains and variable heavy chains that can specifically bind to human CD3. Several implementations Morphologically, A1 is muromonab-CD3 (OKT3), otelixizumab (TRX4), Teplizumab (MGA031), Bicilizumab (Nuvion), SP34, X35, V IT3, BMA030(BW264 / 56), CLB-T3 / 3, CRIS7, YTH1 2.5, F111-409, CLB-T3.4.2, TR-66, WT32, SPv-T 3b, 11D8, XIII-141, XIII-46, XIII-87, 12F6, T3 / RW2-8C8, T3 / RW2-4B6, OKT3D, M-T301, SMC2, F1 01.01, UCHT-1, WT-31, 15865, 15865v12, 15865v Includes a complementarity determination region (CDR) selected from the group consisting of 16 and 15865v19. In some embodiments, if L1 is cleaved by a tumor-specific protease, Polypeptides or polypeptide complexes of formula I bind to effector cells. In this embodiment, L1 is cleaved by a tumor-specific protease, and A1 is an effector molecule. When binding to cells, the polypeptide or polypeptide complex of formula I is transferred to effector cells. They bind. In some embodiments, the effector cells are T cells. Morphologically, A1 is a polypeptide that is part of the TCR-CD3 complex on effector cells. It binds to a polypeptide that is part of the TCR-CD3 complex. In some embodiments, it is part of a polypeptide that is part of the TCR-CD3 complex. This is human CD3ε. In some embodiments, the effector cell antigen includes CD3. scFV has HC-CDR1, HC-CDR2, and as complementary determination regions (CDRs). Including HC-CDR3, scFV's HC-CDR1, HC-CDR2, and HC-C DR3 is sequence number 1 in HC-CDR1, sequence number 2 in HC-CDR2, and HC -CDR3 includes sequence number 3, and scFV is CDR as LC-CDR1, LC-C Includes DR2 and LC-CDR3, and scFV's LC-CDR1, LC-CDR2, LC-CDR3 is sequence number 4 in LC-CDR1 and sequence number 5 in LC-CDR2. , and in LC-CDR3, sequence number 6 is included. In some embodiments, effector details The cytoplasmic antigen contains CD3, and the scFv contains the amino acid sequence according to SEQ ID NO: 7. In some embodiments, the second antigen-recognition molecule includes an antibody or antibody fragment. In the embodiment, the antibody or antibody fragment is a single-chain variable fragment, a single domain The antibody or Fab is included. In some embodiments, the antibody or its antibody fragment The first is a single-chain variable fragment (scFv) of a single-domain antibody derived from the camelid family, and a heavy-chain variable fragment. Main (VH domain), light chain variable domain (VL domain), or variable domain (V Contains HH). In some embodiments, the antibody or antibody fragment is humanized. A is a human. In some embodiments, A2 is Fab. So, Fab uses (a) Fab light chain polypeptide and (b) Fab heavy chain polypeptide. Includes. In some embodiments, Fab is used as the complementarity determination region (CDR) for HC-CD Includes R1, HC-CDR2, and HC-CDR3, and Fab's HC-CDR1, HC- CDR2 and HC-CDR3 are sequence numbers 8 in HC-CDR1 and HC-CDR2. It includes SEQ ID NO: 9, and HC-CDR3 includes SEQ ID NO: 10, and Fab is CDR Includes LC-CDR1, LC-CDR2, and LC-CDR3, and Fab's LC-CDR 1. LC-CDR2 and LC-CDR3 are sequence numbers 11 in LC-CDR1, LC -CDR2 includes sequence number 12, and LC-CDR3 includes sequence number 13. In this embodiment, the Fab light chain polypeptide comprises the amino acid sequence given by SEQ ID NO: 14. In some embodiments, the Fab heavy chain polypeptide is the amino acid sequence according to SEQ ID NO: 15 Includes. In some embodiments, the Fab light chain polypeptide of A2 is a single-chain variable polypeptide of A1. It is bonded to the C-terminus of the lagment (scFv). In some embodiments, the Fab of A2 The heavy chain polypeptide is bound to the C-terminus of the single-chain variable fragment (scFv) of A1. In some embodiments, the Fab light chain polypeptide A2 is a single-chain variable fragment of A1. It is bonded to the N-terminus of (scFv). In some embodiments, the Fab heavy chain poly of A2 The peptide is bound to the N-terminus of the single-chain variable fragment (scFv) of A1. In this embodiment, the Fab heavy chain polypeptide of A2 is converted to the scFv heavy chain polypeptide of A1. They are bonded. In some embodiments, the Fab light chain polypeptide of A2 is bonded to the scF of A1. v is bound to a heavy chain polypeptide. In some embodiments, the Fab heavy chain polypeptide of A2 The cydo is bound to the scFv light chain polypeptide of A1. In some embodiments, A2 The Fab light chain polypeptide of A1 is bound to the scFv light chain polypeptide. In this embodiment, A2 further includes P2 and L2, where P2 is a peptide bonded to A2 It contains L2, which connects A1 to P1 and is a substrate of tumor-specific proteases. Includes a linking portion. In some embodiments, the polypeptide or polypeptide complex is Equation Ia: P2-L2-A2-A1-L1-P1-H1 (Formula Ia) This is due to the following: In some embodiments, the Fab heavy chain polypeptide of A2 is the same as that of A1. The scFv heavy chain polypeptide is bound to L2, and the A2 Fab light chain polypeptide is bound to L2. In some embodiments, the Fab light chain polypeptide of A2 is the scFv heavy chain of A1. Binding to the polypeptide, L2 binds to the Fab heavy chain polypeptide of A2. How many? In that embodiment, the Fab heavy chain polypeptide of A2 is the scFv light chain polypeptide of A1. L2 is bound to the Fab light chain polypeptide of A2. Several embodiments So, the Fab light chain polypeptide of A2 is bound to the scFv light chain polypeptide of A1, L2 is bound to the Fab heavy chain polypeptide of A2. In some embodiments, P1 is , impairing the binding of A1 to effector cell antigens. In some embodiments, P1 is io H interaction, electrostatic interaction, hydrophobic interaction, Pi-stacking interaction, and H It is bonded to A1 by binding interactions, or a combination thereof. In some embodiments, P1 has less than 70% sequence homology to the effector cell antigen. In some embodiments, P2 impairs the coupling of A2 to PSMA. In some embodiments, P2 is involved in ionic interactions, electrostatic interactions, hydrophobic interactions, and Pi-stacking interactions. It is bound to A2 by H-bond interactions, or a combination thereof. In some embodiments, P2 binds to A2 at or near the antigen-binding site. In terms of application, P2 has less than 70% sequence homology to PSMA. In this embodiment, P1 or P2 comprises a peptide sequence having a length of at least 10 amino acids. In some embodiments, P1 or P2 has a length of at least 10 amino acids and 2 It contains a peptide sequence of 0 amino acids or less. In some embodiments, P1 or P2 is long It contains a peptide sequence of at least 16 amino acids. In some embodiments, P1 also P2 contains a peptide sequence with a length of 40 amino acids or less. In some embodiments, P 1 or P2 comprises at least two cysteine ​​amino acid residues. Several embodiments Then, P1 or P2 includes a cyclic peptide or a linear peptide. Several embodiments Then, P1 or P2 contains a cyclic peptide. In some embodiments, P1 or P 2 comprises a linear peptide. In some embodiments, P1 is at least two systo It contains an amino acid residue. In some embodiments, P1 is the same as in SEQ ID NOs. 16-19 or It contains an amino acid sequence of any one of the 78. In some embodiments, L1 is , is bonded to the N-terminus of A1. In some embodiments, L1 is bonded to the C-terminus of A1. In some embodiments, L2 is bonded to the N-terminus of A2. In this configuration, L2 is bonded to the C-terminus of A2. In some embodiments, L1 or L2 This is a peptide sequence having at least 5 to 50 amino acids. Several implementations In this state, L1 or L2 is a peptide sequence having at least 10 to 30 amino acids. In some embodiments, L1 or L2 has at least 10 amino acids. It is a peptide sequence. In some embodiments, L1 or L2 is at least 18 It is a peptide sequence having amino acids. In some embodiments, L1 or L2 is small It is a peptide sequence having at least 26 amino acids. In some embodiments, L1 L2 is (G2S) n The expression contains a function where n is an integer from 1 to 3 (array). Number 118). In some embodiments, L1 is (G2S) n , (GS) n (GSG GS)n (Sequence ID 50), (GGGS) n (Sequence ID 51), (GGGGS) n (array Number 52), and (GSSGGS) n Select an expression from the group consisting of (Sequence No. 53) It has such that n is at least an integer of 1. In some embodiments, L1 is It is cleaved by a ulcer-specific protease, thereby exposing A1 to effector cell antigens. In this case, P1 becomes unbound from A1. In some embodiments, L2 is tumor If A2 is cleaved by a specific protease and thereby exposed to PSMA, then P2 In some embodiments, tumor-specific proteases are released. These include matrix metalloproteinases (MMPs), serine proteases, and cysteine ​​proteases. From the group consisting of theases, threonine proteases, and aspartate proteases Selected. In some embodiments, the matrix metalloproteinase is MMP2, Includes MMP7, MMP9, MMP13, or MMP14. In some embodiments, Serine proteases include matryptase (MTSP1), urokinase, or hepsin. Includes. In some embodiments, L1 or L2 is a urokinase-cleavable amino acid. Row, amino acid sequence cleavable by matrix metalloproteinase, amino acid sequence cleavable by matrix metalloproteinase It comprises an amino acid sequence or a legmaine-cleavable amino acid sequence. In some embodiments, L1 or L2 comprises the amino acid sequence according to SEQ ID NO: 23. In some embodiments, L1 or L2 contains the amino acid sequence of one of sequence numbers 20-49. In some embodiments, L1 or L2 is linker 25 (ISSGLLSGRSD AG) (Sequence No. 45), Linker 26 (AAGLLAPPGGLSGRSDAG) (Distribution Column number 46), linker 27 (SPLGLSGRSDAG) (sequence number 47), or The amino acid sequence of linker 28 (LSGRSDAGSPLGLAG) (SEQ ID NO: 48) Alternatively, the amino acid composition of linker 25, linker 26, linker 27, or linker 28 Amino acid sequences having one, two, or three amino acid substitutions, additions, or deletions in a given column. Includes columns. In some embodiments, H1 comprises a polymer. In some embodiments, The polymer is polyethylene glycol (PEG). In some embodiments, H 1 contains albumin. In some embodiments, H1 contains an Fc domain. In some embodiments, albumin is serum albumin. In some embodiments, Albumin is human serum albumin. In some embodiments, H1 is polypeptide The molecule comprises a cytoplasm, ligand, or small molecule. In some embodiments, polypeptides, ligands, D, or small molecules, include serum proteins or their fragments, and circulating immunoglobulins. It binds to the fragment or CD35 / CR1. In some embodiments, Serum proteins include thyroxine-binding protein, transthyretin, and α1-acid glycoside. Protein (α1-acid glycoprotein), transferrin, tran Spherin receptor or its transferrin-binding site, fibrinogen, or aluminum Contains bumin. In some embodiments, the circulating immunoglobulin molecule is IgG1, IgG 2. Includes IgG3, IgG4, slgA, IgM, or IgD. Several embodiments So, the serum protein is albumin. In some embodiments, the polypeptide is , an antibody. In some embodiments, the antibody is a single-domain antibody, a single-chain variable fragmentation antibody. Includes albumin, or Fab. In some embodiments, the single-domain antibody is albumin It includes a single-domain antibody that binds to . In some embodiments, the single-domain antibody is . It is a mono-antibody or a humanized antibody. In some embodiments, a single-domain antibody is 645 The antibody is gH1gL1. In some embodiments, the single-domain antibody is 645dsgH5. It is gL4. In some embodiments, the single-domain antibody is 23-13-A01-s c02 is used in some embodiments. This is a lagment. In some embodiments, a single-domain antibody is used with DOM7r-31. Yes, in some embodiments, the single-domain antibody is DOM7h-11-15. In some embodiments, the single-domain antibody is Alb-1, Alb-8, or Alb It is -23. In some embodiments, the single-domain antibody is 10E. In this embodiment, the single-domain antibody has HC-CDR1 as the complementarity-determining region (CDR). It includes HC-CDR2 and HC-CDR3, and the single-domain antibody HC-CDR1, HC-CDR2 and HC-CDR3 are sequence numbers 54 and HC-CDR1, respectively. DR2 includes sequence number 55, and HC-CDR3 includes sequence number 56. In the application method, single-domain antibodies use HC-CDR1 and H as complementarity-determining regions (CDRs). Includes C-CDR2 and HC-CDR3, and single-domain antibodies HC-CDR1, HC -CDR2 and HC-CDR3 are sequence numbers 58 in HC-CDR1, HC-CDR It includes SEQ ID NO: 59 in 2 and SEQ ID NO: 60 in HC-CDR3. In some embodiments the single domain antibody is SA21. In some embodiments, the polypeptide or polypeptide complex includes modified amino acids, non-natural amino acids, non-natural modified amino acids, or combinations thereof. In some embodiments, the modified amino acid or non-natural modified amino acid includes post-translational modifications. In some embodiments, H1 includes a linking moiety (L3) that connects H1 to P1. In some embodiments, L3 is a peptide sequence having at least 5 to 50 or fewer amino acids. In some embodiments, L3 is a peptide sequence having at least 10 to 30 or fewer amino acids. In some embodiments, L3 is a peptide sequence having at least 10 amino acids. In some embodiments L3 is a peptide sequence having at least 18 amino acids. In some embodiments L3 is a peptide sequence having at least 26 amino acids. In some embodiments L3 has a formula selected from the group consisting of (G2S) , (GS) n , (GSGGS) n (SEQ ID NO: 50), ( n GGGS) (SEQ ID NO: 51), (GGGGS) n (SEQ ID NO: 52), and (GSSG n GS) (SEQ ID NO: 53), wherein n is an integer of at least n 1. In some embodiments, L3 includes the amino acid sequence according to SEQ ID NO: 22. In some embodiments, the polypeptide or polypeptide complex includes an amino acid sequence having at least 95% sequence identity to SEQ ID NOs: 62 - 77. In some embodiments, the polypeptide or polypeptide complex includes an amino acid sequence having at least 95% sequence identity to SEQ ID NOs: 62 - 77. In some embodiments, the polypeptide or polypeptide complex includes an amino acid sequence having at least 95% sequence identity to SEQ ID NOs: 62 - 77. In one embodiment, the polypeptide or polypeptide complex has at least 95% sequence identity to SEQ ID NO: 72 and comprises an amino acid sequence having at least 95% sequence identity to SEQ ID NO: 72 . In some embodiments, the polypeptide or polypeptide complex comprises an amino acid sequence having at least 95% sequence identity to SEQ ID NO: 73 . In some embodiments, the polypeptide or polypeptide complex comprises an amino acid sequence having at least 95% sequence identity to SEQ ID NO: 62 and SEQ ID NO: 63 . In some embodiments, the polypeptide or polypeptide complex comprises an amino acid sequence having at least 95% sequence identity to SEQ ID NO: 6 4 and SEQ ID NO: 65 . In some embodiments, the polypeptide or polypeptide complex comprises an amino acid sequence having at least 95% sequence identity to SEQ ID NO: 66 and SEQ ID NO: 67 . In some embodiments, the polypeptide or polypeptide complex comprises an amino acid sequence having at least 95% sequence identity to SEQ ID NO: 68 and SEQ ID NO: 69. In some embodiments, the polypeptide or polypeptide complex comprises an amino acid sequence having at least 95% sequence identity to SEQ ID NO: 70 and SEQ ID NO: 71. In some embodiments, the polypeptide or polypeptide complex comprises an amino acid sequence having at least 95% sequence identity to SEQ ID NO: 72 and SEQ ID NO: 73. In some embodiments, the polypeptide or polypeptide complex comprises an amino acid sequence having at least 95% sequence identity to SEQ ID NO: 74 and SEQ ID NO: 75. In some embodiments, the polypeptide or polypeptide complex comprises an amino acid sequence having at least 95% sequence identity to SEQ ID NO: 76 and SEQ ID NO: 77. In some embodiments, the polypeptide or polypeptide complex comprises an amino acid sequence having at least 95% sequence identity to SEQ ID NO: 78 and SEQ ID NO: 79. In some embodiments, the polypeptide or polypeptide complex comprises an amino acid sequence having at least 95% sequence identity to SEQ ID NO: 80 and SEQ ID NO: 81. In some embodiments, the polypeptide or polypeptide complex comprises an amino acid sequence having at least 95% sequence identity to SEQ ID NO: 82 and It contains an amino acid sequence that has at least 95% sequence identity with number 77.

[0004] In certain embodiments herein, (a) isolated polypeptide disclosed herein A pharmaceutical composition comprising (b) a cytoplasm or polypeptide complex and (b) a pharmaceutically acceptable excipient. It will be disclosed.

[0005] In certain embodiments herein, the polypeptide or polypeptide described herein Isolated recombinant nucleic acid molecules encoding the complex are disclosed.

[0006] In certain embodiments of this specification, Formula II: L 1a -P 1a -H 1a (Formula II) A polypeptide or polypeptide complex isolated by the formula, wherein L 1a is, If not blocked, the first antigen-recognition molecule that binds to the effector cell antigen becomes P 1a Connect The first antigen-recognizing molecule includes a linkage portion cleaved by a tumor-specific protease, It is connected to a second antigen recognition molecule that binds to PSMA, P 1a L 1a If the connection is not disconnected The combination includes a peptide that binds to the first antigen recognition molecule, H 1a It contains half-life extension molecules. A polypeptide or polypeptide complex is disclosed. In some embodiments, P1 a L 1a If it is not cleaved, the binding of the first antigen recognition molecule to the effector cell antigen is Damage. In some embodiments, the first antigen recognition molecule is an antibody or antibody fragment. Includes. In some embodiments, the effector cell antigen is anti-CD3 effector cell antigen. In some embodiments, P 1a This is less than 70% against effector cell antigens. It has sequence homology. In some embodiments, P 1a The length is at least 10A Contains a peptide sequence of amino acids. In some embodiments, P 1a is at least 1 It contains a peptide sequence of 0 amino acids and 20 amino acids or less. In some embodiments, P1 a It comprises a peptide sequence having a length of at least 16 amino acids. In some embodiments, P 1a It contains a peptide sequence with a length of 40 amino acids or less. In some embodiments, P 1a It contains at least two cysteine ​​amino acid residues. In some embodiments, P 1a This includes cyclic peptides or linear peptides. In some embodiments, P 1a teeth, Contains cyclic peptides. In some embodiments, P 1a It contains linear peptides. How many In that embodiment, P 1a This is a group consisting of one of sequence numbers 16-19. Includes a selected amino acid sequence. In some embodiments, H 1a It contains polymers. In some embodiments, the polymer is polyethylene glycol (PEG). In one embodiment, H 1a It contains albumin. In some embodiments, H 1a teeth , containing an Fc domain. In some embodiments, albumin is serum albumin. In some embodiments, albumin is human serum albumin. In terms of morphology, H 1a This includes polypeptides, ligands, or small molecules. Several implementation forms In this state, polypeptides, ligands, or small molecules are serum proteins or their flags. Binding to ment, circulating immunoglobulin or its fragments, or CD35 / CR1. In some embodiments, serum proteins are thyroxine-binding proteins, tran Suthyretin, α1-acid glycoprotein, transferrin, and transferrin receptor are also included. It contains either the transferrin binding site, fibrinogen, or albumin. In several embodiments, the circulating immunoglobulin molecules are IgG1, IgG2, IgG3, and Ig Contains G4, slgA, IgM, or IgD. In some embodiments, serum protein The substance is albumin. In some embodiments, the polypeptide is an antibody. In several embodiments, the antibody is a single-domain antibody, a single-chain variable fragment, or Fab This includes. In some embodiments, the antibody includes a single-domain antibody that binds to albumin. In some embodiments, the antibody is a human antibody or a humanized antibody. In the application form, the single-domain antibody is 645gH1gL1. In some embodiments, The single-domain antibody is 645dsgH5gL4. In some embodiments, single The domain antibody is 23-13-A01-sc02. In some embodiments, a single The domain antibody is A10m3 or a fragment thereof. In some embodiments, The single-domain antibody is DOM7r-31. In some embodiments, the single domain The antibody is DOM7h-11-15. In some embodiments, a single-domain antibody is used. These are Alb-1, Alb-8, or Alb-23. In some embodiments, a single The domain antibody is 10E. In some embodiments, the single-domain antibody is complementary. including HC-CDR1, HC-CDR2, and HC-CDR3 as the complementarity-determining regions (CDRs) and the HC-CDR1, HC-CDR2, and HC-CDR3 of the single-domain antibody are H In HC-CDR1, it includes SEQ ID NO: 54, in HC-CDR2, it includes SEQ ID NO: 55, and in HC-CDR 3, it includes SEQ ID NO: 56. In some embodiments, the single-domain antibody includes the complementarity-determining regions (CDRs) as HC-CDR1, HC-CDR2, and HC-CDR3 and the HC-CDR1, HC-CDR2, and HC-CDR3 of the single-domain antibody are HC- In HC-CDR1, it includes SEQ ID NO: 58, in HC-CDR2, it includes SEQ ID NO: 59, and in HC-CDR3 it includes SEQ ID NO: 60. In some embodiments, the single-domain antibody is SA21 . In some embodiments, H 1a is P 1a connected to H 1a by a linker portion (L 1a ) including. In some embodiments, L 1a is a peptide sequence having at least 5 to 50 amino acids . In some embodiments, L 1a is a peptide sequence having at least 10 to 30 amino acids. In some embodiments, L 1a is a peptide sequence having at least 10 amino acids. In some embodiments, L 1a is a peptide sequence having at least [[ID=4३]]18 amino acids. In some embodiments, L 1a is a peptide sequence having at least 26 amino acids. In some embodiments, L 1a is (G2S) n , (GS) n , (GSGGS) n (SEQ ID NO: 50), (GGGS) n (Sequence ID 51), (GGGGS) n (Sequence ID 52), and (GSSGGS) n ( The formula is selected from the group consisting of Sequence ID No. 53, where n is at least 1. It is a number. In some embodiments, L 1a This includes the amino acid sequence according to SEQ ID NO: 23. In some embodiments herein, stereochemical configuration 1:

[0007] [ka] A polypeptide complex comprising a structural arrangement thereof, wherein the polypeptide complex is a light chain variable A single-chain variable fragment (scFv) containing a main and heavy-chain variable domain, Fv further contains peptides that impair the binding of scFv to effector cell antigens. The peptide is a substrate for tumor-specific proteases, and the binding portion allows for variable heavy chain development of scFv. The main molecule is linked, and the peptide further contains half-life extension molecules, scFV and Fab or Fab' that binds to prostate-specific membrane antigen (PSMA), and Fab or Fab' contains Fab light chain polypeptide chains and Fab heavy chain polypeptide chains. The Fab heavy chain polypeptide chain is ligated to the C-terminus of the light chain variable domain of scFv, F A polypeptide complex comprising ab or Fab' is disclosed herein. In some embodiments, three-dimensional configuration 2:

[0008] [ka] A polypeptide complex comprising a structural arrangement thereof, wherein the polypeptide complex is a light chain variable A single-chain variable fragment (scFv) containing a main and heavy-chain variable domain, Fv further contains peptides that impair the binding of scFv to effector cell antigens. The peptide is a substrate for tumor-specific proteases, and the binding portion allows for variable heavy chain development of scFv. The main molecule is linked to the N-terminus, and the peptide further contains half-life extension molecules, sc FV and Fab or Fab' which binds to prostate-specific membrane antigen (PSMA), ab or Fab' contains Fab light chain polypeptide chains and Fab heavy chain polypeptide chains. The Fab light chain polypeptide chain is ligated to the C-terminus of the light chain variable domain of scFv. A polypeptide complex comprising Fab or Fab' is disclosed.

[0009] Reference All publications, patents, and patent applications referred to herein are treated as if they were individual publications. , patents, or patent applications are specifically and individually indicated so as to be invoked by reference, respectively. It is incorporated herein by reference to the same extent as it is incorporated by reference. [Brief explanation of the drawing]

[0010] Novel features of this disclosure are described in particular by the attached claims. Features and advantages are described in the accompanying drawings, illustrating exemplary embodiments in which the principles of this disclosure are utilized. Further understanding can be gained by referring to the detailed explanation below.

[0011] [Figure 1A] This figure illustrates a polypeptide complex of the present disclosure in a normal orientation. [Figure 1B] This figure illustrates a polypeptide complex of the present disclosure in an inverted orientation. [Figure 2A]This figure illustrates titration data for PSMA binding in several polypeptide complexes of this disclosure. [Figure 2B] This figure illustrates titration data for PSMA binding in several polypeptide complexes of this disclosure. [Figure 2C] This figure illustrates titration data for PSMA binding in several polypeptide complexes of this disclosure. [Figure 3A] This figure illustrates titration data for CD3ε binding in several polypeptide complexes of this disclosure. [Figure 3B] This figure illustrates titration data for CD3ε binding in several polypeptide complexes of this disclosure. [Figure 3C] This figure illustrates titration data for CD3ε binding in several polypeptide complexes of this disclosure. [Figure 4] This figure illustrates cell viability data for 22Rv1 tumor cells treated with PC1 or PC2. [Figure 5A] This figure illustrates cell viability data for 22Rv1 tumor cells treated with PC1, PC5, or MTSP1 treated with PC5. [Figure 5B] This figure illustrates cell viability data for 22Rv1 tumor cells treated with PC2, PC4 (with or without MTSP1 treatment), or PC6 (with or without MTSP1 treatment). [Figure 6] This figure illustrates cell viability data for LNCaP tumor cells treated with PC1 or PC2. [Figure 7] This figure illustrates cell viability data for LNCaP tumor cells treated with PC2, PC4, or MTSP1 treated with PC4. [Figure 8A] This figure illustrates how polypeptide complexes mediated the death of 22Rv1 tumor cells in the presence of CD8+ T cells. [Figure 8B] This figure illustrates how polypeptide complexes mediated the death of 22Rv1 tumor cells in the presence of CD8+ T cells. [Figure 9A] This figure illustrates the pharmacokinetics of polypeptide (PSMA TCE) in cynomolgus monkeys after a single IV bolus injection. [Figure 9B] This figure illustrates the pharmacokinetics of polypeptides (PSMA TRACTrs) in cynomolgus monkeys after a single IV bolus injection. [Figure 10A] This figure illustrates cytokine release in cynomolgus monkeys after a single IV bolus of PSMA TCE. [Figure 10B] This figure illustrates cytokine release in cynomolgus monkeys after a single IV bolus of the PSMA polypeptide TRACTr complex. [Figure 10C] This figure illustrates cytokine release in cynomolgus monkeys after a single IV bolus of PSMA TRACTR. [Figure 11A] This figure illustrates serum liver enzymes in cynomolgus monkeys after a single IV bolus of PSMA TCE. [Figure 11B] This figure illustrates serum liver enzymes in cynomolgus monkeys after a single IV bolus of PSMA polypeptide TRACTr complex. [Figure 12A] This figure illustrates the anti-CD3 scFv binding of anti-CD3 scFv peptides A and B by alanine scanning peptides, as measured by ELISA. [Figure 12B] This figure illustrates the anti-CD3 scFv binding of anti-CD3 scFv peptides A and B by alanine scanning peptides as measured by ELISA. [Figure 12C] This figure illustrates the anti-CD3 scFv binding of anti-CD3 scFv peptides A and B by alanine scanning peptides as measured by ELISA. [Figure 12D] This figure illustrates the anti-CD3 scFv binding of anti-CD3 scFv peptides A and B by alanine scanning peptides as measured by ELISA. [Figure 12E]This figure illustrates the anti-CD3 scFv binding of anti-CD3 scFv peptides A and B by alanine scanning peptides as measured by ELISA. [Figure 12F] This figure illustrates the anti-CD3 scFv binding of anti-CD3 scFv peptides A and B by alanine scanning peptides as measured by ELISA. [Figure 13A] This figure illustrates the inhibition of anti-CD3 scFv binding to CD3 by alanine scanning peptides of anti-CD3 scFv peptide-A and peptide-B, as measured by ELISA. [Figure 13B] This figure illustrates the inhibition of anti-CD3 scFv binding to CD3 by alanine scanning peptides of anti-CD3 scFv peptide-A and peptide-B, as measured by ELISA. [Figure 13C] This figure illustrates the inhibition of anti-CD3 scFv binding to CD3 by alanine scanning peptides of anti-CD3 scFv peptide-A and peptide-B, as measured by ELISA. [Figure 13D] This figure illustrates the inhibition of anti-CD3 scFv binding to CD3 by alanine scanning peptides of anti-CD3 scFv peptide-A and peptide-B, as measured by ELISA. [Figure 13E] This figure illustrates the inhibition of anti-CD3 scFv binding to CD3 by alanine scanning peptides of anti-CD3 scFv peptide-A and peptide-B, as measured by ELISA. [Figure 13F] This figure illustrates the inhibition of anti-CD3 scFv binding to CD3 by alanine scanning peptides of anti-CD3 scFv peptide-A and peptide-B, as measured by ELISA. [Figure 14A] This figure illustrates anti-CD3 scFv binding using the sequence of the optimized anti-CD3 scFv peptide-B as measured by ELISA. [Figure 14B]This figure illustrates anti-CD3 scFv binding using the sequence of the optimized anti-CD3 scFv peptide-B as measured by ELISA. [Figure 15A] This figure illustrates the inhibition of anti-CD3 scFv binding to CD3 by the optimized anti-CD3 scFv peptide-B sequence as measured by ELISA. [Figure 15B] This figure illustrates the inhibition of anti-CD3 scFv binding to CD3 by the optimized anti-CD3 scFv peptide-B sequence as measured by ELISA. [Figure 16] This figure illustrates the core sequence motif of the optimized anti-CD3 scFv peptide-B sequence generated using WebLogo3.7.4. [Modes for carrying out the invention]

[0012] Multispecific antibodies combine the benefits of different binding specificities derived from two or more antibodies into a single composition. Combined with: A multispecific antibody to redirect T cells back to cancer, preclinical and clinical It has shown promise in both trials. This method targets tumor-associated antigens or markers. It depends on the binding of one antigen interaction site of the antibody, but another antigen interaction site is connected to the cell It binds to effector cell antigens on T cells, such as differentiation antigen group 3 (CD3), which cause toxic activity. It can be combined.

[0013] One such tumor-associated antigen is PSMA. (Prostate-specific membrane antigen (PSMA)) These are glutamate carboxypeptidase II (GCPII) and N-acetyl-L-acetyl-Axylamine. Partyl-L-glutamate peptidase I (NAALADase I), or NAA Also known as G peptidase, in humans FOLH1 (folate hydrolase 1) It is an enzyme encoded by a gene. PSMA is a zinc metalloenzyme present in membranes. The majority of this enzyme is found in the extracellular fluid space. Human PSMA is found in most other tissues. It is expressed more than 100 times higher in the prostate. In some prostate cancers, PSMA is expressed twice It is the second most upregulated gene product, and is higher than the level of non-cancerous prostate cells. It is also 8 to 12 times higher.

[0014] T-cell enhancer (TCE) therapy has several advantages, including the fact that it is not cell therapy. Therefore, there are existing therapeutic agents that can counter chimeric antigen receptor T cell (CAR T cell) therapy. It can be presented as a treatment. TCE treatment exhibits strong antitumor activity against hematological cancers. However, the development of TCE for treating solid tumors has limitations compared to prior TCE technologies, that is, (i) Excessive activation of the immune system that causes cytokine release syndrome (CRS), (i i) On-target healthy tissue toxicity, and (iii) Tissue drugs that produce a short half-life. We are facing difficulties due to poor physical dynamics (PK). CRS is systemic activation of T cells. It originates from and triggers inflammatory cytokines such as interleukin-6 (IL-6), which threaten life. It may cause a significant increase in the dose limit. Severe acute CRS, which can cause dose-limiting toxicity and death, is a risk factor. Developed using other platforms to treat cancer patients in insufficient clinical trials. This toxicity has been observed when T-cell enhancers are administered. It limits the maximum blood concentration of T cell promoters. The effectiveness of T cell promoters is also limited. This is limited due to the toxicity of the target to healthy tissue. Tumor-specific (tumor-spe Opening up using a platform not designed for activation (cification) The released T-cell enhancer was banned from clinical trials and targeted expression in healthy tissue. T cell engagement is also limited by its short half-life. Because of its short exposure half-life, it is rapidly excreted from the body, and therefore, after administration, the saccharin is quickly eliminated. It reaches a level below therapeutic dose. Therefore, it overcomes the difficulty of a short half-life and the drug is absorbed into the body. To maintain the level of treatment, T-cell engagers such as blinatumomab are typically used. It is administered in low doses via a continuous infusion pump over several weeks. The continuous dosing regimen is for the patient. This would place a significant burden on them.

[0015] To overcome these difficulties related to the effectiveness of T cell promoters, this specification includes This polypeptide contains a binding region that selectively binds to effector cell antigens and PSMA. Alternatively, a polypeptide complex is described, in which one or more binding domains are Selectively activated in the tumor microenvironment, polypeptides or polypeptide complexes are halved. Contains a period-extending molecule. Such modifications enhance the health of CRS and on-target tissue toxicity. The risk of sexual dysfunction is reduced, and the stability of blood flow and serum half-life before activation is improved. The polypeptide or polypeptide complex described herein is active at low levels of target expression. It is easy to manufacture.

[0016] In some embodiments, the polypeptide or polypeptide complex described herein It is used in methods for treating cancer. In some embodiments, cancer expresses PSMA. It has cells that have polypeptides described herein or Polypeptide complexes are used in methods for treating prostate cancer. In some embodiments, The prostate cancer is metastatic castration-resistant prostate cancer (mCRPC). Prostate cancer affects men. It is the second most common cancer in the world, and in the United States alone, more than 3 million men live with prostate cancer. Early diagnosis and effective treatment are effective for the majority of prostate cancer patients, with an average of about 98 percent. This means that they have a 5-year survival rate. However, an estimated 6 percent of prostate cancer patients... The disease develops metastatic disease, which is associated with a 5-year survival rate of approximately 30 percent. In 2020, An estimated 33,000 people die from prostate cancer in the United States.

[0017] In some examples, the polypeptides or polypeptide complexes described herein are solid Used to treat tumors and cancers. In some embodiments, the cancer is in the lung, breast (for example) (HER2+, ER / PR+, TNBC), cervix, ovaries, colon, pancreas, or stomach It is cancer. Some embodiments are methods for treating cancer, which require treatment of cancer Elephants, Formula I: A2-A1-L1-P1-H1 (Formula I) The process includes administering an isolated polypeptide or polypeptide complex, In the formula, A1 comprises a first antigen-recognition molecule that binds to the effector cell antigen, and P1 comprises A1 It contains a peptide that binds to P1, and L1 connects A1 to P1 and tumor-specific protea - It contains a binding portion which is the substrate of the enzyme, H1 contains a half-life extension molecule, and A2 is PSMA This method involves including another antigen-recognizing molecule to bind to.

[0018] In some embodiments of this specification, Formula I: A2-A1-L1-P1-H1 (Formula I) A peptide or peptide complex isolated by, In the formula, A1 comprises a first antigen-recognition molecule that binds to the effector cell antigen, and P1 comprises A1 It contains a peptide that binds to P1, and L1 connects A1 to P1 and tumor-specific protea It contains a binding portion that is a substrate of the enzyme, H1 contains a half-life extension molecule, and A2 is prostate-specific. An isolated peptide or containing a second antigen-recognition molecule that binds to the target membrane antigen (PSMA). A peptide complex is disclosed.

[0019] In some embodiments of this specification, Formula I: A2-A1-L1-P1-H1 (Formula I) A peptide or peptide complex isolated by, In the formula, A1 is the first antigen-recognizing molecule that binds to the effector cell antigen, and P1 is A1 It is a peptide that binds to P1, and L1 attaches A1 to P1 and tumor-specific protea The binding portion is the substrate of the enzyme; H1 is a half-life extension molecule; and A2 is prostate-specific. A second antigen-recognition molecule that binds to the target membrane antigen (PSMA), an isolated peptide or A peptide complex is disclosed.

[0020] In some embodiments of this specification, Formula I: A2-A1-L1-P1-H1 (Formula I) An isolated peptide or peptide complex comprising, In the formula, A1 comprises a first antigen-recognition molecule that binds to the effector cell antigen, and P1 comprises A1 It contains a peptide that binds to P1, and L1 connects A1 to P1 and tumor-specific protea It contains a binding portion that is a substrate of the enzyme, H1 contains a half-life extension molecule, and A2 is prostate-specific. An isolated peptide or containing a second antigen-recognition molecule that binds to the target membrane antigen (PSMA). A peptide complex is disclosed.

[0021] In some embodiments of this specification, Formula I: A2-A1-L1-P1-H1 (Formula I) An isolated peptide or peptide complex comprising, In the formula, A1 is the first antigen-recognizing molecule that binds to the effector cell antigen, and P1 is A1 It is a peptide that binds to P1, and L1 attaches A1 to P1 and tumor-specific protea The binding portion is the substrate of the enzyme; H1 is a half-life extension molecule; and A2 is prostate-specific. A second antigen-recognition molecule that binds to the target membrane antigen (PSMA), an isolated peptide or A peptide complex is disclosed.

[0022] In some embodiments of this specification, Formula I: A2-A1-L1-P1-H1 (Formula I) A peptide or peptide complex isolated by, In the formula, A1 comprises a first antigen-recognizing molecule that binds to prostate-specific membrane antigen (PSMA). P1 contains a peptide that binds to A1, and L1 connects A1 to P1 and also has tumor-specific properties. It contains a binding portion that is a substrate for heteroprotease, H1 contains a half-life extension molecule, and A2 is , an isolated peptide or containing a second antigen-recognition molecule that binds to the effector cell antigen. A peptide complex is disclosed.

[0023] In some embodiments of this specification, Formula I: A2-A1-L1-P1-H1 (Formula I) A peptide or peptide complex isolated by, In the formula, A1 is the first antigen-recognizing molecule that binds to prostate-specific membrane antigen (PSMA). P1 is a peptide that binds to A1, and L1 attaches A1 to P1 and also has tumor-specific properties. The binding portion is the substrate of the heteroprotease, H1 is a half-life extension molecule, and A2 is , an isolated peptide or a second antigen-recognition molecule that binds to effector cell antigens. A peptide complex is disclosed.

[0024] In some embodiments of this specification, Formula I: A2-A1-L1-P1-H1 (Formula I) An isolated peptide or peptide complex comprising, In the formula, A1 comprises a first antigen-recognizing molecule that binds to prostate-specific membrane antigen (PSMA). P1 contains a peptide that binds to A1, and L1 connects A1 to P1 and also has tumor-specific properties. It contains a binding portion that is a substrate for heteroprotease, H1 contains a half-life extension molecule, and A2 is , an isolated peptide or containing a second antigen-recognition molecule that binds to the effector cell antigen. A peptide complex is disclosed.

[0025] In some embodiments of this specification, Formula I: A2-A1-L1-P1-H1 (Formula I) An isolated peptide or peptide complex comprising, In the formula, A1 is the first antigen-recognizing molecule that binds to prostate-specific membrane antigen (PSMA). P1 is a peptide that binds to A1, and L1 attaches A1 to P1 and also has tumor-specific properties. The binding portion is the substrate of the heteroprotease, H1 is a half-life extension molecule, and A2 is , an isolated peptide or a second antigen-recognition molecule that binds to effector cell antigens. A peptide complex is disclosed.

[0026] In some embodiments herein, formula Ia: P2-L2-A2-A1-L1-P1-H1 (Formula Ia) A peptide or peptide complex isolated by, In the formula, A2 further comprises P2 and L2, where P2 comprises a peptide bound to A2. L2 connects A1 to P1 and also has a binding portion that is a substrate for tumor-specific proteases. Isolated peptides or peptide complexes, including those described herein, are disclosed.

[0027] In some embodiments herein, formula Ia: P2-L2-A2-A1-L1-P1-H1 (Formula Ia) A peptide or peptide complex isolated by, In the formula, A2 further comprises P2 and L2, where P2 is a peptide that binds to A2. L1 connects A1 to P1 and has a ligation site that is a substrate for tumor-specific proteases. An isolated peptide or peptide complex is disclosed.

[0028] In some embodiments herein, formula Ia: P2-L2-A2-A1-L1-P1-H1 (Formula Ia) An isolated peptide or peptide complex comprising, In the formula, A2 further comprises P2 and L2, where P2 comprises a peptide bound to A2. L2 connects A1 to P1 and also has a binding portion that is a substrate for tumor-specific proteases. Isolated peptides or peptide complexes, including those described herein, are disclosed.

[0029] In some embodiments herein, formula Ia: P2-L2-A2-A1-L1-P1-H1 (Formula Ia) An isolated peptide or peptide complex comprising, In the formula, A2 further comprises P2 and L2, where P2 is a peptide that binds to A2. L1 connects A1 to P1 and has a ligation site that is a substrate for tumor-specific proteases. An isolated peptide or peptide complex is disclosed.

[0030] In some embodiments herein, Formula II: L 1a -P 1a -H 1a (Formula II) A peptide or peptide complex isolated by, In the formula, L 1a If not cleaved, the first antigen recognition molecule binds to the effector cell antigen. P to child 1a The first includes a connecting portion that is cleaved by a tumor-specific protease, which connects the two. The antigen-recognizing molecule then binds to a second antigen-recognizing molecule that binds to the prostate-specific membrane antigen (PSMA). Continued, P 1a L 1a If it is not cleaved, the peptide that binds to the first antigen recognition molecule Includes H 1a This refers to an isolated peptide or peptide complex containing a half-life extension molecule. It will be disclosed.

[0031] In some embodiments herein, Formula II: L 1a -P 1a -H 1a (Formula II) An isolated peptide or peptide complex comprising, In the formula, L 1a If not cleaved, the first antigen recognition molecule binds to the effector cell antigen. Child P 1a The first includes a connecting portion that is cleaved by a tumor-specific protease, which connects the two. The antigen-recognizing molecule then binds to a second antigen-recognizing molecule that binds to the prostate-specific membrane antigen (PSMA). Continued, P 1a L 1a If it is not cleaved, the peptide that binds to the first antigen recognition molecule Includes H 1aThis refers to an isolated peptide or peptide complex containing a half-life extension molecule. It will be disclosed.

[0032] In some embodiments herein, Formula II: L 1a -P 1a -H 1a (Formula II) A peptide or peptide complex isolated by, In the formula, L 1a If not cleaved, the first antigen recognition molecule binds to the effector cell antigen. Child P 1a The connecting portion is a tumor-specific protease that cleaves the first The antigen-recognizing molecule then binds to a second antigen-recognizing molecule that binds to the prostate-specific membrane antigen (PSMA). Continued, P 1a L 1a If it is not cleaved, the peptide that binds to the first antigen recognition molecule It is D, H 1a This refers to an isolated peptide or peptide complex, which is a half-life extension molecule. It will be disclosed.

[0033] In some embodiments herein, Formula II: L 1a -P 1a -H 1a (Formula II) An isolated peptide or peptide complex comprising, In the formula, L 1a If not cleaved, the first antigen recognition molecule binds to the effector cell antigen. Child P 1a The connecting portion is a tumor-specific protease that cleaves the first The antigen-recognizing molecule then binds to a second antigen-recognizing molecule that binds to the prostate-specific membrane antigen (PSMA). Continued, P 1a L 1a If it is not cleaved, the peptide that binds to the first antigen recognition molecule It is D, H1a This refers to an isolated peptide or peptide complex, which is a half-life extension molecule. It will be disclosed.

[0034] First antigen-recognition molecule (A1) In some embodiments herein, isolated polypeptides or polypeptide complexes are used. In a combination, the first antigen-recognizing molecule binds to the effector cell antigen, and the second antigen-recognizing molecule... An isolated polypeptide or polypeptide complex that binds to PSMA is disclosed. In some embodiments, the effector cell antigen includes CD3. A1 contains a first antigen-recognition molecule that binds to the effector cell antigen.

[0035] In some embodiments, A1 comprises an antibody or antibody fragment. In the application form, A1 contains a human or humanized antibody or antibody fragment. In several embodiments, L1 is bound to the N-terminus of the antibody or antibody fragment. In several embodiments, L1 is bound to the N-terminus of the antibody or antibody fragment, and A2 is , it is bound to the other N-terminus of the antibody or antibody fragment. In some embodiments, A 2 is bound to the C-terminus of the antibody or antibody fragment. In some embodiments, L 1 is bound to the C-terminus of an antibody or antibody fragment. In some embodiments, A 2 is bound to the N-terminus of the antibody or antibody fragment. In some embodiments, The body or antibody fragment is a single-chain variable fragment, a single-domain antibody, or Fab Includes fragments. In some embodiments, A1 is a single-chain variable fragment (scF v) In some embodiments, scFv is scFv heavy chain polypeptide and s Contains cFv light chain polypeptide. In some embodiments, A1 is a single-domain antibody. Yes. In some embodiments, A1 is capable of specifically binding to human CD3. It includes variable light chains and variable heavy chains. In some embodiments, the effector cell antigen is C Includes D3. In some embodiments, A1 includes an anti-CD3e single-chain variable fragment. In some embodiments, A1 is expressed as 1 μM or less of K for CD3 on CD3-expressing cells. D It contains an anti-CD3e single-chain variable fragment having a bond. In some embodiments, A1 These include muromonab-CD3 (OKT3), otelixizumab (TRX4), and teprizumab ( MGA031), Bicilizumab (Nuvion), SP34, X35, VIT3, BMA 030(BW264 / 56), CLB-T3 / 3, CRIS7, YTH12.5, F11 1-409, CLB-T3.4.2, TR-66, WT32, SPv-T3b, 11D8 , XIII-141, XIII-46, XIII-87, 12F6, T3 / RW2-8C 8, T3 / RW2-4B6, OKT3D, M-T301, SMC2, F101.01, U CHT-1, WT-31, 15865, 15865v12, 15865v16, and 1 It includes a complementarity determination region (CDR) selected from the group consisting of 5865v19.

[0036] In some embodiments, A1 includes a first antigen-recognizing molecule that binds to PSMA. In some embodiments, A1 is a variable that can specifically bind to human PSMA. Includes light chains and variable heavy chains.

[0037] In some embodiments, the scFv bound to CD3 has a variable domain in the scFv light chain. It includes the scFv heavy chain variable domain. In some embodiments, the scFv heavy chain variable domain The IN comprises at least one, two, or three complementarity determination regions (CDRs) disclosed in Table 1. , or substantially identical sequences (e.g., at least 90%, 95%, 96%) Includes sequences having 97%, 98%, or 99% sequence identity. Several embodiments So, the scFv light chain variable domains are at least one, two, or three of those disclosed in Table 1. The complementarity determination region (CDR) of the CDR, or substantially identical sequences to it (for example, at least (Also sequences with 90%, 95%, 96%, 97%, 98%, or 99% sequence identity) Includes.

[0038] In some embodiments, the scFv heavy chain variable domain is at least as disclosed in Table 1. 1, 2, or 3 complementarity determination regions (CDRs), or substantially identical distributions thereto Columns (for example, arrays of at least 90%, 95%, 96%, 97%, 98%, or 99%) The scFv light chain variable domains include sequences that are identical to each other, and are disclosed in Table 1 at least as follows: Also, one, two, or three complementary determination regions (CDRs), or substantially identical to them. Arrays (for example, arrays with at least 90%, 95%, 96%, 97%, 98%, or 99% Includes arrays with column identity.

[0039] [Table 1]

[0040] In some embodiments, the scFv heavy chain variable domain is a complementarity determination region (CDR) and It includes HC-CDR1, HC-CDR2, and HC-CDR3, and the scFv heavy chain is variable. The domains HC-CDR1, HC-CDR2, and HC-CDR3 are HC-CDR1 In this case, it includes sequence number 1, in HC-CDR2 sequence number 2, and in HC-CDR3 sequence number 3. The CD-R is at least one of HC-CDR1, HC-CDR2, or HC-CDR3. Each contains 0 to 2 amino acid modifications. In some embodiments, the scFv light chain variable dormancy In, the complementary determination regions (CDRs) are LC-CDR1, LC-CDR2, and LC -Includes CDR3, and scFv light chain variable domains LC-CDR1, LC-CDR2, and LC-CDR3 is sequence number 4 in LC-CDR1, sequence number 5 in LC-CDR2, LC-CDR3 includes sequence number 6, and the CDRs are LC-CDR1, LC-CDR2, and Alternatively, at least one of the LC-CDR3 molecules contains 0 to 2 amino acid modifications.

[0041] In some embodiments, if L1 is cleaved by a tumor-specific protease, the formula Polypeptides or polypeptide complexes of type I bind to effector cells. In this embodiment, L1 is cleaved by a tumor-specific protease, and A1 is an effector cell. When binding, the polypeptide or polypeptide complex of formula I binds to effector cells. Combine. In some embodiments, the effector cell is a T cell. In this state, A1 is a polypeptide that is part of the TCR-CD3 complex on effector cells. They bind. In some embodiments, the polypeptide that is part of the TCR-CD3 complex , human CD3ε. In some embodiments, the effector cell antigen includes CD3, scFV has HC-CDR1, HC-CDR2, and as complementary determination regions (CDRs). Includes HC-CDR3, and scFV versions of HC-CDR1, HC-CDR2, and HC-CD R3 is sequence number 1 in HC-CDR1, sequence number 2 in HC-CDR2, and HC- CDR3 includes sequence number 3, and scFV is used as a CDR for LC-CDR1 and LC-CD. R2 and LC-CDR3 are included, along with scFV's LC-CDR1, LC-CDR2, and LC-CDR3 is sequence number 4 in LC-CDR1, sequence number 5 in LC-CDR2, And LC-CDR3 includes SEQ ID NO: 6. In some embodiments, effector cells The antigen contains CD3, and the scFv contains the amino acid sequence according to SEQ ID NO: 7.

[0042] In some embodiments, the effector cell antigen includes CD3, and A1 is the complementarity determination region. The range (CDR) includes HC-CDR1, HC-CDR2, and HC-CDR3, A HC-CDR1, HC-CDR2, and HC-CDR3 are distributed as follows: Includes column number 1, sequence number 2 in HC-CDR2, and sequence number 3 in HC-CDR3. A1 includes LC-CDR1, LC-CDR2, and LC-CDR3 as CDRs. The A1 LC-CDR1, LC-CDR2, and LC-CDR3 are, in LC-CDR1. This is sequence number 4 in LC-CDR2, sequence number 5 in LC-CDR2, and sequence number 6 in LC-CDR3. Includes. In some embodiments, the effector cell antigen includes CD3, and A1 is Sequence ID No. Includes the amino acid sequence specified by 7.

[0043] In some embodiments, A1 comprises the amino acid sequence given by SEQ ID NO: 7. In this embodiment, A1 has at least 80% sequence identity with respect to sequence number 7. It contains an acid sequence. In some embodiments, A1 is at least 85 relative to SEQ ID NO 7. It contains an amino acid sequence having % sequence identity. In some embodiments, A1 is sequence number It contains an amino acid sequence that has at least 90% sequence identity with number 7. In the application form, A1 is an amino acid having at least 95% sequence identity with SEQ ID NO: 7. Contains an acid sequence. In some embodiments, A1 is at least 99% of SEQ ID NO: 7 It contains an amino acid sequence that has sequence identity.

[0044] In some embodiments, the polypeptide or polypeptide complex has both P1 and L1. Unidentified isolated polypeptides or polypeptide complexes against tumor cell antigens In comparison to the binding affinity, the binding affinity to tumor cell antigens is weak. In some embodiments, Polypeptides or polypeptide complexes have weak binding affinity to tumor cell antigens. Tumor cell antigens in the form of polypeptides or polypeptide complexes that do not possess either P1 or L1. It is at least 5 times higher compared to the binding affinity to polyp Tide or polypeptide complexes have weak binding affinity to tumor cell antigens, and P1 and L1 Binding to tumor cell antigens in the form of polypeptides or polypeptide complexes that do not possess It is at least 8 times higher compared to affinity. In some embodiments, polypeptides or Polypeptide complexes have weak binding affinity to tumor cell antigens and possess both P1 and L1. The binding affinity to tumor cell antigens in the form of polypeptides or polypeptide complexes and It is at least 10 times higher by comparison. In some embodiments, polypeptides or polypept The cytoplasmic reticulum complex has weak binding affinity to tumor cell antigens and lacks both P1 and L1 polynucleotides. Compared to the binding affinity to tumor cell antigens in the form of peptides or polypeptide complexes At least 15 times higher. In some embodiments, polypeptides or polypeptide complexes. The body has a weak binding affinity to tumor cell antigens and does not possess either P1 or L1 polypeptides. Or less than the binding affinity to tumor cell antigens in the form of polypeptide complexes. It is also 20 times higher. In some embodiments, the polypeptide or polypeptide complex is tumor Polypeptides or polypropylenes that have weak binding affinity to ulcer cell antigens and do not possess either P1 or L1. Compared to the binding affinity of the lipeptide complex to tumor cell antigens, this is at least 25 times greater. High. In some embodiments, polypeptides or polypeptide complexes are used against tumor cells. Polypeptides or polypeptides that have weak binding affinity to the source and do not possess either P1 or L1 The binding affinity of the complex to tumor cell antigens is at least 30 times higher compared to the binding affinity of the complex itself. In some embodiments, the polypeptide or polypeptide complex is effective against tumor cell antigens. Polypeptides or polypeptide complexes that have weak binding affinity and lack both P1 and L1 groups. It is at least 35 times higher compared to the binding affinity to tumor cell antigens of some morphologies. In the embodiment, the polypeptide or polypeptide complex is a binding parent to tumor cell antigens. Polypeptides or polypeptide complexes with weak affinity and lacking both P1 and L1 It is at least 40 times higher compared to the binding affinity to tumor cell antigens. Therefore, polypeptides or polypeptide complexes have weak binding affinity to tumor cell antigens. Tumor cells in the form of polypeptides or polypeptide complexes that do not possess either P1 or L1. It is at least 45 times higher compared to the binding affinity to the antigen. In some embodiments, Lipeptide or polypeptide complexes have weak binding affinity to tumor cell antigens, P1 Against tumor cell antigens in the form of polypeptides or polypeptide complexes that do not have L1, The binding affinity is at least 50 times higher compared to [another method]. In some embodiments, the polypeptide The polypeptide or polypeptide complex has weak binding affinity to tumor cell antigens, and both P1 and L1... Binding to tumor cell antigens in the form of polypeptides or polypeptide complexes that do not possess this ability. It is at least 55 times higher compared to affinity. In some embodiments, polypeptide or Polypeptide complexes have weak binding affinity to tumor cell antigens and possess both P1 and L1. The binding affinity to tumor cell antigens in the form of polypeptides or polypeptide complexes and It is at least 60 times higher by comparison. In some embodiments, polypeptides or polypeptide The cytoplasmic reticulum complex has weak binding affinity to tumor cell antigens and lacks both P1 and L1 polynucleotides. Compared to the binding affinity to tumor cell antigens in the form of peptides or polypeptide complexes At least 65 times higher. In some embodiments, polypeptides or polypeptide complexes. The body has a weak binding affinity to tumor cell antigens and does not possess either P1 or L1 polypeptides. Or less than the binding affinity to tumor cell antigens in the form of polypeptide complexes. It is also 70 times higher. In some embodiments, the polypeptide or polypeptide complex is tumor Polypeptides or polypropylenes that have weak binding affinity to ulcer cell antigens and do not possess either P1 or L1. Compared to the binding affinity of the lipeptide complex to tumor cell antigens, this is at least 75 times greater. High. In some embodiments, polypeptides or polypeptide complexes are used against tumor cells. Polypeptides or polypeptides that have weak binding affinity to the source and do not possess either P1 or L1 The binding affinity of the complex to tumor cell antigens is at least 80 times higher compared to the binding affinity of the complex itself. In some embodiments, the polypeptide or polypeptide complex is effective against tumor cell antigens. Polypeptides or polypeptide complexes that have weak binding affinity and lack both P1 and L1 groups. It is at least 85 times higher compared to the binding affinity to tumor cell antigens of some morphologies. In the embodiment, the polypeptide or polypeptide complex is a binding parent to tumor cell antigens. Polypeptides or polypeptide complexes with weak affinity and lacking both P1 and L1 It is at least 90 times higher compared to the binding affinity to tumor cell antigens. Therefore, polypeptides or polypeptide complexes have weak binding affinity to tumor cell antigens. Tumor cells in the form of polypeptides or polypeptide complexes that do not possess either P1 or L1. It is at least 95 times higher compared to the binding affinity to the antigen. In some embodiments, Lipeptide or polypeptide complexes have weak binding affinity to tumor cell antigens, P1 Against tumor cell antigens in the form of polypeptides or polypeptide complexes that do not have L1, It is at least 100 times higher compared to the binding affinity. In some embodiments, the polyp Tide or polypeptide complexes have weak binding affinity to tumor cell antigens, and P1 and L1 Binding to tumor cell antigens in the form of polypeptides or polypeptide complexes that do not possess It is at least 120 times higher compared to affinity. In some embodiments, polypeptides Alternatively, polypeptide complexes have weak binding affinity to tumor cell antigens and possess both P1 and L1. Binding affinity to tumor cell antigens in the form of polypeptides or polypeptide complexes that do not bind to the antigen. It is at least 1000 times higher compared to sex.

[0045] In some embodiments, the polypeptide or polypeptide complex is tumor-specific in L1. Tumor cells containing polypeptides or polypeptide complexes cleaved by specific proteases The binding affinity to tumor cell antigens is weaker compared to the binding affinity to antigens. In this embodiment, the polypeptide or polypeptide complex binds to tumor cell antigens. Polypeptides with weak affinity and L1 cleaved by tumor-specific proteases or The binding affinity of the polypeptide complex to tumor cell antigens is at least five times higher. In some embodiments, the polypeptide or polypeptide complex is opposed to tumor cell antigens. Polypeptides with weak binding affinity and L1 cleavage by tumor-specific proteases Compared to the binding affinity of the d or polypeptide complex to tumor cell antigens, at least 8 It is twice as high. In some embodiments, the polypeptide or polypeptide complex is used in tumor cells The antigen has weak binding affinity, and L1 is cleaved by tumor-specific proteases. The binding affinity of the lipeptide or polypeptide complex to tumor cell antigens is low compared to the binding affinity of the lipeptide or polypeptide complex to tumor cell antigens. At least 10 times higher. In some embodiments, polypeptides or polypeptide complexes are Furthermore, it has weak binding affinity to tumor cell antigens, and L1 is cleaved by tumor-specific proteases. The binding affinity and ratio of the polypeptide or polypeptide complex to tumor cell antigens. It is at least 15 times higher by comparison. In some embodiments, polypeptides or polypeptides The complex has weak binding affinity to tumor cell antigens, and L1 is linked to tumor-specific proteases. Binding of more cleaved polypeptides or polypeptide complexes to tumor cell antigens It is at least 20 times higher compared to affinity. In some embodiments, polypeptide or Polypeptide complexes have weak binding affinity to tumor cell antigens, and L1 is tumor-specific antigen. The polypeptide or polypeptide complex cleaved by the thase is used as a tumor cell antigen. It is at least 25 times higher compared to the binding affinity for the opposite. In some embodiments, polyp The cytoplasm or polypeptide complex has weak binding affinity to tumor cell antigens, and L1 is tumor Tumors of polypeptides or polypeptide complexes cleaved by specific proteases It is at least 30 times higher compared to the binding affinity to the cell antigen. In some embodiments, Polypeptides or polypeptide complexes have weak binding affinity to tumor cell antigens. L1 is a polypeptide or polypeptide complex that is cleaved by a tumor-specific protease. The binding affinity to the combined tumor cell antigen is at least 35 times higher. In application, polypeptides or polypeptide complexes have binding affinity to tumor cell antigens. Polypeptides or polypropylenes with weak properties, where L1 is cleaved by tumor-specific proteases. The binding affinity of the peptide complex to tumor cell antigens is at least 40 times higher. In some embodiments, the polypeptide or polypeptide complex is effective against tumor cell antigens. Polypeptides with weak binding affinity and L1 cleavage by tumor-specific proteases. Or at least 45 compared to the binding affinity of the polypeptide complex to tumor cell antigens. It is twice as high. In some embodiments, the polypeptide or polypeptide complex is used in tumor cells The antigen has weak binding affinity, and L1 is cleaved by tumor-specific proteases. The binding affinity of the lipeptide or polypeptide complex to tumor cell antigens is low compared to the binding affinity of the lipeptide or polypeptide complex to tumor cell antigens. At least 50 times higher. In some embodiments, polypeptides or polypeptide complexes are Furthermore, it has weak binding affinity to tumor cell antigens, and L1 is cleaved by tumor-specific proteases. The binding affinity and ratio of the polypeptide or polypeptide complex to tumor cell antigens. It is at least 55 times higher by comparison. In some embodiments, polypeptides or polypeptides The complex has weak binding affinity to tumor cell antigens, and L1 is linked to tumor-specific proteases. Binding of more cleaved polypeptides or polypeptide complexes to tumor cell antigens It is at least 60 times higher compared to affinity. In some embodiments, polypeptide or Polypeptide complexes have weak binding affinity to tumor cell antigens, and L1 is tumor-specific antigen. The polypeptide or polypeptide complex cleaved by the thase is used as a tumor cell antigen. It is at least 65 times higher compared to the binding affinity for the opposite. In some embodiments, polyp The cytoplasm or polypeptide complex has weak binding affinity to tumor cell antigens, and L1 is tumor Tumors of polypeptides or polypeptide complexes cleaved by specific proteases It is at least 70 times higher compared to the binding affinity to the cell antigen. In some embodiments, Polypeptides or polypeptide complexes have weak binding affinity to tumor cell antigens. L1 is a polypeptide or polypeptide complex that is cleaved by a tumor-specific protease. The binding affinity to the tumor cell antigen of the combined organism is at least 75 times higher. In application, polypeptides or polypeptide complexes have binding affinity to tumor cell antigens. Polypeptides or polypropylenes with weak properties, where L1 is cleaved by tumor-specific proteases. The binding affinity of the peptide complex to tumor cell antigens is at least 80 times higher. In some embodiments, the polypeptide or polypeptide complex is effective against tumor cell antigens. Polypeptides with weak binding affinity and L1 cleavage by tumor-specific proteases. Or at least 85 compared to the binding affinity of the polypeptide complex to tumor cell antigens. It is twice as high. In some embodiments, the polypeptide or polypeptide complex is used in tumor cells The antigen has weak binding affinity, and L1 is cleaved by tumor-specific proteases. The binding affinity of the lipeptide or polypeptide complex to tumor cell antigens is low compared to the binding affinity of the lipeptide or polypeptide complex to tumor cell antigens. At least 90 times higher. In some embodiments, polypeptides or polypeptide complexes are Furthermore, it has weak binding affinity to tumor cell antigens, and L1 is cleaved by tumor-specific proteases. The binding affinity and ratio of the polypeptide or polypeptide complex to tumor cell antigens. It is at least 95 times higher by comparison. In some embodiments, polypeptides or polypeptides The complex has weak binding affinity to tumor cell antigens, and L1 is linked to tumor-specific proteases. Binding of more cleaved polypeptides or polypeptide complexes to tumor cell antigens It is at least 100 times higher compared to affinity. In some embodiments, polypeptides also Polypeptide complexes have weak binding affinity to tumor cell antigens, and L1 is tumor-specific Tumor cell antigens of polypeptides or polypeptide complexes cleaved by rotease It is at least 120 times higher compared to the binding affinity to [the target]. In some embodiments, poly The peptide or polypeptide complex has a weak binding affinity for tumor cell antigens, and L1 is The IFNγ-releasing T cell activity of an isolated polypeptide or polypeptide complex that has been cleaved by a tumor-specific protease is at least 1000-fold higher compared to the binding affinity for tumor cell antigens.

[0046] In some embodiments, the polypeptide or polypeptide complex has an elevated EC in an IFNγ-releasing T cell activation assay compared to the IFNγ-releasing T cell activity of an isolated polypeptide or polypeptide complex that has neither P1 nor L1 The IFNγ-releasing T cell activity of an isolated polypeptide or polypeptide complex that has neither P1 nor L1 in the EC in the IFNγ-releasing T cell activation assay 50 and the E in the IFNγ-releasing T cell activation assay C​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​ The peptide or polypeptide complex has an increased EC50 in the IFNγ release T cell activation assay, and is at least 40-fold higher than that of a polypeptide or polypeptide complex having neither P1 nor L1 in the IFNγ release T cell activation assay in the form of. In some embodiments, the peptide or polypeptide complex has an increased EC50 in the IFNγ release 50 T cell activation assay, and is at least 50-fold higher than that of a polypeptide or polypeptide complex having neither P1 nor L1 in the IFNγ release T cell activation assay in the form of. In some embodiments, the peptide or polypeptide complex has an increased EC50 in the IFNγ release 50 T cell activation assay, and is at least 60-fold higher than that of a polypeptide or polypeptide complex having neither P1 nor L1 in the IFNγ release T cell activation assay in the form of. In some embodiments, the peptide or polypeptide complex has an increased EC50 in the IFNγ release 50 T cell activation assay, and is at least 70-fold higher than that of a polypeptide or polypeptide complex having neither P1 nor L1 in the IFNγ release T cell activation assay 50 in the form of. In some embodiments, the peptide or polypeptide complex has an increased EC 50 in the IFNγ release T cell activation assay, and is at least 80-fold higher than that of a polypeptide or polypeptide complex having neither P1 nor L1 in the IFNγ release T cell activation assay 50 in the form of. In some embodiments, the peptide or polypeptide complex has an increased EC50 in the IFNγ release T cell activation assay, and is at least 80-fold higher than that of a polypeptide or polypeptide complex having neither P1 nor L1 in the IFNγ release T cell activation assay 50 in the form of. In some embodiments, the peptide or polypeptide complex has an increased EC50 in the IFNγ release T cell activation assay, and is at least 80-fold higher than that of a polypeptide or polypeptide complex having neither P1 nor L1 in the IFNγ release T cell activation assay in the form of. In some embodiments, the peptide or polypeptide complex has an increased EC50 in the IFNγ release 50 T cell activation assay, and is at least 80-fold higher than that of a polypeptide or polypeptide complex having neither P1 nor L's in the IFNγ release T cell activation assay in the form of. In some embodiments, the peptide or polypeptide complex has an increased EC50 in the IFNγ release 50 T cell activation assay, and is at least 80-fold In the embodiment, the polypeptide or polypeptide complex activates IFNγ-releasing T cells. E-commerce in Issay 50 The number is rising, and polypeptides that do not have either P1 or L1 or EC in IFNγ-releasing T cell activation assays of polypeptide complex morphology 50 Compare and at least 90 times higher. In some embodiments, polypeptide or polypeptide The complex is used in IFNγ-releasing T cell activation assays. 50 P1 is rising, and P1 is also IFNγ-releasing T cells in the form of polypeptides or polypeptide complexes that do not have L1 EC in activation assays 50 It is at least 100 times higher compared to several implementations. In this state, polypeptides or polypeptide complexes are used in IFNγ-releasing T cell activation assays. EC in 50 The number of polypeptides or polypeptides that do not have either P1 or L1 is increasing. EC in IFNγ-releasing T cell activation assays of the morphology of plutide complexes 50 Compared to less Even without it, it's 1000 times more expensive.

[0047] In some embodiments, the polypeptide or polypeptide complex is tumor-specific in L1. IFNγ of polypeptides or polypeptide complexes cleaved by the target protease EC in releasing T cell activation assay 50 Compared to IFNγ-releasing T cell activation EC in Sey 50 The amount is increasing. In some embodiments, polypeptides or poly The lipeptide complex is used in IFNγ-releasing T cell activation assays. 50 It is rising Furthermore, L1 is cleaved by tumor-specific proteases, which are polypeptides or polypeptides. EC in the IFNγ-releasing T cell activation assay of the EC complex 50 Compared to at least 1 0 times higher. In some embodiments, the polypeptide or polypeptide complex is IFN EC in γ-releasing T cell activation assay 50 The levels are rising, and L1 is tumor-specific protein IFNγ-releasing T cells of polypeptides or polypeptide complexes cleaved by ase EC in cellular activation assays 50 It is at least 20 times higher compared to several implementations. In this state, polypeptides or polypeptide complexes are used in IFNγ-releasing T cell activation assays. EC in 50 The levels are elevated, and L1 is being cleaved by tumor-specific proteases. E in IFNγ-releasing T cell activation assays of polypeptides or polypeptide complexes C 50 It is at least 30 times higher compared to. In some embodiments, polypeptide or Polypeptide complexes are used in IFNγ-releasing T cell activation assays. 50 It is rising Furthermore, L1 is cleaved by tumor-specific proteases, and polypeptides or polypeptides EC in IFNγ-releasing T cell activation assay of tide complex 50 Compared to at least 40 times higher. In some embodiments, the polypeptide or polypeptide complex is IF EC in Nγ-releasing T cell activation assay 50 The levels are rising, and L1 is tumor-specific protein IFNγ release of polypeptides or polypeptide complexes cleaved by thease EC in cell activation assays 50 It is at least 50 times higher compared to several implementations. Morphologically, polypeptides or polypeptide complexes are IFNγ-releasing T cell activation aggregates. EC in I 50 The levels are elevated, and L1 is being cleaved by tumor-specific proteases. In an IFNγ-releasing T cell activation assay of polypeptides or polypeptide complexes EC 50 It is at least 60 times higher compared to. In some embodiments, polypeptides also Polypeptide complexes are used in IFNγ-releasing T cell activation assays. 50 It is rising L1 is cleaved by tumor-specific proteases, and polypeptides or polypeptides EC in the IFNγ-releasing T cell activation assay of the plutide complex 50 Compared to It is also 70 times higher. In some embodiments, the polypeptide or polypeptide complex is I EC in FNγ-releasing T cell activation assay 50 The levels are rising, and L1 is tumor-specific IFNγ release of polypeptides or polypeptide complexes cleaved by rotease EC in T cell activation assays 50 It is at least 80 times higher compared to several actual In the application form, polypeptides or polypeptide complexes enhance IFNγ-releasing T cell activation. EC in Sey 50 The levels are elevated, and L1 is cleaved by tumor-specific proteases. In an IFNγ-releasing T cell activation assay of polypeptides or polypeptide complexes ru EC 50 It is at least 90 times higher compared to. In some embodiments, polypeptides Alternatively, polypeptide complexes are used in IFNγ-releasing T cell activation assays. 50 rising L1 is cleaved by tumor-specific proteases, and polypeptides or poly EC in IFNγ-releasing T cell activation assay of peptide complexes 50 Compared to less Both are 100 times higher. In some embodiments, polypeptides or polypeptide complexes are , EC in IFNγ-releasing T cell activation assay 50 The levels are rising, and L1 is tumor-specific. IFNγ of polypeptides or polypeptide complexes cleaved by the target protease EC in releasing T cell activation assay 50 It is at least 1000 times higher compared to [the other].

[0048] In some embodiments, the polypeptide or polypeptide complex has both P1 and L1. In T cell lysis assays of isolated polypeptides or polypeptide complexes that have not been isolated. keru EC 50 In comparison, EC in IFNγ-releasing T cell lysis assay 50 It is rising In some embodiments, polypeptides or polypeptide complexes are used to lyse T cells. EC in assays 50 The number of polypeptides that do not have either P1 or L1 is increasing, and EC in the form of polypeptide complexes in T cell lysis assays 50 Compared to It is also 10 times higher. In some embodiments, the polypeptide or polypeptide complex is T EC in cell lysis assays 50 Polypeptide, which does not have P1 or L1, is on the rise. EC in T cell lysis assays in the form of tide or polypeptide complexes 50 Compared At least 20 times higher. In some embodiments, polypeptides or polypeptide complexes. The body is involved in EC in T cell lysis assays. 50 It is rising and does not have P1 or L1. EC in T cell lysis assays in the form of polypeptides or polypeptide complexes 50 and It is at least 30 times higher by comparison. In some embodiments, polypeptides or polypept Tydos complexes are involved in EC in T cell lysis assays. 50 It is rising, and both P1 and L1 are E in T cell lysis assays in the form of polypeptides or polypeptide complexes that are not present. C 50 It is at least 40 times higher compared to. In some embodiments, polypeptide or Polypeptide complexes are used in EC in T cell lysis assays. 50 It is rising, and P1 is also L In T cell lysis assays that do not contain 1 polypeptide or polypeptide complex EC 50 It is at least 50 times higher compared to. In some embodiments, polypeptide The ion or polypeptide complex is used in EC in T cell lysis assays. 50 It is rising, T cell lysis in the form of polypeptides or polypeptide complexes that do not possess either P1 or L1 E-commerce in Issay 50 It is at least 60 times higher compared to that. In some embodiments, Lipeptide or polypeptide complexes in T cell lysis assays 50 It is rising Furthermore, the T-cells are in the form of polypeptides or polypeptide complexes that do not possess either P1 or L1. EC in cytolysis assays 50 It is at least 70 times higher compared to several embodiments. So, polypeptides or polypeptide complexes in T cell lysis assays 50 The amount is increasing, and the shape of polypeptides or polypeptide complexes that do not have either P1 or L1 is increasing. EC in T cell lysis assay 50 It is at least 80 times higher compared to several In the embodiment, the polypeptide or polypeptide complex is used in the T cell lysis assay. EC 50 The number of polypeptides that do not have either P1 or L1, or polypeptide complexes, are increasing. EC in a T cell lysis assay of aggregative morphology 50 It is at least 90 times higher compared to [another factor]. In some embodiments, the polypeptide or polypeptide complex is used in a T cell lysis assay. EC in 50 The number of polypeptides or polypeptides that do not have either P1 or L1 is increasing. EC in T cell lysis assays of the morphology of plutide complexes 50 Compared to at least 100 It is twice as high. In some embodiments, polypeptides or polypeptide complexes are T cell lysate. EC in solution assays 50 The number is rising, and polypeptides that do not possess either P1 or L1 are increasing. or polypeptide complex morphology in T cell lysis assays 50 Compared to less Both are 1000 times higher.

[0049] In some embodiments, the polypeptide or polypeptide complex is tumor-specific in L1. T cell lysis of polypeptides or polypeptide complexes cleaved by specific proteases EC in solution assays 50 In comparison with EC in T cell lysis assays 50 It is rising In some embodiments, the polypeptide or polypeptide complex is used to dissolve T cells. EC in solution assays 50 The levels are elevated, and L1 is cleaved by tumor-specific proteases. EC5 in T cell lysis assay of polypeptides or polypeptide complexes It is at least 10 times higher compared to 0. In some embodiments, polypeptides or poly The peptide complex is used in EC in T cell lysis assays. 50The levels are rising, and L1 is tumor-specific. T cells containing polypeptides or polypeptide complexes cleaved by heteroproteases EC in dissolution assays 50 It is at least 20 times higher compared to that. In some embodiments Polypeptides or polypeptide complexes are involved in EC in T cell lysis assays. 50 but The level is elevated, and L1 is cleaved by tumor-specific proteases in polypeptides or EC in T cell lysis assay of polypeptide complexes 50 Compared to that, at least 30 times High. In some embodiments, polypeptides or polypeptide complexes are used to lyse T cells. EC in assays 50 The levels are elevated, and L1 is cleaved by tumor-specific proteases. EC in T cell lysis assays of polypeptides or polypeptide complexes 50 It is at least 40 times higher compared to [another method]. In some embodiments, polypeptides or polypeptides The plutide complex is used in EC in T cell lysis assays. 50 The levels are rising, and L1 is tumor-specific. T cell lysis of polypeptides or polypeptide complexes cleaved by specific proteases EC in solution assays 50 It is at least 50 times higher compared to [the other method]. In some embodiments, it is at least 50 times higher. Polypeptides or polypeptide complexes are used in EC in T cell lysis assays. 50 Up The L1 molecule is elevated, and the polypeptide or poly(L1) is cleaved by a tumor-specific protease. EC in T cell lysis assay of lipeptide complexes 50 Compared to that, it is at least 60 times higher i. In some embodiments, polypeptides or polypeptide complexes are used to lyse T cells. E-commerce in Issay 50The levels are elevated, and L1 is cleaved by tumor-specific proteases. EC in T cell lysis assays of polypeptides or polypeptide complexes 50 and It is at least 70 times higher by comparison. In some embodiments, polypeptides or polypept Tydos complexes are involved in EC in T cell lysis assays. 50 The levels are elevated, and L1 is tumor-specific. T cell lysis of polypeptides or polypeptide complexes cleaved by proteases EC in assays 50 It is at least 80 times higher compared to. In some embodiments, Polypeptides or polypeptide complexes are used in EC in T cell lysis assays. 50 rising L1 is cleaved by tumor-specific proteases, and polypeptides or poly EC in T cell lysis assay of peptide complexes 50 Compared to that, it is at least 90 times higher In some embodiments, polypeptides or polypeptide complexes enhance T cell lysis. EC in Sey 50 The levels are elevated, and L1 is cleaved by tumor-specific proteases. EC in T cell lysis assays of polypeptides or polypeptide complexes 50 and comparison It is at least 100 times higher in comparison. In some embodiments, polypeptides or polypept Tydos complexes are involved in EC in T cell lysis assays. 50 The levels are elevated, and L1 is tumor-specific. T cell lysis of polypeptides or polypeptide complexes cleaved by proteases EC in assays 50 It is at least 1000 times higher compared to [the other].

[0050] In some embodiments, polypeptides or polypeptide complexes P2-L2-A2- A1-L1-P1-H1 (Formula Ia) is a formula that does not have P1, L1, P2, or L2. The binding parent of isolated polypeptides or polypeptide complexes of Ia to tumor cell antigens Compared to the ion type, it has weaker binding affinity to tumor cell antigens. In some embodiments, Lipeptide or polypeptide complex P2-L2-A2-A1-L1-P1-H1 (Formula I a) has a weak binding affinity to tumor cell antigens and possesses P1, L1, P2, or L2. For tumor cell antigens in the form of polypeptides or polypeptide complexes of formula Ia that are not present. It is at least 10 times higher compared to binding affinity. In some embodiments, polypeptides Alternatively, the polypeptide complex P2-L2-A2-A1-L1-P1-H1 (formula Ia) is a tumor Formula I has weak binding affinity to cell antigens and does not possess P1, L1, P2, or L2. The binding affinity of a polypeptide or polypeptide complex to tumor cell antigens and It is at least 50 times higher by comparison. In some embodiments, polypeptides or polypept The cytoplasmic complex P2-L2-A2-A1-L1-P1-H1 (formula Ia) is used against tumor cell antigens. Polypeptides of formula Ia that have weak binding affinity and do not have P1, L1, P2, or L2 The binding affinity to tumor cell antigens in the form of tide or polypeptide complexes is low compared to other forms. At least 75 times higher. In some embodiments, polypeptide or polypeptide complex P 2-L2-A2-A1-L1-P1-H1 (Formula Ia) has binding affinity to tumor cell antigens. Polypeptides of formula Ia that have weak properties and do not have P1, L1, P2, or L2 Compared to the binding affinity of the lipeptide complex to tumor cell antigens, at least 100 Twice as high. In some embodiments, polypeptides or polypeptide complexes P2-L2- A2-A1-L1-P1-H1 (Formula Ia) has weak binding affinity to tumor cell antigens. polypeptides of formula Ia that do not have P1, L1, P2, or L2 The binding affinity to tumor cell antigens in the form of the complex is at least 120 times higher. In some embodiments, polypeptide or polypeptide complex P2-L2-A2-A1 -L1-P1-H1 (Formula Ia) has weak binding affinity to tumor cell antigens, and P1, L1 Polypeptides or polypeptide complexes of formula Ia that do not have P2 or L2. It is at least 200 times higher compared to the binding affinity of the tumor cell antigen. In terms of application form, polypeptides or polypeptide complexes P2-L2-A2-A1-L1-P 1-H1 (formula Ia) has weak binding affinity to tumor cell antigens, and P1, L1, P2, and or tumor cells in the form of polypeptides or polypeptide complexes of formula Ia that do not have L2 The binding affinity is at least 300 times higher compared to the binding affinity to the cytoplasmic antigen. In some embodiments, , polypeptide or polypeptide complex P2-L2-A2-A1-L1-P1-H1( Formula Ia) has weak binding affinity to tumor cell antigens, and P1, L1, P2, or L2 For tumor cell antigens in the form of polypeptides or polypeptide complexes of formula Ia that do not have formula Ia It is at least 400 times higher compared to the binding affinity. In some embodiments, the polyp The cytoplasmic or polypeptide complex P2-L2-A2-A1-L1-P1-H1 (formula Ia) is , has weak binding affinity to tumor cell antigens and does not possess P1, L1, P2, or L2. The binding parent of the polypeptide or polypeptide complex of formula Ia to tumor cell antigens It is at least 500 times higher compared to the saturation. In some embodiments, polypeptides or The polypeptide complex P2-L2-A2-A1-L1-P1-H1 (formula Ia) is found in tumor cells. Formula Ia has weak binding affinity to the antigen and does not have P1, L1, P2, or L2. Comparison with the binding affinity of polypeptides or polypeptide complexes to tumor cell antigens. And it is at least 600 times higher. In some embodiments, polypeptides or polypeptides The complex P2-L2-A2-A1-L1-P1-H1 (formula Ia) is effective against tumor cell antigens. Polypeptides of formula Ia that have weak binding affinity and do not have P1, L1, P2, or L2 The binding affinity to tumor cell antigens in the form of a nucleotide or polypeptide complex is lower compared to other forms. Both are 700 times higher. In some embodiments, polypeptide or polypeptide complex P 2-L2-A2-A1-L1-P1-H1 (Formula Ia) has binding affinity to tumor cell antigens. Polypeptides of formula Ia that have weak properties and do not have P1, L1, P2, or L2 Compared to the binding affinity to tumor cell antigens in the form of a lipeptide complex, at least 800 Twice as high. In some embodiments, polypeptides or polypeptide complexes P2-L2- A2-A1-L1-P1-H1 (Formula Ia) has weak binding affinity to tumor cell antigens. polypeptides of formula Ia that do not have P1, L1, P2, or L2 The binding affinity to tumor cell antigens in the form of a complex is at least 900 times higher. In some embodiments, polypeptide or polypeptide complex P2-L2-A2-A1 -L1-P1-H1 (Formula Ia) has weak binding affinity to tumor cell antigens, and P1, L1 Polypeptides or polypeptide complexes of formula Ia that do not have P2 or L2. It is at least 1000 times higher than the binding affinity to tumor cell antigens of the state. In one embodiment, polypeptides or polypeptide complexes P2-L2-A2-A1-L1- P1-H1 (Formula Ia) has weak binding affinity to tumor cell antigens, and P1, L1, P2, Or tumors in the form of polypeptides or polypeptide complexes of formula Ia that do not have L2. It is at least 10,000 times higher than the binding affinity to the cell antigen.

[0051] In some embodiments, polypeptides or polypeptide complexes P2-L2-A2- A1-L1-P1-H1 (Formula Ia) is a molecule in which L1 and L2 are cleaved by tumor-specific proteases. The binding of polypeptides or polypeptide complexes of formula Ia to tumor cell antigens. Compared to affinity, the binding affinity to tumor cell antigens is weak. In some embodiments, Polypeptide or polypeptide complex P2-L2-A2-A1-L1-P1-H1 (formula) Ia) has weak binding affinity to tumor cell antigens, and L1 and L2 are tumor-specific proteas Tumor cell antigens of polypeptide or polypeptide complex of formula Ia cleaved by ze It is at least 10 times higher compared to the binding affinity to the polyp Plitter or polypeptide complex P2-L2-A2-A1-L1-P1-H1 (Formula Ia) It has a weak binding affinity to tumor cell antigens, and L1 and L2 are tumor-specific proteases The cleaved polypeptide or polypeptide complex of formula Ia against tumor cell antigens The binding affinity is at least 50 times higher compared to other binding affinities. In some embodiments, polypeptides Alternatively, the polypeptide complex P2-L2-A2-A1-L1-P1-H1 (formula Ia) is a tumor It has weak binding affinity to tumor cell antigens, and L1 and L2 are cleaved by tumor-specific proteases. The binding of polypeptides or polypeptide complexes of formula Ia to tumor cell antigens. It is at least 75 times higher compared to affinity. In some embodiments, polypeptide or The polypeptide complex P2-L2-A2-A1-L1-P1-H1 (formula Ia) is found in tumor cells. The binding affinity to the antigen is weak, and L1 and L2 are cleaved by tumor-specific proteases. Binding affinity of polypeptides or polypeptide complexes of formula Ia to tumor cell antigens It is at least 100 times higher compared to [another method]. In some embodiments, polypeptides or poly The peptide complex P2-L2-A2-A1-L1-P1-H1 (formula Ia) is a tumor cell antigen. It has weak binding affinity to and L1 and L2 are cleaved by tumor-specific proteases. Binding affinity and ratio of polypeptides or polypeptide complexes of formula Ia to tumor cell antigens It is at least 120 times higher in comparison. In some embodiments, polypeptides or polypept The cytoplasmic complex P2-L2-A2-A1-L1-P1-H1 (formula Ia) is used against tumor cell antigens. Formula I has weak binding affinity and L1 and L2 are cleaved by tumor-specific proteases. Compared to the binding affinity of polypeptide a or polypeptide complex to tumor cell antigens. and at least 200 times higher. In some embodiments, polypeptide or polypeptide The complex P2-L2-A2-A1-L1-P1-H1 (Formula Ia) is effective against tumor cell antigens. Formula Ia has weak binding affinity, and L1 and L2 are cleaved by tumor-specific proteases. The binding affinity of polypeptides or polypeptide complexes to tumor cell antigens is small. At least 300 times higher. In some embodiments, polypeptides or polypeptide complexes are used. Body P2-L2-A2-A1-L1-P1-H1 (Formula Ia) binds to tumor cell antigens. Polypodium Ia, which has weak affinity and whose L1 and L2 are cleaved by tumor-specific proteases. Less than the binding affinity of the peptide or polypeptide complex to tumor cell antigens. Both are 400 times higher. In some embodiments, polypeptide or polypeptide complex P 2-L2-A2-A1-L1-P1-H1 (Formula Ia) has binding affinity to tumor cell antigens. Polypeptide of formula Ia, which has weak properties and is cleaved by tumor-specific proteases at L1 and L2. Compared to the binding affinity of the thiosulfate or polypeptide complex to tumor cell antigens, at least 500 times higher. In some embodiments, polypeptide or polypeptide complex P2- L2-A2-A1-L1-P1-H1 (Formula Ia) has binding affinity to tumor cell antigens. A polypeptide of formula Ia, in which L1 and L2 are weakly cleaved by tumor-specific proteases. Or at least 60 compared to the binding affinity of the polypeptide complex to tumor cell antigens. 0 times higher. In some embodiments, polypeptide or polypeptide complex P2-L2 -A2-A1-L1-P1-H1 (Formula Ia) has a weak binding affinity to tumor cell antigens. , a polypeptide of formula Ia in which L1 and L2 are cleaved by a tumor-specific protease This is at least 700 times greater than the binding affinity of the polypeptide complex to tumor cell antigens. High. In some embodiments, polypeptide or polypeptide complex P2-L2-A 2-A1-L1-P1-H1 (Formula Ia) has weak binding affinity to tumor cell antigens, L Polypeptide of formula Ia or poly(I) 1 and L2 are cleaved by tumor-specific proteases. The binding affinity of the lipeptide complex to tumor cell antigens is at least 800 times higher. In some embodiments, polypeptides or polypeptide complexes P2-L2-A2- A1-L1-P1-H1 (Formula Ia) has weak binding affinity to tumor cell antigens, and L1 and L2 is cleaved by a tumor-specific protease. The binding affinity of the ptide complex to tumor cell antigens is at least 900 times higher. In some embodiments, polypeptide or polypeptide complex P2-L2-A2-A1 -L1-P1-H1 (Formula Ia) has weak binding affinity to tumor cell antigens, and L1 and L2 polypeptides of formula Ia that are cleaved by tumor-specific proteases The binding affinity of the complex to tumor cell antigens is at least 1000 times higher. In several embodiments, polypeptides or polypeptide complexes P2-L2-A2-A1- L1-P1-H1 (Formula Ia) has weak binding affinity to tumor cell antigens, and L1 and L2 polypeptides of formula Ia or polypeptides cleaved by tumor-specific proteases The binding affinity of the complex to tumor cell antigens is at least 10,000 times higher.

[0052] In some embodiments, polypeptides or polypeptide complexes P2-L2-A2- A1-L1-P1-H1 (Formula Ia) is a formula that does not have P1, L1, P2, or L2. Ia isolated polypeptide or polypeptide complex IFNγ-releasing T cell activation E-commerce in Issay 50 In comparison, EC5 in the IFNγ-releasing T cell activation assay 0 is increasing. In some embodiments, polypeptide or polypeptide complex P 2-L2-A2-A1-L1-P1-H1 (Formula Ia) is an IFNγ-releasing T cell activation enhancer. EC in Sey 50 The expression is rising and does not have P1, L1, P2, or L2. Ia polypeptide or polypeptide complex form IFNγ-releasing T cell activation assay EC in I 50 It is at least 10 times higher compared to. In some embodiments, polyp Plitter or polypeptide complex P2-L2-A2-A1-L1-P1-H1 (Formula Ia) This refers to EC in IFNγ-releasing T cell activation assays. 50 P1, L1, Forms of polypeptides or polypeptide complexes of formula Ia that do not have P2 or L2 EC in IFNγ-releasing T cell activation assay 50 Compared to that, it is at least 50 times higher In some embodiments, polypeptides or polypeptide complexes P2-L2-A2- A1-L1-P1-H1 (Formula Ia) is used in EC in IFNγ-releasing T cell activation assays. 50 The amount is increasing, and the amount of the polypeptide of formula Ia that does not have P1, L1, P2, or L2 EC5 in IFNγ-releasing T cell activation assays in the form of a polypeptide or polypeptide complex It is at least 75 times higher compared to 0. In some embodiments, polypeptides or poly The peptide complex P2-L2-A2-A1-L1-P1-H1 (formula Ia) releases IFNγ. EC in T cell activation assays 50 The numbers are rising, and P1, L1, P2, or L2 IFNγ-releasing T in the form of a polypeptide or polypeptide complex of formula Ia that does not have EC in cell activation assays 50It is at least 100 times higher compared to several actual In terms of application form, polypeptides or polypeptide complexes P2-L2-A2-A1-L1-P 1-H1 (Formula Ia) is used in IFNγ-releasing T cell activation assays. 50 It is rising Polypeptides of formula Ia that do not have P1, L1, P2, or L2 EC in IFNγ-releasing T cell activation assays of the morphology of plutide complexes 50 Compared to less At least 200 times higher. In some embodiments, polypeptides or polypeptide complexes are used. P2-L2-A2-A1-L1-P1-H1 (Formula Ia) activates IFNγ-releasing T cells EC in assays 50 It is rising and does not have P1, L1, P2, or L2 IFNγ-releasing T cell activation in the form of polypeptide or polypeptide complex of formula Ia E-commerce in Issay 50 It is at least 300 times higher compared to. In some embodiments, Polypeptide or polypeptide complex P2-L2-A2-A1-L1-P1-H1 (formula) Ia) EC in IFNγ-releasing T cell activation assays 50 P1 is rising, Polypeptides or polypeptide complexes of formula Ia that lack L1, P2, or L2. EC in IFNγ-releasing T cell activation assays 50 Compared to at least 40 0 times higher. In some embodiments, polypeptide or polypeptide complex P2-L2 -A2-A1-L1-P1-H1 (Formula Ia) is used in IFNγ-releasing T cell activation assays. keru EC 50 The value is rising, and the value of formula Ia does not have P1, L1, P2, or L2. In IFNγ-releasing T cell activation assays using the form of lipeptides or polypeptide complexes. ru EC 50 It is at least 500 times higher compared to [another factor]. In some embodiments, polypeptides Alternatively, the polypeptide complex P2-L2-A2-A1-L1-P1-H1 (formula Ia) is I EC in FNγ-releasing T cell activation assay 50 The following are rising: P1, L1, P2, Or IF in the form of a polypeptide or polypeptide complex of formula Ia that does not have L2. EC in Nγ-releasing T cell activation assay 50 It is at least 600 times higher compared to [another factor]. In some embodiments, polypeptide or polypeptide complex P2-L2-A2-A1 -L1-P1-H1 (Formula Ia) is used in EC in IFNγ-releasing T cell activation assays. 50 The amount is increasing, and polypeptides of formula Ia that do not have P1, L1, P2, or L2 are increasing. or polypeptide complex morphology in IFNγ-releasing T cell activation assays 50 and It is at least 700 times higher by comparison. In some embodiments, polypeptides or polyp The ptide complex P2-L2-A2-A1-L1-P1-H1 (formula Ia) is an IFNγ-releasing T EC in cell activation assays 50 It is rising, and P1, L1, P2, or L2 IFNγ-releasing T cells in the form of polypeptides or polypeptide complexes of formula Ia that do not have formula Ia EC in cellular activation assays 50 It is at least 800 times higher compared to several implementations. Morphologically, it refers to polypeptides or polypeptide complexes P2-L2-A2-A1-L1-P1 -H1 (Formula Ia) is used in IFNγ-releasing T cell activation assays. 50 It is rising Polypeptides of formula Ia that do not have P1, L1, P2, or L2 EC in IFNγ-releasing T cell activation assays of tide complex morphology 50 Compared to At least 900 times higher. In some embodiments, polypeptides or polypeptide complexes. P2-L2-A2-A1-L1-P1-H1 (Formula Ia) is an IFNγ-releasing T cell activation protein. E-commerce in Issay 50 It is rising and does not have P1, L1, P2, or L2. IFNγ-releasing T cell activation in the form of polypeptide or polypeptide complex of formula Ia EC in Sey 50 It is at least 1000 times higher compared to. In some embodiments, Polypeptide or polypeptide complex P2-L2-A2-A1-L1-P1-H1 (formula) Ia) EC in IFNγ-releasing T cell activation assays 50 P1 is rising, Polypeptides or polypeptide complexes of formula Ia that lack L1, P2, or L2. EC in IFNγ-releasing T cell activation assays 50 Compared to at least 10 It's 000 times more expensive.

[0053] In some embodiments, polypeptides or polypeptide complexes P2-L2-A2- A1-L1-P1-H1 (Formula Ia) is a molecule in which L1 and L2 are cleaved by tumor-specific proteases. In a T cell lysis assay of polypeptides or polypeptide complexes of formula Ia, ru EC 50 In comparison, the binding affinity to tumor cell antigens is increased. In terms of application form, polypeptides or polypeptide complexes P2-L2-A2-A1-L1-P 1-H1 (Formula Ia) is the EC in T cell lysis assays. 50 L1 and L are rising. 2 is a polypeptide of formula Ia that is cleaved by a tumor-specific protease. EC in T cell lysis assay of tide complex 50 It is at least 10 times higher compared to [another factor]. In some embodiments, polypeptide or polypeptide complex P2-L2-A2-A1 -L1-P1-H1 (Formula Ia) is used in EC in T cell lysis assays. 50 It is rising , a polypeptide of formula Ia in which L1 and L2 are cleaved by a tumor-specific protease EC in T cell lysis assay of polypeptide complexes 50 Compared to at least 50 Twice as high. In some embodiments, polypeptides or polypeptide complexes P2-L2- A2-A1-L1-P1-H1 (Formula Ia) is used in EC in T cell lysis assays. 50 Up The L1 and L2 cells are elevated, and the polyp of formula Ia is cleaved by tumor-specific proteases. EC in T cell lysis assays of peptides or polypeptide complexes 50 Compared to At least 75 times higher. In some embodiments, polypeptide or polypeptide complex P 2-L2-A2-A1-L1-P1-H1 (Formula Ia) is used in E cell lysis assays. C 50 Equation I is elevated, and L1 and L2 are cleaved by tumor-specific proteases. EC in T cell lysis assay of polypeptide or polypeptide complex a 50 and comparison It is at least 100 times higher in comparison. In some embodiments, polypeptides or polypept The T-cell complex P2-L2-A2-A1-L1-P1-H1 (formula Ia) is a T-cell lysing agent. EC in I 50 The levels are elevated, and L1 and L2 are cleaved by tumor-specific proteases. In T cell lysis assays of polypeptides or polypeptide complexes of formula Ia EC 50 It is at least 200 times higher compared to [another factor]. In some embodiments, polypeptides Alternatively, the polypeptide complex P2-L2-A2-A1-L1-P1-H1 (formula Ia) is T EC in cytolysis assays 50 The levels are elevated, and L1 and L2 are tumor-specific proteases. T cell lysis of polypeptides or polypeptide complexes of formula Ia that have been cleaved by EC in Sey 50 It is at least 300 times higher compared to. In some embodiments, Lipeptide or polypeptide complex P2-L2-A2-A1-L1-P1-H1 (Formula I a) EC in T cell lysis assays 50 The levels are elevated, and L1 and L2 are tumor-specific. A polypeptide or polypeptide complex of formula Ia that has been cleaved by a protease. EC in cell lysis assays 50 It is at least 400 times higher compared to several implementations. Morphologically, it refers to polypeptides or polypeptide complexes P2-L2-A2-A1-L1-P1 -H1 (Formula Ia) is used in EC in T cell lysis assays. 50 L1 and L2 are rising. polypeptides of formula Ia that are cleaved by tumor-specific proteases EC in T cell lysis assay of the EC complex 50 It is at least 500 times higher compared to [another factor]. In some embodiments, polypeptide or polypeptide complex P2-L2-A2-A1 -L1-P1-H1 (Formula Ia) is used in EC in T cell lysis assays. 50 It is rising , a polypeptide of formula Ia in which L1 and L2 are cleaved by a tumor-specific protease EC in T cell lysis assay of polypeptide complexes 50 Compared to at least 60 0 times higher. In some embodiments, polypeptide or polypeptide complex P2-L2 -A2-A1-L1-P1-H1 (Formula Ia) is used in EC in T cell lysis assays. 50 but The polymorphism of formula Ia is elevated, and L1 and L2 are cleaved by tumor-specific proteases. EC in T cell lysis assays of peptides or polypeptide complexes 50 Compared to less At least 700 times higher. In some embodiments, polypeptides or polypeptide complexes are used. The P2-L2-A2-A1-L1-P1-H1 (formula Ia) is used in T cell lysis assays. ru EC 50 The levels are elevated, and L1 and L2 are being cleaved by tumor-specific proteases. EC in T cell lysis assay of polypeptides or polypeptide complexes of formula Ia 50 It is at least 800 times higher compared to [another method]. In some embodiments, polypeptides or poly The peptide complex P2-L2-A2-A1-L1-P1-H1 (formula Ia) is used in T cell lysis. E-commerce in Issay 50 The levels are elevated, and L1 and L2 are being cut by tumor-specific proteases. In T cell lysis assays of polypeptides or polypeptide complexes of formula Ia that have been interrupted keru EC 50 It is at least 900 times higher compared to. In some embodiments, polypeptide The peptide complex P2-L2-A2-A1-L1-P1-H1 (formula Ia) is, EC in T cell lysis assay 50 The levels are rising, and L1 and L2 are tumor-specific proteas. T cell lysis of polypeptides or polypeptide complexes of formula Ia that have been cleaved by -ase. EC in assays50 It is at least 1000 times higher compared to. In some embodiments This refers to polypeptides or polypeptide complexes P2-L2-A2-A1-L1-P1-H1 (Formula Ia) shows the EC in T cell lysis assays. 50 The levels are rising, and L1 and L2 are tumors. polypeptides or polypeptide complexes of formula Ia that are cleaved by specific proteases. EC in the body's T cell lysis assay 50 It is at least 10,000 times higher compared to [the other].

[0054] In some embodiments, polypeptides or polypeptide complexes P2-L2-A2- A1-L1-P1-H1 (Formula Ia) is a single unit that does not have P1, L1, P2, or L2. EC in T cell lysis assay of separated polypeptides or polypeptide complexes 50 In comparison with EC in T cell lysis assays 50 It is rising. Several embodiments So, polypeptides or polypeptide complexes P2-L2-A2-A1-L1-P1-H 1 (Equation Ia) shows EC in T cell lysis assay. 50 P1, L1, P 2, or a polypeptide or polypeptide complex of formula Ia that does not have L2 EC in T cell lysis assay 50 It is at least 10 times higher compared to several implementations. Morphologically, it refers to polypeptides or polypeptide complexes P2-L2-A2-A1-L1-P1 -H1 (Formula Ia) is used in EC in T cell lysis assays. 50 P1, L1 Polypeptides or polypeptide complexes of formula Ia that do not have P2 or L2. EC in T cell lysis assay 50 It is at least 50 times higher compared to several In one embodiment, polypeptides or polypeptide complexes P2-L2-A2-A1-L1- P1-H1 (Formula Ia) is used in EC in T cell lysis assays. 50 P1 is rising, Polypeptides or polypeptide complexes of formula Ia that lack L1, P2, or L2. EC in morphological T cell lysis assays 50 It is at least 75 times higher compared to that. In one embodiment, polypeptide or polypeptide complex P2-L2-A2-A1-L 1-P1-H1 (Formula Ia) is used in EC in T cell lysis assays. 50 P is rising, 1, L1, P2, or isolated polypeptides that do not have L2 EC in T cell lysis assay of the complex 50 It is at least 100 times higher compared to In several embodiments, polypeptides or polypeptide complexes P2-L2-A2-A1- L1-P1-H1 (Formula Ia) is used in EC in T cell lysis assays. 50 It is rising, Isolated polypeptides or polypeptides that do not have P1, L1, P2, or L2 EC in T cell lysis assay of the EC complex 50 It is at least 200 times higher compared to [another factor]. In some embodiments, polypeptide or polypeptide complex P2-L2-A2-A1 -L1-P1-H1 (Formula Ia) is used in EC in T cell lysis assays. 50 It is rising , isolated polypeptides or polypeptides that do not have P1, L1, P2, or L2 EC in T cell lysis assay of tide complex 50 It is at least 300 times higher compared to [the other]. In some embodiments, polypeptides or polypeptide complex P2-L2-A2-A 1-L1-P1-H1 (Formula Ia) is used in EC in T cell lysis assays. 50 It is rising Isolated polypeptides or polypeptides that do not have P1, L1, P2, or L2 EC in T cell lysis assay of plutide complex 50 Compared to that, it is at least 400 times higher. In some embodiments, polypeptides or polypeptide complexes P2-L2-A2- A1-L1-P1-H1 (Formula Ia) is used in EC in T cell lysis assays. 50 It is rising Therefore, isolated polypeptides or poly(P1, L1, P2, or L2) EC in T cell lysis assay of peptide complexes 50 Compared to that, it is at least 500 times higher i. In some embodiments, polypeptides or polypeptide complex P2-L2-A2 -A1-L1-P1-H1 (Formula Ia) is used in EC in T cell lysis assays. 50 It is rising Isolated polypeptides or polypeptides that do not have P1, L1, P2, or L2 EC in T cell lysis assay of lipeptide complexes 50 Compared to that, at least 600 times High. In some embodiments, polypeptide or polypeptide complex P2-L2-A 2-A1-L1-P1-H1 (Formula Ia) is used in EC in T cell lysis assays. 50 rising Isolated polypeptides that do not have P1, L1, P2, or L2 EC in T cell lysis assay of polypeptide complexes 50 Compared to at least 700 Twice as high. In some embodiments, polypeptides or polypeptide complexes P2-L2- A2-A1-L1-P1-H1 (Formula Ia) is used in EC in T cell lysis assays. 50 Up Isolated polypeptides that are elevated and do not have P1, L1, P2, or L2 EC in T cell lysis assay of polypeptide complexes 50 Compared to at least 80 0 times higher. In some embodiments, polypeptide or polypeptide complex P2-L2 -A2-A1-L1-P1-H1 (Formula Ia) is used in EC in T cell lysis assays. 50 but The levels are elevated, and isolated polypeptides that do not have P1, L1, P2, or L2 are... or EC in T cell lysis assays of polypeptide complexes 50 Compared to at least 9 00 times higher. In some embodiments, polypeptide or polypeptide complex P2-L 2-A2-A1-L1-P1-H1 (Formula Ia) is used in EC in T cell lysis assays. 50 Isolated polypeptides that have elevated levels and lack P1, L1, P2, or L2 or EC in T cell lysis assays of polypeptide complexes 50 Compared to at least 1000 times higher. In some embodiments, polypeptide or polypeptide complex P2 -L2-A2-A1-L1-P1-H1 (Formula Ia) is used in EC in T cell lysis assays. 50 The levels are elevated, and isolated polypeptides that do not have P1, L1, P2, or L2 are elevated. EC in T cell lysis assays of tide or polypeptide complexes 50 Compared to less Both are 10,000 times higher.

[0055] In some embodiments, polypeptides or polypeptide complexes P2-L2-A2- A1-L1-P1-H1 (Formula Ia) is a molecule in which L1 and L2 are cleaved by tumor-specific proteases. In a T cell lysis assay of polypeptides or polypeptide complexes of formula Ia, ru EC 50 In comparison, the binding affinity to tumor cell antigens is increased. In terms of application form, polypeptides or polypeptide complexes P2-L2-A2-A1-L1-P 1-H1 (Formula Ia) is the EC in T cell lysis assays. 50 L1 and L are rising. 2 is a polypeptide of formula Ia that is cleaved by a tumor-specific protease. EC in T cell lysis assay of tide complex 50 It is at least 10 times higher compared to [another factor]. In some embodiments, polypeptide or polypeptide complex P2-L2-A2-A1 -L1-P1-H1 (Formula Ia) is used in EC in T cell lysis assays. 50 It is rising , a polypeptide of formula Ia in which L1 and L2 are cleaved by a tumor-specific protease EC in T cell lysis assay of polypeptide complexes 50 Compared to at least 50 Twice as high. In some embodiments, polypeptides or polypeptide complexes P2-L2- A2-A1-L1-P1-H1 (Formula Ia) is used in EC in T cell lysis assays. 50 Up The L1 and L2 cells are elevated, and the polyp of formula Ia is cleaved by tumor-specific proteases. EC in T cell lysis assays of peptides or polypeptide complexes 50 Compared to At least 75 times higher. In some embodiments, polypeptide or polypeptide complex P 2-L2-A2-A1-L1-P1-H1 (Formula Ia) is used in E cell lysis assays. C 50 Equation I is elevated, and L1 and L2 are cleaved by tumor-specific proteases. EC in T cell lysis assay of polypeptide or polypeptide complex a 50 and comparison It is at least 100 times higher in comparison. In some embodiments, polypeptides or polypept The T-cell complex P2-L2-A2-A1-L1-P1-H1 (formula Ia) is a T-cell lysing agent. EC in I 50 The levels are elevated, and L1 and L2 are cleaved by tumor-specific proteases. In T cell lysis assays of polypeptides or polypeptide complexes of formula Ia EC 50 It is at least 200 times higher compared to [another factor]. In some embodiments, polypeptides Alternatively, the polypeptide complex P2-L2-A2-A1-L1-P1-H1 (formula Ia) is T EC in cytolysis assays 50 The levels are elevated, and L1 and L2 are tumor-specific proteases. T cell lysis of polypeptides or polypeptide complexes of formula Ia that have been cleaved by EC in Sey 50 It is at least 300 times higher compared to. In some embodiments, Lipeptide or polypeptide complex P2-L2-A2-A1-L1-P1-H1 (Formula I a) EC in T cell lysis assays 50 The levels are elevated, and L1 and L2 are tumor-specific. A polypeptide or polypeptide complex of formula Ia that has been cleaved by a protease. EC in cell lysis assays 50 It is at least 400 times higher compared to several implementations. Morphologically, it refers to polypeptides or polypeptide complexes P2-L2-A2-A1-L1-P1 -H1 (Formula Ia) is used in EC in T cell lysis assays. 50 L1 and L2 are rising. polypeptides of formula Ia that are cleaved by tumor-specific proteases EC in T cell lysis assay of the EC complex 50 It is at least 500 times higher compared to [another factor]. In some embodiments, polypeptide or polypeptide complex P2-L2-A2-A1 -L1-P1-H1 (Formula Ia) is used in EC in T cell lysis assays. 50 It is rising , a polypeptide of formula Ia in which L1 and L2 are cleaved by a tumor-specific protease EC in T cell lysis assay of polypeptide complexes 50 Compared to at least 60 0 times higher. In some embodiments, polypeptide or polypeptide complex P2-L2 -A2-A1-L1-P1-H1 (Formula Ia) is used in EC in T cell lysis assays. 50 but The polymorphism of formula Ia is elevated, and L1 and L2 are cleaved by tumor-specific proteases. EC in T cell lysis assays of peptides or polypeptide complexes 50 Compared to less At least 700 times higher. In some embodiments, polypeptides or polypeptide complexes are used. The P2-L2-A2-A1-L1-P1-H1 (formula Ia) is used in T cell lysis assays. ru EC 50 The levels are elevated, and L1 and L2 are being cleaved by tumor-specific proteases. EC in T cell lysis assay of polypeptides or polypeptide complexes of formula Ia 50 It is at least 800 times higher compared to [another method]. In some embodiments, polypeptides or poly The peptide complex P2-L2-A2-A1-L1-P1-H1 (formula Ia) is used in T cell lysis. E-commerce in Issay 50 The levels are elevated, and L1 and L2 are being cut by tumor-specific proteases. In T cell lysis assays of polypeptides or polypeptide complexes of formula Ia that have been interrupted keru EC50 It is at least 900 times higher compared to. In some embodiments, polypeptide The peptide complex P2-L2-A2-A1-L1-P1-H1 (formula Ia) is, EC in T cell lysis assay 50 The levels are rising, and L1 and L2 are tumor-specific proteas. T cell lysis of polypeptides or polypeptide complexes of formula Ia that have been cleaved by -ase. EC in assays 50 It is at least 1000 times higher compared to. In some embodiments This refers to polypeptides or polypeptide complexes P2-L2-A2-A1-L1-P1-H1 (Formula Ia) shows the EC in T cell lysis assays. 50 The levels are rising, and L1 and L2 are tumors. polypeptides or polypeptide complexes of formula Ia that are cleaved by specific proteases. EC in the body's T cell lysis assay 50 It is at least 10,000 times higher compared to [the other].

[0056] Second antigen-recognition molecule (A2) In some embodiments, A2 comprises an antibody or antibody fragment. In terms of application form, the antibody or antibody fragment is a single-chain variable fragment, single domain The antibody comprises an antibody, Fab, or Fab'. In some embodiments, the antibody or its antibody f Ragment is a single-chain variable fragment (scFv) of a single-domain antibody derived from camelids, heavy Chain variable domain (VH domain), light chain variable domain (VL domain), or variable domain Contains (VHH). In some embodiments, the antibody or antibody fragment is Humanized or human. In some embodiments, A2 is Fab or Fab'. In some embodiments, Fab or Fab' is (a) Fab light chain polypeptide. and (b) comprising a Fab heavy chain polypeptide. In some embodiments, an antibody or The antibody fragment contains a PSMA-binding region.

[0057] In some embodiments, an antigen-binding fragment (Fab) that binds to PSMA or 'Fab' includes Fab light chain polypeptide chains and Fab heavy chain polypeptide chains. In some embodiments, the Fab light chain polypeptide includes Fab light chain variable domains. In some embodiments, the Fab heavy chain polypeptide includes Fab heavy chain variable domains. In this embodiment, the Fab heavy chain variable domain is at least 1, 2, or if disclosed in Table 2. or three complementarity-determining regions (CDRs), or substantially identical sequences to them (for example, Having at least 90%, 95%, 96%, 97%, 98%, or 99% sequence identity It includes a sequence. In some embodiments, the Fab light chain variable domain is disclosed in Table 2. at least one, two, or three complementarity determination regions (CDRs), or substantially relative to them. Similar sequences (for example, at least 90%, 95%, 96%, 97%, 98%, or It includes sequences with 99% sequence identity.

[0058] In some embodiments, the Fab heavy chain variable domain is at least one of the domains disclosed in Table 2. , 2 or 3 complementarity-determining regions (CDRs), or substantially identical sequences to them. (For example, at least 90%, 95%, 96%, 97%, 98%, or 99% of the sequence The Fab light chain variable domain includes a sequence having uniformity, and at least one of the sequences disclosed in Table 2 is included. , 2 or 3 complementarity-determining regions (CDRs), or substantially identical sequences to them. (For example, at least 90%, 95%, 96%, 97%, 98%, or 99% of the sequence Includes sequences that have uniformity.

[0059] [Table 2]

[0060] In some embodiments, Fab uses HC-CDR1 as the complementary determination region (CDR). This includes HC-CDR2 and HC-CDR3, as well as Fab's HC-CDR1 and HC-CD R2 and HC-CDR3 are sequence numbers 8 in HC-CDR1 and distribution in HC-CDR2. Column number 9, and in HC-CDR3, sequence number 10, are included, and Fab is LC as CDR. -Includes CDR1, LC-CDR2, and LC-CDR3, Fab's LC-CDR1, LC-CDR2 and LC-CDR3 are sequence numbers 11 and 11 respectively for LC-CDR1. DR2 includes sequence number 12, and LC-CDR3 includes sequence number 13. In the application configuration, Fab uses HC-CDR1 and HC-CDR as complementarity-determining regions (CDRs). 2, and HC-CDR3, including Fab's HC-CDR1, HC-CDR2, and H C-CDR3 is sequence number 8 in HC-CDR1, sequence number 9 in HC-CDR2, and HC-CDR3 includes sequence number 10, and the CDRs are HC-CDR1, HC-CDR2 , or containing 0-2 amino acid modifications in at least one of the HC-CDR3s, Fab This includes LC-CDR1, LC-CDR2, and LC-CDR3 as CDRs, Fa LC-CDR1, LC-CDR2, and LC-CDR3 of b are distributed in LC-CDR1 Column number 11, sequence number 12 in LC-CDR2, and sequence number 13 in LC-CDR3 The CD-R includes one of the following: LC-CDR1, LC-CDR2, or LC-CDR3. Each molecule contains 0 to 2 amino acid modifications.

[0061] In some embodiments, A2 is HC-CDR1 as the complementarity determination region (CDR), Includes HC-CDR2 and HC-CDR3, and A2 HC-CDR1, HC-CDR2 , and HC-CDR3 is sequence number 8 in HC-CDR1 and sequence number 8 in HC-CDR2. Number 9, and in HC-CDR3, sequence number 10 is included, and A2 is LC-CD as CDR. Includes R1, LC-CDR2, and LC-CDR3, and A2's LC-CDR1, LC-C DR2 and LC-CDR3 are sequence numbers 11 in LC-CDR1 and LC-CDR2. This includes sequence number 12, and sequence number 13 in LC-CDR3. In some embodiments, A2 has HC-CDR1, HC-CDR2, and as complementary determination regions (CDRs). Includes HC-CDR3, and A2 HC-CDR1, HC-CDR2, and HC-CDR3 This is represented by sequence number 8 in HC-CDR1, sequence number 9 in HC-CDR2, and HC-CD R3 includes sequence number 10, and the CDR is HC-CDR1, HC-CDR2, or HC - At least one of the CDR3s contains 0 to 2 amino acid modifications, and A2 is CDR This includes LC-CDR1, LC-CDR2, and LC-CDR3, and A2 LC-CDR 1. LC-CDR2 and LC-CDR3 are sequence numbers 11 in LC-CDR1, LC -CDR2 includes sequence number 12, and LC-CDR3 includes sequence number 13, and CDR is , LC-CDR1, LC-CDR2, or LC-CDR3, at least one of them, 0~ It contains two amino acid modifications.

[0062] In some embodiments, the Fab light chain polypeptide is configured according to the amino acid sequence of SEQ ID NO: 14. Includes a column. In some embodiments, the Fab light chain polypeptide is used for SEQ ID NO: 14 It contains an amino acid sequence having at least 80% sequence identity. In some embodiments, The Fab light chain polypeptide has at least 85% sequence identity with SEQ ID NO: 14. It contains an amino acid sequence. In some embodiments, the Fab light chain polypeptide is sequence number It contains an amino acid sequence that has at least 90% sequence identity with 14. In the application form, the Fab light chain polypeptide has at least 95% of the sequence of SEQ ID NO: 14. It contains an identical amino acid sequence. In some embodiments, the Fab light chain polypeptide It contains an amino acid sequence that has at least 99% sequence identity with SEQ ID NO: 14.

[0063] In some embodiments, the Fab heavy chain polypeptide is configured according to the amino acid sequence of SEQ ID NO: 15. Includes a column. In some embodiments, the Fab heavy chain polypeptide is used for SEQ ID NO: 15 It contains an amino acid sequence having at least 80% sequence identity. In some embodiments, The Fab heavy chain polypeptide has at least 85% sequence identity with SEQ ID NO: 15. It contains an amino acid sequence. In some embodiments, the Fab heavy chain polypeptide is sequence number It contains an amino acid sequence that has at least 90% sequence identity with 15. In the application form, the Fab heavy chain polypeptide has at least 95% of the sequence of SEQ ID NO: 15. It contains an amino acid sequence that is identical. In some embodiments, it is a Fab heavy chain polypeptide. It contains an amino acid sequence that has at least 99% sequence identity with SEQ ID NO: 15.

[0064] In some embodiments, the Fab light chain polypeptide A2 is a single-chain variable fragment of A1. It is bonded to the C-terminus of the Fab heavy chain of A2. In some embodiments, the Fab heavy chain of A2 The lipeptide is bound to the C-terminus of the single-chain variable fragment (scFv) of A1. In that embodiment, the Fab light chain polypeptide of A2 is a single-chain variable fragment (s It is bonded to the N-terminus of cFv. The d is bound to the N-terminus of the single-chain variable fragment (scFv) of A1. Several implementations Morphologically, the Fab heavy chain polypeptide of A2 is bound to the scFv heavy chain polypeptide of A1. In some embodiments, the Fab light chain polypeptide of A2 is the scFv heavy chain of A1. It is bound to the polypeptide. In some embodiments, the Fab heavy chain polypeptide of A2 is , is bound to the scFv light chain polypeptide of A1. In some embodiments, Fa of A2 The b light chain polypeptide is bound to the scFv light chain polypeptide of A1.

[0065] In some embodiments, A2 further includes P2 and L2, where P2 is coupled to A2. It contains a peptide, and L2 ligates A1 to P1 and tumor-specific protease It contains a substrate linking portion. In some embodiments, the Fab heavy chain polypeptide of A2 is L2 is bound to the scFv heavy chain polypeptide of A1, and L2 is bound to the Fab light chain polypeptide of A2. It is bonded to the Fab light chain polypeptide of A2. In some embodiments, the Fab light chain polypeptide of A1 is bonded to the sc Fv is bound to the heavy chain polypeptide, and L2 is bound to the Fab heavy chain polypeptide of A2. In some embodiments, the Fab heavy chain polypeptide of A2 is the scFv light chain polypeptide of A1. Binding to the peptide, L2 binds to the Fab light chain polypeptide of A2. In this embodiment, the Fab light chain polypeptide of A2 is bonded to the scFv light chain polypeptide of A1. Combined, L2 is bound to the Fab heavy chain polypeptide of A2.

[0066] In some embodiments, the Fab heavy chain polypeptide of A2 is the scFv heavy chain polypeptide of A1. Binding to the peptide, L2 binds to the Fab light chain polypeptide of A2. In this embodiment, the Fab light chain polypeptide of A2 is bonded to the scFv heavy chain polypeptide of A1. Combined, L2 is bonded to the Fab heavy chain polypeptide of A2. In some embodiments, The Fab heavy chain polypeptide of A2 is bound to the scFv light chain polypeptide of A1, L2 It is bound to the Fab light chain polypeptide of A2. In some embodiments, the Fa of A2 The b light chain polypeptide is bound to the scFv light chain polypeptide of A1, and L2 is bound to the F of A2. It is bound to the ab heavy chain polypeptide.

[0067] Peptides (P1, P2, and P 1a ) In some embodiments, P1, P2, or P 1a The sequences are disclosed in Table 3, and In contrast, a substantially identical sequence (for example, having 0, 1, or 2 amino acid modifications) Includes arrays.

[0068] [Table 3]

[0069] In some embodiments, P1 impairs the binding of A1 to the first target antigen. In some embodiments, P1 impairs the binding of A1 to the effector cell antigen. Several implementations Morphologically, P1 is involved in ionic interactions, electrostatic interactions, hydrophobic interactions, and Pi-stacks. It binds to A1 through ligation interactions and H-bond interactions, or a combination thereof. In some embodiments, P1 binds to A1 at or near the antigen-binding site. In some embodiments, L1 is cleaved by a tumor-specific protease, thereby A1 When exposed to effector cell antigens, P1 becomes unbound from A1. In some embodiments, the protease is a matrix metalloprotease (MMP) or It contains serine protease. In some embodiments, matrix metalloprotein Ze includes MMP2, MMP7, MMP9, MMP13, or MMP14. In this embodiment, the serine protease is matryptase (MTSP1), urokinase , or hepsin. In some embodiments, P1 is against effector cell antigens It has less than 70% sequence identity. In some embodiments, P1 is an effector It has less than 75% sequence identity with respect to the cell antigen. In some embodiments, P1 It has less than 80% sequence identity with effector cell antigens. Several implementations Morphologically, P1 exhibits less than 85% sequence identity with effector cell antigens. In some embodiments, P1 has less than 90% sequence identity to the effector cell antigen. It has. In some embodiments, P1 is 95% unaffected by effector cell antigens. It has complete sequence identity. In some embodiments, P1 is the effector cell antigen. In contrast, it has less than 98% sequence identity. In some embodiments, P1 is effervescent. It has less than 99% sequence identity with the kuta cell antigen. In some embodiments, P1 is a de novo cell that shares less than 10% sequence identity with effector cell antigens. It contains an amino acid sequence. In some embodiments, P1 is one of the sequence numbers 16-19. It contains an amino acid sequence consisting of one of the following: In some embodiments, P1 is sequence number 16 It includes the amino acid sequence of SEQ ID NO: 17. In some embodiments, P1 is the amino acid sequence of SEQ ID NO: 17. It includes. In some embodiments, P1 includes the amino acid sequence of SEQ ID NO: 18. In one embodiment, P1 comprises the amino acid sequence of SEQ ID NO: 19. In some embodiments, P1 contains the amino acid sequence of SEQ ID NO: 78.

[0070] In some embodiments, P1 is Z1-Z2-C-Z4-P-Z6-Z7-Z8-Z 9-Z 10 -Z 11 -Z 12 -CZ 14 The amino acid sequence includes, and Z1 is D, Y, Selected from F, I, N, V, H, L, A, T, S, and P, Z2 is D, Y, L, F , selected from I, N, A, V, H, T, and S, Z4 selected from G and W, Z6 is selected from E, D, V, and P, and Z7 is selected from W, L, F, V, G, M, I, and Z8 is selected from E, D, P, and Q, and Z9 is selected from E, D, and Y. Selected from V, F, W, P, L, and Q, Z 10 S, D, Y, T, I, F, V Selected from N, A, P, L, and H, Z 11 is I, Y, F, V, L, T, N, S Selected from D, A, and H, Z 12 are F, D, Y, L, I, V, A, N, T, P Selected from , S, and H, Z14 are D, Y, N, F, I, P, V, A, T, H, L Selected from , and S. In some embodiments, Z1 is selected from D, Y, F, I, and N is selected, Z2 is selected from D, Y, L, F, I, and N, Z4 is G and Z6 is selected from W, Z7 is selected from E and D, and Z7 is selected from W, L, F, and V. Z8 is selected from E and D, and Z9 is selected from E, D, Y, and V. And, Z 10 is selected from S, D, Y, T, and I, and Z 11 is I, Y, F, V, Selected from L and T, Z 12 The following can be selected from F, D, Y, L, I, V, A, and N. And, Z 14 This is selected from D, Y, N, F, I, and P. In some embodiments Z1 is selected from D, Y, and F, and Z2 is selected from D, Y, L, and F. The Z4 is available in G and W, the Z6 is available in E and D, and the Z7 is available in W. Z8 is selected from L and F, Z9 is selected from E and D Selected, Z 10 The following are selected from S, D, and Y, and Z 11 These are from I, Y, and F. Selected, Z 12 The following are selected from F, D, Y, and L, and Z 14 D, Y, and N Selected from.

[0071] In some embodiments, P1 is U1-U2-C-U4-P-U6-U7-U8-U 9-U 10 -U 11 -U 12 -CU 14 The amino acid sequence includes, and U1 is D, Y, Selected from F, I, N, V, H, L, A, T, S, and P, U2 is D, Y, L, F , selected from I, N, A, V, H, T, and S, U4 is selected from G and W, U6 is selected from E, D, V, and P, and U7 is selected from W, L, F, V, G, M, I, and U8 is selected from E, D, P, and Q, and U9 is selected from E, D, and Y. Selected from V, F, W, P, L, and Q, U 10 S, D, Y, T, I, F, V Selected from N, A, P, L, and H, U 11 is I, Y, F, V, L, T, N, S Selected from D, A, and H, U 12 are F, D, Y, L, I, V, A, N, T, P Selected from S, G, and H, U 14 are D, Y, N, F, I, P, V, A, T, H Selected from L, M, and S. In some embodiments, U1 is D, Y, F, I U2 is selected from D, Y, L, F, I, and N, U 4 is selected from G and W, U6 is selected from E and D, and U7 is selected from W, L, and F U8 is selected from G and V, U9 is selected from E and D, and selected from V, U 10 It is selected from S, D, Y, T, and I, U 11 teeth, Selected from I, Y, F, V, L, and T, U 12 F, D, Y, L, I, V, A, Selected from G and N, U 14 The following are selected from D, Y, N, F, I, M, and P. In some embodiments, U1 is selected from D, Y, V, and F, and U2 is D U4 is selected from Y, L, and F, U4 is selected from G and W, U6 is selected from E and U7 is selected from D, U7 is selected from W, L, G, and F, and U8 is selected from E and D. Selected, U9 is selected from E and D, U 10 The options are S, D, T, and Y. And, U 11 is selected from I, Y, V, L, and F, U 12 F, D, Y, G, Selected from A and L, U 14 This is selected from D, Y, M, and N.

[0072] In some embodiments, P1 is one of sequence numbers 79 to 105. It includes an amino acid sequence. In some embodiments, P1 is one of the sequences in Table 20. It contains an amino acid sequence consisting of one. In some embodiments, P1 is sequence numbers 106-1 It contains an amino acid sequence consisting of one of the 17.

[0073] In some embodiments, P1 is the amino acid sequence according to Sequence ID No. 18, or sequence number Peptide with 1, 2, or 3 amino acid substitutions, additions, or deletions compared to No. 18 Contains a cydoid sequence.

[0074] In some embodiments, P1 is the amino acid sequence according to Sequence ID No. 19, or sequence number Peptide with one, two, or three amino acid substitutions, additions, or deletions compared to number 19 Contains a cydoid sequence.

[0075] In some embodiments, P1 is the amino acid sequence according to SEQ ID NO: 116, or the sequence Number 116 has one, two, or three amino acid substitutions, additions, or deletions. Contains a peptide sequence.

[0076] In some embodiments, P1 includes the amino acid sequence given by SEQ ID NO: 18.

[0077] In some embodiments, P1 includes the amino acid sequence given by SEQ ID NO: 19.

[0078] In some embodiments, P1 comprises the amino acid sequence given by SEQ ID NO: 116.

[0079] In some embodiments, P2 impairs the binding of A2 to a second target antigen. In some embodiments, P2 impairs the coupling of A2 to PSMA. In some embodiments, P2 is involved in ionic interactions, electrostatic interactions, hydrophobic interactions, and Pi-stacking interactions. It is bound to A2 by H-bond interactions, or a combination thereof. In some embodiments, P2 binds to A2 at or near the antigen-binding site. In this morphology, L2 is cleaved by tumor-specific proteases, thereby converting A2 to PSMA. When exposed to, P2 becomes uncoupled from A2. In some embodiments, Proteases are matrix metalloproteinases (MMPs) or serine proteases. Includes. In some embodiments, the matrix metalloproteinase is MMP2, MM Includes P7, MMP9, MMP13, or MMP14. In some embodiments, cerium Proteases include matryptase (MTSP1), urokinase, or hepsin. In some embodiments, P2 has less than 70% sequence identity with respect to PSMA. In some embodiments, P2 has less than 75% sequence identity to PSMA. It has. In some embodiments, P2 has less than 80% sequence identity with respect to PSMA. It has properties. In some embodiments, P2 is less than 85% of the sequence relative to PSMA. They have identity. In some embodiments, P2 is less than 90% of PSMA. It has sequence identity. In some embodiments, P2 is 95% unidentified with respect to PSMA. It has complete sequence identity. In some embodiments, P2 is 98 relative to PSMA. It has less than % sequence identity. In some embodiments, P2 is relative to PSMA. It has less than 99% sequence identity. In some embodiments, P2 is opposite to PSMA. It contains de novo amino acid sequences that share less than 10% sequence identity.

[0080] In some embodiments, P 1a L 1a If the first part is not cleaved, the first part of the target antigen It impairs the binding of the antigen-recognition molecule. In some embodiments, the antigen-recognition molecule is an antibody or anti It contains a body fragment. In some embodiments, the target antigen is anti-CD3 effector cells. It is an antigen. In some embodiments, the target antigen is prostate-specific membrane antigen (PSMA). Yes. In some embodiments, P 1a This indicates a sequence identity of less than 70% for the target antigen. It has. In some embodiments, P 1a This is a sequence that is less than 75% identical to the target antigen. In some embodiments, P 1a This is less than 80% against the target antigen. It has sequence identity. In some embodiments, P 1a 85% against the target antigen It has less than sequence identity. In some embodiments, P 1a against the target antigen It has less than 90% sequence identity. In some embodiments, P 1a is the target antigen In contrast, it has less than 95% sequence identity. In some embodiments, P 1ais, It has less than 98% sequence identity with respect to the antigen. In some embodiments, P 1a teeth , it has less than 99% sequence identity with respect to the target antigen. In some embodiments, P 1a This is a de novo amino acid that shares less than 10% sequence identity with the second target antigen. Contains an acid sequence.

[0081] In some embodiments, P 1a is Z1-Z2-C-Z4-P-Z6-Z7-Z8- Z9-Z 10 -Z 11 -Z 12 -CZ 14 The amino acid sequence includes the following, where Z1 is D, Y , selected from F, I, N, V, H, L, A, T, S, and P, Z2 is D, Y, L, The following are available: F, I, N, A, V, H, T, and S, and Z4 is available in G and W. Z6 is selected from E, D, V, and P, and Z7 is selected from W, L, F, V, G, M, I, Z8 is selected from E, D, P, and Q, and Z9 is selected from E, D, Selected from Y, V, F, W, P, L, and Q, Z 10 S, D, Y, T, I, F, Selected from V, N, A, P, L, and H, Z 11 are I, Y, F, V, L, T, N, Selected from S, D, A, and H, Z 12 are F, D, Y, L, I, V, A, N, T, Selected from P, S, and H, Z 14 are D, Y, N, F, I, P, V, A, T, H, Selected from L and S. In some embodiments, Z1 is D, Y, F, I, and Z2 is selected from N, Z4 is selected from D, Y, L, F, I, and N, and Z4 is G Z6 is selected from W, Z7 is selected from E and D, and Z7 is selected from W, L, F, and V Z8 is selected from E and D, and Z9 is selected from E, D, Y, and V. Selected, Z 10 is selected from S, D, Y, T, and I, and Z 11 I, Y, F, V Selected from , L, and T, Z 12 The following can be selected from F, D, Y, L, I, V, A, and N. Selected, Z 14 This is selected from D, Y, N, F, I, and P. Several embodiments Then, Z1 is selected from D, Y, and F, and Z2 is selected from D, Y, L, and F. The Z4 is available in G and W, the Z6 in E and D, and the Z7 in W Z8 is selected from L and F, Z9 is selected from E and D or Selected from, Z 10 The following are selected from S, D, and Y, and Z 11 are I, Y, and F Selected from, Z 12 The following are selected from F, D, Y, and L, and Z 14 D, Y, and Selected from N.

[0082] In some embodiments, P 1a U1-U2-C-U4-P-U6-U7-U8- U9-U 10 -U 11 -U 12 -CU 14 The amino acid sequence includes, where U1 is D, Y , selected from F, I, N, V, H, L, A, T, S, and P, U2 is D, Y, L, U4 is selected from F, I, N, A, V, H, T, and S, and U4 is selected from G and W. U6 is selected from E, D, V, and P, and U7 is selected from W, L, F, V, G, M, I, U8 is selected from E, D, P, and Q, and U9 is selected from E, D, Selected from Y, V, F, W, P, L, and Q, U 10 S, D, Y, T, I, F, Selected from V, N, A, P, L, and H, U 11 are I, Y, F, V, L, T, N, Selected from S, D, A, and H, U 12 are F, D, Y, L, I, V, A, N, T, Selected from P, S, G, and H, U 14 are D, Y, N, F, I, P, V, A, T, Selected from H, L, M, and S. In some embodiments, U1 is D, Y, F, I, V, and N are selected, and U2 is selected from D, Y, L, F, I, and N. U4 is selected from G and W, U6 is selected from E and D, and U7 is selected from W, L, U8 is selected from F, G, and V, U9 is selected from E and D, and U9 is selected from E, D, and Y. Selected from V, U 10 It is selected from S, D, Y, T, and I, U 11 teeth Selected from I, Y, F, V, L, and T, U 12 F, D, Y, L, I, V, A Selected from , G, and N, U 14 The following can be selected from D, Y, N, F, I, M, and P. In some embodiments, U1 is selected from D, Y, V, and F, and U2 is U4 is selected from D, Y, L, and F, U4 is selected from G and W, U6 is selected from E and U7 is selected from W, L, G, and F, and U8 is selected from E and D. U9 is selected from E and D, U 10 The following can be selected from S, D, T, and Y. Selected, U 11 is selected from I, Y, V, L, and F, U 12F, D, Y, G Selected from A, L, and U 14 This is selected from D, Y, M, and N.

[0083] In some embodiments, P 1a This is based on one of the sequence numbers 79-105. It contains the amino acid sequence.

[0084] In some embodiments, P 1a This is an amino acid derived from one of the sequences in Table 20. Contains acid sequences.

[0085] In some embodiments, P 1a This corresponds to one of the sequence numbers 106-117. It contains the amino acid sequence.

[0086] In some embodiments, P 1a This is the amino acid sequence according to Sequence ID No. 18, or the sequence A pedia with one, two, or three amino acid substitutions, additions, or deletions in place of number 18 Includes a ptido sequence.

[0087] In some embodiments, P 1a This is the amino acid sequence according to Sequence ID No. 19, or the sequence A pedia with one, two, or three amino acid substitutions, additions, or deletions in place of number 19 Includes a ptido sequence.

[0088] In some embodiments, P 1a This is the amino acid sequence according to SEQ ID NO: 116, or the combined Column number 116 has one, two, or three amino acid substitutions, additions, or deletions. It contains a peptide sequence.

[0089] In some embodiments, P 1a This includes the amino acid sequence according to SEQ ID NO: 18.

[0090] In some embodiments, P 1a This includes the amino acid sequence according to SEQ ID NO: 19.

[0091] In some embodiments, P 1a This includes the amino acid sequence according to SEQ ID NO: 116.

[0092] In some embodiments, P1, P2, or P 1a It has a length of at least 5 amino acids. It contains the peptide sequence of P1, P2, or P 1a The length is It contains a peptide sequence of at least 6 amino acids. In some embodiments, P1, P2, Taha P 1a It contains a peptide sequence with a length of at least 10 amino acids. Several implementations In this state, P1, P2, or P 1a It is a length of at least 10 amino acids and 20 amino acids The following peptide sequence is included. In some embodiments, P1, P2, or P 1a is, The peptide sequence contains at least 16 amino acids. In some embodiments, P1, P 2, or P 1a It contains peptide sequences with a length of 40 amino acids or less. Several implementation forms In this state, P1, P2, or P 1a It contains at least two cysteine ​​amino acid residues. In some embodiments, P1, P2, or P 1a These are cyclic peptides or linear peptides. Contains cydo. In some embodiments, P1, P2, or P 1a It contains cyclic peptides. In some embodiments, P1, P2, or P 1a It contains linear peptides.

[0093] In some embodiments, P1, P2, or P 1a, or P1, P2, and P 1a This refers to modified amino acids, unnatural amino acids, unnatural modified amino acids, or combinations thereof. Includes. In some embodiments, modified amino acids or unnaturally modified amino acids are post-translational. Includes modifications. In some embodiments, P1, P2, or P 1a , or P1, P2 , and P 1a This involves acetylation, acylation, ADP-ribosylation, amidation, and flavin synthesis. Bonded, covalent bond of the heme portion, covalent bond of nucleotide or nucleotide derivative, lipid Covalent bonding of lipids or lipid derivatives, covalent bonding of phosphatidylinositol, crosslinking, and cyclization. Disulfide bond formation, demethylation, covalent crosslinking, cystine formation, pyrogly Formation of tamate, formylation, gamma carboxylation, glycosylation, GPI anchor type Formulation, hydroxylation, iodization, methylation, myristoylation, oxidation, proteolysis treatment Amino acids undergo processes such as phosphorylation, prenylation, racemization, selenoylation, sulfated and arginylated reactions. This includes, but is not limited to, transfer RNA-mediated addition to proteins and ubiquitination. Includes modifications without modifications. Modifications include the peptide backbone, amino acid side chains, and endpoints, P1, P2 , or P 1a , or P1, P2, and P 1a It will be done everywhere.

[0094] In some embodiments, P1, P2, or P 1a is albumin or albuminf It does not include a lagment. In some embodiments, P1, P2, or P 1a is, Alb It does not contain a min-binding domain.

[0095] Connecting parts (L1, L2, L3, and L1a ) In some embodiments, L1, L2, L3, or L 1a At least 5 to 50 The peptide sequence has the following amino acids. In some embodiments, L1, L2, L3 , or L 1a This is a peptide sequence having at least 10 to 30 amino acids. In some embodiments, L1, L2, L3, or L 1a is at least 10 amino acids It is a peptide sequence containing acid. In some embodiments, L1, L2, L3, or L 1a This is a peptide sequence having at least 18 amino acids. In some embodiments, is L1, L2, L3, or L 1a This is a peptide compound having at least 26 amino acids. These are columns. In some embodiments, they are L1, L2, L3, or L 1a (G2S) n It has an expression containing n, where n is an integer between 1 and 3 (sequence number 118). In the embodiment, L1, L2, L3, or L 1a (G2S) n It has an expression that includes In the formula, n is an integer of at least 1. In some embodiments, L1, L2, L3, or L 1a (G2S) n , (GS) n (GSGGS) n (Sequence ID 50), (G GGS) n (Sequence ID 51), (GGGGS) n (Sequence ID 52), and (GSSGG S) n The formula has an expression selected from the group consisting of (Sequence No. 53), where n is at least It is also an integer of 1. In some embodiments, tumor-specific proteases are metalloproteins Aase, serine protease, cysteine ​​protease, threonine protease, and Selected from the group consisting of biaspartate proteases. In some embodiments, L 1, L2, L3, or L 1a This is a urokinase cleavable amino acid sequence, matryptase Cleavable amino acid sequence, legmine-cleavable amino acid sequence, or matrix metallop Contains a rotease-cleavable amino acid sequence. In some embodiments, the matrix metallo The protease includes MMP2, MMP7, MMP9, MMP13, or MMP14. In some embodiments, serine protease is matryptase (MTSP1), Contains rokinase or hepsin.

[0096] In some embodiments, L1, L2, L3, or L 1a The sequence is disclosed in Table 4. , or substantially identical sequences (e.g., 0, 1, or 2 amino acid modifications) Includes (array containing).

[0097] In some embodiments, L1 includes an array of linkers 25 (sequence number 45). In some embodiments, L1 includes an array of linkers 26 (sequence number 46). In the embodiment, L1 includes an array of linkers 27 (sequence number 47). Several implementations In this state, L1 contains the linker 28 sequence (sequence number 48).

[0098] In some embodiments, L2 includes an array of linkers 25 (sequence number 45). In some embodiments, L2 includes an array of linkers 26 (sequence number 46). In the embodiment, L2 includes an array of linkers 27 (sequence number 47). Several embodiments In this state, L2 contains the linker 28 sequence (sequence number 48).

[0099] [Table 4]

[0100] In some embodiments, L1 is bonded to the N-terminus of A1. In some embodiments, L1 is bonded to the C-terminus of A1. In some embodiments, L2 is bonded to the N-terminus of A2. It is bonded to the end. In some embodiments, L2 is bonded to the C-terminus of A2. In that embodiment, L1 is cleaved by a tumor-specific protease, thereby A1 is When exposed to feta cell antigens, P1 becomes unbound from A1. In this embodiment, L2 is cleaved by a tumor-specific protease, thereby converting A2 to PS When exposed to MA, P2 will be decoupled from A2.

[0101] In some embodiments, L1, L2, L3, or L 1a These are modified amino acids, unnatural This includes amino acids, unnaturally modified amino acids, or combinations thereof. In some embodiments, Modified amino acids, or unnaturally modified amino acids, include post-translational modifications. Several implementation forms In state, L1, L2, L3, or L 1a This involves acetylation, acylation, and ADP-ribosylation. amidation, covalent bonding of flavin, covalent bonding of the heme moiety, nucleotide or nucleo Covalent bonding of lipid derivatives, covalent bonding of lipids or lipid derivatives, phosphatidylinositol Covalent bonding, crosslinking, cyclization, disulfide bond formation, demethylation, covalent crosslinking, Cystine formation, pyroglutamate formation, formylation, gamma carboxylation, glyco Silation, GPI anchor formation, hydroxylation, iodation, methylation, myristoylation, Oxidation, proteolysis, phosphorylation, prenylation, racemization, selenoylation, sulfation, Transfer RNA-mediated addition of amino acids to proteins, such as arginylation, and ubiquitination. This includes, but is not limited to, modifications to the peptide backbone or amino acid side chains. L1, L2, L3, or L 1a It will be done everywhere.

[0102] In some embodiments, the cleavable linker is cleavable by a protease. In some embodiments, the protease is used in the microenvironment of a diseased state, not in healthy tissue or It is present in the disease state microenvironment at a higher level compared to the level in the microenvironment. In some embodiments, the protease includes a tumor-specific protease. Proteases are matrix metalloproteinases (MMPs) or serine proteases. -ase is included. In some embodiments, the matrix metalloproteinase is MMP2, Includes MMP7, MMP9, MMP13, or MMP14. In some embodiments, Matrix metalloproteinases include MMP2, MMP7, MMP9, MMP13, and Selected from the group consisting of and MMP14. In some embodiments, matrix metallo The protease includes MMP2. In some embodiments, matrix metalloprotein The ase contains MMP7. In some embodiments, it is a matrix metalloproteinase. This includes MMP9. In some embodiments, the matrix metalloproteinase is M Includes MP13. In some embodiments, matrix metalloproteinase is MMP Includes 14. In some embodiments, serine protease is matryptase (MTS P1) comprises urokinase or hepsin. In some embodiments, serine proteo A ase is a group consisting of matryptase (MTSP1), urokinase, and hepsin. They are selected. In some embodiments, serine protease is matryptase (MT Includes SP1). In some embodiments, the serine protease includes urokinase. In some embodiments, the serine protease includes hepsin. In this state, the cleavable linker is cleaved by various proteases. Several implementations In terms of configuration, the severable linkers are at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 It is cleaved by 15, 20, or more than 20 different proteases.

[0103] Half-life extension molecule (H1 and H 1a ) In some embodiments, H1 does not interfere with A1 binding to effector cell antigens. In some embodiments, H1 includes a connecting portion (L3) that connects H1 to P1. In that embodiment, H 1a This interferes with the binding of the first antigen-recognition molecule to the effector cell antigen. No. In some embodiments, H 1a P 1a H 1a The connecting part (L3 ) contains. In some embodiments, half-life extension molecules (H1 or H 1a ) is antigen recognition It does not have binding affinity to the molecule. In some embodiments, the half-life extension molecule (H 1 or H 1a ) does not have binding affinity to effector cell antigens. In this embodiment, the half-life extending molecule (H1 or H1a ) is from effector cell antigen to antigen It does not block the recognition molecule. In some embodiments, the half-life extension molecule (H1 or H 1a ) It is not directly linked to the antigen-recognizing molecule.

[0104] In some embodiments, H1 or H 1a This refers to the sequences disclosed in Table 5, or those related thereto. In contrast, substantially identical sequences (for example, at least 90%, 95%, 96%, 97%, 98%) Includes sequences that have 99% sequence identity.

[0105] [Table 5]

[0106] In some embodiments, H1 or H 1a This is an amino acid combination having a repeating sequence motif. Includes columns. In some embodiments, H1 or H 1a This is a highly ordered secondary structure It contains an amino acid sequence that has a structure. The term "highly ordered secondary structure" is used in this context. When H1 or H 1a At least about 50%, about 70%, and about 80% of the amino acid residues , or about 90% are spectrophotometric methods (for example, in the "far ultraviolet" spectral region (190-2) (by circular dichroism spectroscopy at 50 nm) and the Chou-Fasman algorithm The Garnier-Osguthorpe-Robson ("GOR") algorithm means including, but not limited to, computer programs or algorithms such as Therefore, when measured or determined, it means that it contributes to the secondary structure.

[0107] In some embodiments, H1 or H 1aThis includes polymers. Several embodiments In this case, the polymer is polyethylene glycol (PEG). In some embodiments, , H1 or H 1a It contains albumin. In some embodiments, H1 or H 1a It includes an Fc domain. In some embodiments, albumin is serum albumin. In some embodiments, albumin is human serum albumin. In terms of application method, H1 or H 1a This includes polypeptides, ligands, or small molecules. In several embodiments, the polypeptide, ligand, or small molecule is a serum protein or Its fragment, circulating immunoglobulin or its fragment, or CD35 / It binds to CR1. In some embodiments, serum proteins bind to thyroxine-binding proteins. Protein, transthyretin, α1-acid glycoprotein, transferrin, transfer Phosphate receptors or their transferrin-binding sites, fibrinogen, or albumin Includes. In some embodiments, the circulating immunoglobulin molecules include IgG1, IgG2, and I It comprises gG3, IgG4, slgA, IgM, or IgD. In some embodiments, The serum protein is albumin. In some embodiments, the polypeptide is an antibody. In some embodiments, the antibody is a single-domain antibody, a single-chain variable fragment, or includes Fab. In some embodiments, a single-domain antibody is bound to albumin. It includes a single-domain antibody. In some embodiments, the single-domain antibody is a human antibody. Alternatively, it is a humanized antibody. In some embodiments, the single-domain antibody is 645gH1 gL1, 645dsgH5gL4, 23-13-A01-sc02, A10m3 or so Fragments of DOM7r-31, DOM7h-11-15, Alb-1, Alb-8 Selected from the group consisting of Alb-23, 10G, 10E, and SA21. In this embodiment, the single-domain antibody has HC-CDR1 as the complementarity-determining region (CDR). It includes HC-CDR2 and HC-CDR3, and the single-domain antibody HC-CDR1, HC-CDR2 and HC-CDR3 are sequence numbers 54 and HC-CDR1, respectively. DR2 includes sequence number 55, and HC-CDR3 includes sequence number 56. In the application method, single-domain antibodies use HC-CDR1 and H as complementarity-determining regions (CDRs). Includes C-CDR2 and HC-CDR3, and single-domain antibodies HC-CDR1, HC -CDR2 and HC-CDR3 are sequence numbers 54 in HC-CDR1, HC-CDR 2 includes sequence number 55, HC-CDR3 includes sequence number 56, and CDR is HC-CDR 1. 0 to 2 amino acids in at least one of HC-CDR2 or HC-CDR3 Includes modifications. In some embodiments, a single-domain antibody has a complementarity-determining region (CDR). This includes HC-CDR1, HC-CDR2, and HC-CDR3, and is a single-domain antimicrobial agent. The HC-CDR1, HC-CDR2, and HC-CDR3 of the body are distributed in HC-CDR1. Column number 58, sequence number 59 in HC-CDR2, and sequence number 60 in HC-CDR3 This includes. In some embodiments, a single-domain antibody is used as the complementarity-determining region (CDR). The single-domain antibody comprises HC-CDR1, HC-CDR2, and HC-CDR3. HC-CDR1, HC-CDR2, and HC-CDR3 are, in HC-CDR1, sequence number Number 58, HC-CDR2 includes sequence number 59, and HC-CDR3 includes sequence number 60, C DR is at least one of HC-CDR1, HC-CDR2, or HC-CDR3. It contains 0 to 2 amino acid modifications.

[0108] In some embodiments, H1 comprises the amino acid sequence given by SEQ ID NO: 57. In this embodiment, H1 has at least 80% sequence identity with respect to sequence number 57. Includes an amino acid sequence. In some embodiments, H1 is at least as shown in SEQ ID NO: 57 It also contains an amino acid sequence with 85% sequence identity. In some embodiments, H1 is It contains an amino acid sequence that has at least 90% sequence identity with SEQ ID NO: 57. In one embodiment, H1 has at least 95% sequence identity with respect to sequence number 57. It contains the amino acid sequence. In some embodiments, H1 is less than in SEQ ID NO: 57 It contains amino acid sequences with at least 99% sequence identity.

[0109] In some embodiments, H 1a This includes the amino acid sequence according to SEQ ID NO: 57. How many? In that embodiment, H 1a It has at least 80% sequence identity with sequence number 57. It includes the amino acid sequence. In some embodiments, H 1a This is less than the value of sequence number 57. It contains an amino acid sequence having at least 85% sequence identity. In some embodiments, H 1a This contains an amino acid sequence that has at least 90% sequence identity with SEQ ID NO: 57. In some embodiments, H 1a This is at least 95% of the sequence for sequence number 57. It contains an amino acid sequence that has identity. In some embodiments, H 1a This is sequence number 57 It contains an amino acid sequence that has at least 99% sequence identity with respect to [the given sequence].

[0110] In some embodiments, H1 comprises the amino acid sequence given by SEQ ID NO: 61. In this embodiment, H1 has at least 80% sequence identity with respect to sequence number 61. Includes an amino acid sequence. In some embodiments, H1 is at least as shown in SEQ ID NO: 61 It also contains an amino acid sequence with 85% sequence identity. In some embodiments, H1 is It contains an amino acid sequence that has at least 90% sequence identity with SEQ ID NO: 61. In one embodiment, H1 has at least 95% sequence identity with respect to sequence number 61. It contains the amino acid sequence. In some embodiments, H1 is less than in SEQ ID NO: 61 It contains amino acid sequences with at least 99% sequence identity.

[0111] In some embodiments, H 1a This includes the amino acid sequence according to SEQ ID NO: 61. How many? In that embodiment, H 1a It has at least 80% sequence identity with respect to sequence number 61. It includes the amino acid sequence. In some embodiments, H 1a This is less than the value of sequence number 61. It contains an amino acid sequence having at least 85% sequence identity. In some embodiments, H 1a This includes an amino acid sequence that has at least 90% sequence identity with SEQ ID NO: 61. In some embodiments, H 1a This is at least 95% of the sequence for sequence number 61. It contains an amino acid sequence that has identity. In some embodiments, H 1a This is sequence number 61 It contains an amino acid sequence that has at least 99% sequence identity with respect to [the given sequence].

[0112] In some embodiments, H1 or H 1a , or H1 and H 1a is a modified mesh Includes ano acids, unnatural amino acids, unnaturally modified amino acids, or combinations thereof. In this embodiment, the modified amino acid or unnaturally modified amino acid includes post-translational modifications. In some embodiments, H1 or H 1a , or H1 and H 1a acetylation, a Silation, ADP-ribosylation, amidation, covalent bonding of flavin, covalent bonding of heme moiety, nucleation Covalent bonding of rheotides or nucleotide derivatives, covalent bonding of lipids or lipid derivatives, Covalent bonding, crosslinking, cyclization, disulfide bond formation, and demethylation of phosphatidylinositol Formation, covalent crosslinking, cystine formation, pyroglutamate formation, formylation, cancer Maccarboxylation, glycosylation, GPI anchor formation, hydroxylation, iodation, Chillation, myristoylation, oxidation, proteolysis, phosphorylation, prenylation, racemization RNA-mediated transfer of amino acids to proteins, such as selenoylation, sulfated transfer, and arginylation. Modifications include, but are not limited to, addition and ubiquitination. Modifications include peptides. H1 or H including the backbone, amino acid side chains, and terminus 1a , or H1 and H 1a It will be done everywhere.

[0113] In some embodiments, H1 includes a connecting portion (L3) that connects H1 to P1. In some embodiments, L3 is a peptide compound having at least 5 to 50 amino acids. It is a row. In some embodiments, L3 has at least 10 to 30 amino acids. This is a peptide sequence. In some embodiments, L3 is at least 10 amino acids It is a peptide sequence having . In some embodiments, L3 is at least 18 amines It is a peptide sequence having an acid. In some embodiments, L3 is at least 26 It is a peptide sequence having amino acids. In some embodiments, L3 is (G2S) n , (GS) n (GSGGS) n (Sequence ID 50), (GGGS) n (Sequence ID 51) (GGGGS) n (Sequence ID 52), and (GSSGGS) n (Sequence number 53) It has an expression selected from the group, where n is an integer of at least 1. In this embodiment, L3 includes the amino acid sequence according to SEQ ID NO: 22.

[0114] In some embodiments, H 1a P 1a H 1a The connecting part (L 1a )of Includes. In some embodiments, L 1a It has at least 5 to 50 amino acids. It is a peptide sequence. In some embodiments, L 1a at least 10 to 30 or less It is a peptide sequence having amino acids. In some embodiments, L 1a At least It is a peptide sequence having 10 amino acids. In some embodiments, L 1a There are few It is a peptide sequence having at least 18 amino acids. In some embodiments, L 1a teeth , a peptide sequence having at least 26 amino acids. In some embodiments, L 1a (G2S) n , (GS) n (GSGGS) n (Sequence ID 50), (GGGS) n (Sequence ID 51), (GGGGS) n (Sequence ID 52), and (GSSGGS) n ( The formula is selected from the group consisting of Sequence ID No. 53, where n is at least 1. It is a number. In some embodiments, L 1a This includes the amino acid sequence according to SEQ ID NO: 22. .

[0115] PSMA and CD3-binding antibodies In some embodiments, polypeptides or polypeptide complexes are disclosed in Table 6. an amino acid sequence, or a sequence substantially identical thereto (for example, at least 90%, 9 Includes sequences with 5%, 96%, 97%, 98%, or 99% sequence identity. In several embodiments, the polypeptide or polypeptide complex is represented by SEQ ID NOs. 62-77. It contains an amino acid sequence that has at least 95% sequence identity with any one of them. In several embodiments, the polypeptide or polypeptide complex is used in relation to SEQ ID NO: 72 It contains an amino acid sequence having at least 95% sequence identity. In some embodiments, The polypeptide or polypeptide complex contains at least 95% of the polypeptide in SEQ ID NO: 73. It contains amino acid sequences that have sequence identity.

[0116] [Table 6-1]

[0117] [Table 6-2]

[0118] [Table 6-3]

[0119] [Table 6-4]

[0120] In some embodiments, the polypeptide or polypeptide complex is as shown in SEQ ID NO: 62 It includes the amino acid sequence by sequence number 63. In some embodiments, the polypeptide Alternatively, the polypeptide complex is at least 90% of SEQ ID NO: 62 and SEQ ID NO: 63 It contains an amino acid sequence having sequence identity. In some embodiments, the polypeptide also The polypeptide complex contains at least 95% of the polysaccharides in SEQ ID NO: 62 and SEQ ID NO: 63. It contains amino acid sequences having sequence identity. In some embodiments, polypeptides or The polypeptide complex contains at least 99% of the components of SEQ ID NO: 62 and SEQ ID NO: 63. It contains amino acid sequences that have sequence identity.

[0121] In some embodiments, the polypeptide or polypeptide complex is represented by SEQ ID NO: 64 It includes the amino acid sequence by SEQ ID NO: 65. In some embodiments, the polypeptide Alternatively, the polypeptide complex is at least 90% of SEQ ID NO: 64 and SEQ ID NO: 65 It contains an amino acid sequence having sequence identity. In some embodiments, the polypeptide also The polypeptide complex contains at least 95% of the values ​​for SEQ ID NOs. 64 and 65. It contains amino acid sequences having sequence identity. In some embodiments, polypeptides or The polypeptide complex contains at least 99% of the components of SEQ ID NO: 64 and SEQ ID NO: 65. It contains amino acid sequences that have sequence identity.

[0122] In some embodiments, the polypeptide or polypeptide complex is represented by SEQ ID NO: 66 It includes the amino acid sequence by SEQ ID NO: 67. In some embodiments, the polypeptide Alternatively, the polypeptide complex is at least 90% of SEQ ID NO: 66 and SEQ ID NO: 67 It contains an amino acid sequence having sequence identity. In some embodiments, the polypeptide also The polypeptide complex contains at least 95% of the polypeptide complex for SEQ ID NO: 66 and SEQ ID NO: 67. It contains amino acid sequences having sequence identity. In some embodiments, polypeptides or The polypeptide complex contains at least 99% of the components of SEQ ID NO: 66 and SEQ ID NO: 67. It contains amino acid sequences that have sequence identity.

[0123] In some embodiments, the polypeptide or polypeptide complex is represented by SEQ ID NO: 68 It includes the amino acid sequence by SEQ ID NO: 69. In some embodiments, the polypeptide Alternatively, the polypeptide complex is at least 90% of SEQ ID NO: 68 and SEQ ID NO: 69 It contains an amino acid sequence having sequence identity. In some embodiments, the polypeptide also The polypeptide complex contains at least 95% of the polysaccharides in SEQ ID NO: 68 and SEQ ID NO: 69. It contains amino acid sequences having sequence identity. In some embodiments, polypeptides or The polypeptide complex contains at least 99% of the components of SEQ ID NO: 68 and SEQ ID NO: 69. It contains amino acid sequences that have sequence identity.

[0124] In some embodiments, the polypeptide or polypeptide complex is represented by SEQ ID NO: 70 It includes the amino acid sequence by SEQ ID NO: 71. In some embodiments, the polypeptide Alternatively, the polypeptide complex is at least 90% of SEQ ID NO: 70 and SEQ ID NO: 71 It contains an amino acid sequence having sequence identity. In some embodiments, the polypeptide also The polypeptide complex contains at least 95% of the polypeptide complex for SEQ ID NOs. 70 and 71. It contains amino acid sequences having sequence identity. In some embodiments, polypeptides or The polypeptide complex contains at least 99% of the polypeptides for SEQ ID NOs. 70 and 71. It contains amino acid sequences that have sequence identity.

[0125] In some embodiments, the polypeptide or polypeptide complex is as shown in SEQ ID NO: 72 It includes the amino acid sequence by SEQ ID NO: 73. In some embodiments, the polypeptide Alternatively, the polypeptide complex is at least 90% of SEQ ID NO: 72 and SEQ ID NO: 73 It contains an amino acid sequence having sequence identity. In some embodiments, the polypeptide also The polypeptide complex contains at least 95% of the polypeptide complex for SEQ ID NOs. 72 and 73. It contains amino acid sequences having sequence identity. In some embodiments, polypeptides or The polypeptide complex contains at least 99% of the components of SEQ ID NO: 72 and SEQ ID NO: 73. It contains amino acid sequences that have sequence identity.

[0126] In some embodiments, the polypeptide or polypeptide complex is represented by SEQ ID NO: 74 It includes the amino acid sequence by SEQ ID NO: 75. In some embodiments, the polypeptide Alternatively, the polypeptide complex is at least 90% of SEQ ID NO: 74 and SEQ ID NO: 75 It contains an amino acid sequence having sequence identity. In some embodiments, the polypeptide also The polypeptide complex contains at least 95% of the polypeptide complex for SEQ ID NOs. 74 and 75. It contains amino acid sequences having sequence identity. In some embodiments, polypeptides or The polypeptide complex contains at least 99% of the components of SEQ ID NO: 74 and SEQ ID NO: 75. It contains amino acid sequences that have sequence identity.

[0127] In some embodiments, the polypeptide or polypeptide complex is represented by SEQ ID NO: 76 It includes the amino acid sequence by SEQ ID NO: 77. In some embodiments, the polypeptide Alternatively, the polypeptide complex is at least 90% of SEQ ID NO: 76 and SEQ ID NO: 77 It contains an amino acid sequence having sequence identity. In some embodiments, the polypeptide also The polypeptide complex contains at least 95% of the polypeptide complex for SEQ ID NOs. 76 and 77. It contains amino acid sequences having sequence identity. In some embodiments, polypeptides or The polypeptide complex contains at least 99% of the components of SEQ ID NO: 76 and SEQ ID NO: 77. It contains amino acid sequences that have sequence identity.

[0128] Polypeptides or polypeptide complexes are, in some embodiments, as specified in Table 6. The sequence includes a sequence of 62-77. In some embodiments, the sequence is one of the sequence numbers 62-77. In contrast, at least or about 70%, 80%, 85%, 90%, 91%, 92%, 93%, Includes sequence identity of 94%, 95%, 96%, 97%, 98%, 99%, or 100%. In some embodiments, the sequence is less than one of sequence numbers 62-77. It includes both or approximately 95% sequence identity. In some embodiments, the sequence is sequence number 6 Contains at least or approximately 97% sequence identity for one of 2 to 77. In that embodiment, the sequence is at least one of sequence numbers 62-77 This includes approximately 99% sequence identity. In some embodiments, the sequences are sequence numbers 62-77. Contains at least or approximately 100% sequence identity for any one of the following. So the array is one of the following sequence numbers: 62, 65, 66, 69, 70, 73, 75, or 77 Any one of the following, at least or about 10, 20, 30, 40, 50, 60, 70, 80, 9 0, 100, 110, 120, 130, 140, 150, 160, 170, 180, 19 Contains at least a portion having 0, 200, 210, or more than 210 amino acids. In some examples, the array contains at least one of either sequence number 63 or 64. The numbers are approximately 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, and 12. 0, 130, 140, 150, 160, 170, 180, 190, 200, 210, 22 0, 230, 240, 250, 260, 270, 280, 290, 300, 310, 32 0, 330, 340, 350, 360, 370, 380, 390, 400, 410, 42 At least one having 0, 430, 440, 450, or more than 450 amino acids Includes parts. In some examples, the sequence is sequence numbers 67, 68, 71, 72, 74, or At least one of 76, at least 10, 20, 30, 40, 50, 60, 70 , 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 1 80, 190, 200, 210, 220, 230, 240, 250, 260, 270, 2 80, 290, 300, 310, 320, 330, 340, 350, 360, 370, 3 80, 390, 400, 410, 420, 430, 440, 450, 460, 470, 4 80, 490, 500, 510, 520, 530, 540, 550, 560, 570, 5 80, 590, 600, 610, 620, 630, 640, or more than 640 It contains at least a portion of amino acids.

[0129] When used herein, the term "amino acid sequence identity percentage (%)" refers to the sequence. The term refers to a specific sequence after aligning the sequence as needed and introducing gaps. It is identical to the amino acid residues within it, achieving the highest sequence identity percentage, but sequence identity Conservative substitutions are not considered as part of the definition; it is defined as the proportion of amino acid residues in the candidate sequence. Alignment for the purpose of determining amino acid sequence identity percentage is, for example, EMB. OSS MATCHER, EMBOSS WATER, EMBOSS STRETCHE R, EMBOSS NEEDLE, EMBOSS LALIGN, BLAST, BLAS Software such as T-2, ALIGN, or Megalign (DNASTAR), Using publicly available computer software, various things within the scope of what a person skilled in the art could imagine This can be achieved in the following way. Those skilled in the art will know the maximum length that exceeds the full length of the sequences being compared. To measure alignment, including all the algorithms necessary to achieve the sequence. Appropriate parameters can be determined. ALIGN-2 is used for amino acid sequence comparison. In situations where this is the case, (to, with, or against) a given amino acid sequence A ) The amino acid sequence of a given amino acid sequence B (or, a specific amino acid sequence B) It can be represented as a given amino acid sequence A that has or contains an amino acid sequence identity percentage. The result is calculated as 100 × fraction X / Y, where X is the result of programs A and B. In alignment, the sequence alignment program ALIGN-2 was used to ensure identical matching. This is the number of amino acid residues that are scored, and Y is the total number of amino acid residues in B. If the length of amino acid sequence A is not equal to the length of amino acid sequence B, then the amino acids of A and B It is recognized that the percentage of acid sequence identity is not equal to the percentage of amino acid sequence identity between B and A. Unless otherwise specified, all amino acid sequence identity % used herein is A This is obtained using the LIGN-2 computer program as described in the previous paragraph.

[0130] In some embodiments herein, stereochemical configuration 1:

[0131] [ka] An isolated polypeptide or polypeptide complex comprising a structural arrangement by poly The peptide complex is a single-chain variable fragment containing a light chain variable domain and a heavy chain variable domain. (scFv) is a compound that impairs the binding of scFv to effector cell antigens. It further contains peptides, which are binding sites that are substrates for tumor-specific proteases. It is linked to the heavy chain variable domain of scFv, and the peptide is a half-life extension molecule. It contains scFV and Fab or that binds to prostate-specific membrane antigen (PSMA). Fab', where Fab or Fab' is a Fab light chain polypeptide chain and Fab heavy chain. It contains a chain polypeptide chain, and the Fab heavy chain polypeptide chain is a light chain variable domain of scFv. Isolated polypeptide containing Fab or Fab' ligated to the C-terminus of 'in' A cytoplasmic or polypeptide complex is disclosed.

[0132] In some embodiments herein, stereochemical configuration 2:

[0133] [ka] An isolated polypeptide or polypeptide complex comprising a structural arrangement by poly The peptide complex is a single-chain variable fragment containing a light chain variable domain and a heavy chain variable domain. (scFv) is a compound that impairs the binding of scFv to effector cell antigens. It further contains peptides, which are binding sites that are substrates for tumor-specific proteases. It is linked to the N-terminus of the heavy chain variable domain of scFv, and the peptide has an extended half-life. The molecule further contains scFV and Fa, which binds to prostate-specific membrane antigen (PSMA). b or Fab', where Fab or Fab' is a Fab light chain polypeptide chain and It contains a Fab heavy chain polypeptide chain, and the Fab light chain polypeptide chain is the light chain of scFv. Isolated containing Fab or Fab', which is ligated to the C-terminus of the variable domain. A polypeptide or polypeptide complex is disclosed.

[0134] In some embodiments described herein, stereochemical configuration 3:

[0135] [ka] An isolated polypeptide or polypeptide complex comprising a structural arrangement by poly Peptide or polypeptide complexes include light chain variable domains and heavy chain variable domains. A single-chain variable fragment (scFv), where the scFv is sc to effector cell antigens. It is linked to a peptide (P1) that impairs Fv binding, and P1 is a tumor-specific protea The L1 ligation site, which is the substrate of ze, is linked to the N-terminus of the light chain variable domain of scFv. Furthermore, P1 is linked to a half-life extension molecule, scFV, and prostate-specific membrane inhibitors. Fab that binds to the progenitor (PSMA), wherein Fab is a Fab light chain polypeptide and F The ab heavy chain polypeptide is contained in the scFv heavy chain variable domain. The C-terminus is linked to Fab, which is linked to P2 and L2, and P2 is linked to Fab to PSMA. It contains a peptide that impairs the binding, and when L2 connects the Fab light chain polypeptide to P2, The isolated Fab contains a binding portion which is a substrate for tumor-specific proteases. A polypeptide or polypeptide complex is disclosed.

[0136] In some embodiments herein, stereochemical configuration 4:

[0137] [ka] An isolated polypeptide or polypeptide complex comprising a structural arrangement by poly Peptide or polypeptide complexes include light chain variable domains and heavy chain variable domains. A single-chain variable fragment (scFv), where the scFv is sc to effector cell antigens. It is linked to a peptide that disrupts Fv binding, and the peptide is a tumor-specific protease. The substrate is linked to the light chain variable domain of scFv, and the peptide is semi-transparent. Further linked to the time-extending molecule are scFV and prostate-specific membrane antigen (PSMA). The binding Fab is a Fab light chain polypeptide chain and a Fab heavy chain polypeptide chain. It contains a butyl chain, and the Fab heavy chain polypeptide chain is the C of the heavy chain variable domain of scFv. An isolated polypeptide or polypeptide containing Fab, which is ligated to the terminal. The complex is revealed.

[0138] In some embodiments herein, stereochemical configuration 5:

[0139] [ka] An isolated polypeptide or polypeptide complex comprising a structural arrangement by poly Peptide or polypeptide complexes include light chain variable domains and heavy chain variable domains. A single-chain variable fragment (scFv), where the scFv is sc to effector cell antigens. It is linked to a peptide (P1) that impairs Fv binding, and P1 is a tumor-specific protea The substrate of ze is linked to the N-terminus of the light chain variable domain of scFv, P1 is further linked to a half-life extension molecule, scFV, and prostate-specific membrane antigen (PS). Fab that binds to MA), wherein Fab is a Fab light chain polypeptide and a Fab heavy chain The polypeptide contains the Fab light chain polypeptide at the C-terminus of the heavy chain variable domain of scFv. The Fab is linked to P2 and L2, and P2 links the Fab to the PSMA. Including the destructive peptide, L2 ligates the Fab heavy chain polypeptide to P2 and tumor Isolated polypropylene containing a Fab containing a binding portion which is a substrate for a specific protease. Lipeptide or polypeptide complexes are disclosed.

[0140] In some embodiments described herein, stereochemical configuration 6:

[0141] [ka] An isolated polypeptide or polypeptide complex comprising a structural arrangement by poly Peptide or polypeptide complexes include light chain variable domains and heavy chain variable domains. A single-chain variable fragment (scFv), where the scFv is sc to effector cell antigens. It is further linked to a peptide that impairs Fv binding, and the peptide is tumor-specific protea The -ase substrate is linked to the N-terminus of the light chain variable domain of scFv. The peptide is further linked to a half-life extension molecule, scFV, and prostate-specific membrane antigen. Fab that binds to (PSMA), wherein Fab is a Fab light chain polypeptide chain and F It contains an ab heavy chain polypeptide chain, and the Fab light chain polypeptide chain is a scFv heavy chain polypeptide. An isolated polypeptide containing Fab, which is linked to the C-terminus of the variant domain. A polypeptide complex is disclosed.

[0142] In some embodiments herein, stereochemical configuration 7:

[0143] [ka] An isolated polypeptide or polypeptide complex comprising a structural arrangement by poly Peptide or polypeptide complexes include light chain variable domains and heavy chain variable domains. A single-chain variable fragment (scFv), where the scFv is sc to effector cell antigens. It is linked to a peptide (P1) that impairs Fv binding, and P1 is a tumor-specific protea The binding portion (L1), which is the substrate of ze, is linked to the N-terminus of the heavy chain variable domain of scFv. Furthermore, P1 is linked to a half-life extension molecule, scFV, and prostate-specific membrane inhibitors. Fab that binds to the progenitor (PSMA), wherein Fab is a Fab light chain polypeptide and F The ab heavy chain polypeptide is contained in the scFv light chain variable domain. The C-terminus is linked to Fab, which is linked to P2 and L2, and P2 is linked to Fab to PSMA. It contains a peptide that impairs the binding, and when L2 connects the Fab light chain polypeptide to P2, The isolated Fab contains a binding portion which is a substrate for tumor-specific proteases. A polypeptide or polypeptide complex is disclosed.

[0144] In some embodiments herein, stereochemical configuration 8:

[0145] [ka] An isolated polypeptide or polypeptide complex comprising a structural arrangement by poly Peptide or polypeptide complexes include light chain variable domains and heavy chain variable domains. A single-chain variable fragment (scFv), where the scFv is sc to effector cell antigens. It is linked to a peptide (P1) that impairs Fv binding, and P1 is a tumor-specific protea The binding portion (L1), which is the substrate of ze, is linked to the N-terminus of the heavy chain variable domain of scFv. Furthermore, P1 is linked to a half-life extension molecule, scFV, and prostate-specific membrane inhibitors. Fab that binds to the progenitor (PSMA), wherein Fab is a Fab light chain polypeptide and F The ab heavy chain polypeptide contains the Fab light chain polypeptide, which is the light chain variable domain of scFv. The C-terminus is linked to Fab, which is linked to P2 and L2, and P2 is linked to Fab to PSMA. It contains a peptide that impairs the binding, and when L2 connects the Fab heavy chain polypeptide to P2, The isolated Fab contains a binding portion which is a substrate for tumor-specific proteases. A polypeptide or polypeptide complex is disclosed.

[0146] In some embodiments herein, stereochemical configuration 9:

[0147] [ka] An isolated polypeptide or polypeptide complex comprising a structural arrangement by poly Peptides or polypeptide complexes bind to prostate-specific membrane antigen (PSMA). b, wherein Fab comprises a Fab light chain polypeptide and a Fab heavy chain polypeptide. Fab is linked to a peptide (P1) that impairs the binding of Fab to PSMA, and P1 is The Fab light chain polypeptide is converted by the ligation region (L1), which is a substrate for tumor-specific proteases. Fab and effect A single-chain variable fragment (scFv) that binds to a cell antigen, wherein the scFv is a light chain variable fragment. It includes a variable domain and a heavy chain variable domain, and the heavy chain variable domain of scFv is the Fab heavy chain The scFv is linked to the N-terminus of the polypeptide, and is linked to P2 and L2, and P2 is The peptide contains a peptide that impairs the binding of scFv to feta cell antigens, and L2 impairs the binding of scFv to P2. The light chain variable domain is connected, and the binding portion is a substrate for tumor-specific proteases. An isolated polypeptide or polypeptide complex containing scFv is opened It will be shown.

[0148] In some embodiments of this specification, stereochemical configuration 10:

[0149] [ka] An isolated polypeptide or polypeptide complex comprising a structural arrangement by poly Peptides or polypeptide complexes bind to prostate-specific membrane antigen (PSMA). b, wherein Fab comprises a Fab light chain polypeptide and a Fab heavy chain polypeptide. Fab is linked to a peptide that impairs the binding of Fab to PSMA, and the peptide is tumor The N-terminus of the Fab light chain polypeptide is formed by the ligation site (L1), which is a substrate for a specific protease. The peptide is linked to the end and further linked to a half-life extension molecule, Fab and Effect A single-chain variable fragment (scFv) that binds to a cell antigen, wherein the scFv is a light chain variable fragment. It includes a variable domain and a heavy chain variable domain, and the heavy chain variable domain of scFv is the Fab heavy chain Isolated polypeptide containing scFv, which is ligated to the N-terminus of the polypeptide. Alternatively, polypeptide complexes are disclosed.

[0150] In some embodiments herein, stereochemical configuration 11:

[0151] [ka] An isolated polypeptide or polypeptide complex comprising a structural arrangement by poly Peptides or polypeptide complexes bind to prostate-specific membrane antigen (PSMA). b, wherein Fab comprises a Fab light chain polypeptide and a Fab heavy chain polypeptide. Fab is linked to a peptide (P1) that impairs the binding of Fab to PSMA, and P1 is The Fab heavy chain polypeptide is converted by the ligation region (L1), which is a substrate for tumor-specific proteases. Fab and effect A single-chain variable fragment (scFv) that binds to a cell antigen, wherein the scFv is a light chain variable fragment. It includes a variable domain and a heavy chain variable domain, and the heavy chain variable domain of scFv is the Fab light chain The scFv is ligated to the N-terminus of the polypeptide, and is further ligated to P2 and L2, P2 It contains a peptide that impairs the binding of scFv to effector cell antigens, and L2 is s to P2. The light chain variable domain of cFv is linked, and the ligation is a substrate for tumor-specific proteases. Isolated polypeptide or polypeptide complex containing scFv, including a portion of The body is revealed.

[0152] In some embodiments herein, stereochemical configuration 12:

[0153] [ka] An isolated polypeptide or polypeptide complex comprising a structural arrangement by poly Peptides or polypeptide complexes bind to prostate-specific membrane antigen (PSMA). b, wherein Fab comprises a Fab light chain polypeptide and a Fab heavy chain polypeptide. Fab is linked to a peptide that impairs the binding of Fab to PSMA, and the peptide is tumor The N-terminus of the Fab heavy chain polypeptide is formed by the ligation region (L1), which is a substrate for a specific protease. The peptide is linked to the end and further linked to a half-life extension molecule, Fab and Effect A single-chain variable fragment (scFv) that binds to a cell antigen, wherein the scFv is a light chain variable fragment. It includes a variable domain and a heavy chain variable domain, and the heavy chain variable domain of scFv is the Fab light chain Isolated polypeptide containing scFv, which is ligated to the N-terminus of the polypeptide. Alternatively, polypeptide complexes are disclosed.

[0154] In some embodiments of this specification, stereochemical configuration 13:

[0155] [ka] An isolated polypeptide or polypeptide complex comprising a structural arrangement by poly Peptides or polypeptide complexes bind to prostate-specific membrane antigen (PSMA). b, wherein Fab comprises a Fab light chain polypeptide and a Fab heavy chain polypeptide. Fab is linked to a peptide (P1) that impairs the binding of Fab to PSMA, and P1 is tumor The ligation site (L1), which is a substrate for ulcer-specific proteases, is the N1 of the Fab light chain polypeptide. Fab and effector are linked at the end, and P1 is further linked to the half-life extension molecule. A single-chain variable fragment (scFv) that binds to a cell antigen, wherein the scFv is a light chain variable fragment. The domain and heavy chain variable domain are included, and the light chain variable domain of scFv is Fab heavy chain The scFv is linked to the N-terminus of the lipeptide, and is linked to P2 and L2, with P2 being the F It contains a peptide that impairs the binding of scFv to the ectocellular antigen, and L2 impairs the binding of scFv to P2. It contains a linking portion that connects heavy chain variable domains and is a substrate for tumor-specific proteases. An isolated polypeptide or polypeptide complex containing scFv is disclosed. It will be done.

[0156] In some embodiments herein, stereochemical configuration 14:

[0157] [ka] An isolated polypeptide or polypeptide complex comprising a structural arrangement by poly Peptides or polypeptide complexes bind to prostate-specific membrane antigen (PSMA). b, wherein Fab comprises a Fab light chain polypeptide and a Fab heavy chain polypeptide. Fab is linked to a peptide that impairs the binding of Fab to PSMA, and the peptide is tumor The Fab light chain polypeptide is ligated to the N-terminus by a ligation site that is a substrate of a specific protease. Furthermore, the peptide is linked to a half-life extension molecule, Fab, and effector cell anti A single-chain variable fragment (scFv) that binds to the original, where the scFv is a light-chain variable domain. The scFv light chain variable domain includes a heavy chain variable domain, and the Fab heavy chain polypeptide An isolated polypeptide or polypeptide containing scFv, which is linked to the N-terminus of the polypeptide. A lipeptide complex is disclosed.

[0158] In some embodiments of this specification, stereochemical configuration 15:

[0159] [ka] An isolated polypeptide or polypeptide complex comprising a structural arrangement by poly Peptides or polypeptide complexes bind to prostate-specific membrane antigen (PSMA). b, wherein Fab comprises a Fab light chain polypeptide and a Fab heavy chain polypeptide. Fab is linked to (P1), which impairs the binding of Fab to PSMA, and P1 is tumor-specific. The protease substrate (L1) is linked to the N-terminus of the Fab heavy chain polypeptide. P1 is further linked to the half-life extension molecule, Fab, and effector cell antigen. A single-chain variable fragment (scFv) that binds to a light chain variable domain, wherein the scFv is a light chain variable domain and includes a heavy chain variable domain, and the light chain variable domain of scFv is Fab light chain polypeptide The N-terminus of the do is connected, and scFv is connected to P2 and L2, and P2 is the effector details It contains a peptide that impairs the binding of scFv to the cytoplasmic antigen, and L2 modulates the heavy chain of scFv to P2. The sc contains a ligation portion that connects domains and is a substrate for tumor-specific proteases. An isolated polypeptide or polypeptide complex containing Fv is disclosed.

[0160] In some embodiments herein, stereochemical configuration 16:

[0161] [ka] An isolated polypeptide or polypeptide complex comprising a structural arrangement by poly Peptides or polypeptide complexes bind to prostate-specific membrane antigen (PSMA). b, wherein Fab comprises a Fab light chain polypeptide and a Fab heavy chain polypeptide. Fab is linked to a peptide that impairs the binding of Fab to PSMA, and the peptide is tumor The N-terminus of the Fab heavy chain polypeptide is formed by the ligation region (L1), which is a substrate for a specific protease. The peptide is linked to the end and further linked to a half-life extension molecule, Fab and Effect A single-chain variable fragment (scFv) that binds to a cell antigen, wherein the scFv is a light chain variable fragment. It includes a variable domain and a heavy chain variable domain, and the light chain variable domain of scFv is the Fab light chain Isolated polypeptide containing scFv, which is ligated to the N-terminus of the polypeptide. Alternatively, polypeptide complexes are disclosed.

[0162] Polynucleotides that encode polypeptides or polypeptide complexes In some embodiments herein, the polypeptide or polypropylene disclosed herein is used. Isolated recombinant nucleic acid molecules encoding lipeptide complexes are disclosed. Several implementations Morphologically, a polypeptide or polypeptide complex contains an antibody or antibody fragment. In some embodiments, the polypeptide or polypeptide complex is Fab and Includes single-chain variable fragments (scFv).

[0163] In some embodiments of this specification, Formula I: A2-A1-L1-P1-H1 (Formula I) Isolated recombinant nucleic acid molecules encoding polypeptides or polypeptide complexes by There is, In the formula, A1 comprises a first antigen-recognition molecule that binds to the effector cell antigen, and P1 comprises A1 It contains a peptide that binds to P1, and L1 connects A1 to P1 and tumor-specific protea It contains a binding portion that is a substrate of the enzyme, H1 contains a half-life extension molecule, and A2 is prostate-specific. Isolated recombinant nucleic acid molecules containing a second antigen-recognition molecule that binds to the target membrane antigen (PSMA). It will be disclosed.

[0164] In some embodiments of this specification, Formula I: A2-A1-L1-P1-H1 (Formula I) Isolated recombinant nucleic acid molecules encoding polypeptides or polypeptide complexes by There is, In the formula, A1 is the first antigen-recognizing molecule that binds to the effector cell antigen, and P1 is A1 It is a peptide that binds to P1, and L1 attaches A1 to P1 and tumor-specific protea The binding portion is the substrate of the enzyme; H1 is a half-life extension molecule; and A2 is prostate-specific. Isolated recombinant nucleic acid molecules, which are a second antigen-recognition molecule that binds to the target membrane antigen (PSMA). It will be disclosed.

[0165] In some embodiments of this specification, Formula I: A2-A1-L1-P1-H1 (Formula I) Isolated recombinant nucleic acid molecules encoding polypeptides or polypeptide complexes containing There is, In the formula, A1 comprises a first antigen-recognition molecule that binds to the effector cell antigen, and P1 comprises A1 It contains a peptide that binds to P1, and L1 connects A1 to P1 and tumor-specific protea It contains a binding portion that is a substrate of the enzyme, H1 contains a half-life extension molecule, and A2 is prostate-specific. Isolated recombinant nucleic acid molecules containing a second antigen-recognition molecule that binds to the target membrane antigen (PSMA). It will be disclosed.

[0166] In some embodiments of this specification, Formula I: A2-A1-L1-P1-H1 (Formula I) Isolated recombinant nucleic acid molecules encoding polypeptides or polypeptide complexes containing There is, In the formula, A1 is the first antigen-recognizing molecule that binds to the effector cell antigen, and P1 is A1 It is a peptide that binds to P1, and L1 attaches A1 to P1 and tumor-specific protea The binding portion is the substrate of the enzyme; H1 is a half-life extension molecule; and A2 is prostate-specific. Isolated recombinant nucleic acid molecules, which are a second antigen-recognition molecule that binds to the target membrane antigen (PSMA). It will be disclosed.

[0167] In some embodiments herein, formula Ia: P2-L2-A2-A1-L1-P1-H1 (Formula Ia) Isolated recombinant nucleic acid molecules encoding polypeptides or polypeptide complexes It will be disclosed.

[0168] In some embodiments herein, Formula II: L 1a -P 1a -H 1a (Formula II) Isolated recombinant nucleic acid molecules encoding polypeptides or polypeptide complexes by There is, In the formula, L 1a If not cleaved, the first antigen recognition molecule binds to the effector cell antigen. Child P 1a The first includes a connecting portion that is cleaved by a tumor-specific protease, which connects the two. The antigen-recognizing molecule then binds to a second antigen-recognizing molecule that binds to the prostate-specific membrane antigen (PSMA). Continued, P 1a L 1a If it is not cleaved, the peptide that binds to the first antigen recognition molecule Includes H 1a This discloses isolated recombinant nucleic acid molecules, including half-life extension molecules.

[0169] In some embodiments herein, Formula II: L 1a -P 1a -H 1a (Formula II) Isolated recombinant nucleic acid molecules encoding polypeptides or polypeptide complexes containing There is, In the formula, L 1a If not cleaved, the first antigen recognition molecule binds to the effector cell antigen. Child P 1a The first includes a connecting portion that is cleaved by a tumor-specific protease, which connects the two. The antigen-recognizing molecule then binds to a second antigen-recognizing molecule that binds to the prostate-specific membrane antigen (PSMA). Continued, P 1a L 1a If it is not cleaved, the peptide that binds to the first antigen recognition molecule Includes H 1a This discloses isolated recombinant nucleic acid molecules, including half-life extension molecules.

[0170] In some embodiments herein, Formula II: L 1a -P 1a -H 1a (Formula II) Isolated recombinant nucleic acid molecules encoding polypeptides or polypeptide complexes by There is, In the formula, L 1a If not cleaved, the first antigen recognition molecule binds to the effector cell antigen. Child P 1a The connecting portion is a tumor-specific protease that cleaves the first The antigen-recognizing molecule then binds to a second antigen-recognizing molecule that binds to the prostate-specific membrane antigen (PSMA). Continued, P 1a L 1a If it is not cleaved, the peptide that binds to the first antigen recognition molecule It is D, H 1a This discloses isolated recombinant nucleic acid molecules that are half-life extension molecules.

[0171] In some embodiments herein, Formula II: L 1a -P 1a -H 1a (Formula II) Isolated recombinant nucleic acid molecules encoding polypeptides or polypeptide complexes containing There is, In the formula, L 1a If not cleaved, the first antigen recognition molecule binds to the effector cell antigen. Child P 1a The connecting portion is a tumor-specific protease that cleaves the first The antigen-recognizing molecule then binds to a second antigen-recognizing molecule that binds to the prostate-specific membrane antigen (PSMA). Continued, P 1a L 1a If it is not cleaved, the peptide that binds to the first antigen recognition molecule It is D, H 1a This discloses isolated recombinant nucleic acid molecules that are half-life extension molecules.

[0172] In some embodiments herein, stereochemical configuration 1:

[0173] [ka] Isolated nuclei encoding polypeptides or polypeptide complexes including structural arrangements by The acid molecule, a polypeptide complex, contains a light chain variable domain and a heavy chain variable domain. A single-chain variable fragment (scFv), where the scFv is a single-chain variable fragment that interacts with effector cell antigens. It further contains peptides that impair cFv binding, and these peptides affect tumor-specific proteas. The substrate of ze is linked to the heavy chain variable domain of scFv, and the peptide It further contains half-life-extending molecules, scFV and prostate-specific membrane antigen (PSMA). The binding is Fab or Fab', where Fab or Fab' is Fab light chain polypeptide It contains a tide chain and a Fab heavy chain polypeptide chain, and the Fab heavy chain polypeptide chain is s The cFv light chain variable domain contains Fab or Fab', which is ligated to the C-terminus. Isolated nucleic acid molecules are disclosed.

[0174] In some embodiments herein, stereochemical configuration 2:

[0175] [ka] Isolated nuclei encoding polypeptides or polypeptide complexes including structural arrangements by The acid molecule, a polypeptide complex, contains a light chain variable domain and a heavy chain variable domain. A single-chain variable fragment (scFv), where the scFv is a single-chain variable fragment that interacts with effector cell antigens. It further contains peptides that impair cFv binding, and these peptides affect tumor-specific proteas. The substrate of ze is linked to the N-terminus of the heavy chain variable domain of scFv, The peptide further contains half-life extension molecules, scFV and prostate-specific membrane antigen (PS). Fab or Fab' that binds to MA, wherein Fab or Fab' is a Fab light chain It contains polypeptide chains and Fab heavy chain polypeptide chains, and Fab light chain polypeptide chains. The chain contains Fab or Fab', which is ligated to the C-terminus of the light chain variable domain of scFv. And then, isolated nucleic acid molecules are disclosed.

[0176] Pharmaceutical composition In some embodiments herein, (a) the polypeptides disclosed herein A pharmaceutical composition comprising (b) a polypeptide complex and (b) a pharmaceutically acceptable excipient is disclosed. It will be done.

[0177] In some embodiments, the pharmaceutical composition is (a) Formula I: A2-A1-L1-P1-H1 (Formula I) A polypeptide or polypeptide complex comprising, In the formula, A1 comprises a first antigen-recognition molecule that binds to the effector cell antigen, and P1 comprises A1 It contains a peptide that binds to P1, and L1 connects A1 to P1 and tumor-specific protea It contains a binding portion that is a substrate of the enzyme, H1 contains a half-life extension molecule, and A2 is prostate-specific. A polypeptide or polypeptide containing a second antigen-recognition molecule that binds to the target membrane antigen (PSMA). (b) a butylated complex and (b) a pharmaceutically acceptable excipient.

[0178] In some embodiments, the pharmaceutical composition is (a) Formula I: A2-A1-L1-P1-H1 (Formula I) A polypeptide or polypeptide complex comprising, In the formula, A1 is the first antigen-recognizing molecule that binds to the effector cell antigen, and P1 is A1 It is a peptide that binds to P1, and L1 attaches A1 to P1 and tumor-specific protea The binding portion is the substrate of the enzyme; H1 is a half-life extension molecule; and A2 is prostate-specific. A polypeptide or polypeptide is a second antigen-recognition molecule that binds to the target membrane antigen (PSMA). (b) a butylated complex and (b) a pharmaceutically acceptable excipient.

[0179] In some embodiments, the pharmaceutical composition is (a) Formula I: A2-A1-L1-P1-H1 (Formula I) A polypeptide or polypeptide complex comprising, In the formula, A1 comprises a first antigen-recognition molecule that binds to the effector cell antigen, and P1 comprises A1 It contains a peptide that binds to P1, and L1 connects A1 to P1 and tumor-specific protea It contains a binding portion that is a substrate of the enzyme, H1 contains a half-life extension molecule, and A2 is prostate-specific. A polypeptide or polypeptide containing a second antigen-recognition molecule that binds to the target membrane antigen (PSMA). (b) a butylated complex and (b) a pharmaceutically acceptable excipient.

[0180] In some embodiments, the pharmaceutical composition is (a) Formula I: A2-A1-L1-P1-H1 (Formula I) A polypeptide or polypeptide complex comprising, In the formula, A1 is the first antigen-recognizing molecule that binds to the effector cell antigen, and P1 is A1 It is a peptide that binds to P1, and L1 attaches A1 to P1 and tumor-specific protea The binding portion is the substrate of the enzyme; H1 is a half-life extension molecule; and A2 is prostate-specific. A polypeptide or polypeptide is a second antigen-recognition molecule that binds to the target membrane antigen (PSMA). (b) a butylated complex and (b) a pharmaceutically acceptable excipient.

[0181] In some embodiments, the pharmaceutical composition is (a) Formula Ia: P2-L2-A2-A1-L1-P1-H1 (Formula Ia) (b) a polypeptide or polypeptide complex and a pharmaceutically acceptable excipient include.

[0182] In some embodiments, the pharmaceutical composition is (a) Formula II: L 1a -P 1a -H 1a (Formula II) A polypeptide or polypeptide complex comprising, In the formula, L 1a If not cleaved, the first antigen recognition molecule binds to the effector cell antigen. Child P 1a The first includes a connecting portion that is cleaved by a tumor-specific protease, which connects the two. The antigen-recognizing molecule then binds to a second antigen-recognizing molecule that binds to the prostate-specific membrane antigen (PSMA). Continued, P 1a L 1a If it is not cleaved, the peptide that binds to the first antigen recognition molecule Includes H 1a This includes polypeptides or polypeptide complexes containing half-life extension molecules. (b) a pharmaceutically acceptable excipient.

[0183] In some embodiments, the pharmaceutical composition is (a) Formula II: L 1a -P 1a -H 1a (Formula II) A polypeptide or polypeptide complex comprising, In the formula, L 1a If not cleaved, the first antigen recognition molecule binds to the effector cell antigen. Child P 1a The first includes a connecting portion that is cleaved by a tumor-specific protease, which connects the two. The antigen-recognizing molecule then binds to a second antigen-recognizing molecule that binds to the prostate-specific membrane antigen (PSMA). Continued, P 1a L 1a If it is not cleaved, the peptide that binds to the first antigen recognition molecule Includes H 1a This includes polypeptides or polypeptide complexes containing half-life extension molecules. (b) a pharmaceutically acceptable excipient.

[0184] In some embodiments, the pharmaceutical composition is (a) Formula II: L 1a -P 1a -H 1a (Formula II) A polypeptide or polypeptide complex comprising, In the formula, L 1a If not cleaved, the first antigen recognition molecule binds to the effector cell antigen. Child P 1a The connecting portion is a tumor-specific protease that cleaves the first The antigen-recognizing molecule then binds to a second antigen-recognizing molecule that binds to the prostate-specific membrane antigen (PSMA). Continued, P 1a L 1a If it is not cleaved, the peptide that binds to the first antigen recognition molecule It is D, H 1a These are half-life extension molecules, polypeptides or polypeptide complexes. (b) a pharmaceutically acceptable excipient.

[0185] In some embodiments, the pharmaceutical composition is (a) Formula II: L 1a -P 1a -H 1a (Formula II) A polypeptide or polypeptide complex comprising, In the formula, L 1a If not cleaved, the first antigen recognition molecule binds to the effector cell antigen. Child P 1a The connecting portion is a tumor-specific protease that cleaves the first The antigen-recognizing molecule then binds to a second antigen-recognizing molecule that binds to the prostate-specific membrane antigen (PSMA). Continued, P 1a L 1a If it is not cleaved, the peptide that binds to the first antigen recognition molecule It is D, H 1a These are half-life extension molecules, polypeptides or polypeptide complexes. (b) a pharmaceutically acceptable excipient.

[0186] In some embodiments herein, the pharmaceutical composition is (a) stereostructure 1:

[0187] [ka] A polypeptide or polypeptide complex comprising a structural arrangement by the polypeptide complex The fusion is a single-chain variable fragment (scF) containing a light-chain variable domain and a heavy-chain variable domain. v) scFv has a peptide that impairs the binding of scFv to effector cell antigens. Furthermore, it contains peptides that are substrates for tumor-specific proteases via a binding portion. The peptide is linked to the heavy chain variable domain of cFv and further contains a half-life extension molecule. scFV, as well as Fab or F that bind to prostate-specific membrane antigen (PSMA), are present. ab', where Fab or Fab' is the Fab light chain polypeptide chain and the Fab heavy chain It contains polypeptide chains, and the Fab heavy chain polypeptide chain is a light chain variable domain of scFv. A polypeptide or polypeptide containing Fab or Fab', which is linked to the C-terminus of (b) a complex and (b) a pharmaceutically acceptable excipient.

[0188] In some embodiments herein, the pharmaceutical composition is (a) stereoconformation 2:

[0189] [ka] A polypeptide or polypeptide complex comprising a structural arrangement by the polypeptide complex The fusion is a single-chain variable fragment (scF) containing a light-chain variable domain and a heavy-chain variable domain. v) scFv has a peptide that impairs the binding of scFv to effector cell antigens. Furthermore, it contains peptides that are substrates for tumor-specific proteases via a binding portion. It is linked to the N-terminus of the heavy chain variable domain of cFv, and the peptide further extends the half-life of the molecule. The scFV contained in it binds to prostate-specific membrane antigen (PSMA) each time. or Fab', where Fab or Fab' is a Fab light chain polypeptide chain and F It contains an ab heavy chain polypeptide chain, and the Fab light chain polypeptide chain is a scFv light chain polypeptide. A polypeptide or polypeptide containing Fab or Fab' is ligated to the C-terminus of the variant domain. The present invention comprises (b) a lipeptide complex and (b) a pharmaceutically acceptable excipient.

[0190] In some embodiments, the polypeptide or polypeptide complex is a detectable label. The further comprises a therapeutic agent or a pharmacokinetic modification moiety. In some embodiments, detectable Labeling includes fluorescent labels, radioactive labels, enzymes, nucleic acid probes, or contrast agents.

[0191] In administration to the target, the polypeptide or polypeptide complex disclosed herein The body is incorporated into a pharmaceutical composition together with one or more pharmaceutically acceptable carriers or additives. It may be provided as such. The term "pharmaceutically acceptable carrier" refers to the bioactivity of the component. Any carrier that does not interfere with efficacy and is non-toxic to the target patient can be mentioned. However, it is not limited to these. Examples of suitable pharmaceutical carriers are well known in the art, such as phosphate buffered carriers. Examples include physiological saline solution, water, emulsions such as oil / water emulsions, various types of humectants, and sterile solutions. Such carriers can be formulated using conventional methods and administered to the target at a suitable dose. This is possible. Preferably, the composition is sterile. These compositions contain preservatives, emulsifiers, and It may also contain adjuvants such as dispersants. Prevention of microbial activity is possible with various antimicrobial agents. They may be secured by inclusion bodies containing fungal and antifungal agents.

[0192] The pharmaceutical composition may be in any preferred form (depending on the desired method of administration). When the drug composition is provided in unit dosage form, in a sealed container, and as part of a kit... Such kits may include instructions for use. The kit contains the unit agent mentioned above. It may contain multiple shapes.

[0193] The pharmaceutical composition includes any of the parenteral (e.g., subcutaneous, intramuscular, or intravenous) routes. It may be suitable for administration via an appropriate route. Such compositions may, for example, have their active ingredients sterilized. By mixing with a carrier or excipient under certain conditions, any of the methods known in the pharmaceutical arts can be produced. It may be prepared by law.

[0194] The dosage of the substance disclosed herein is determined based on the disease or disorder to be treated and the treatment to be performed. The results may vary widely depending on the age and condition of the individual, and a physician should consult a doctor. The appropriate dosage will ultimately be determined.

[0195] Treatment method Some embodiments are methods for treating cancer in subjects requiring cancer treatment, and the present invention The process involves administering the isolated polypeptide or polypeptide complex described in the details. This method includes the following steps. In some embodiments, cancer has cells that express PSMA. In some cases, cancer is a solid tumor carcinoma. In some embodiments, cancer is lung cancer. , breast (e.g., HER2+, ER / PR+, TNBC), cervix, ovaries, colon, pancreas , or stomach cancer.

[0196] Some embodiments describe a method for treating prostate cancer in a subject requiring treatment for prostate cancer. The isolated polypeptide or polypeptide complex described herein is included. The method includes the step of administering to. Some embodiments describe metastatic castration-resistant prostate cancer ( A method for treating mCRPCs (mCRPCs) that require treatment, as specified herein The process includes administering the isolated polypeptide or polypeptide complex described. This is one way to do it.

[0197] In some embodiments herein, isolated polypeptides having a long half-life are used. Or polypeptide complexes are described. In some examples, polypeptides or polypeptide complexes are described. The half-life of the tide complex is at least or about 12 hours, 24 hours, 36 hours, 48 ​​hours, 60 hours, 72 hours, 84 hours, 96 hours, 100 hours, 108 hours, 119 hours, 12 0 hours, 140 hours, 160 hours, 180 hours, 200 hours, or more than 200 hours In some cases, the half-life of a polypeptide or polypeptide complex is approximately 12 hours. Approximately 300 hours, approximately 20 hours to approximately 280 hours, approximately 40 hours to approximately 240 hours, approximately 60 hours to approximately It's 200 hours, or in the range of approximately 80 to 140 hours.

[0198] In some embodiments herein, polypeptides or polypeptides are administered once a week. A polypeptide complex is described. In some embodiments, polypeptides or polypeptide complexes are described. The compound is administered once a week intravenously, intramuscularly, intra-injured, topically, subcutaneously, by infusion, or orally. In some embodiments, polypeptides or polypeptide complexes are administered via bolus injection. It is administered once a week. In some embodiments, polypeptides or polypeptide complexes are used. The compound is administered once a week by continuous infusion. In some embodiments, polypeptide Alternatively, polypeptide complexes are administered once a week to the target as a continuous infusion lasting 60 minutes or less. In some embodiments, the polypeptide or polypeptide complex is given for 60 minutes. The following is administered to the subject once a week as a continuous intravenous infusion over the period below. In some embodiments, Polypeptides or polypeptide complexes are administered by continuous intravenous infusion for a period of at least 10 minutes. It is administered to the target group once a week.

[0199] In some embodiments, polypeptides or polypeptide complexes are administered to the subject once a week. The half-life of the given polypeptide or polypeptide complex is at least 30 hours. In some embodiments, the polypeptide or polypeptide complex is administered to the subject once a week. The half-life of the given polypeptide or polypeptide complex is at least 50 hours. In some embodiments, the polypeptide or polypeptide complex is administered to the subject once a week. The half-life of the given polypeptide or polypeptide complex is at least 60 hours. In some embodiments, the polypeptide or polypeptide complex is administered to the subject once a week. The half-life of the given polypeptide or polypeptide complex is at least 70 hours. In some embodiments, the polypeptide or polypeptide complex is administered to the subject once a week. The half-life of the given polypeptide or polypeptide complex is at least 80 hours. In some embodiments, the polypeptide or polypeptide complex is administered to the subject once a week. The half-life of the given polypeptide or polypeptide complex is at least 90 hours. In some embodiments, the polypeptide or polypeptide complex is administered to the subject once a week. The half-life of the given polypeptide or polypeptide complex is at least 100 hours. In some embodiments, the polypeptide or polypeptide complex is administered to the target once a week. The half-life of the administered polypeptide or polypeptide complex is at least 110 hours. Yes. In some embodiments, polypeptides or polypeptide complexes are applied to the target once a week. A single dose is administered, and the half-life of the polypeptide or polypeptide complex is at least 115 hours. In some embodiments, the polypeptide or polypeptide complex is used weekly. A single dose is administered, and the half-life of the polypeptide or polypeptide complex is at least 119 hours. It is in between.

[0200] Production of antibodies that bind to PSMA and CD3 In some embodiments, polypeptides described herein (e.g., antibodies and The binding fragment is useful in the synthesis of polypeptides (e.g., antibodies), according to the technology. It is produced by any method known in the field, specifically by chemosynthesis or recombinant expression. Preferably, it is produced by recombinant expression techniques.

[0201] In some cases, the antibody or its conjugated fragment is expressed by recombination, and the antibody The nucleic acid encoding that binding fragment is a chemically synthesized oligonucleotide (e.g.) For example, Kutmeier et al.'s 1994 "BioTechniques 17:2 Assembled from those described in "42", and containing multiple parts of the sequence encoding the antibody. Synthesis of duplicate oligonucleotides, annealing and lithography of those oligonucleotides This requires ligation and subsequent PCR amplification of the ligated oligonucleotides. Let's assume that.

[0202] Alternatively, nucleic acid molecules encoding antibodies can be hybridized at the 3' and 5' ends of the sequence. PCR amplification using synthetic primers that can be adjusted, or specific to a particular gene sequence. By creating clones using oligonucleotide probes, suitable sources (e.g.) can be used. For example, an antibody cDNA library, or any tissue expressing immunoglobulins. It is generated from a cDNA library (generated from cells) by arbitrary selection.

[0203] In some cases, the antibody or its conjugated fragment generates a polyclonal antibody. To do this, animals such as mice are immunized, or more preferably, for example, K Ohler and Milstein (1975, Nature 256:495-497) ) or Kozbor et al. (1983, Immunology Today 4) :72) or Cole et al. (1985 in Monoclonal Antibodies and Cancer Therapy,Alan R.Li By generating monoclonal antibodies as described in ss, Inc., pp. 77-96) , optionally generated. Alternatively, a clone encoding at least the Fab portion of the antibody. However, for clones of Fab fragments that bind to specific antigens, the Fab expression library By screening (for example, Huse et al.'s 1989 Science (As described in e 246:1275-1281), or by scripting an antibody library By doing so (for example, Clackson et al.'s 1991 Nature) 352:624, Proc.Natl.Acad.Sc by Hane et al., 1997 i. See USA 94:4937), obtainable by choice.

[0204] In some embodiments, along with genes from suitable bioactive human antibody molecules, By splicing genes from mouse antibody molecules with specific antigens, a "chimera" can be created. A technique developed to produce "antibodies" (Pro by Morrison et al. in 1984) c.Natl.Acad.Sci.81:851-855, by Neuberger et al. Nature 312:604-608, 1984; Takeda et al., 1985. The Nature 314:452-454 report is used. The chimeric antibody is mouse mono A variable region derived from a clonal antibody and a constant region of human immunoglobulins. However, various parts of these molecules originate from various animal species.

[0205] In some embodiments, techniques for producing single-chain antibodies are described (U.S. 4, 6). Specification No. 94,778, by Bird, Science 242:423- 42. Proc.Natl.Acad.Sci.US, 1988, by Huston et al. A 85:5879-5883, and Nature 3, 1989 by Ward et al. 34:544-54) is suitable for producing single-chain antibodies. Single-chain antibodies are amino acid chains. By connecting the heavy chain fragment and light chain fragment of the Fv region via a bridge, Formed, yielding a single-chain polypeptide. Assembly of functional Fv fragments in E. coli. Techniques related to this are also used at will (Skerra et al.'s 1988 Science (CE 242:1038-1041).

[0206] In some embodiments, an expression vector containing the nucleotide sequence of an antibody, or an antibody Nucleotide sequences are obtained using conventional techniques (e.g., electroporation, liposome transfusion). It is transferred to the host cell by infection and calcium phosphate precipitation reaction, and then Transfected cells are cultured using conventional techniques for antibody production. In certain embodiments, antibody expression is controlled by a constitutive promoter, an inducible promoter, or tissue It is regulated by a specific promoter.

[0207] In some embodiments, various host expression vector systems are used with the antibodies described herein. This is used to express the binding fragment. Such a host expression system is used for antibodies It not only represents a vehicle that produces and subsequently purifies the coding sequence of appropriate nucleotides, but also the appropriate nucleotide code. When transformed or transfected by a sequence, an antibody can be detected in situ. This represents cells that express the binding fragment. These include antibodies or their binding flags. Recombinant bacteriophage DNA, plasmid DNA, and the coding sequence of the ment. or bacteria transformed by cosmid DNA expression vectors (e.g., Escherichia coli and Bacillus subtilis) Recombinant yeast containing microorganisms such as bacteria, antibodies, or their binding fragments. Yeast transformed with a bacterial expression vector (e.g., Saccharomyces picia), antibody or recombinant viral expression vectors containing the coding sequence of the binding fragment (for example) Insect cell lines infected with baculovirus, recombinant virus expression vectors (for example, Cauliflower mosaic virus (CaMV) and tobacco mosaic virus (T Infected with MV)) or containing the coding sequence of an antibody or its binding fragment Plant cell systems transformed with recombinant plasmid expression vectors (e.g., Ti plasmids) , or the genome of mammalian cells (e.g., metallothionein promoter) or mammary Animal viruses (e.g., late promoter adenovirus, vaccinia virus 7.5K) Mammalian cell lines containing recombinant expression constructs that include promoters derived from romomota For example, COS, CHO, BH, 293, 293T, and 3T3 cells can be cited, but this It is not limited to them.

[0208] For long-term high-yield production of recombinant proteins, stable expression is preferable. In that example, cell lines that stably express antibodies are manipulated by arbitrary selection. Virus replication Rather than using an expression vector that includes the starting point, the host cell uses the appropriate expression regulatory element. (For example, promoters, enhancers, sequences, transcription terminators, polyadenylation sites, etc.) and transformed with DNA controlled by selectable markers. After introduction, the manipulated cells are grown in enriched medium for 1-2 days, then switched to selective medium. Selectable markers in recombinant plasmids confer resistance to selection, allowing cells to plasmin The focus that stably integrates into those chromosomes, is cloned, and is extended into cell lines. This allows for growth in which the antibody or its conjugated fragments can form. It can be advantageously used to manipulate cell lines that express this gene.

[0209] In some cases, herpes simplex virus thymidine kinase (by Wigler et al.) Cell 11:223 of 977), hypoxanthine guanine phosphoribosyltrans Ferase (by Szybalska and Szybalski, 192, Proc. Nat) (l.Acad.Sci.USA 48:202), and adenine phosphoribosyltran It includes the spherase gene (Cell 22:817 by Lowy et al., 1980). Many selection systems are used, not limited to these, such as tk-, hgprt-, Alternatively, it is used in APRT cells. It is also used in antimetabolites (antimetabolite resistance). (ITE resistance) also involves the following gene: dh which gives resistance to methotrexate. fr(Proc.Natl.Acad.Sci.US, 1980 by Wigler et al.) A 77:357, Proc.Natl.Acad. 1981 by O'Hare et al. Sci.USA 78:1527), gpt (Mull) that gives resistance to mycophenolic acid. Proc.Natl.Acad.Sci.USA, 1981 by Igan and Berg 78:2072), neo(Clinic) that gives resistance to aminoglycoside G-418 al Pharmacy 12:488-505, Bio by Wu and Wu, 1991 Therapy 3:87-95, Ann. R. Tolstoshev, 1993. ev.Pharmacol.Toxicol.32:573-596, Mulligan Science 260:926-932, 1993, by Morgan and A. Anderson's 1993 Ann. Rev. Biochem. 62:191-2 17; May 1993, TIB TECH 11(5):155-215), and H Hygro (Gen, developed by Santare et al. in 1984) gives resistance to glomycin. Used as a selection criterion for e 30:147). Available recombinant DNA technology A well-known method in the field of law is Ausubel et al. (eds.), Curren, 1993. t Protocols in Molecular Biology,John Wi ley & Sons, NY, Gene Tran by Kriegler, 1990 sfer and Expression,A Laboratory Manual, Stockton Press, NY, edited by Dracopoli et al., 1994 Current Protocols in Human Genetics,John Chapters 12 and 13 of Wiley & Sons, NY, Colberre-Gar This is described in J.Mol.Biol.150:1) by Apin et al. in 1981.

[0210] In some cases, antibody expression levels increase with vector amplification (regarding commentary). This is "the use of vec" by Bebbington and Hentschel. tors based on gene amplification for the expression of cloned genes in mammalian cells in DNA cloning” Vol.3.(Academic Pr See Ess, New York, 1987). Markers in antibody-expressing vector systems If amplification is possible, the marker gene can be amplified by an increase in the level of the inhibitor present in the host cell culture. The number of copies of the offspring increases. The amplified region is related to the antibody's nucleotide sequence. Therefore, antibody production also increases (Mol. Ce by Crouse et al., 1983). (Ill. Biol. 3:257).

[0211] In some cases, any method known in the field of antibody purification, for example, Mathematics (for example, ion exchange, affinity, specifically specific antibodies after protein A) Affinity for the raw material, and size column chromatography (sizing column chromatography) Lumn chromatography, centrifugation, differential solubility (Urgent solubility), or other standard techniques for protein purification It is used as such.

[0212] Expression vector In some embodiments, the vector is supplied from either a eukaryote or a prokaryote. This includes any suitable vectors that may result from this. In some cases, the vector may be a bacterium (e.g., E. coli). ), insects, yeast (e.g., Pichia pastris), algae, or mammalian sources Examples of bacterial vectors include pACYC177, pASK75, and pBAD vectors. - Series, pBADM Vector Series, pET Vector Series, pETM Vector Series, pGEX Vector Series, pHAT, pHAT2, pMal-c2, pMa l-p2, pQE vector series, pRSET A, pRSET B, pRSET C pTrcHis2 series, pZA31-Luc, pZE21-MCS-1, pFLA G ATS, pFLAG CTS, pFLAG MAC, pFLAG Shift-12 Examples include c, pTAC-MAT-1, pFLAG CTC, or pTAC-MAT-2. It can be done.

[0213] Examples of insect vectors include pFastBac1, pFastBac DUAL, and pF astBac ET, pFastBac HTa, pFastBac HTb, pFas tBac HTc, pFastBac M30a, pFastBact M30b, pF astBac, M30c, pVL1392, pVL1393, pVL1393 M10, pVL1393 M11, pVL1393 M12, pPolh-FLAG1 or p FLAG vectors such as Polh-MAT 2, or pPolh-MAT1 or p Examples of MAT vectors include Polh-MAT2.

[0214] In some cases, yeast vectors may contain Gateway(registered trademark) pDEST(trademark)1 4 Vector, Gateway (Registered Trademark) pDEST (Trademark) 15 Vector, Gateway ay(registered trademark) pDEST(trademark) 17 Vector, Gateway(registered trademark) pDE ST(trademark)24 vector, Gateway(registered trademark)pYES-DEST52 vector - pBAD-DEST49 Gateway (registered trademark) destination vector pAO815 Pichia vector, pFLD1 Pichia pastris vector, pGAPZA, B,&C Pichia Pastris vectors, pPIC3.5K Pichia vectors, pPIC6 A B,&C Pichia vectors, pPIC9K Pichia vectors, pTEF1 / Zeo, pYE S2 yeast vector, pYES2 / CT yeast vector, pYES2 / NT A,B,& Examples include C yeast vectors or pYES3 / CT yeast vectors.

[0215] Examples of algal vectors include the pChlamy-4 vector or the MCS vector. It is possible.

[0216] Examples of mammalian vectors include transient expression vectors and stable expression vectors. Mammalian transient expression vectors include pRK5, p3xFLAG-CMV 8, and p FLAG-Myc-CMV 19, pFLAG-Myc-CMV 23, pFLAG-C MV 2, pFLAG-CMV 6a,b,c, pFLAG-CMV 5.1, pFLA G-CMV 5a,b,c, p3xFLAG-CMV 7.1, pFLAG-CMV 2 0, p3xFLAG-Myc-CMV 24, pCMV-FLAG-MAT1, pCMV -FLAG-MAT2, pBICEP-CMV 3, or hpBICEP-CMV 4 Examples include pFLAG-CMV, a stable expression vector for mammals. 3, p3xFLAG-CMV 9, p3xFLAG-CMV 13, pFLAG-Myc -CMV 21, p3xFLAG-Myc-CMV 25, pFLAG-CMV 4, p 3xFLAG-CMV 10, p3xFLAG-CMV 14, pFLAG-Myc-C MV 22, p3xFLAG-Myc-CMV 26, pBICEP-CMV 1, also pBICEP-CMV 2 may be one example.

[0217] In some cases, cell-free systems are obtained from a mixture of cytoplasmic and / or nuclear components from cells. It exists and is used for nucleic acid synthesis in vitro. In some cases, cell-free systems are used for prokaryotic nucleic acid synthesis. It utilizes either cellular or eukaryotic components. Sometimes, nucleic acid synthesis is performed, for example, in ginger. Obtained from cell-free systems based on Uva cells, Xenopus eggs, or HeLa cells. (Example) Cell-free systems include the E. coli S30 extract system, the E. coli T7 S30 system, or PURExpre Examples include, but are not limited to, ss (registered trademark).

[0218] host cell In some embodiments, the host cells are either naturally occurring or genetically modified. Any suitable cell type can be mentioned, such as a host cell. In some cases, the host cell is a production host cell. It is a principal cell. In some cases, the host cell is a eukaryotic cell. In other cases, the host cell These are prokaryotic cells. In some cases, eukaryotic cells are fungi (e.g., yeast), animal cells, or This includes plant cells. In some cases, prokaryotic cells are bacterial cells. An example of a bacterial cell is Gram. Examples include Gram-positive bacteria and Gram-negative bacteria. Sometimes, Gram-negative bacteria are anaerobic, rod-shaped, and It is both.

[0219] In some cases, Gram-positive bacteria include Actinobacteria, Firmicutes, and Examples include Tenericutes. In some cases, Gram-negative bacteria include Aquifex. Phylum, Deinococcus thermus phylum, Fibrobacter phylum - Chlorobium phylum / Bacteroides Phylum s (FCB group), Phylum fusobacterium, Phylum gemmatimonas, Phylum nitrospira, Plan Phylum Cotomyces - Phylum Vercomicrobium / Phylum Chlamydia (PVC group), Proteobacteria Examples include the phyla A, Spirochetes, or Synergistes. Other bacteria include Acidobacteria. Phylum: *Lium*, Phylum: *Chloroflexus*, Phylum: *Chrysiogenes*, Phylum: *Cyanobacteria*, Phylum: *Deferibacterium* Taa phylum, Dictyoglomi phylum, Thermodesulfobacter It could also be from the Lia phylum or the Thermotoga phylum. The bacterial cells include E. coli, Clostridium botulinum, and It could also be E. coli (Coli bacilli).

[0220] Examples of prokaryotic host cells include BL21, Mach1(trademark), and DH10B(trademark). ), TOP10, DH5α, DH10Bac (trademark), OmniMax (trademark), Meg aX(trademark), DH12S(trademark), INV110, TOP10F', INVαF, TO P10 / P3, ccdB Survival, PIR1, PIR2, Stbl2(trademark) Examples include, but are not limited to, Stbl3 (trademark) or Stbl4 (trademark). stomach.

[0221] In some examples, animal cells include cells from vertebrates or invertebrates. As a result of the fusion, animal cells include marine invertebrates, fish, insects, amphibians, reptiles, or mammals. Examples include the cells of various organisms. In some cases, fungal cells include those of brewer's yeast, baker's yeast, or wine. Examples include yeast cells, such as those of yeast.

[0222] Fungi include yeasts, molds, filamentous fungi, basidiomycetes, and ascomycetes such as zygomycetes. In some cases, yeasts belong to the Ascomycota or Basidiomycota phyla. The Ascomycota includes the Saccharomyces subphylum (true yeasts, such as Saccharomyces cerevisiae). Examples include baker's yeast or the Taflina subphylum (for example, the Schizosaccharomyces class (fission yeast)). In some cases, the Basidiomycota may include the Agaricus subphylum (e.g., Tremella class) or Subphylum Sabikini (e.g., Microbotryomycetes) ) are some examples.

[0223] Exemplary yeasts or filamentous fungi include, for example, the genera: Saccharomyces, Schizosaccharomyces. Genus Cetus, Candida, Pichia, Hanzenula, Kruyweromyces, Tygosaccharo Myces, Yarrowia, Trichosporon, Rhodosporidium Examples include the genera Aspergillus, Fusarium, or Trichoderma (idi). Typical yeasts or filamentous fungi include, for example, the following species: Saccharomyces cerevisiae, Schizosaccharomyces. Romyces pombe, Candida euchylis, Candida boi Candida albicans, Candida tropicalis, Candida stellatoi Candida glabrata, Candida crusae, Candida parasylosis, Candida Dida Gilliermondi, Candida Biswanati, Candida Lusitaniye, Rhodotorula • Pichia metanolica, Pichia Pichia angusta, Pichia pastris, Pichia anoma La, Hansenula polymorpha, Cluiveromyces lactis, Tygosaccharomyces • Luxii, Yarouia liporitica, Trichosporon pullulan, Rhodosporidium Rhodosporidium toru-Aspergillus (black aspergillus) Aspergillus niger, Aspergillus nidurans, Aspergillus awamo Aspergillus oryzae, Trichoderma liesei, Yarouia lipopolitica, Brettanomyces bruccellensis, Candida stellata, Schizosaccharomyces p Mbe, Torraspora delbrückii, Zigosaccharomyces baili, Cryptococcus S. neoformans, Cryptococcus gattii, or Saccharomyces braude One example is [this].

[0224] Exemplary yeast host cells include GS115, KM71H, SMD1168, and SMD11 Pichia pastris yeast strains such as 68H and X-33, as well as INVSc1, etc. Examples include, but are not limited to, the Saccharomyces cerevisiae yeast strains.

[0225] In some instances, the additional animal cells include mollusks, arthropods, annelids, or marine animals. Examples include cells obtained from cottony animals. In some cases, additional animal cells may be used, for example, primates. These are mammalian cells from apes, horses, cattle, pigs, dogs, cats, or rodents. More specifically, rodents include mice, rats, hamsters, gerbils, and chins. Examples include chirpers, fancy rats, or guinea pigs.

[0226] Exemplary mammalian host cells include the 293A cell line, the 293FT cell line, and the 293F cell line. 293H cells, CHO DG44 cells, CHO-S cells, CHO-K1 cells, FUT8 KO CHOK1, Expi293F(TM) cells, Flp-In(TM) T-REx( Trademark)293 cell line, Flp-In(trademark)-293 cell line, Flp-In(trademark)-3 T3 cell line, Flp-In(TM)-BHK cell line, Flp-In(TM)-CHO cells strain, Flp-In(TM)-CV-1 cell line, Flp-In(TM)-Jurkat cells strain, FreeStyle(TM) 293-F cells, FreeStyle(TM) CHO- S cells, GripTite™ 293 MSR cell line, GS-CHO cell line, Hep aRG(TM) cells, T-REx(TM) Jurkat cell line, Per.C6 cells, T- REx(TM)-293 cell line, T-REx(TM)-CHO cell line, and T-REx (Trademark) - Examples include, but are not limited to, HeLa cell lines.

[0227] In some cases, mammalian host cells become stable cell lines, i.e., the genetic material of the desired species. It incorporates this into its own genome, and after multiple generations of cell division, it maintains the ability to express the genetic material product. It is a cell line that possesses it. In some cases, mammalian host cells are transient cell lines, i.e. The target genetic material is not incorporated into its own genome, and the genetic material is not incorporated after multiple generations of cell division. This is a cell line that does not possess the ability to express qualitative products.

[0228] Exemplary insect host cells include Drosophila S2 cells, Sf9 cells, Sf21 cells, Examples include High Five® cells and expressSF+® cells. However, it is not limited to these.

[0229] In some examples, plant cells include algal cells. Exemplary insect cell lines include: Chlamydomonas reinhaltii 137c, or Synechococcus elongatus PC 7942 is listed.

[0230] manufactured goods In another aspect of the present invention, a material useful for treating, preventing, and / or diagnosing the above-mentioned disorders is provided. A manufactured product containing the product is provided. This manufactured product is provided in a container and on or attached to the container. It is equipped with a label or accompanying document. Suitable containers include, for example, bottles, vials. Examples include bottles, syringes, and IV solution bags. The containers are made of glass or plastic. It may be formed from any variety of materials. The container may be used on its own or to treat and prevent disease. , and / or retain one composition in combination with another composition effective for diagnosis, It may have a sterile access port (for example, the container has a stopper through which a subcutaneous injection needle can pass). (This may be an intravenous injection bag or vial.) At least one active in the composition The drug has a first antigen-binding site that specifically binds to CD3, and is defined herein as It is a bispecific antibody that contains a second antigen-binding site that specifically binds to PSMA.

[0231] The label or accompanying leaflet indicates that the composition is used to treat the selected disease. Furthermore, the manufactured product comprises (a) a composition containing the bispecific antibody of the present invention. (b) a container and a second containing a composition comprising a cytotoxic or other therapeutic agent The container may be provided. The product manufactured in this embodiment of the present invention is a composition that is effective against a specific disease. The package may also include additional documentation indicating that it is available for use in treating [condition].

[0232] Alternatively or additionally, the product may contain bacteriostatic distilled water for injection (BWFI), phosphate saturates. Pharmaceutically acceptable buffers such as physiological saline, Ringer's solution, and dextrose solution. A second (or third) container containing the following may further be provided. Other materials desirable from a commercial and user perspective, including agents, filters, needles, and syringes. It may also be equipped with.

[0233] Specific definition The terms used herein are intended to describe only specific cases and are limited to those described herein. It is not intended to be used in this specification. When used herein, the singular forms "a", "an", and Unless otherwise specified in the context, "the" is intended to include plural forms as well. Furthermore, "including", "includes", "having", "has", "with", and The terms such as those variations refer to the forms and / or claims for carrying out the invention. The term "comprising" is used to the extent that it is used in any of the surrounding contexts. They are intended to be similarly included.

[0234] The term "antibody" is used in its broadest sense, referring to a fully assembled organism capable of binding to an antigen. Antibodies, antibody fragments, e.g., Fab, F(ab')2, Fv, single-chain antibodies (s cFv), diabodies, antibody chimeras, hybrid antibodies, dual This includes specific antibodies, etc.

[0235] The term "complementarity-determining region" or "CDR" refers to the region to which an antibody binds, and the remainder of the variable region. A segment of the antibody's variable region that is structurally complementary to an epitope that is more variable. Therefore, CDR is sometimes referred to as having a highly variable area. The variable area is equivalent to three CDs. Contains R. CDR peptides are used to construct the gene that encodes the CDR of the target antibody. Such genes can be obtained, for example, from RNA of antibody-producing cells. The region is prepared by using a polymerase chain reaction to synthesize it. For example, Methods: A Companion to Method by Larrick et al. s in Enzymology 2:106 (1991), Courtenay-Lu "Genetic Manipulation of Monoclonal" by ck Antibodies”, in Monoclonal Antibodies:Pr oduction,Engineering and Clinical Applic ation, Ritter et al. (eds.), pp. 166-179 (Cambridge University) (City Press 1995), and "Genetic M" by Ward et al. anipulation and expression of antibodies ",in Monoclonal Antibodies:Principles an d Applications, Birch et al. (eds.), pp. 137-185 (Wiley- See Liss, Inc. (1995).

[0236] The term "Fab" includes the constant region of the light chain and the first constant region (CH1) of the heavy chain. It refers to the protein that contains it. A Fab fragment is one or more of the antibody hinge region. By adding a small number of residues at the carboxyl terminus of the cysteine-containing heavy chain CH1 domain... Furthermore, it differs from Fab' fragment. Fab'-SH, in this specification, refers to the constant region. The Fab' flag is a designation term for Fab', which refers to a cysteine ​​residue that carries a free thiol group. Ment is produced by reducing the heavy chain disulfide bridges of the F(ab')2 fragment. Other chemical couplings of antibody fragments are also known.

[0237] "Single-chain variable fragments (scFv)" are short linker peptides consisting of 10 to approximately 25 amino acids. A fusion protein of the variable regions of the antibody's heavy chain (VH) and light chain (VL) that are linked to the cydoplasm. Linkers are usually rich in glycine for flexibility, and also for solubility. It is also rich in serine or threonine, and the N-terminus of VH and the C-terminus of VL, or vice versa, are connected. This can continue. This protein is involved in the removal of the constant region and the introduction of the linker. Furthermore, it retains the specificity of the original antibody. scFv antibodies are, for example, from Houston, JS. , as described in Methods in Enzymol. 203 (1991) 46-96 In addition, the antibody fragment is assembled together with the VL domain to the functional antigen-binding site. The characteristic of a VH domain is that it is possible, or that a VL domain can be assembled together with a VH domain. It contains a single-chain polypeptide with the characteristics of , thereby enabling the antigenic binding of full-length antibodies. It provides optimal characteristics.

[0238] Preferred embodiments of the present disclosure have been shown and described herein, but those skilled in the art will find them useful. Therefore, it will be clear that such embodiments are provided as merely examples. Numerous variations, alterations, and substitutions will be conceived by those skilled in the art without deviation. Various alternatives to the embodiments of this disclosure described herein may be used in the implementation of this disclosure. It should be understood that this may be done. The following claims define the scope of this disclosure. Therefore, methods and structures within the scope of these claims and their equivalents are thereby encompassed. That is the intention.

[0239] Embodiment Embodiment 1 uses an isolated polypeptide according to formula I:A2-A1-L1-P1-H1. or is a polypeptide complex, in which A1 is a first that binds to the effector cell antigen. It contains an antigen recognition molecule, P1 contains a peptide that binds to A1, and L1 binds A1 to P1. It continues to contain a linking portion that is a substrate for tumor-specific proteases, and H1 has a prolonged half-life. It contains long molecules, and A2 is a second antigen-recognizing molecule that binds to prostate-specific membrane antigen (PSMA). Includes isolated polypeptides or polypeptide complexes containing [the specified substance].

[0240] Embodiment 2 is an embodiment in which the first antigen recognition molecule comprises an antibody or antibody fragment. The isolated polypeptide or polypeptide complex described in 1.

[0241] Embodiment 3 is an antibody or antibody flag in which the first antigen recognition molecule is human or humanized. The isolated polypeptide or polypeptide complex described in Embodiment 1, comprising ment include.

[0242] Embodiment 4 is an embodiment in which L1 is bound to the N-terminus of the first antigen recognition molecule, as in Embodiments 1 to 3. The isolated polypeptide or polypeptide complex described in any one of the following:

[0243] Embodiment 5 is an embodiment in which A2 is bound to the C-terminus of the first antigen recognition molecule, as in Embodiments 1 to 3. The isolated polypeptide or polypeptide complex described in any one of the following:

[0244] Embodiment 6 is an embodiment in which L1 is bound to the C-terminus of the first antigen recognition molecule, as in Embodiments 1 to 3. The isolated polypeptide or polypeptide complex described in any one of the following:

[0245] Embodiment 7 is an embodiment in which A2 is bound to the N-terminus of the first antigen recognition molecule, as in Embodiments 1 to 3. The isolated polypeptide or polypeptide complex described in any one of the following:

[0246] Embodiment 8 describes an antibody or antibody fragment as a single-chain variable fragment, a single domain Isolation according to any one of Embodiments 2 to 7, comprising an antibody or Fab fragment. Contains a polypeptide or polypeptide complex.

[0247] Embodiment 9 is described in Embodiment 8, wherein A1 is a single-chain variable fragment (scFv). Contains isolated polypeptides or polypeptide complexes.

[0248] Embodiment 10 is an embodiment in which scFv is a heavy chain polypeptide and scFv light chain polypeptide Includes an isolated polypeptide or polypeptide complex as described in Embodiment 9, which contains cydo. nothing.

[0249] Embodiment 11 is an isolated antibody as described in Embodiment 8, wherein A1 is a single-domain antibody. Contains lipeptides or polypeptide complexes.

[0250] Embodiment 12 is a single-domain antibody derived from a camelid family, in which the antibody or antibody fragment is a single-domain antibody derived from a camelid family. Chain variable fragment (scFv), heavy chain variable domain (VH domain), light chain variable domain The isolated substance according to Embodiment 8, comprising a (VL domain) or a variable domain (VHH) Contains a polypeptide or polypeptide complex.

[0251] Embodiment 13 is an embodiment in which A1 includes an anti-CD3e single-chain variable fragment, from Embodiment 1 to 1. The comprising an isolated polypeptide or polypeptide complex as described in any one of 2.

[0252] Embodiment 14 is a state in which A1 has a K content of 1 μM or less relative to CD3 on CD3-expressing cells. D Join One of embodiments 1 to 12 includes an anti-CD3e single-chain variable fragment. Contains isolated polypeptides or polypeptide complexes.

[0253] Embodiment 15 is any of Embodiments 1 to 14, wherein the effector cell antigen includes CD3. One comprises an isolated polypeptide or polypeptide complex as described in one.

[0254] Embodiment 16 is a variable light chain in which A1 can each specifically bind to human CD3. The isolated polypeptide or polypeptide composite described in Embodiment 1, including a variable heavy chain. Includes fusion.

[0255] Embodiment 17 is in which A1 is muromonab-CD3 (OKT3), otelixizumab (TR X4), Teplizumab (MGA031), Bicilizumab (Nuvion), SP34, X 35, VIT3, BMA030(BW264 / 56), CLB-T3 / 3, CRIS7, YTH12.5, F111-409, CLB-T3.4.2, TR-66, WT32, S Pv-T3b, 11D8, XIII-141, XIII-46, XIII-87, 12F 6, T3 / RW2-8C8, T3 / RW2-4B6, OKT3D, M-T301, SMC 2, F101.01, UCHT-1, WT-31, 15865, 15865v12, 15 Complementarity determination region (CD) selected from the group consisting of 865v16 and 15865v19. Includes an isolated polypeptide or polypeptide complex as described in Embodiment 1, comprising R). nothing.

[0256] Embodiment 18 describes how, when L1 is cleaved by a tumor-specific protease, the poly of formula I is produced. The single peptide or polypeptide complex binds to effector cells, as described in Embodiment 1. Contains separated polypeptides or polypeptide complexes.

[0257] Embodiment 19 is in which L1 is cleaved by a tumor-specific protease and A1 is an effector molecule. When binding to cells, the polypeptide or polypeptide complex of formula I is transferred to effector cells. The conjugate includes the isolated polypeptide or polypeptide complex described in Embodiment 1. .

[0258] Embodiment 20 is an isolated pharmacology described in Embodiment 19, wherein the effector cells are T cells. Contains lipeptides or polypeptide complexes.

[0259] Embodiment 21 is a poly(A1) which is part of the TCR-CD3 complex on effector cells. A peptide is bound to the isolated polypeptide or polypeptide compound described in Embodiment 1. Includes fusion.

[0260] Embodiment 22 describes a polypeptide that is part of the TCR-CD3 complex, which is in human CD3ε. This includes an isolated polypeptide or polypeptide complex as described in Embodiment 21.

[0261] Embodiment 23 is an effector cell antigen comprising CD3, and scFV is a complementarity-determining region ( The CD-Rs include HC-CDR1, HC-CDR2, and HC-CDR3, and scF V's HC-CDR1, HC-CDR2, and HC-CDR3 are distributed in HC-CDR1 Includes column number 1, sequence number 2 in HC-CDR2, and sequence number 3 in HC-CDR3. scFV uses LC-CDR1, LC-CDR2, and LC-CDR3 as CDRs. Including scFV's LC-CDR1, LC-CDR2, and LC-CDR3, LC-C In DR1, it's sequence number 4; in LC-CDR2, it's sequence number 5; and in LC-CDR3, it's the sequence. The isolated polypeptide or polypeptide complex described in Embodiment 1, including number 6 include.

[0262] Embodiment 24 is an effector cell antigen comprising CD3, and scFv is according to Sequence ID No. 7. An isolated polypeptide or polypeptide composite according to Embodiment 1, comprising an amino acid sequence. Includes fusion.

[0263] Embodiment 25 is an embodiment in which the second antigen recognition molecule comprises an antibody or antibody fragment. The isolated polypeptide or polypeptide complex described in any one of states 1 to 24. Includes.

[0264] Embodiment 26 is an antibody or antibody fragment that is a single-chain variable fragment, single domain An antibody, or an isolated polypeptide or polypropylene according to Embodiment 25, comprising Fab. Contains peptide complexes.

[0265] Embodiment 27 is a single-domain antibody derived from a camelid family, in which the antibody or antibody fragment is Single-chain variable fragment (scFv), heavy-chain variable domain (VH domain), light-chain variable domain Embodiment 27, which includes a main (VL domain) or a variable domain (VHH). Contains isolated polypeptides or polypeptide complexes.

[0266] Embodiment 28 is an embodiment in which the antibody or antibody fragment is human or humanized. The present invention comprises an isolated polypeptide or polypeptide complex as described in Form 25.

[0267] Embodiment 29 is the isolated polypeptide described in Embodiment 26, wherein A2 is Fab. or containing polypeptide complexes.

[0268] Embodiment 30 is a combination of (a) Fab light chain polypeptide and (b) Fab heavy chain polypeptide. The isolated polypeptide or polypeptide composite described in Embodiment 29, comprising a lipeptide. Includes fusion.

[0269] Embodiment 31 is an embodiment in which Fab uses HC-CDR1 and HC- as complementarity determination regions (CDRs). Includes CDR2 and HC-CDR3, Fab's HC-CDR1, HC-CDR2, HC-CDR3 is sequence number 8 in HC-CDR1 and sequence number 9 in HC-CDR2. , and HC-CDR3 includes sequence number 10, and Fab is calling it LC-CDR as a CDR. 1. Includes LC-CDR2 and LC-CDR3, and Fab's LC-CDR1, LC-C DR2 and LC-CDR3 are sequence numbers 11 in LC-CDR1 and LC-CDR2. The embodiment described in Embodiment 29 includes Sequence ID 12, and Sequence ID 13 in LC-CDR3. Contains isolated polypeptides or polypeptide complexes.

[0270] Embodiment 32 is a Fab light chain polypeptide comprising the amino acid sequence of SEQ ID NO: 14 , comprising the isolated polypeptide or polypeptide complex described in Embodiment 30.

[0271] Embodiment 33 comprises a Fab heavy chain polypeptide containing the amino acid sequence of SEQ ID NO: 15. , comprising the isolated polypeptide or polypeptide complex described in Embodiment 30.

[0272] Embodiment 34 is an embodiment in which the Fab light chain polypeptide of A2 is a single-chain variable fragment (s The isolated polypeptide or poly(cFv) described in Embodiment 30 is bound to the C-terminus of cFv. Contains lipeptide complexes.

[0273] Embodiment 35 is an embodiment in which the Fab heavy chain polypeptide of A2 is a single chain variable fragment (s The isolated polypeptide or poly(cFv) described in Embodiment 30 is bound to the C-terminus of cFv. Contains lipeptide complexes.

[0274] Embodiment 36 is a Fab light chain polypeptide of A2 that is a single chain variable fragment (s The isolated polypeptide or poly(cFv) described in Embodiment 30 is bound to the N-terminus of cFv. Contains lipeptide complexes.

[0275] Embodiment 37 is a Fab heavy chain polypeptide of A2 that is a single chain variable fragment (s The isolated polypeptide or poly(cFv) described in Embodiment 30 is bound to the N-terminus of cFv. Contains lipeptide complexes.

[0276] Embodiment 38 is an embodiment in which the Fab heavy chain polypeptide of A2 is replaced with the scFv heavy chain polypeptide of A1. The isolated polypeptide or polypeptide complex described in Embodiment 30 is bound to the isolated polypeptide or polypeptide complex described in Embodiment 30. Includes.

[0277] Embodiment 39 is an embodiment in which the Fab light chain polypeptide of A2 is replaced with the scFv heavy chain polypeptide of A1. The isolated polypeptide or polypeptide complex described in Embodiment 30 is bound to the isolated polypeptide or polypeptide complex described in Embodiment 30. Includes.

[0278] Embodiment 40 is a combination of the Fab heavy chain polypeptide of A2 and the scFv light chain polypeptide of A1. The isolated polypeptide or polypeptide complex described in Embodiment 30 is bound to the isolated polypeptide or polypeptide complex described in Embodiment 30. Includes.

[0279] Embodiment 41 is an embodiment in which the Fab light chain polypeptide of A2 is replaced with the scFv light chain polypeptide of A1. The isolated polypeptide or polypeptide complex described in Embodiment 30 is bound to the isolated polypeptide or polypeptide complex described in Embodiment 30. Includes.

[0280] Embodiment 42 further comprises P2 and L2, wherein P2 is a pepto bonded to A2. It contains tide, and L2 connects A1 to P1 and is a substrate of tumor-specific proteases. An isolated polypeptide according to any one of Embodiments 1 to 41, including a connecting portion. or containing polypeptide complexes.

[0281] Embodiment 43 is a polypeptide or polypeptide complex of formula Ia:P2-L2-A The isolated polypectin described in Embodiment 42 is due to 2-A1-L1-P1-H1. Contains a peptide or polypeptide complex.

[0282] Embodiment 44 is an embodiment in which the Fab heavy chain polypeptide of A2 is replaced with the scFv heavy chain polypeptide of A1. Embodiment 43 describes how L2 is bound to the Fab light chain polypeptide of A2. Contains isolated polypeptides or polypeptide complexes.

[0283] Embodiment 45 is an embodiment in which the Fab light chain polypeptide of A2 is replaced with the scFv heavy chain polypeptide of A1. Embodiment 43 describes how L2 is bound to the Fab heavy chain polypeptide of A2. Contains isolated polypeptides or polypeptide complexes.

[0284] Embodiment 46 is a combination of the Fab heavy chain polypeptide of A2 and the scFv light chain polypeptide of A1. Embodiment 43 describes how L2 is bound to the Fab light chain polypeptide of A2. Contains isolated polypeptides or polypeptide complexes.

[0285] Embodiment 47 is an embodiment in which the Fab light chain polypeptide of A2 is replaced with the scFv light chain polypeptide of A1. Embodiment 43 describes how L2 is bound to the Fab heavy chain polypeptide of A2. Contains isolated polypeptides or polypeptide complexes.

[0286] Embodiment 48 is an embodiment in which P1 impairs the binding of A1 to the effector cell antigen, as in Embodiment 1. comprising an isolated polypeptide or polypeptide complex as described in any one of the 47s .

[0287] Embodiment 49 is an embodiment in which P1 is ionic interaction, electrostatic interaction, hydrophobic interaction, Pi- A1 is formed by stacking interactions, H-bond interactions, or a combination thereof. The isolated polypeptide or polypeptide described in any one of Embodiments 1 to 48 is combined. Contains lipeptide complexes.

[0288] Embodiment 50 has less than 70% sequence homology to the effector cell antigen. The isolated polypeptide or poly(poly) described in any one of Embodiments 1 to 48 Contains peptide complexes.

[0289] Embodiment 51 is an embodiment in which P2 impairs the coupling of A2 to PSMA, as in Embodiments 1 to 50. The comprising any one of the isolated polypeptides or polypeptide complexes described herein.

[0290] Embodiment 52 is an embodiment in which P2 is involved in ionic interaction, electrostatic interaction, hydrophobic interaction, and Pi- A2 is formed by stacking interactions, H-bond interactions, or a combination thereof. The isolated polypeptide or polypropylene described in any one of Embodiments 1 to 50 is combined. Contains lipeptide complexes.

[0291] Embodiment 53 is an embodiment in which P2 is bound to A2 at or near the antigen-binding site. An isolated polypeptide or polypeptide complex described in any one of 1 to 50 include.

[0292] Embodiment 54 is an embodiment in which P2 has less than 70% sequence homology to PSMA. An isolated polypeptide or polypeptide composite described in any one of the application forms 1 to 50 Includes fusion.

[0293] Embodiment 55 is a peptide sequence in which P1 or P2 has a length of at least 10 amino acids. Including isolated polypeptides or polypeptides described in any one of Embodiments 1 to 54. Contains butyl complex.

[0294] Embodiment 56 is a configuration in which P1 or P2 has a length of at least 10 amino acids and a length of 20 amino acids. The isolated peptide sequence described in any one of Embodiments 1 to 54, comprising a peptide sequence below the mino acid Contains polypeptides or polypeptide complexes.

[0295] Embodiment 57 is a peptide sequence in which P1 or P2 has a length of at least 16 amino acids. Including isolated polypeptides or polypeptides described in any one of Embodiments 1 to 54. Contains butyl complex.

[0296] Embodiment 58 is a peptide sequence in which P1 or P2 has a length of 40 amino acids or less. Isolated polypeptide or polypeptide as described in any one of Embodiments 1 to 54 Includes complexes.

[0297] Embodiment 59 is a configuration in which P1 or P2 comprises at least two cysteine ​​amino acid residues. , isolated polypeptide or polypeptide as described in any one of Embodiments 1 to 54 Includes the D complex.

[0298] Embodiment 60 is an embodiment in which P1 or P2 comprises a cyclic peptide or a linear peptide. The isolated polypeptide or polypeptide complex described in any one of states 1 to 54. Includes.

[0299] Embodiment 61 is any of Embodiments 1 to 54, wherein P1 or P2 comprises a cyclic peptide. The comprising one of the isolated polypeptides or polypeptide complexes described herein.

[0300] Embodiment 62 is any of Embodiments 1 to 54, wherein P1 or P2 comprises a linear peptide. The comprising one of the isolated polypeptides or polypeptide complexes described herein.

[0301] Embodiment 63 is an embodiment in which P1 comprises at least two cysteine ​​amino acid residues. An isolated polypeptide or polypeptide complex as described in any one of 1 to 54 include.

[0302] Embodiment 64 is an amino acid by any one of SEQ ID NOs: 16-19 or 78. An isolated polypeptide according to any one of Embodiments 1 to 54, including an acid sequence, or It contains polypeptide complexes.

[0303] Embodiment 65 is one of any 1 to 64 embodiments in which L1 is bonded to the N-terminus of A1. The following comprises an isolated polypeptide or polypeptide complex described above.

[0304] Embodiment 66 is any of Embodiments 1 to 64, wherein L1 is bonded to the C-terminus of A1. One comprises an isolated polypeptide or polypeptide complex as described in one.

[0305] Embodiment 67 is any of Embodiments 1 to 66, wherein L2 is bonded to the N-terminus of A2. One comprises an isolated polypeptide or polypeptide complex as described in one.

[0306] Embodiment 68 is one of Embodiments 1 to 66, wherein L2 is bonded to the C-terminus of A2. One comprises an isolated polypeptide or polypeptide complex as described in one.

[0307] Embodiment 69 is a peptide in which L1 or L2 has at least 5 to 50 amino acids. The isolated polypeptide described in any one of Embodiments 1 to 68 is a cytoplasmic sequence. or includes polypeptide complexes.

[0308] Embodiment 70 is a pair in which L1 or L2 has at least 10 to 30 amino acids. A plutide sequence, an isolated polypeptide according to any one of Embodiments 1 to 68 or containing polypeptide complexes.

[0309] Embodiment 71 is a peptide sequence in which L1 or L2 has at least 10 amino acids. The isolated polypeptide or poly(poly) described in any one of Embodiments 1 to 68 is Contains peptide complexes.

[0310] Embodiment 72 is a peptide sequence in which L1 or L2 has at least 18 amino acids. The isolated polypeptide or poly(poly) described in any one of Embodiments 1 to 68 is Contains peptide complexes.

[0311] Embodiment 73 is a peptide sequence in which L1 or L2 has at least 26 amino acids. The isolated polypeptide or poly(poly) described in any one of Embodiments 1 to 68 is Contains peptide complexes.

[0312] Embodiment 74 is such that L1 or L2 is (G2S) n The expression contains n, where n is A single integer from 1 to 3 (sequence number 118), as described in any one of embodiments 1 to 68. Contains separated polypeptides or polypeptide complexes.

[0313] Embodiment 75 is (G2S) n , (GS) n (GSGGS) n (Sequence 5) 0), (GGGS) n (Sequence ID 51), (GGGGS) n (Sequence ID 52), and ( GSSGGS) n The formula has an expression selected from the group consisting of (Sequence No. 53), where n The isolated port according to any one of embodiments 1 to 68, wherein the integer is at least 1. Contains lipeptides or polypeptide complexes.

[0314] Embodiment 76 is characterized in that L1 is cleaved by a tumor-specific protease, thereby A1 is In one embodiment, when exposed to feta cell antigen, P1 is debound from A1. An isolated polypeptide or polypeptide complex as described in any one of 1 to 68 include.

[0315] Embodiment 77 is characterized by L2 being cleaved by a tumor-specific protease, thereby converting A2 to P When exposed to SMA, P2 becomes uncoupled from A2, Embodiments 1 to 68 The isolated polypeptide or polypeptide complex described in any one of the following:

[0316] Embodiment 78 describes a tumor-specific protease as a matrix metalloproteinase (MM P), serine protease, cysteine ​​protease, threonine protease, and Any of Embodiments 1 to 68 selected from the group consisting of aspartic acid proteases One comprises an isolated polypeptide or polypeptide complex as described in one.

[0317] Embodiment 79 is a matrix metalloproteinase that comprises MMP2, MMP7, and MMP9 Isolated polypeptide according to Embodiment 78, comprising MMP13 or MMP14 or containing polypeptide complexes.

[0318] Embodiment 80 is a serine protease that is a matryptase (MTSP1), urokinase An isolated polypeptide or polypeptide according to Embodiment 78, comprising ze or hepsin. Contains butyl complex.

[0319] Embodiment 81 is a urokinase-cleavable amino acid sequence, matrix Amino acid sequences that can be cleaved by matrix metalloproteinases, or as described in any one of Embodiments 1 to 68, which includes a legmine-cleavable amino acid sequence. Contains isolated polypeptides or polypeptide complexes.

[0320] Embodiment 82 is an embodiment in which L1 or L2 includes the amino acid sequence according to SEQ ID NO: 23. The isolated polypeptide or polypeptide complex described in any one of states 1 to 68. Includes.

[0321] Embodiment 83 is such that L1 or L2 corresponds to one of sequence numbers 20-49 or 78. The isolated polypectomy according to any one of Embodiments 1 to 68, comprising the amino acid sequence thereof Contains a peptide or polypeptide complex.

[0322] Embodiment 84 is a linker 25 (ISSGLLSGRSDAG) ( Sequence ID 45), Linker 26 (AAGLLAPPGGLSGRSDAG) (Sequence ID 4 6) Linker 27 (SPLGLSGRSDAG) (Sequence ID 47), or Linker The amino acid sequence of 28(LSGRSDAGSPLGLAG)(SEQ ID NO: 48), or For the amino acid sequence of linker 25, linker 26, linker 27, or linker 28 Includes an amino acid sequence having one, two, or three amino acid substitutions, additions, or deletions. , isolated polypeptide or polypeptide as described in any one of Embodiments 1 to 68 Includes the D complex.

[0323] Embodiment 85 is described in any one of Embodiments 1 to 83, wherein H1 comprises a cyclic peptide. Contains isolated polypeptides or polypeptide complexes.

[0324] Embodiment 86 is an embodiment of Embodiment 1 in which the polymer contains polyethylene glycol (PEG). The isolated polypeptide or polypeptide complex described in any one of the 83 items is included. .

[0325] Embodiment 87 is described in any one of Embodiments 1 to 83, wherein H1 contains albumin. Contains isolated polypeptides or polypeptide complexes.

[0326] Embodiment 88 is described in any one of Embodiments 1 to 83, wherein H1 includes an Fc domain. Contains isolated polypeptides or polypeptide complexes.

[0327] Embodiment 89 is the isolated method described in Embodiment 87, wherein the albumin is serum albumin. Contains polypeptides or polypeptide complexes.

[0328] Embodiment 90 is the isolation described in Embodiment 87, wherein the albumin is human serum albumin. Contains a polypeptide or polypeptide complex.

[0329] Embodiment 91 is an embodiment in which H1 comprises a polypeptide, ligand, or small molecule, as in Embodiment 1. Includes an isolated polypeptide or polypeptide complex as described in any one of 83 from nothing.

[0330] Embodiment 92 describes a polypeptide, ligand, or small molecule as a serum protein or The fragment, circulating immunoglobulin or its fragment, or CD35 / C The isolated polypeptide or polypeptide complex described in Embodiment 91, which binds to R1. Including the body.

[0331] Embodiment 93 is a serum protein that is a thyroxine-binding protein, transthyretin , α1-acid glycoprotein, transferrin, transferrin receptor or its Embodiment 88 includes a lanceferrin binding moiety, fibrinogen, or albumin. Contains isolated polypeptides or polypeptide complexes.

[0332] Embodiment 94 is a circulating immunoglobulin molecule consisting of IgG1, IgG2, IgG3, and IgG 4. Isolated polypeptide according to Embodiment 88, comprising slgA, IgM, or IgD Contains cytoplasm or polypeptide complexes.

[0333] Embodiment 95 is the isolated embodiment described in Embodiment 92, wherein the serum protein is albumin. Contains polypeptides or polypeptide complexes.

[0334] Embodiment 96 is an isolated polypeptide as described in Embodiment 91, wherein the polypeptide is an antibody. Contains a peptide or polypeptide complex.

[0335] Embodiment 97 describes an antibody that is a single-domain antibody, a single-chain variable fragment, or a Fab. This includes, but is not limited to, the isolated polypeptide or polypeptide complex described in Embodiment 96.

[0336] Embodiment 98 includes a single-domain antibody that binds to albumin. , comprising the isolated polypeptide or polypeptide complex described in Embodiment 97.

[0337] Embodiment 99 is an embodiment in which the single-domain antibody is a human antibody or a humanized antibody. The isolated polypeptide or polypeptide complex described in 7.

[0338] Embodiment 100 is described in Embodiment 97, in which the single-domain antibody is 645gH1gL1. Contains isolated polypeptides or polypeptide complexes.

[0339] Embodiment 101 is an embodiment in which the single-domain antibody is 645dsgH5gL4, and Embodiment 97 Contains the isolated polypeptide or polypeptide complex described above.

[0340] Embodiment 102 is an embodiment in which the single-domain antibody is 23-13-A01-sc02. The isolated polypeptide or polypeptide complex described in Form 97.

[0341] Embodiment 103 is a single-domain antibody in which A10m3 or a fragment thereof This includes the isolated polypeptide or polypeptide complex described in Embodiment 97.

[0342] Embodiment 104 is described in Embodiment 97, wherein the single-domain antibody is DOM7r-31. Contains isolated polypeptides or polypeptide complexes.

[0343] Embodiment 105 is an embodiment of Embodiment 97 in which the single-domain antibody is DOM7h-11-15. Contains the isolated polypeptide or polypeptide complex described above.

[0344] Embodiment 106 is a single-domain antibody that is Alb-1, Alb-8, or Alb-2 3, comprising the isolated polypeptide or polypeptide complex described in Embodiment 97 nothing.

[0345] Embodiment 107 is the isolated antibody described in Embodiment 97, wherein the single-domain antibody is 10E. Contains a polypeptide or polypeptide complex.

[0346] Embodiment 108 is an embodiment in which a single-domain antibody is used as the complementarity-determining region (CDR) for HC-CD Includes R1, HC-CDR2, and HC-CDR3, and is a single-domain antibody HC-CDR 1. HC-CDR2 and HC-CDR3 are sequence numbers 54 in HC-CDR1, HC -CDR2 includes sequence number 55, and HC-CDR3 includes sequence number 56, Embodiments Contains an isolated polypeptide or polypeptide complex as described in 97.

[0347] Embodiment 109 is an HC-CD antibody with a complementation-determining region (CDR). Includes R1, HC-CDR2, and HC-CDR3, and is a single-domain antibody HC-CDR 1. HC-CDR2 and HC-CDR3 are sequence numbers 58 in HC-CDR1, HC -CDR2 includes sequence number 59, and HC-CDR3 includes sequence number 60, Embodiments Contains an isolated polypeptide or polypeptide complex as described in 97.

[0348] Embodiment 110 is the isolated antibody described in Embodiment 97, wherein the single-domain antibody is SA21. Contains a polypeptide or polypeptide complex.

[0349] Embodiment 111 is a polypeptide or polypeptide complex in which modified amino acids, non-natural Embodiments 1 to 110 include amino acids, unnaturally modified amino acids, or combinations thereof. The isolated polypeptide or polypeptide complex described in any one of the following:

[0350] Embodiment 112 describes a modified amino acid or a non-naturally modified amino acid that includes post-translational modifications. This includes the isolated polypeptide or polypeptide complex described in Embodiment 111.

[0351] Embodiment 113 is an embodiment in which H1 includes a connecting portion (L3) that connects H1 to P1. Isolated polypeptide or polypeptide complex as described in any one of 1 to 112 Includes.

[0352] Embodiment 114 is a peptide sequence in which L3 has at least 5 to 50 amino acids. This includes the isolated polypeptide or polypeptide complex described in Embodiment 113. nothing.

[0353] Embodiment 115 is a peptide compound in which L3 has at least 10 to 30 amino acids. The column is the isolated polypeptide or polypeptide complex described in Embodiment 113. include.

[0354] Embodiment 116 is a peptide sequence in which L3 has at least 10 amino acids. This includes the isolated polypeptide or polypeptide complex described in Embodiment 113.

[0355] Embodiment 117 is a peptide sequence in which L3 has at least 18 amino acids. This includes the isolated polypeptide or polypeptide complex described in Embodiment 113.

[0356] Embodiment 118 is a peptide sequence in which L3 has at least 26 amino acids. This includes the isolated polypeptide or polypeptide complex described in Embodiment 113.

[0357] Embodiment 119 is such that L3 is (G2S) n , (GS) n (GSGGS) n (Sequence 50), (GGGS) n (Sequence ID 51), (GGGGS) n (Sequence ID 52), and (GSSGGS)n The formula has an expression selected from the group consisting of (Sequence ID 53), in which, The isolated polypeptide according to Embodiment 113, wherein n is an integer of at least 1 or Contains polypeptide complexes.

[0358] Embodiment 120 is an embodiment in which L3 includes the amino acid sequence according to SEQ ID NO: 22, as in Embodiment 113. Contains the isolated polypeptide or polypeptide complex described above.

[0359] Embodiment 121 is a polypeptide or polypeptide complex that corresponds to SEQ ID NOs. 62-77. In contrast, the embodiment 1 includes an amino acid sequence having at least 95% sequence identity. Contains isolated polypeptides or polypeptide complexes.

[0360] Embodiment 122 is a polypeptide or polypeptide complex that corresponds to Sequence ID No. 72. The isolated amino acid sequence according to Embodiment 1, having at least 95% sequence identity Contains a polypeptide or polypeptide complex.

[0361] Embodiment 123 is a polypeptide or polypeptide complex that corresponds to SEQ ID NO: 73 The isolated amino acid sequence according to Embodiment 1, having at least 95% sequence identity Contains a polypeptide or polypeptide complex.

[0362] Embodiment 124 is a polypeptide or polypeptide complex, with Sequence ID No. 62 and The implementation includes an amino acid sequence having at least 95% sequence identity with sequence number 63. The material comprises an isolated polypeptide or polypeptide complex described in Form 1.

[0363] Embodiment 125 is an embodiment in which a polypeptide or polypeptide complex is represented by Sequence ID No. 64 and The implementation includes an amino acid sequence having at least 95% sequence identity with sequence number 65. The material comprises an isolated polypeptide or polypeptide complex described in Form 1.

[0364] Embodiment 126 is a polypeptide or polypeptide complex, with Sequence ID No. 66 and The implementation includes an amino acid sequence having at least 95% sequence identity with sequence number 67. The material comprises an isolated polypeptide or polypeptide complex described in Form 1.

[0365] Embodiment 127 is a polypeptide or polypeptide complex, with Sequence ID No. 68 and The implementation includes an amino acid sequence having at least 95% sequence identity with sequence number 69. The material comprises an isolated polypeptide or polypeptide complex described in Form 1.

[0366] Embodiment 128 is a polypeptide or polypeptide complex, with Sequence ID No. 70 and The implementation includes an amino acid sequence having at least 95% sequence identity with sequence number 71. The material comprises an isolated polypeptide or polypeptide complex described in Form 1.

[0367] Embodiment 129 is a polypeptide or polypeptide complex, with Sequence ID No. 72 and The implementation includes an amino acid sequence having at least 95% sequence identity with sequence number 73. The material comprises an isolated polypeptide or polypeptide complex described in Form 1.

[0368] Embodiment 130 is a polypeptide or polypeptide complex, with SEQ ID NO: 74 and The implementation includes an amino acid sequence having at least 95% sequence identity with sequence number 75. The material comprises an isolated polypeptide or polypeptide complex described in Form 1.

[0369] Embodiment 131 is a polypeptide or polypeptide complex, with Sequence ID No. 76 and The implementation includes an amino acid sequence having at least 95% sequence identity with sequence number 77. The material comprises an isolated polypeptide or polypeptide complex described in Form 1.

[0370] Embodiment 132 is (a) a polypeptide described in any one of Embodiments 1 to 131. or a pharmaceutical composition comprising (b) a polypeptide complex and (b) a pharmaceutically acceptable excipient. nothing.

[0371] Embodiment 133 is a polypeptide or as described in any one of Embodiments 1 to 131 Contains isolated recombinant nucleic acid molecules encoding polypeptide complexes.

[0372] Embodiment 134 is based on formula II:L 1a -P 1a -H 1a Polypeptides isolated by or is a polypeptide complex, in which L 1a If it is not disconnected, the effector details The first antigen-recognition molecule that binds to the cytoplasmic antigen is P 1a By tumor-specific proteases that connect them Including a cleaved linkage, the first antigen-recognizing molecule binds to the second antigen-recognizing molecule that binds to PSMA. Connected to the sensory molecule, P 1a L 1a If it is not cleaved, it binds to the first antigen recognition molecule. It contains a peptide that H 1a This includes isolated polypeptides containing half-life extension molecules or Contains polypeptide complexes.

[0373] Embodiment 135 is L 1aIf it is not disconnected, P 1a However, the second of the effector cell antigens The isolated polypeptide described in Embodiment 134 impairs the binding of the antigen recognition molecule 1. It contains polypeptide complexes.

[0374] Embodiment 136 is an embodiment in which the first antigen recognition molecule comprises an antibody or antibody fragment. The material comprises an isolated polypeptide or polypeptide complex as described in Form 134.

[0375] Embodiment 137 is an embodiment in which the effector cell antigen is an anti-CD3 effector cell antigen. The isolated polypeptide or polypeptide complex described in Form 134.

[0376] Embodiment 138 is P 1a It has less than 70% sequence homology to the effector cell antigen. The isolated polypeptide or polypeptide complex described in Embodiment 134 is being used. include.

[0377] Embodiment 139 is P 1a However, it contains a peptide sequence of at least 10 amino acids in length. This includes the isolated polypeptide or polypeptide complex described in Embodiment 134.

[0378] Embodiment 140 is P 1a However, the length is at least 10 amino acids and the length is 20 amino acids The isolated polypeptide or poly(P) described in Embodiment 134, comprising the following peptide sequence Contains peptide complexes.

[0379] Embodiment 141 is P 1a However, it contains a peptide sequence of at least 16 amino acids in length. This includes the isolated polypeptide or polypeptide complex described in Embodiment 134.

[0380] Embodiment 142 is P 1a However, the implementation includes peptide sequences with a length of 40 amino acids or less. The isolated polypeptide or polypeptide complex described in Form 134.

[0381] Embodiment 143 is P 1a However, it contains at least two cysteine ​​amino acid residues. The material comprises an isolated polypeptide or polypeptide complex as described in Form 134.

[0382] Embodiment 144 is P 1a Embodiment 13 includes a cyclic peptide or a linear peptide. The isolated polypeptide or polypeptide complex described in 4.

[0383] Embodiment 145 is the isolation described in Embodiment 134, wherein the albumin is serum albumin. Contains a polypeptide or polypeptide complex.

[0384] Embodiment 146 is P 1a The isolated peptide described in Embodiment 134 is a linear peptide. Contains polypeptides or polypeptide complexes.

[0385] Embodiment 147 is P 1a However, select from the group consisting of one of sequence numbers 16-19. The isolated polypeptide or poly(P) described in Embodiment 134, comprising the amino acid sequence Contains peptide complexes.

[0386] Embodiment 148 is H 1a any one of embodiments 132 to 145, which includes a polymer. The following comprises an isolated polypeptide or polypeptide complex described above.

[0387] Embodiment 149 is an embodiment of Embodiment 1 in which the polymer contains polyethylene glycol (PEG). Contains the isolated polypeptide or polypeptide complex described in 48.

[0388] Embodiment 150 is H 1a Any of embodiments 134 to 147, wherein the albumin is included. One comprises an isolated polypeptide or polypeptide complex as described in one.

[0389] Embodiment 151 is H 1a Any of embodiments 134 to 147, which include an Fc domain The comprising one of the isolated polypeptides or polypeptide complexes described herein.

[0390] Embodiment 152 is the isolation described in Embodiment 150, wherein the albumin is serum albumin. Contains a polypeptide or polypeptide complex.

[0391] Embodiment 153 is the same as Embodiment 152, wherein the albumin is human serum albumin. Contains isolated polypeptides or polypeptide complexes.

[0392] Embodiment 154 is H 1a However, the implement includes polypeptides, ligands, or small molecules. An isolated polypeptide or polypeptide as described in any one of forms 134 to 147 Includes complexes.

[0393] Embodiment 155 describes a polypeptide, ligand, or small molecule as a serum protein or Its fragment, circulating immunoglobulin or its fragment, or CD35 / The isolated polypeptide or polypeptide described in Embodiment 154 that binds to CR1. Includes complexes.

[0394] Embodiment 156 is a serum protein that is a thyroxine-binding protein, transthyreti n, α1-acid glycoprotein, transferrin, transferrin receptor or its Embodiment 155, comprising a transferrin binding moiety, fibrinogen, or albumin. Contains the isolated polypeptide or polypeptide complex described above.

[0395] Embodiment 157 is a circulating immunoglobulin molecule consisting of IgG1, IgG2, IgG3, and Ig The isolated poly(Isolated Poly(IgM) described in Embodiment 155 comprises G4, slgA, IgM, or IgD. Contains peptides or polypeptide complexes.

[0396] Embodiment 158 ​​is the isolation described in Embodiment 153, wherein the serum protein is albumin. Contains a polypeptide or polypeptide complex.

[0397] Embodiment 159 is an isolated polypeptide as described in Embodiment 154, wherein the polypeptide is an antibody. Contains lipeptides or polypeptide complexes.

[0398] Embodiment 160 is an antibody that is a single-domain antibody, a single-chain variable fragment, or a Fab Includes an isolated polypeptide or polypeptide complex as described in Embodiment 159, which includes nothing.

[0399] Embodiment 161 includes a single-domain antibody that binds to albumin. The method includes, for example, the isolated polypeptide or polypeptide complex described in Embodiment 159.

[0400] Embodiment 162 is an embodiment in which the single-domain antibody is a human antibody or a humanized antibody. Contains isolated polypeptides or polypeptide complexes as described in 159.

[0401] Embodiment 163 is an embodiment of Embodiment 160 in which the single-domain antibody is 645gH1gL1. Contains the isolated polypeptide or polypeptide complex described.

[0402] Embodiment 164 is an embodiment in which the single-domain antibody is 645dsgH5gL4. Contains the isolated polypeptide or polypeptide complex described in 0.

[0403] Embodiment 165 is an embodiment in which the single-domain antibody is 23-13-A01-sc02. The isolated polypeptide or polypeptide complex described in Form 160.

[0404] Embodiment 166 is a single-domain antibody which is A10m3 or a fragment thereof. This includes the isolated polypeptide or polypeptide complex described in Embodiment 160.

[0405] Embodiment 167 is described in Embodiment 160, wherein the single-domain antibody is DOM7r-31. Contains isolated polypeptides or polypeptide complexes.

[0406] Embodiment 168 is an embodiment in which the single-domain antibody is DOM7h-11-15. Contains the isolated polypeptide or polypeptide complex described in 0.

[0407] Embodiment 169 is a single-domain antibody that is Alb-1, Alb-8, or Alb-2 3. The isolated polypeptide or polypeptide complex described in Embodiment 160 include.

[0408] Embodiment 170 is the isolation described in Embodiment 160, wherein the single-domain antibody is 10E. Contains a polypeptide or polypeptide complex.

[0409] Embodiment 171 is an embodiment in which a single-domain antibody is used as the complementarity-determining region (CDR) for HC-CD Includes R1, HC-CDR2, and HC-CDR3, and is a single-domain antibody HC-CDR 1. HC-CDR2 and HC-CDR3 are sequence numbers 54 in HC-CDR1, HC -CDR2 includes sequence number 55, and HC-CDR3 includes sequence number 56, Embodiments Contains the isolated polypeptide or polypeptide complex described in 160.

[0410] Embodiment 172 is an embodiment in which a single-domain antibody is used as the complementarity-determining region (CDR) for HC-CD Includes R1, HC-CDR2, and HC-CDR3, and is a single-domain antibody HC-CDR 1. HC-CDR2 and HC-CDR3 are sequence numbers 58 in HC-CDR1, HC -CDR2 includes sequence number 59, and HC-CDR3 includes sequence number 60, Embodiments Contains isolated polypeptides or polypeptide complexes as described in 158.

[0411] Embodiment 173 is the isolation described in Embodiment 160, wherein the single-domain antibody is SA21. Contains a polypeptide or polypeptide complex.

[0412] Embodiment 174 is H 1a However, P 1a H 1a The connecting part (L 1a ) including, Isolated polypeptide or polypeptide according to any one of embodiments 134 to 173 Contains butyl complex.

[0413] Embodiment 175 is L 1a However, peptides having at least 5 to 50 amino acids The column is the isolated polypeptide or polypeptide complex described in Embodiment 174. include.

[0414] Embodiment 176 is L 1a However, peptides having at least 10 to 30 amino acids The sequence is an isolated polypeptide or polypeptide complex as described in Embodiment 174. Includes.

[0415] Embodiment 177 is L 1a However, it is a peptide sequence having at least 10 amino acids. , comprising an isolated polypeptide or polypeptide complex as described in Embodiment 174.

[0416] Embodiment 178 is L 1a However, it is a peptide sequence having at least 18 amino acids. , comprising an isolated polypeptide or polypeptide complex as described in Embodiment 174.

[0417] Embodiment 179 is L 1a However, it is a peptide sequence having at least 26 amino acids. , comprising an isolated polypeptide or polypeptide complex as described in Embodiment 174.

[0418] Embodiment 180 is L 1a However, (G2S) n , (GS) n (GSGGS) n (arrangement number) No. 50), (GGGS) n (Sequence ID 51), (GGGGS) n (Sequence number 52), oyo bi (GSSGGS) n The formula has an expression selected from the group consisting of (Sequence No. 53), in the formula , an isolated polypeptide according to Embodiment 174, where n is at least an integer of 1 or It contains polypeptide complexes.

[0419] Embodiment 181 is L1a However, Embodiment 17 includes the amino acid sequence according to Sequence ID No. 23. The isolated polypeptide or polypeptide complex described in 4.

[0420] Embodiment 182 is P 1a But Z1-Z2-C-Z4-P-Z6-Z7-Z8-Z9- Z 10 -Z 11 -Z 12 -CZ 14 The amino acid sequence includes, where Z1 is D, Y, F, Selected from I, N, V, H, L, A, T, S, and P, Z2 is D, Y, L, F, I Selected from N, A, V, H, T, and S, Z4 is selected from G and W, Z6 However, the following are selected from E, D, V, and P, and Z7 is W, L, F, V, G, M, I, and Y is selected, Z8 is selected from E, D, P, and Q, and Z9 is selected from E, D, Y, V Selected from F, W, P, L, and Q, Z 10 But, S, D, Y, T, I, F, V, N Selected from A, P, L, and H, Z 11 But I, Y, F, V, L, T, N, S, D Selected from A, H, and Z 12 But, F, D, Y, L, I, V, A, N, T, P, S Selected from H and Z 14 But, D, Y, N, F, I, P, V, A, T, H, L, o An isolated port selected from and S, as described in any one of embodiments 134 to 181 Contains lipeptides or polypeptide complexes.

[0421] Embodiment 183 is characterized in that Z1 is selected from D, Y, F, I, and N, and Z2 is selected from D, Y Selected from L, F, I, and N, Z4 is selected from G and W, Z6 is selected from E Z7 is selected from D, and Z8 is selected from W, L, F, and V, and Z8 is selected from E and D Selected from, Z9 is selected from E, D, Y, and V, Z 10 However, S, D, Y, T Selected from I and Z 11 However, it is selected from I, Y, F, V, L, and T, Z1 2 is selected from F, D, Y, L, I, V, A, and N, Z 14 However, D, Y, N, F An isolated polypeptide according to Embodiment 182, selected from I, and P or Contains polypeptide complexes.

[0422] Embodiment 184 is characterized in that Z1 is selected from D, Y, and F, and Z2 is selected from D, Y, L, and Z4 is selected from F, Z6 from G and W, and Z6 from E and D. Then, Z7 is selected from W, L, and F, Z8 is selected from E and D, and Z9 is Selected from E and D, Z 10 However, it is selected from S, D, and Y, and Z 11 However, I, Selected from Y and F, Z 12 However, it is selected from F, D, Y, and L, and Z 14 but, A single unit as described in any one of embodiments 182 to 183, selected from D, Y, and N. Contains separated polypeptides or polypeptide complexes.

[0423] Embodiment 185 is P 1a However, U1-U2-C-U4-P-U6-U7-U8-U9- U 10 -U 11 -U 12 -CU 14 The amino acid sequence includes U1, D, Y, F, Selected from I, N, V, H, L, A, T, S, and P, U2 is D, Y, L, F, I U4 is selected from N, A, V, H, T, and S, and U6 is selected from G and W. However, U7 is selected from E, D, V, and P, and W, L, F, V, G, M, I, and U8 is selected from Y, and U9 is selected from E, D, P, and Q, and U9 is selected from E, D, Y, and V Selected from F, W, P, L, and Q, U 10 But, S, D, Y, T, I, F, V, N Selected from A, P, L, and H, U 11 But I, Y, F, V, L, T, N, S, D Selected from A, H, and U 12 But, F, D, Y, L, I, V, A, N, T, P, S Selected from , G, and H, U 14 But D, Y, N, F, I, P, V, A, T, H, L Isolation according to any one of embodiments 134 to 181, selected from M and S Contains a polypeptide or polypeptide complex.

[0424] Embodiment 186 is one in which U1 is selected from D, Y, F, I, V, and N, and U2 is D Selected from Y, L, F, I, and N, U4 is selected from G and W, U6 is selected from U7 is selected from E and D, and U8 is selected from W, L, F, G, and V. And selected from D, U9 is selected from E, D, Y, and V, U 10 However, S, D Selected from Y, T, and I, U 11 However, you can choose from I, Y, F, V, L, and T. Re, U 12 However, it is selected from F, D, Y, L, I, V, A, G, and N, U 14 However, D Isolated poles selected from Y, N, F, I, M, and P, as described in Embodiment 185 Contains lipeptides or polypeptide complexes.

[0425] Embodiment 187 is one in which U1 is selected from D, Y, V, and F, and U2 is one in which D, Y, L U4 is selected from G and W, U6 is selected from E and D Selected, U7 is chosen from W, L, G, and F, and U8 is chosen from E and D. U9 is selected from E and D, U 10 However, it is selected from S, D, T, and Y, U 11 However, it is selected from I, Y, V, L, and F, U 12 But F, D, Y, G, A, oyo Select...

Claims

1. A pharmaceutical composition for the treatment of prostate cancer, Formula I: comprising a polypeptide or polypeptide complex represented by A2-A1-L1-P1-H1, In formula I, A1 comprises a first antigen recognition molecule that binds to an effector cell antigen, A1 comprises an anti-CD3 binding molecule, and comprises HC-CDR1, HC-CDR2, and HC-CDR3 as complementarity-determining regions (CDRs), wherein HC-CDR1, HC-CDR2, and HC-CDR3 of A1 each contain SEQ ID NO: 1 for HC-CDR1, SEQ ID NO: 2 for HC-CDR2, and SEQ ID NO: 3 for HC-CDR3, A1 comprises LC-CDR1, LC-CDR2, and LC-CDR3 as CDRs, wherein LC-CDR1, LC-CDR2, and LC-CDR3 of A1 each contain SEQ ID NO: 4 for LC-CDR1, SEQ ID NO: 5 for LC-CDR2, and SEQ ID NO: 6 for LC-CDR3. P1 contains a peptide that binds to A1, and P1 contains the amino acid sequence U1-U2-C-U4-P-U6-U7-U8-U9-U10-U11-U12-C-U14, where U1 is selected from D, Y, F, I, N, V, H, L, A, T, S, and P; U2 is selected from D, Y, L, F, I, N, A, V, H, T, and S; U4 is selected from G and W; U6 is selected from E, D, V, and P; U7 is selected from W, L, F, V, G, M, I, and Y; U8 is selected from E, D, P, and Q; and U9 U10 is selected from E, D, Y, V, F, W, P, L, and Q, U11 is selected from I, Y, F, V, T, N, S, D, A, and H, U12 is selected from F, D, Y, L, I, V, A, N, T, P, S, G, and H, and U14 is selected from D, Y, N, F, I, P, V, A, T, H, L, M, and S. L1 connects A1 to P1 and includes a binding portion that is a substrate for tumor-specific proteases. H1 contains a half-life extension molecule. A2 contains a second antigen-recognizing molecule that binds to prostate-specific membrane antigen (PSMA). The aforementioned pharmaceutical composition.

2. The pharmaceutical composition according to claim 1, wherein the prostate cancer is metastatic castration-resistant prostate cancer (mCRPC).

3. The pharmaceutical composition according to claim 1, characterized in that the polypeptide or polypeptide complex is administered once a week.

4. The pharmaceutical composition according to claim 1, characterized in that the polypeptide or polypeptide complex is administered intravenously, intramuscularly, intra-injured, topically, subcutaneously, or orally.

5. The pharmaceutical composition according to claim 1, characterized in that the polypeptide or polypeptide complex is administered by continuous infusion or bolus injection.

6. The pharmaceutical composition according to claim 1, characterized in that the polypeptide or polypeptide complex is administered once a week by continuous intravenous infusion.

7. The pharmaceutical composition according to claim 1, wherein A2 comprises HC-CDR1, HC-CDR2, and HC-CDR3 as complementarity determining regions (CDRs), and HC-CDR1, HC-CDR2, and HC-CDR3 each include SEQ ID NO: 8 for HC-CDR1, SEQ ID NO: 9 for HC-CDR2, and SEQ ID NO: 10 for HC-CDR3, and A2 comprises LC-CDR1, LC-CDR2, and LC-CDR3 as CDRs, and LC-CDR1, LC-CDR2, and LC-CDR3 of A2 each include SEQ ID NO: 11 for LC-CDR1, SEQ ID NO: 12 for LC-CDR2, and SEQ ID NO: 13 for LC-CDR3.

8. The pharmaceutical composition according to claim 1, wherein the effector cell antigen comprises differentiation antigen group 3 (CD3).

9. The pharmaceutical composition according to claim 1, wherein A1 comprises a single-chain variable fragment and an antibody form selected from Fab or Fab' fragments.

10. The pharmaceutical composition according to claim 1, wherein A2 comprises an antibody form selected from a single-chain variable fragment, a single-domain antibody, and a Fab or Fab' fragment.

11. The pharmaceutical composition according to claim 1, wherein A1 comprises an antibody form of a single-chain variable fragment (scFv), and A2 comprises an antibody form of Fab or Fab'.

12. The pharmaceutical composition according to claim 1, wherein when L1 is cleaved by a tumor-specific protease, and A1 is thereby exposed to the effector cell antigen, P1 is decoupled from A1.

13. The pharmaceutical composition according to claim 1, wherein the tumor-specific protease is selected from the group consisting of matrix metalloproteinase (MMP), serine protease, cysteine ​​protease, threonine protease, and aspartate protease.

14. The pharmaceutical composition according to claim 13, wherein the matrix metalloproteinase comprises MMP2, MMP7, MMP9, MMP13, or MMP14.

15. The pharmaceutical composition according to claim 13, wherein the serine protease comprises matryptase (MTSP1), urokinase, or hepsin.

16. The pharmaceutical composition according to claim 1, wherein L1 comprises a urokinase-cleavable amino acid sequence, a matryptase-cleavable amino acid sequence, a matrix metalloproteinase-cleavable amino acid sequence, or a regmine-cleavable amino acid sequence.

17. The pharmaceutical composition according to claim 1, wherein L1 comprises the amino acid sequence according to SEQ ID NO:

23.

18. The pharmaceutical composition according to claim 1, wherein L1 comprises an amino acid sequence according to any one of SEQ ID NOs: 20 to 49.

19. The pharmaceutical composition according to claim 1, wherein L1 comprises the amino acid sequence of linker 25 (ISSGLLSGRSDAG) (SEQ ID NO: 45), linker 26 (AAGLLAAPPGGLSGRSDAG) (SEQ ID NO: 46), linker 27 (SPLGLLSGRSDAG) (SEQ ID NO: 47), or linker 28 (LSGRSDAGSPLGLAG) (SEQ ID NO: 48), or an amino acid sequence having one, two, or three amino acid substitutions, additions, or deletions to the amino acid sequence of linker 25, linker 26, linker 27, or linker 28.

20. The pharmaceutical composition according to claim 1, wherein H1 comprises serum albumin.

21. The pharmaceutical composition according to claim 20, wherein the serum albumin is human serum albumin.

22. The pharmaceutical composition according to claim 1, wherein H1 comprises a single-domain antibody.

23. The pharmaceutical composition according to claim 22, wherein H1 includes HC-CDR1, HC-CDR2, and HC-CDR3 as complementarity determining regions (CDRs), and HC-CDR1, HC-CDR2, and HC-CDR3 of H1 include SEQ ID NO: 54 for HC-CDR1, SEQ ID NO: 55 for HC-CDR2, and SEQ ID NO: 56 for HC-CDR3.

24. The pharmaceutical composition according to claim 22, wherein H1 includes HC-CDR1, HC-CDR2, and HC-CDR3 as complementarity determining regions (CDRs), and HC-CDR1, HC-CDR2, and HC-CDR3 of H1 include SEQ ID NO: 58 for HC-CDR1, SEQ ID NO: 59 for HC-CDR2, and SEQ ID NO: 60 for HC-CDR3.

25. The pharmaceutical composition according to claim 1, wherein U1 is selected from D, Y, F, I, V, and N; U2 is selected from D, Y, L, F, I, and N; U4 is selected from G and W; U6 is selected from E and D; U7 is selected from W, L, F, G, and V; U8 is selected from E and D; U9 is selected from E, D, Y, and V; U10 is selected from S, D, Y, T, and I; U11 is selected from I, Y, F, V, L, and T; U12 is selected from F, D, Y, L, I, V, A, G, and N; and U14 is selected from D, Y, N, F, I, M, and P.

26. The pharmaceutical composition according to claim 25, wherein U1 is selected from D, Y, V, and F, U2 is selected from D, Y, L, and F, U4 is selected from G and W, U6 is selected from E and D, U7 is selected from W, L, G, and F, U8 is selected from E and D, U9 is selected from E and D, U10 is selected from S, D, T, and Y, U11 is selected from I, Y, V, L, and F, U12 is selected from F, D, Y, G, A, and L, and U14 is selected from D, Y, M, and N.

27. ​​The pharmaceutical composition according to claim 1, wherein P1 comprises an amino acid sequence according to any one of SEQ ID NOs: 93-95 and 102-105.

28. The pharmaceutical composition according to claim 1, wherein P1 comprises an amino acid sequence according to any one of SEQ ID NOs: 106 and 108-117.

29. The pharmaceutical composition according to claim 1, wherein P1 comprises the amino acid sequence according to Sequence ID No.

19.

30. The pharmaceutical composition according to claim 1, wherein P1 comprises the amino acid sequence according to SEQ ID NO:

116.

31. The pharmaceutical composition according to claim 1, comprising the amino acid sequences of SEQ ID NO: 72 and SEQ ID NO: 73.

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