Methods and uses of Anti-CD38 t-cell adapters in the treatment of peripheral t-cell lymphoma

Abstract

The present disclosure provides methods of treating peripheral T cell lymphoma (PTCL) using anti-CD38 T cell adapters, as well as uses, compositions, and kits related thereto.

Description

Cross Reference to Related Applications

[0001] This application claims priority benefit of U.S. Provisional Application Serial No. 63 / 529,942, filed July 31, 2023; and EP Application No. EP24315126.3, filed April 8, 2024; each of which is incorporated by reference herein in its entirety. Reference to Electronic Sequence Listing

[0002] The contents of the electronic sequence listing (183952035240seqlist.xml; size: 274,177 bytes; and date created: July 24, 2024) are incorporated herein by reference in their entirety. TECHNICAL FIELD

[0003] The present disclosure provides methods of treating peripheral T-cell lymphoma (PTCL) using anti-CD38 T-cell engagers, as well as uses, compositions, and kits related thereto. BACKGROUND

[0004] Peripheral T-cell lymphoma (PTCL) is a complex group of clinicopathological entities that are generally associated with aggressive clinical courses. Despite the introduction of new chemotherapy regimens and a large number of new agents, the progression-free survival of PTCL patients remains to be improved.

[0005] In recent years, immunotherapy has been established as a major advance in the treatment of a variety of cancers. Trispecific binding proteins targeting CD38, CD3, and CD28 have been developed for the treatment of multiple myeloma and chronic viral infections. See, e.g., International Publication Nos. WO 2019074973 and WO 2020076853 and U.S. Patent Nos. 11,186,649 and 11,530,268.

[0006] Accordingly, there remains a need for improved treatments for PTCL. SUMMARY

[0007] In one aspect, provided herein is a method for treating peripheral T-cell lymphoma (PTCL) in an individual in need thereof, the method comprising administering to the individual an effective amount of an anti-CD38 T-cell engager.

[0008] In some embodiments according to any of the embodiments described herein, the anti-CD38 T cell engager is a bispecific binding protein comprising a first antigen binding site that specifically binds a CD38 polypeptide and a second antigen binding site that specifically binds a CD3 polypeptide. In some embodiments, the anti-CD38 T cell engager is a trispecific binding protein comprising a first antigen binding site that specifically binds a CD38 polypeptide, a second antigen binding site that specifically binds a CD28 polypeptide, and a third antigen binding site that specifically binds a CD3 polypeptide.

[0009] In some embodiments according to any of the embodiments described herein, the first antigen binding site that binds to a CD38 polypeptide comprises: (a) an antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence comprising the amino acid sequence of GYTFTSFN (SEQ ID NO: 31) or GYTFTSYA (SEQ ID NO: 37), a CDR-H2 sequence comprising the amino acid sequence of IYPGNGGT (SEQ ID NO: 32) or IYPGQGGT (SEQ ID NO: 38), and a CDR-H3 sequence comprising the amino acid sequence of ARTGGLRRAYFTY (SEQ ID NO: 33); and (b) an antibody light chain variable (VL) domain comprising a CDR-L1 sequence comprising the amino acid sequence of ESVDSYGNGF (SEQ ID NO: 34) or QSVSSYGQGF (SEQ ID NO: 39), a CDR-L2 sequence comprising the amino acid sequence of LAS or GAS, and a CDR-L3 sequence comprising the amino acid sequence of QQNKEDPWT (SEQ ID NO: 36). In some embodiments, the first antigen binding site comprises: (a) an antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence comprising the amino acid sequence of GYTFTSYA (SEQ ID NO: 37), a CDR-H2 sequence comprising the amino acid sequence of IYPGQGGT (SEQ ID NO: 38), and a CDR-H3 sequence comprising the amino acid sequence of ARTGGLRRAYFTY (SEQ ID NO: 33); and (b) an antibody light chain variable (VL) domain comprising a CDR-L1 sequence comprising the amino acid sequence of QSVSSYGQGF (SEQ ID NO: 39), a CDR-L2 sequence comprising the amino acid sequence of GAS, and a CDR-L3 sequence comprising the amino acid sequence of QQNKEDPWT (SEQ ID NO: 36). In some embodiments, the first antigen binding site comprises a VH domain comprising the amino acid sequence of SEQ ID NO: 13 and a VL domain comprising the amino acid sequence of SEQ ID NO: 14.In some embodiments, the first antigen-binding site comprises: (a) an antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence containing the amino acid sequence GYTFTSFN (SEQ ID NO: 31), a CDR-H2 sequence containing the amino acid sequence IYPGNGGT (SEQ ID NO: 32), and a CDR-H3 sequence containing the amino acid sequence ARTGGLRRAYFTY (SEQ ID NO: 33); and (b) an antibody light chain variable (VL) domain comprising a CDR-L1 sequence containing the amino acid sequence ESVDSYGNGF (SEQ ID NO: 34), a CDR-L2 sequence containing the amino acid sequence LAS, and a CDR-L3 sequence containing the amino acid sequence QQNKEDPWT (SEQ ID NO: 36). In some embodiments, the first antigen-binding site comprises a VH domain and a VL domain, the VH domain comprising the amino acid sequence of SEQ ID NO: 5, and the VL domain comprising the amino acid sequence of SEQ ID NO: 6. In some embodiments, the first antigen-binding site comprises a VH domain and a VL domain, wherein the VH domain comprises the amino acid sequence of SEQ ID NO: 17 and the VL domain comprises the amino acid sequence of SEQ ID NO: 18. In some embodiments, the first antigen-binding site comprises a VH domain and a VL domain, wherein the VH domain comprises the amino acid sequence of SEQ ID NO: 21 and the VL domain comprises the amino acid sequence of SEQ ID NO: 18. In some embodiments, the first antigen-binding site comprises a VH domain and a VL domain, wherein the VH domain comprises the amino acid sequence of SEQ ID NO: 23 and the VL domain comprises the amino acid sequence of SEQ ID NO: 18. In some embodiments, the first antigen-binding site binding to the CD38 peptide comprises: (a) an antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence containing the amino acid sequence GFTFSSYG (SEQ ID NO: 41), a CDR-H2 sequence containing the amino acid sequence IWYDGSNK (SEQ ID NO: 42), and a CDR-H3 sequence containing the amino acid sequence ARMFRGAFDY (SEQ ID NO: 43); and (b) an antibody light chain variable (VL) domain comprising a CDR-L1 sequence containing the amino acid sequence QGIRND (SEQ ID NO: 44), a CDR-L2 sequence containing the amino acid sequence AAS, and a CDR-L3 sequence containing the amino acid sequence LQDYIYYPT (SEQ ID NO: 46).In some embodiments, the first antigen-binding site comprises a VH domain and a VL domain, the VH domain comprising the amino acid sequence of SEQ ID NO: 9 and the VL domain comprising the amino acid sequence of SEQ ID NO: 10. In some embodiments, the first antigen-binding site binding the CD38 peptide comprises: (a) an antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence containing the amino acid sequence GYTLTEFS (SEQ ID NO: 2), a CDR-H2 sequence containing the amino acid sequence FDPEDGET (SEQ ID NO: 3), and a CDR-H3 sequence containing the amino acid sequence TTGRFFDWF (SEQ ID NO: 4); and (b) an antibody light chain variable (VL) domain comprising a CDR-L1 sequence containing the amino acid sequence QSVISRF (SEQ ID NO: 7), a CDR-L2 sequence containing the amino acid sequence GAS, and a CDR-L3 sequence containing the amino acid sequence QQDSNLPIT (SEQ ID NO: 11). In some embodiments, the first antigen-binding site comprises a VH domain and a VL domain, the VH domain comprising the amino acid sequence of SEQ ID NO: 108 and the VL domain comprising the amino acid sequence of SEQ ID NO: 109. In some embodiments, the first antigen-binding site binding the CD38 peptide comprises: (a) an antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence containing the amino acid sequence GYAFTTYL (SEQ ID NO: 12), a CDR-H2 sequence containing the amino acid sequence INPGSGST (SEQ ID NO: 15), and a CDR-H3 sequence containing the amino acid sequence ARYAYGY (SEQ ID NO: 16); and (b) an antibody light chain variable (VL) domain comprising a CDR-L1 sequence containing the amino acid sequence QNVGTA (SEQ ID NO: 19), a CDR-L2 sequence containing the amino acid sequence SAS, and a CDR-L3 sequence containing the amino acid sequence QQYSTYPFT (SEQ ID NO: 22). In some embodiments, the first antigen-binding site comprises a VH domain and a VL domain, wherein the VH domain comprises the amino acid sequence of SEQ ID NO: 110 and the VL domain comprises the amino acid sequence of SEQ ID NO: 111. In some embodiments, the first antigen-binding site comprises a VH domain and a VL domain, wherein the VH domain comprises the amino acid sequence of SEQ ID NO: 116 and the VL domain comprises the amino acid sequence of SEQ ID NO: 117.In some embodiments, the first antigen-binding site binding to the CD38 peptide comprises: (a) an antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence containing the amino acid sequence GYSFTNYA (SEQ ID NO: 24), a CDR-H2 sequence containing the amino acid sequence ISPHYGDT (SEQ ID NO: 25), and a CDR-H3 sequence containing the amino acid sequence ARFEGFYYSMDY (SEQ ID NO: 26); and (b) an antibody light chain variable (VL) domain comprising a CDR-L1 sequence containing the amino acid sequence QSLVHSNGNTY (SEQ ID NO: 27), a CDR-L2 sequence containing the amino acid sequence KVS, and a CDR-L3 sequence containing the amino acid sequence SQSTHVPLT (SEQ ID NO: 29). In some embodiments, the first antigen-binding site comprises a VH domain and a VL domain, wherein the VH domain comprises the amino acid sequence of SEQ ID NO: 112 and the VL domain comprises the amino acid sequence of SEQ ID NO: 113. In some embodiments, the first antigen-binding site comprises a VH domain and a VL domain, wherein the VH domain comprises the amino acid sequence of SEQ ID NO: 118 and the VL domain comprises the amino acid sequence of SEQ ID NO: 119. In some embodiments, the first antigen-binding site binding to the CD38 peptide comprises: (a) an antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence containing the amino acid sequence GFTFSSYG (SEQ ID NO: 41), a CDR-H2 sequence containing the amino acid sequence IWYDGSNK (SEQ ID NO: 42), and a CDR-H3 sequence containing the amino acid sequence ARDPGLRYFDGGMDV (SEQ ID NO: 106); and (b) an antibody light chain variable (VL) domain comprising a CDR-L1 sequence containing the amino acid sequence QGISSY (SEQ ID NO: 107), a CDR-L2 sequence containing the amino acid sequence AAS, and a CDR-L3 sequence containing the amino acid sequence QQLNSFPYT (SEQ ID NO: 229). In some embodiments, the first antigen binding site comprises a VH domain and a VL domain, the VH domain comprising the amino acid sequence of SEQ ID NO: 114 and the VL domain comprising the amino acid sequence of SEQ ID NO: 115.

[0010] In some embodiments according to any of the embodiments described herein, the antigen-binding site of the CD3 peptide comprises: (a) an antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence containing the amino acid sequence GFTFTKAW (SEQ ID NO: 120), a CDR-H2 sequence containing the amino acid sequence IKDKSNSYAT (SEQ ID NO: 121), and a CDR-H3 sequence containing the amino acid sequence RGVYYALSPFDY (SEQ ID NO: 122); and (b) an antibody light chain variable (VL) domain comprising a CDR-L1 sequence containing the amino acid sequence QSLVHNNGNTY (SEQ ID NO: 218), a CDR-L2 sequence containing the amino acid sequence KVS, and a CDR-L3 sequence containing the amino acid sequence GQGTQYPFT (SEQ ID NO: 130). In some embodiments, the third antigen binding site comprises a VH domain and a VL domain, the VH domain comprising the amino acid sequence of SEQ ID NO: 84 and the VL domain comprising the amino acid sequence of SEQ ID NO: 85. In some embodiments, the antigen-binding site of the CD3 peptide comprises: (a) an antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence containing the amino acid sequence GFTFTKAW (SEQ ID NO: 120), a CDR-H2 sequence containing the amino acid sequence IKDKSNSYAT (SEQ ID NO: 121), and a CDR-H3 sequence containing the amino acid sequence RGVYYALSPFDY (SEQ ID NO: 122); and (b) an antibody light chain variable (VL) domain comprising a CDR-L1 sequence containing the amino acid sequence QSLVHX1NX2X3TY (where X1 is E or Q, X2 is A or L, and X3 is Q, R, or F) (SEQ ID NO: 131), a CDR-L2 sequence containing the amino acid sequence KVS, and a CDR-L3 sequence containing the amino acid sequence GQGTQYPFT (SEQ ID NO: 130). In some embodiments, the CDR-L1 sequence of the VL domain of the antigen-binding site of the CD3 polypeptide is selected from the group consisting of: QSLVHNNANTY (SEQ ID NO: 123), QSLVHQNAQTY (SEQ ID NO: 124), QSLVHENLQTY (SEQ ID NO: 125), QSLVHENLFTY (SEQ ID NO: 126), QSLVHENLRTY (SEQ ID NO: 127), and QSLVHDNAQTY (SEQ ID NO: 128).In some embodiments, the VH domain of the antigen-binding site of the CD3 polypeptide comprises the amino acid sequence of SEQ ID NO: 53 or 138, and the VL domain of the antigen-binding site of the CD3 polypeptide comprises the amino acid sequence of SEQ ID NO: 54, 133, 134, 135, 136 or 137.

[0011] In some embodiments according to any of the embodiments described herein, the antigen-binding site binding to the CD28 peptide comprises: (a) an antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence containing the amino acid sequence GYTFTSYY (SEQ ID NO: 139), a CDR-H2 sequence containing the amino acid sequence IYPGNVNT (SEQ ID NO: 140), and a CDR-H3 sequence containing the amino acid sequence TRSHYGLDWNFDV (SEQ ID NO: 141); and (b) an antibody light chain variable (VL) domain comprising a CDR-L1 sequence containing the amino acid sequence QNIYVW (SEQ ID NO: 142), a CDR-L2 sequence containing the amino acid sequence KAS, and a CDR-L3 sequence containing the amino acid sequence QQGQTYPY (SEQ ID NO: 144). In some embodiments, the antigen-binding site of the CD28 peptide comprises a VH domain and a VL domain, wherein the VH domain comprises the amino acid sequence of SEQ ID NO: 49 and the VL domain comprises the amino acid sequence of SEQ ID NO: 50. In some embodiments, the antigen-binding site of the CD28 peptide comprises: (a) an antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence containing the amino acid sequence GFSLSDYG (SEQ ID NO: 212), a CDR-H2 sequence containing the amino acid sequence IWAGGGT (SEQ ID NO: 213), and a CDR-H3 sequence containing the amino acid sequence ARDKGYSYYYSMDY (SEQ ID NO: 214); and (b) an antibody light chain variable (VL) domain comprising a CDR-L1 sequence containing the amino acid sequence ESVEYYVTSL (SEQ ID NO: 215), a CDR-L2 sequence containing the amino acid sequence AAS, and a CDR-L3 sequence containing the amino acid sequence QQSRKVPYT (SEQ ID NO: 217). In some embodiments, the antigen-binding site of the CD28 peptide comprises a VH domain and a VL domain, wherein the VH domain comprises the amino acid sequence of SEQ ID NO: 51 and the VL domain comprises the amino acid sequence of SEQ ID NO: 52. In some embodiments, the antigen-binding site of the CD28 peptide comprises a VH domain and a VL domain, wherein the VH domain comprises the amino acid sequence of SEQ ID NO: 49 and the VL domain comprises the amino acid sequence of SEQ ID NO: 50; the antigen-binding site of the CD3 peptide comprises a VH domain and a VL domain, wherein the VH domain comprises the amino acid sequence of SEQ ID NO: 53 and the VL domain comprises the amino acid sequence of SEQ ID NO: 54.

[0012] In some embodiments according to any of the examples described herein, the trispecific binding protein comprises four polypeptide chains forming the three antigen-binding sites, wherein the first polypeptide chain comprises a structure represented by the following formula: V L2 -L1-V L1 -L2-C L [I] Furthermore, the second polypeptide chain contains a structure represented by the following formula: V H1 -L3-V H2 -L4-C H1 -Hinge-C H2 -C H3 [II] Furthermore, the third polypeptide chain contains a structure represented by the following formula: V H3 -C H1 -Hinge-C H2 -C H3 [III] Furthermore, the fourth polypeptide chain contains a structure represented by the following formula: V L3 -C L [IV] in: V L1 It is the variable domain of the first immunoglobulin light chain; V L2 It is the variable domain of the second immunoglobulin light chain; V L3 It is the variable domain of the third immunoglobulin light chain; V H1 It is the variable domain of the first immunoglobulin heavy chain; V H2 It is the variable domain of the second immunoglobulin heavy chain; V H3 It is the variable domain of the third immunoglobulin heavy chain; C L It is the constant structural domain of the immunoglobulin light chain; C H1 It is immunoglobulin C H1 Heavy chain constant structural domain; C H2 It is immunoglobulin C H2 Heavy chain constant structural domain; C H3 It is immunoglobulin C H3 Heavy chain constant structural domain; The hinge is the connection of C H1 and C H2The immunoglobulin hinge region of the structural domain; and L1, L2, L3, and L4 are amino acid linkers; The polypeptide having Formula I and the polypeptide having Formula II form a cross-linked light chain-heavy chain pair; wherein V H1 and V L1 Forming one of these three antigen-binding sites; among which V H2 and V L2 Forming another of these three antigen-binding sites; and where V H3 and V L3 This forms another of the three antigen-binding sites. In some embodiments, V H1 and V L1 Forms a second antigen-binding site for the CD28 polypeptide, V H2 and V L2 Forming a third antigen-binding site for the CD3 polypeptide, and V H3 and V L3 Forming the first antigen-binding site for the CD38 polypeptide. In some embodiments, V H1 and V L1 Forms a third antigen-binding site for the CD3 polypeptide, V H2 and V L2 Forming a second antigen-binding site that binds to the CD28 peptide, and V H3 and V L3 Forming the first antigen-binding site for the CD38 polypeptide. In some embodiments, V H1 and V L1 Forms a second antigen-binding site for the CD28 polypeptide, V H2 and V L2 Forming the first antigen-binding site for the CD38 polypeptide, and V H3 and V L3 This forms a third antigen-binding site for the CD3 polypeptide. In some embodiments, V H1 and V L1 Forms a third antigen-binding site for the CD3 polypeptide, V H2 and V L2 Forming the first antigen-binding site for the CD38 polypeptide, and V H3 and V L3 A second antigen-binding site is formed for binding to the CD28 polypeptide. In some embodiments, V H1 and V L1 Forms the first antigen-binding site for the CD38 polypeptide, V H2 and V L2 Forming a third antigen-binding site for the CD3 polypeptide, and V H3 and V L3A second antigen-binding site is formed for binding to the CD28 polypeptide. In some embodiments, V H1 and V L1 Forms the first antigen-binding site for the CD38 polypeptide, V H2 and V L2 Forming a second antigen-binding site that binds to the CD28 peptide, and V H3 and V L3 A third antigen-binding site is formed for binding to the CD3 polypeptide. In some embodiments, L1 comprises the sequence GQPKAAP (SEQ ID NO: 58), L2 comprises the sequence TKGPS (SEQ ID NO: 57), L3 comprises amino acid S, and L4 comprises the sequence RT. In some embodiments, L1 comprises the sequence GGGGSGGGGS (SEQ ID NO: 55), L2 comprises the sequence GGGGSGGGGS (SEQ ID NO: 55), L3 is 0 amino acids long, and L4 is 0 amino acids long. In some embodiments, L1 comprises the sequence GGSGSSGSGG (SEQ ID NO: 59), L2 comprises the sequence GGSGSSGSGG (SEQ ID NO: 59), L3 is 0 amino acids long, and L4 is 0 amino acids long. In some embodiments, L1 comprises the sequence GGGGSGGGGSGGGGS (SEQ ID NO: 56), L2 is 0 amino acids long, L3 comprises the sequence GGGGSGGGGSGGGGS (SEQ ID NO: 56), and L4 is 0 amino acids long. In some embodiments, at least one of L1, L2, L3, or L4 comprises the sequence DKTHT (SEQ ID NO: 147). In some embodiments, L1, L2, L3, and L4 comprise the sequence DKTHT (SEQ ID NO: 147). In some embodiments, the hinge-C of the second and third polypeptide chains... H2 -C H3 The structural domain is the human IgG4 hinge-C H2 -C H3 Structural domain, and in which these hinges - C H2 -C H3 Each of the domains contains amino acid substitutions at positions 234 and 235 corresponding to human IgG4 according to the EU index, wherein these amino acid substitutions are F234A and L235A. In some embodiments, the hinge-C of the second and third polypeptide chains... H2 -C H3 The structural domain is the human IgG4 hinge-C H2 -C H3 Structural domain, and in which these hinges - C H2 -C H3Each domain contains an amino acid substitution at positions 233-236 corresponding to human IgG4 according to the EU index, wherein these amino acid substitutions are E233P, F234V, L235A, and a deletion at position 236. In some embodiments, the hinge-C of the second and third polypeptide chains... H2 -C H3 The structural domain is the human IgG4 hinge-C H2 -C H3 Structural domain, and in which these hinges - C H2 -C H3 Each of the domains contains amino acid substitutions at positions 228 and 409 corresponding to human IgG4 according to the EU index, wherein these amino acid substitutions are S228P and R409K. In some embodiments, the hinge-C of the second and third polypeptide chains... H2 -C H3 The structural domain is the human IgG1 hinge-C H2 -C H3 Structural domain, and in which these hinges - C H2 -C H3 Each of the domains contains an amino acid substitution at positions 234, 235, and 329 corresponding to human IgG1 according to the EU index, wherein these amino acid substitutions are L234A, L235A, and P329A. In some embodiments, the hinge-C of the second and third polypeptide chains... H2 -C H3 The structural domain is the human IgG1 hinge-C H2 -C H3 Structural domain, and in which these hinges - C H2 -C H3 Each of the domains contains an amino acid substitution at positions 298, 299, and 300 corresponding to human IgG1 according to the EU index, wherein these amino acid substitutions are S298N, T299A, and Y300S. In some embodiments, the hinge-CH2-CH3 domain of the second polypeptide chain contains an amino acid substitution at positions 349, 366, 368, and 407 corresponding to human IgG1 or IgG4 according to the EU index, wherein these amino acid substitutions are Y349C, T366S, L368A, and Y407V; and wherein the hinge-CH2-CH3 domain of the third polypeptide chain contains an amino acid substitution at positions 349, 366, 368, and 407 corresponding to human IgG1 or IgG4 according to the EU index ... H2 -C H3 The domain contains amino acid substitutions at positions 354 and 366 corresponding to human IgG1 or IgG4 according to the EU index, wherein these amino acid substitutions are S354C and T366W. In some embodiments, the hinge-C of the second polypeptide chain... H2 -C H3The domain contains amino acid substitutions at positions 354 and 366 corresponding to human IgG1 or IgG4 according to the EU index, wherein these amino acid substitutions are S354C and T366W; and wherein the hinge of the third polypeptide chain is C H2 -C H3 The domain contains amino acid substitutions at positions 349, 366, 368, and 407 corresponding to human IgG1 or IgG4 according to the EU index, wherein these amino acid substitutions are Y349C, T366S, L368A, and Y407V. In some embodiments, V H1 and V L1 Forms a second antigen-binding site for the CD28 polypeptide, V H2 and V L2 Forming a third antigen-binding site for the CD3 polypeptide, and V H3 and V L3 Forms the first antigen-binding site for the CD38 polypeptide; V H1 The CDR-H1 sequence contains the amino acid sequence GYTFTSYY (SEQ ID NO: 139), the CDR-H2 sequence contains the amino acid sequence IYPGNVNT (SEQ ID NO: 140), and the CDR-H3 sequence contains the amino acid sequence TRSHYGLDWNFDV (SEQ ID NO: 141). L1 The CDR-L1 sequence contains the amino acid sequence QNIYVW (SEQ ID NO: 142), the CDR-L2 sequence contains the amino acid sequence KAS, and the CDR-L3 sequence contains the amino acid sequence QQGQTYPY (SEQ ID NO: 144); V H2 The CDR-H1 sequence contains the amino acid sequence GFTFTKAW (SEQ ID NO: 120), the CDR-H2 sequence contains the amino acid sequence IKDKSNSYAT (SEQ ID NO: 121), and the CDR-H3 sequence contains the amino acid sequence RGVYYALSPFDY (SEQ ID NO: 122). L2 The CDR-L1 sequence contains the amino acid sequence QSLVHNNANTY (SEQ ID NO: 123), the CDR-L2 sequence contains the amino acid sequence KVS, and the CDR-L3 sequence contains the amino acid sequence GQGTQYPFT (SEQ ID NO: 130); and V H3The CDR-H1 sequence contains the amino acid sequence GYTFTSYA (SEQ ID NO: 37), the CDR-H2 sequence contains the amino acid sequence IYPGQGGT (SEQ ID NO: 38), and the CDR-H3 sequence contains the amino acid sequence ARTGGLRRAYFTY (SEQ ID NO: 33). L3 The CDR-L1 sequence comprises an amino acid sequence containing QSVSSYGQGF (SEQ ID NO: 39), the CDR-L2 sequence comprises an amino acid sequence containing GAS, and the CDR-L3 sequence comprises an amino acid sequence containing QQNKEDPWT (SEQ ID NO: 36). In some embodiments, V H1 and V L1 Forms a second antigen-binding site for the CD28 polypeptide, V H2 and V L2 Forming a third antigen-binding site for the CD3 polypeptide, and V H3 and V L3 Forming the first antigen-binding site for the CD38 polypeptide; wherein V H1 Contains the amino acid sequence of SEQ ID NO: 49, and V L1 Contains the amino acid sequence of SEQ ID NO: 50; wherein V H2 Contains the amino acid sequence of SEQ ID NO: 53, and V L2 Contains the amino acid sequence of SEQ ID NO: 54; and wherein V H3 Contains the amino acid sequence of SEQ ID NO: 13, and V L3The polypeptide chain comprises the amino acid sequence of SEQ ID NO: 14. In some embodiments, the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 61, the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 60, the third polypeptide chain comprises the amino acid sequence of SEQ ID NO: 62, and the fourth polypeptide chain comprises the amino acid sequence of SEQ ID NO: 63. In some embodiments, the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 61, the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 64, the third polypeptide chain comprises the amino acid sequence of SEQ ID NO: 65, and the fourth polypeptide chain comprises the amino acid sequence of SEQ ID NO: 63. In some embodiments, the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 61, the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 66, the third polypeptide chain comprises the amino acid sequence of SEQ ID NO: 67, and the fourth polypeptide chain comprises the amino acid sequence of SEQ ID NO: 63. In some embodiments, the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 61, the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 60, the third polypeptide chain comprises the amino acid sequence of SEQ ID NO: 68, and the fourth polypeptide chain comprises the amino acid sequence of SEQ ID NO: 69. In some embodiments, the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 61, the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 64, the third polypeptide chain comprises the amino acid sequence of SEQ ID NO: 70, and the fourth polypeptide chain comprises the amino acid sequence of SEQ ID NO: 69. In some embodiments, the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 61, the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 66, the third polypeptide chain comprises the amino acid sequence of SEQ ID NO: 71, and the fourth polypeptide chain comprises the amino acid sequence of SEQ ID NO: 69. In some embodiments, the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 148, the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 149, the third polypeptide chain comprises the amino acid sequence of SEQ ID NO: 150, and the fourth polypeptide chain comprises the amino acid sequence of SEQ ID NO: 151. In some embodiments, the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 152, the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 153, the third polypeptide chain comprises the amino acid sequence of SEQ ID NO: 154, and the fourth polypeptide chain comprises the amino acid sequence of SEQ ID NO: 155.In some embodiments, the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 156, the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 157, the third polypeptide chain comprises the amino acid sequence of SEQ ID NO: 158, and the fourth polypeptide chain comprises the amino acid sequence of SEQ ID NO: 159. In some embodiments, the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 160, the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 161, the third polypeptide chain comprises the amino acid sequence of SEQ ID NO: 162, and the fourth polypeptide chain comprises the amino acid sequence of SEQ ID NO: 163. In some embodiments, the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 164, the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 165, the third polypeptide chain comprises the amino acid sequence of SEQ ID NO: 166, and the fourth polypeptide chain comprises the amino acid sequence of SEQ ID NO: 167. In some embodiments, the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 168, the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 169, the third polypeptide chain comprises the amino acid sequence of SEQ ID NO: 170, and the fourth polypeptide chain comprises the amino acid sequence of SEQ ID NO: 171. In some embodiments, the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 172, the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 173, the third polypeptide chain comprises the amino acid sequence of SEQ ID NO: 174, and the fourth polypeptide chain comprises the amino acid sequence of SEQ ID NO: 175. In some embodiments, the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 176, the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 177, the third polypeptide chain comprises the amino acid sequence of SEQ ID NO: 178, and the fourth polypeptide chain comprises the amino acid sequence of SEQ ID NO: 179.

[0013] In some embodiments according to any of the embodiments described herein, the CD38 peptide is a human CD38 peptide. In some embodiments, the CD3 peptide is a human CD3 peptide. In some embodiments, the CD28 peptide is a human CD28 peptide. In some embodiments, PTCL is angioimmunoblastic T-cell lymphoma (AITL), hepatosplenic T-cell lymphoma (HSTL), adult T-cell leukemia / lymphoma (ATLL), extranodal NK / T-cell lymphoma (ENKTCL), mycosis fungoides (MF), Cezari syndrome (SS), anaplastic large cell ALK-lymphoma (ALCL), PTCL-nospecific (NOS), enteropathy-type T-cell lymphoma (EATL), monomorphic epithelial intestinal T-cell lymphoma (MEITL), primary cutaneous CD4+ lymphoproliferative disorder, subcutaneous panniculitis-like T-cell lymphoma (SCTCL), or primary cutaneous γδ T-cell lymphoma. In some embodiments, PTCL is HSTL, SS, or MF. In some embodiments, mycosis fungoides is transformed mycosis fungoides (tMF or MFt). In some embodiments, PTCL cells express CD38 and / or CD28. In some embodiments, the individual is human.

[0014] On the other hand, this article provides an anti-CD38 T-cell binder for use in a method of treating peripheral T-cell lymphoma (PTCL) in an individual in need, the method comprising administering to the individual an effective amount of the anti-CD38 T-cell binder according to any one of the above embodiments.

[0015] On the other hand, this document provides the use of the anti-CD38 T-cell binder according to any one of the above embodiments in the manufacture of a medicament for treating peripheral T-cell lymphoma (PTCL) in individuals in need.

[0016] On the other hand, this document provides a kit comprising an anti-CD38 T-cell binder (e.g., an effective amount thereof, optionally in combination with a pharmaceutically acceptable carrier) and instructions for administering an effective amount of the anti-CD38 T-cell binder to an individual suffering from peripheral T-cell lymphoma (PTCL) according to the method described in any one of the above embodiments.

[0017] It should be understood that one, some, or all of the features of the various embodiments described herein can be combined to form other embodiments of the invention. These and other aspects of the invention will become apparent to those skilled in the art. These and other embodiments of the invention are further described in the following detailed description. Attached Figure Description

[0018] Figures 1A-1EThe expression and distribution of CD38 (left side of each subtype map) and CD28 (right side of each subtype map) in each PTCL subtype are shown by immunochemistry (IHC). Data for each entity are presented according to IHC scores. Only cases with CD38 and CD28 scores on TC were considered. Figure 1A The distribution of CD38 (left) and CD28 (right) expression differed significantly between PTCLs with cytotoxic and non-cytotoxic phenotypes. Figure 1B Distribution of CD38 and CD28 target expression patterns in tumor cells ( Figure 1C ).exist Figure 1C In the figure, the letters A through J at the top show the following patterns of CD38 and CD28 expression for each tumor type (read from bottom to top): A: CD38+CD28+ and CD38-CD28+; B: CD38-CD28- and CD38-CD28+; C: CD38+CD28+, CD38-CD28-, and CD38-CD28+; D: CD38+CD28+ and CD38+CD28-; E: CD38+CD28+, CD38- ... 8+CD28- and CD38-CD28+; F: CD38+CD28+, CD38-CD28- and CD38+CD28-; G: CD38-CD28- and CD38+CD28-; H: CD38+CD28+, CD38-CD28-, CD38+CD28- and CD38+CD28-; I: CD38-CD28-, CD38+CD28- and CD38+CD28-; J: CD38-CD28-. PTCL-TFH tumor cells (n=8) Figure 1D ) and Cézari syndrome (n=6) or mycosis fungoides tumor cells (n=4) Figure 1E Whole blood flow cytometry analysis of CD28 and CD38 surface expression in [a specific cell type]. Figure 1D In the image, letters A through C on the left side show the following patterns of CD38 and CD28 expression for each patient (read from left to right): A: CD38-CD28+, CD38+CD28-, CD38+CD28+, CD38-CD28-; B: CD38-CD28+, CD38+CD28+, CD38-CD28-; C: CD38-CD28+, CD38-CD28-. Figure 1EIn the figure, letters A to D on the left side show the following patterns of CD38 and CD28 expression for each patient (read from left to right): A: CD38-CD28+, CD38+CD28-, CD38+CD28+, CD38-CD28-; B: CD38-CD28+, CD38-CD28-; C: CD38-CD28+, CD38+CD28+, CD38-CD28-; D: CD38-CD28+, CD38-CD28-.

[0019] Figures 2A-2D IHC analysis of CD38 and CD28 in four T-cell lymphoma subtypes is shown, along with multiplex immunofluorescence of CD38, CD28, and tumor cell markers such as PD-1. A representative example of angioimmunoblastic T-cell lymphoma (AITL) is also presented. Figure 2A ); Sezari syndrome (SS) Figure 2B Anaplastic large cell ALK+ lymphoma (ALCL) Figure 2C ); extranodal NK / T-cell lymphoma with cutaneous manifestations (ENKTCL) Figure 2D The sub-images, from left to right, show CD28 IHC, CD38 IHC, and multiplex immunofluorescence IHC (targets as labeled). All sub-images are shown at 20× magnification.

[0020] Figures 3A-3F The anti-CD38 / CD28xCD3 trispecific binding protein was shown in different T lymphoma cell lines ( Figures 3A-3C ) or T leukemia cell line ( Figures 3D-3F Cytotoxic effects in ) . Using human PBMCs (E:T = 10:1) as effector cells and at indicated concentrations against the SEAX cell line (N=3) ( Figure 3A H9 (N=6) Figure 3B ), DERL2 (N=5) Figure 3C MOLT4 (N=5) Figure 3D ), JURKAT (N=4) Figure 3E ) and YT (N=5) Figure 3F In similar assays, indicated wild-type trispecific binding proteins (round) and null mutant trispecific binding proteins (trispecific binding proteins containing null mutant CD38 binding sites, square; trispecific binding proteins containing null mutant CD28 binding sites, triangle; and trispecific binding proteins containing null mutant CD38 and CD28 binding sites, inverted triangle) were used to assess cell lysis in different cell lines. Lysis was assessed by the percentage of CFSE+ apoptosis-active dye-positive (VD+) cells. Mean values ​​from data from different experiments conducted on different donors are shown.

[0021] Figures 4A-4E This presentation summarizes the IHC staining results for CD3, CD38, and CD28 in 261 solid T-cell lymphoma cases. IHC staining results for each marker (expressed as the percentage of positive cells in the sample) are shown for each case, along with intensity, percentage (%), and CD38 score. Cases are grouped by diagnosis; cutaneous anaplastic large cell lymphoma (cALCL) is also shown. Figure 4A (Above image), lymphoma-like papules ( Figure 4A (Above image), mycosis fungoides ( Figure 4A (middle image), transformed mycosis fungoides (MFt); Figure 4A (See image below), Cézari syndrome ( Figure 4B Primary cutaneous CD4+ TCL ( Figure 4B SCTCL (subcutaneous T-cell lymphoma, panniculitis-like) Figure 4B Primary cutaneous γδ T-cell lymphoma ( Figure 4B Extranodal NK / T-cell lymphoma (ENKTCL); Figure 4C (Above image) Enteropathy-associated T-cell lymphoma ( Figure 4C (middle image), MEITL (monomorphic epithelial intestinal T-cell lymphoma) Figure 4C (middle image), hepatosplenic T-cell lymphoma (HSTL); Figure 4C (See image below) Angioimmunoblastic T-cell lymphoma (AITL); Figure 4D (See above), PTCL is not specifically specified ( Figure 4D (See image below) Anaplastic lymphoma kinase (ALK) negative anaplastic large cell lymphoma (ALCL); Figure 4E ), anaplastic lymphoma kinase (ALK) positive ALCL ( Figure 4E ), ALCL breast implants ( Figure 4E ) and adult T-cell leukemia / lymphoma (ATLL; Figure 4E ).

[0022] Figure 5A and 5B It shows two different T leukemia cell lines (YTLT; Figure 5A and MOLT4LT; Figure 5B Cytotoxic effects of anti-CD38 / CD28xCD3 trispecific binding protein were assessed using sorted CD3+ T lymphocytes as effector cells. Sorted CD3+ T lymphocytes were used as effector cells. In a similar assay using sorted human lymphocytes (E:T = 10:1), cell lysis in different cell lines was also assessed using wild-type and null mutant trispecific binding proteins. Detailed Implementation

[0023] This disclosure provides, in particular, methods for treating PTCL using anti-CD38 T cell binders, as well as related uses, compositions, and kits. These are based, at least in part, on the evidence presented herein that anti-CD38 T cell binders (e.g., anti-CD38 / CD28xCD3 trispecific binding proteins) can induce cytotoxic lysis in various PTCL cell lines, including those derived from Cezari syndrome, MF (e.g., transformed MF), and hepatosplenic T-cell lymphoma, in the presence of human T cells. I. General Definition

[0024] As used in this disclosure, unless otherwise stated, the following terms should be understood to have the following meanings. Furthermore, unless the context requires otherwise, singular terms will include plural and plural terms will include singular. As used in this specification and the appended claims, unless the context clearly indicates otherwise, the singular forms “a / an” and “the” include plural references. Thus, for example, reference to “molecule” optionally includes a combination of two or more such molecules, etc.

[0025] It should be understood that the aspects and embodiments described herein include “comprising aspects and embodiments,” “consisting of aspects and embodiments,” and “substantially consisting of aspects and embodiments.”

[0026] As used herein, the term "polynucleotide" refers to a single-stranded or double-stranded nucleic acid polymer having a length of at least 10 nucleotides. In some embodiments, the nucleotides constituting the polynucleotide may be ribonucleotides or deoxyribonucleotides, or both of these in modified form. Such modifications include base modifications (such as bromouridine), ribose modifications (such as arabinoside and 2',3'-dideoxyribose), and internucleotide linking modifications, such as thiophosphates, dithiophosphates, selenophosphates, diselenophosphates, phenylaminothiophosphates, phenylaminophosphates, and aminophosphates. The term "polynucleotide" specifically includes both single-stranded and double-stranded forms of DNA.

[0027] "Separated polynucleotide" is a polynucleotide of the genome, cDNA, or synthetic origin or some combination thereof, which: (1) is not associated with all or part of the polynucleotide of the separated polynucleotide found in nature, (2) is linked with a polynucleotide in nature that is not linked to it, or (3) does not appear in nature as part of a larger sequence.

[0028] "Isolated polypeptide" means that the polypeptide: (1) does not contain at least some other polypeptides that are normally present with it, (2) is substantially free of other polypeptides from the same source (e.g., from the same species), (3) is expressed by cells from a different species, (4) has been isolated from at least about 50% of the polynucleotides, lipids, carbohydrates, or other substances associated with it in nature, (5) is not associated with any part of the polypeptides associated with the "isolated polypeptide" in nature (through covalent or non-covalent interactions), (6) is operatively associated with polypeptides not associated with it in nature (through covalent or non-covalent interactions), or (7) is not present in nature. Such isolated polypeptides may be encoded by genomic DNA, cDNA, mRNA, or other synthetically derived RNA or any combination thereof. Preferably, the isolated polypeptide is substantially free of polypeptides or other contaminants present in its natural environment that would interfere with its use (therapeutic, diagnostic, preventative, research, or otherwise).

[0029] Naturally occurring antibodies typically comprise tetramers. Each such tetramer usually consists of two pairs of identical polypeptide chains, each pair having a full-length “light” chain (typically with a molecular weight of about 25 kDa) and a full-length “heavy” chain (typically with a molecular weight of about 50–70 kDa). As used herein, the terms “heavy chain” and “light chain” refer to any immunoglobulin polypeptide with a sufficient sequence of variable domains to confer specificity against a target antigen. The amino-terminal portion of each light and heavy chain typically contains a variable domain of about 100 to 110 or more amino acids, which is typically responsible for antigen recognition. The carboxyl-terminal portion of each chain typically defines a constant domain responsible for effector cell function. Thus, in naturally occurring antibodies, the full-length heavy chain immunoglobulin polypeptide includes a variable domain (V... H ) and three constant structural domains (C H1 C H2 and C H3 ), where V H The domain is located at the amino terminus of the polypeptide and C H3 The domain is located at the carboxyl terminus, and the full-length light chain immunoglobulin polypeptide includes a variable domain (V). L ) and a constant structural domain (C L ), where V L The domain is located at the amino terminus of the polypeptide and C L The structural domain is located at the end of the carboxyl group.

[0030] Human light chains are typically classified as κ and λ light chains, and human heavy chains are typically classified as μ, δ, γ, α, or ε, with antibody isotypes defined as IgM, IgD, IgG, IgA, and IgE, respectively. IgG has several subclasses, including but not limited to IgG1, IgG2, IgG3, and IgG4. IgM has multiple subclasses, including but not limited to IgM1 and IgM2. IgA is similarly subdivided into multiple subclasses, including but not limited to IgA1 and IgA2. Within both the full-length light and heavy chains, variable and constant domains are typically linked by a “J” region of about 12 or more amino acids, and the heavy chain also includes a “D” region of about 10 or more amino acids. See, for example, Fundamental Immunology (edited by Paul, W., Raven Press, 2nd ed., 1989), which is incorporated herein by reference in its entirety for all purposes. The variable region of each light / heavy chain pair typically forms an antigen-binding site. Naturally occurring antibody variable domains typically exhibit the same overall structure as relatively conserved framework regions (FRs) linked by three hypervariable regions (also known as complementarity-determining regions or CDRs). The CDRs of each pair of chains are usually arranged within the framework regions, which allows for binding to specific epitopes. Both the light chain and heavy chain variable domains typically contain domains FR1, CDR1, FR2, CDR2, FR3, CDR3, and FR4, from the amino terminus to the carboxyl terminus.

[0031] The term "CDR set" refers to a group of three CDRs present in a single variable region capable of binding an antigen. The exact boundaries of these CDRs have been defined differently according to various systems. The system described by Kabat (Kabat et al., Sequences of Proteins of Immunological Interest (National Institutes of Health, Bethesda, Md., 1987 and (1991))) not only provides a definitive residue numbering system applicable to any variable region of an antibody but also provides precise residue boundaries defining three CDRs. These CDRs may be referred to as Kabat CDRs. Chothia and colleagues (Chothia and Lesk, 1987, J. Mol. Biol. [Journal of Molecular Biology] 196: 901-17; Chothia et al., 1989, Nature [Nature] 342: 877-83) discovered that, despite significant diversity at the amino acid sequence level, certain subregions within the Kabat CDR adopt nearly identical peptide skeleton architectures. These subregions are designated L1, L2, and L3 or H1, H2, and H3, where "L" and "H" designate the light chain and heavy chain regions, respectively. These regions may be referred to as the Chothia CDR, whose boundaries overlap with the Kabat CDR. Other boundaries defining CDRs overlapping with the Kabat CDR have been described in the following literature: Padlan, 1995, FASEB J. [Journal of the Federation for Experimental Biology] 9: 133-39; MacCallum, 1996, J. Mol. Biol. [Journal of Molecular Biology] 262(5): 732-45; and Lefranc, 2003, Dev. Comp. Immunol. [Developmental and Comparative Immunology] 27: 55-77. There are other CDR boundary definitions that may not strictly follow one of the systems described herein, but will still overlap with the Kabat CDR, although they may be shortened or lengthened depending on whether a particular residue or group of residues or even the entire CDR significantly affects the prediction or experimental findings of antigen binding. The methods used herein can utilize CDRs defined according to any of these systems, but some embodiments use CDRs defined by Kabat or Chothia.The use of amino acid sequence identification of predicted CDRs is well-known in this field, as illustrated in, for example, the following literature: Martin, AC "Protein sequence and structure analysis of antibody variable domains", in Antibody Engineering, Vol. 2. Edited by Kontermann R. and Dübel S., Springer-Verla, Berlin, pp. 33-51 (2010). The amino acid sequence of the heavy chain and / or light chain variable domains can also be examined to identify CDR sequences using other conventional methods, such as comparing them with known amino acid sequences of other heavy and light chain variable regions to identify regions with sequence hypervariability. Numbered sequences can be aligned visually or using alignment programs such as one of the CLUSTAL program suites, as described in Thompson, 1994, Nucleic Acids Res. 22: 4673-80. Molecular models are often used to accurately depict the framework region and CDR region, thereby correcting sequence-based allocation.

[0032] In some embodiments, the CDR / FR definition in the immunoglobulin light or heavy chain will be determined based on the IMGT definition (Lefranc et al., Dev. Comp. Immunol. [Developmental and Comparative Immunology], 2003, 27(1):55-77; www.imgt.org).

[0033] As used herein, the term "Fc" refers to a molecule (whether in monomeric or multimeric form) containing a sequence of a non-antigen-binding fragment produced by antibody digestion or other means, and may contain a hinge region. The original immunoglobulin source of natural Fc is preferably human, but can be any immunoglobulin. Fc molecules are composed of monomeric polypeptides that can be covalently (i.e., disulfide bonds) and non-covalently linked into dimers or multimers. The number of intermolecular disulfide bonds between the monomeric subunits of a natural Fc molecule ranges from 1 to 4, depending on the class (e.g., IgG, IgA, and IgE) or subclass (e.g., IgG1, IgG2, IgG3, IgA1, IgGA2, and IgG4). One example of Fc is a disulfide-bonded dimer produced by papain digestion of IgG. As used herein, the term "natural Fc" is used generally for monomeric, dimer, and multimer forms.

[0034] F(ab) segments typically consist of a light chain and a heavy chain V. Hand C H1 Structural domain, where the V of the F(ab) segment H -C H1 The heavy chain portion cannot form disulfide bonds with another heavy chain polypeptide. As used herein, the F(ab) fragment may also comprise a light chain containing two variable domains separated by amino acid linkers, and a C-chain containing two variable domains separated by amino acid linkers. H1 A heavy chain of a structural domain.

[0035] F(ab') segments typically consist of a light chain and a portion of a heavy chain containing more constant regions (in C... H1 With C H2 (between structural domains), which allows interchain disulfide bonds to form between two heavy chains to form the F(ab')2 molecule.

[0036] "Recombinant" molecules are molecules that are prepared, expressed, created, or isolated through recombination.

[0037] As used herein, the term "anti-CD38 T cell binder" refers to a multispecific (e.g., bispecific or trispecific) binding molecule capable of specifically binding to CD38 (e.g., expressed on the surface of target cells) and one or more surface proteins expressed by T cells (including, but not limited to, CD3 and / or CD28).

[0038] As used herein, the term “binding protein” refers to a non-naturally occurring (or recombinant or engineered) molecule that specifically binds to at least one target antigen.

[0039] The term "bispecific binding protein" refers to a binding protein that specifically binds to two different antigen targets. In some embodiments, a bispecific binding protein binds to two different antigens. For example, a bispecific binding protein may contain two antigen-binding sites, each of which specifically binds to a different target antigen (e.g., CD38 and CD3).

[0040] The term "trispecific binding protein" refers to a binding protein that specifically binds to three different antigen targets. In some embodiments, a trispecific binding protein binds to three different antigens. For example, a trispecific binding protein may contain three antigen-binding sites, each of which specifically binds to a different target antigen (e.g., CD38, CD28, and CD3).

[0041] The term "trivalent binding protein" refers to a binding protein having three antigen-binding sites. In a particular embodiment, a trivalent binding protein may bind to three antigen targets. For example, a trivalent binding protein may contain three antigen-binding sites, each of which specifically binds to a different target antigen (e.g., CD38, CD28, and CD3). In some embodiments, the binding protein may be trispecific and trivalent.

[0042] One embodiment of this disclosure provides a binding protein with biological and immunological specificity to two or three target antigens. Another embodiment of this disclosure provides nucleic acid molecules comprising a nucleotide sequence encoding a polypeptide chain forming such a binding protein. Yet another embodiment of this disclosure provides an expression vector comprising nucleic acid molecules comprising a nucleotide sequence encoding a polypeptide chain forming such a binding protein. Still another embodiment of this disclosure provides a host cell expressing such a binding protein (i.e., a nucleic acid molecule or vector comprising a polypeptide chain encoding such a binding protein).

[0043] As used herein, the term "interchangeability" refers to the interchangeability of variable domains within the binding protein formulation while preserving folding and final binding affinity. "Full interchangeability" refers to the interchangeability of V domains in either Formula I or Formula II polypeptide chains while maintaining the full functionality of the binding protein (as demonstrated by the preservation of binding affinity). H1 and V H2 The order of the two structural domains, and therefore the interchange of V. L1 and V L2 The ability to determine the order of structural domains (i.e., to reverse that order). Furthermore, it should be noted that the name V... H and V L This refers only to the location of a domain on a specific protein chain in the final format. For example, V H1 and V H2 V can be derived from parental antibodies L1 and V L2 The domain is located within the V-binding protein. H1 and V H2 Location. Similarly, V L1 and V L2 V can be derived from parental antibodies H1 and V H2 The domain is located within the V-binding protein. H1 and V H2 Location. Therefore, the V H and V L The name refers to the current position within the parent antibody, not the original position. Therefore, V H and V L The structural domain is "commutative".

[0044] As used herein, the terms “antigen” or “target antigen” or “antigen target” refer to a molecule or part of a molecule that can be bound by a binding protein and can also be used in animals to generate antibodies that can bind to an epitope of that antigen. A target antigen may have one or more epitopes. For each target antigen recognized by a binding protein, that binding protein can compete with an intact antibody that recognizes that target antigen.

[0045] "CD38" is the differentiation cluster 38 polypeptide, a glycoprotein found on the surface of many immune cells. In some embodiments, the anti-CD38 T cell binder disclosed herein binds to one or more extracellular domains of the CD38 polypeptide. Exemplary CD38 extracellular domain polypeptide sequences include, but are not limited to, the extracellular domain of human CD38 (e.g., as shown in SEQ ID NO: 1) and the extracellular domain of cynomolgus monkey CD38 (e.g., as shown in SEQ ID NO: 30). In some embodiments, the CD38 polypeptide is the human CD38 polypeptide.

[0046] "CD28" is the differentiation cluster 28 polypeptide, a T cell surface protein that provides co-stimulatory signals for T cell activation and survival. CD28 polynucleotide and polypeptide sequences are known in the art; see, for example, NCBI gene IDs 940 and NP_001230006. In some embodiments, the CD28 polypeptide is the human CD28 polypeptide.

[0047] "CD3" is differentiation cluster factor 3 polypeptide, a T cell surface protein, typically part of the T cell receptor (TCR) complex formed by the TCR α / β or γ / δ heterodimer and CD3-ε, -γ, -δ, and -ζ. CD3 polynucleotide and polypeptide sequences are known in the art; see, for example, NCBI gene IDs 915 and NP_000723 (CD3-δ), NCBI gene IDs 916 and NP_000724 (CD3-ε), NCBI gene IDs 917 and NP_000064 (CD3-γ), and NCBI gene IDs 919 and NP_000725 (CD3-ζ). In some embodiments, the CD3 polypeptide is the human CD3 polypeptide.

[0048] "Isolated" binding proteins refer to binding proteins that have been identified, isolated, and / or recovered from components of their native environment. Contaminating components of their native environment are substances that can interfere with the diagnostic or therapeutic use of the binding protein and may include enzymes, hormones, and other protein or non-protein solutes. In some embodiments, the binding protein is purified to: (1) greater than 95% by weight of the antibody, and most preferably greater than 99% by weight, as determined by the Lowry method; (2) to a degree sufficient to obtain at least 15 residues of the N-terminal or internal amino acid sequence using a rotary cup sequencer; or (3) based on homogeneity under reducing or non-reducing conditions using Coomassie blue or preferably silver staining. Isolated binding proteins include recombinant intracellular in situ binding proteins, since at least one component of the binding protein's native environment will be absent.

[0049] As used herein, the terms "substantially pure" or "substantially purified" refer to the dominant compounds or species present (i.e., more than any other single species in the composition, based on molar units). In some embodiments, a substantially purified fraction is a composition in which these species constitute at least about 50% (on a molar basis) of all macromolecular species present. In other embodiments, a substantially pure composition would contain more than about 80%, 85%, 90%, 95%, or 99% of all macromolecular species present in the composition. In still other embodiments, the species are purified to substantially homogeneity (contaminant species in the composition cannot be detected by conventional detection methods), wherein the composition consists essentially of a single macromolecular species.

[0050] The term "epitope" includes any determinant cluster (preferably a polypeptide determinant cluster) capable of specifically binding to immunoglobulins or T-cell receptors. In some embodiments, epitope determinants comprise chemically active surface clusters of molecules (such as amino acids, sugar side chains, phosphoryl groups, or sulfonyl groups), and in some embodiments, may have specific three-dimensional structural features and / or specific charge features. An epitope is an antigenic region bound by an antibody or binding protein. In some embodiments, when a binding protein preferentially recognizes a target antigen in a complex mixture of proteins and / or macromolecules, it is called a protein-specific binding antigen. In some embodiments, when the equilibrium dissociation constant is ≤ 10... -8 More preferably, when the equilibrium dissociation constant is ≤ 10, M is used. -9 When M, and most preferably when the dissociation constant ≤ 10 -10 In case M, it is believed that the binding protein specifically binds to the antigen.

[0051] The dissociation constant (K) of the bound protein DThis can be determined, for example, by surface plasmon resonance (SPR). Typically, SPR analysis uses a BIAcore system (Pharmacia Biosensor; Piscatave, NJ) to measure the real-time binding interaction between the ligand (target antigen on the biosensor matrix) and the analyte (binding protein in solution) via surface plasmon resonance (SPR). SPR analysis can also be performed by immobilizing the analyte (binding protein on the biosensor matrix) and presenting the ligand (target antigen). As used herein, the term "K" is used... D "" refers to the dissociation constant of the interaction between a specific binding protein and a target antigen.

[0052] As used herein, the term "binding" in relation to binding proteins refers to a binding protein or its antigen-binding fragment with a binding force of at least about 1 × 10⁻⁶. -6 M, 1 × 10 -7 M, 1 × 10 -8 M, 1 × 10 -9 M, 1 × 10 -10 M, 1 × 10 -11 M, 1 × 10 -12 The ability of M or higher Kd to bind to antigens containing epitopes, and / or the ability to bind to epitopes with an affinity at least twice that of the binding protein or its antigen-binding fragment for nonspecific antigens. In some embodiments, the binding protein disclosed herein binds two or more antigens, such as human and cynomolgus monkey CD38 peptides.

[0053] In some embodiments, the antigen-binding domains and / or binding proteins disclosed herein are “cross-reactive” with human and cynomolgus CD38 peptides (e.g., CD38 extracellular domains, such as SEQ ID NO: 1 (human CD38 isotype A), SEQ ID NO: 105 (human CD38 isotype E), and SEQ ID NO: 30 (cynomolgus CD38)). When the EC50 for both antigens is within a similar range, the binding protein binding to antigen 1 (Ag1) is “cross-reactive” with antigen 2 (Ag2). In this application, when the affinity ratio of Ag2 to Ag1 is equal to or less than 10 (e.g., 5, 2, 1, or 0.5), the binding protein binding to Ag1 is cross-reactive with Ag2, and the affinity for both antigens is measured using the same method.

[0054] When the affinities for two antigens are very different, the binding protein that binds to Ag1 does not exhibit significant cross-reactivity with Ag2. If the binding response is too low, the affinity for Ag2 may not be measurable. In this application, under the same experimental conditions and at the same antibody concentration, when the binding response of the Ag1-binding protein to Ag2 is less than 5% of the binding response of the same binding protein to Ag1, the binding protein does not exhibit significant cross-reactivity with Ag2. In practice, the concentration of the binding protein used may be EC50 or the concentration required to reach the saturation plateau obtained with Ag1.

[0055] As used herein, the term "linker" refers to one or more amino acid residues inserted between immunoglobulin domains, thereby providing sufficient mobility for the light and heavy chain domains to fold into a cross-linked, dual-variable-region immunoglobulin. At the sequence level, linkers are inserted into transition regions between variable domains or between variable and constant domains, respectively. Since the approximate sizes of immunoglobulin domains are well understood, transition regions between domains can be identified. The precise location of the domain transition can be determined by locating peptide extensions that do not form (as shown by experimental data or hypothesized by techniques such as modeling or secondary structure prediction) secondary structural elements (such as β-sheets or α-helices). The linker described herein is referred to as L1, which is located on the V-shaped region of the light chain. L2 C-terminus and V-terminus of the structural domain L1 Between the N-termini of the structural domains; and L2, which lies on the V-terminus of the light chain. L1 The C-terminus of the domain and C L Between the N-termini of the structural domains. The relink head is called L3, which is located at V. H1 C-terminus and V-terminus of the structural domain H2 Between the N-termini of the structural domains; and L4, which is located in V. H2 The C-terminus of the domain and C H1 Between the N-termini of the structural domain.

[0056] As used herein, the term "vector" refers to any molecule (e.g., nucleic acid, plasmid, or virus) used to transfer coding information to a host cell. The term "vector" includes nucleic acid molecules capable of transporting another nucleic acid molecule linked to them. One type of vector is the "plasmid," which refers to a circular double-stranded DNA molecule into which an additional DNA segment can be inserted. Another type of vector is a viral vector, in which an additional DNA segment can be inserted into the viral genome. Some vectors are capable of autonomous replication in the host cells into which they are introduced (e.g., bacterial vectors with bacterial origins of replication and free-living mammalian vectors). Other vectors (e.g., non-free-living mammalian vectors) can integrate into the host cell's genome after introduction, thereby replicating along with the host genome. Furthermore, some vectors are capable of directing the expression of genes to which they are operatively linked. Such vectors are referred to herein as "recombinant expression vectors" (or simply "expression vectors"). Typically, expression vectors useful in recombinant DNA technology are in the form of plasmids. The terms "plasmid" and "vector" are used interchangeably herein because plasmids are the most commonly used form of vector. However, this disclosure is intended to include other forms of expression vectors that perform equivalent functions, such as viral vectors (e.g., replication-defective retroviruses, adenoviruses, and adeno-associated viruses).

[0057] As used herein, the phrase “recombinant host cell” (or “host cell”) refers to a cell in which a recombinant expression vector has been introduced. Recombinant host cell or host cell means not only the specific test cell but also the progeny of such cells. Such progeny may actually differ from the parent cell because certain modifications can occur in the progeny due to mutations or environmental influences, but such cells are still included within the scope of the term “host cell” as used herein. A wide variety of host cell expression systems can be used to express binding proteins, including bacterial, yeast, baculovirus, and mammalian expression systems (as well as phage display expression systems). An example of a suitable bacterial expression vector is pUC19. To recombinantly express the binding protein, host cells are transformed or transfected with one or more recombinant expression vectors carrying a DNA fragment encoding a polypeptide chain of the binding protein, such that the polypeptide chain is expressed in the host cells and preferably secreted into the culture medium from which the binding protein can be recovered.

[0058] As used herein, the term "transformation" refers to an alteration of the genetic characteristics of a cell, and a cell is considered transformed when it is modified to contain new DNA. For example, a cell is transformed from its native state through genetic modification. After transformation, the transformed DNA can either physically integrate into the cell's chromosome and recombine with the cell's DNA, or it can be temporarily maintained as a non-replicating free element, or it can replicate independently as a plasmid. A cell is considered stably transformed when this DNA replicates with cell division. As used herein, the term "transfection" refers to the uptake of foreign or exogenous DNA by a cell, and a cell is considered "transfected" when the exogenous DNA has been introduced into the cell membrane. Many transfection techniques are well known in the art. Such techniques can be used to introduce one or more exogenous DNA molecules into a suitable host cell.

[0059] As used herein and applied to objects, the term "naturally occurring" means the fact that the object exists in nature and has not been artificially manipulated. For example, polynucleotides or polypeptides that exist in organisms (including viruses) and can be isolated from natural sources and have not been intentionally modified by humans are naturally occurring. Similarly, as used herein, "non-naturally occurring" means an object that has not been found in nature or that has been artificially modified or synthesized.

[0060] As used herein, the twenty common amino acids and their abbreviations follow conventional usage. Stereoisomers of the twenty common amino acids (e.g., D-amino acids); non-natural amino acids and analogues (such as α-, α-disubstituted amino acids, N-alkyl amino acids, lactic acid, and other unconventional amino acids) may also be suitable components of the polypeptide chain of the protein. Examples of unconventional amino acids include: 4-hydroxyproline, γ-carboxyglutamic acid, ε-N,N,N-trimethyllysine, ε-N-acetyllysine, O-phosphoserine, N-acetylserine, N-formylmethionine, 3-methylhistidine, 5-hydroxylysine, σ-N-methylarginine, and other similar amino acids and imino acids (e.g., 4-hydroxyproline). In the polypeptide symbols used herein, according to standard usage and convention, the left-hand direction is the amino-terminal direction, and the right-hand direction is the carboxyl-terminal direction.

[0061] Naturally occurring residues can be classified based on their common side-chain characteristics: (1) Hydrophobicity: Met, Ala, Val, Leu, Ile, Phe, Trp, Tyr, Pro; (2) Polar hydrophilicity: Arg, Asn, Asp, Gln, Glu, His, Lys, Ser, Thr; (3) Aliphatic: Ala, Gly, Ile, Leu, Val, Pro; (4) Aliphatic hydrophobicity: Ala, Ile, Leu, Val, Pro; (5) Neutral hydrophilicity: Cys, Ser, Thr, Asn, Gln; (6) Acidic: Asp, Glu; (7) Alkaline: His, Lys, Arg; (8) Residues affecting chain orientation: Gly, Pro; (9) Aromatic compounds: His, Trp, Tyr, Phe; and (10) Aromatic hydrophobicity: Phe, Trp, Tyr.

[0062] Conservative amino acid substitutions may involve the exchange of a member from one of these classes with another member from the same class. Non-conservative substitutions may involve the exchange of a member from one of these classes with a member from another class.

[0063] Those skilled in the art will be able to use well-known techniques to identify suitable variants of the polypeptide chain that binds to the protein. For example, those skilled in the art can identify suitable regions of the polypeptide chain that can be altered without destroying activity by targeting regions considered unimportant to activity. Alternatively, those skilled in the art can identify conserved residues and molecular moieties in similar polypeptides. Furthermore, it is even possible to subject regions that may be important for biological activity or structure to conserved amino acid substitutions without destroying biological activity or adversely affecting the polypeptide structure.

[0064] As used in this article, the terms “patient,” “individual,” or “subject” include both people and animals (e.g., mammals such as dogs, pigs, horses, cats, cows, etc.).

[0065] As used herein, the term "treatment" refers to both therapeutic treatment and preventative or preventative measures. Those requiring treatment include those who have a condition, those who are susceptible to a condition, or those for whom prevention is desired. In a particular embodiment, the binding protein may be used to treat a person with cancer or a person susceptible to cancer, or to improve cancer in human subjects. The binding protein may also be used to prevent cancer in human patients. In a particular embodiment, the cancer is PTCL.

[0066] As used herein, the terms “pharmaceutical composition” or “therapeutic composition” refer to a compound or composition that, when properly administered to a patient, can induce a desired therapeutic effect.

[0067] As used herein, the terms “pharmaceutically acceptable carrier” or “physiologically acceptable carrier” refer to one or more formulations suitable for achieving or enhancing the delivery of binding proteins.

[0068] When referring to the use of pharmaceutical compositions containing one or more binding proteins, the terms "effective amount" and "therapeutic effective amount" mean an amount or dose sufficient to produce the desired therapeutic outcome. More specifically, a therapeutic effective amount is an amount of binding protein sufficient to inhibit one or more clinically defined pathological processes associated with the treated condition for a period of time. Effective amounts can vary depending on the specific binding protein used and also on a variety of factors and conditions related to the patient being treated and the severity of the disorder. For example, if the binding protein is to be administered in vivo, factors such as the patient's age, weight, and health status, as well as dose-response curves and toxicity data obtained in preclinical animal work, would be among those considered. Determining the effective amount or therapeutic effective amount of a given pharmaceutical composition is entirely within the capabilities of those skilled in the art.

[0069] The embodiments disclosed herein provide a pharmaceutical composition comprising a pharmaceutically acceptable carrier and a therapeutically effective amount of a binding protein. II. Methods and Applications

[0070] Certain aspects of this disclosure relate to methods for treating peripheral T-cell lymphoma (PTCL) in, for example, an individual in need. In some embodiments, these methods include administering an effective amount of the anti-CD38 T-cell binder of this disclosure to the individual. In some embodiments, the individual is a human being.

[0071] In some embodiments, an individual has PTCL or has been diagnosed with PTCL. In some embodiments, PTCL is angioimmunoblastic T-cell lymphoma (AITL), hepatosplenic T-cell lymphoma (HSTL), adult T-cell leukemia / lymphoma (ATLL), extranodal NK / T-cell lymphoma (ENKTCL), mycosis fungoides (MF), Cézari syndrome (SS), anaplastic large cell ALK-lymphoma (ALCL), PTCL-nospecific (NOS), enteropathy-type T-cell lymphoma (EATL), monomorphic epithelial eosinophilic intestinal T-cell lymphoma (MEITL), primary cutaneous CD4+ lymphoproliferative disorder, subcutaneous panniculitis-like T-cell lymphoma (SCTCL), or primary cutaneous γδ T-cell lymphoma. In some embodiments, MF is transformed MF. In some embodiments, PTCL is HSTL, Cézari syndrome, or MF (e.g., transformed MF).

[0072] In some embodiments, PTCL cells express CD38 and / or CD28. In some embodiments, PTCL cells express both CD38 and CD28. In some embodiments, PTCL cells express CD38. In some embodiments, PTCL cells express CD28. In some embodiments, the expression of CD38 and / or CD28 by PTCL cells can be measured in a sample from PTCL (e.g., a sample containing PTCL cells). In some embodiments, a sample obtained from an individual's PTCL contains cells expressing CD38 and / or CD28 (e.g., PTCL cells). In some embodiments, the method disclosed herein further includes measuring the expression of CD38 and / or CD28 in a sample obtained from an individual (e.g., a sample containing PTCL cells). In some embodiments, the method disclosed herein further includes obtaining a sample from an individual, such as a sample containing PTCL cells. In some embodiments, the sample is from a tumor biopsy. A variety of methods known in the art are suitable for measuring the expression of CD38 and / or CD28 in a sample, including but not limited to immunohistochemistry (IHC), immunofluorescence (IF) microscopy, and flow cytometry.

[0073] Any anti-CD38 T cell binder, bispecific binding protein, or trispecific binding protein described herein may be used in the methods, uses, kits, and compositions disclosed herein. Anti-CD38 T cell engager (TCE)

[0074] Certain aspects of this disclosure relate to anti-CD38 T-cell conjugates. In some embodiments, the anti-CD38 T-cell conjugate binds, for example, human CD38 expressed on the surface of cells (e.g., PTCL cells). In some embodiments, the anti-CD38 T-cell conjugate binds, for example, human CD28 expressed on the surface of cells (e.g., PTCL cells). In some embodiments, the anti-CD38 T-cell conjugate binds, for example, human CD3 expressed on the surface of cells (e.g., PTCL cells). In some embodiments, the anti-CD38 T-cell conjugate binds both human CD38 and human CD3. In some embodiments, the anti-CD38 T-cell conjugate binds human CD38, human CD28, and human CD3. Exemplary human CD38, CD28, and CD3 peptides are known in the art and are disclosed herein.

[0075] In some embodiments, the anti-CD38 T cell conjugate is a bispecific binding protein (e.g., a bispecific antibody) comprising a first antigen-binding site that specifically binds to a CD38 peptide and a second antigen-binding site that specifically binds to a CD3 peptide. For example, the anti-CD38 T cell conjugate may be a bispecific antibody comprising at least a first antigen-binding domain that specifically binds to a CD38 peptide and at least a second antigen-binding domain that specifically binds to a CD3 peptide.

[0076] In some embodiments, the anti-CD38 T cell conjugate is a trispecific binding protein. In some embodiments, the binding protein includes an antigen-binding site that specifically binds a CD38 peptide, an antigen-binding site that specifically binds a CD28 peptide, and an antigen-binding site that specifically binds a CD3 peptide. In some embodiments, the binding protein induces apoptosis in CD38+ cells. In some embodiments, the binding protein recruits T cells to CD38+ cells and optionally activates T cells (e.g., via TCR stimulation and / or co-stimulation). Any anti-CD38 T cell conjugate described herein can be used in the methods, uses, kits, and compositions disclosed herein. Anti-CD38 antigen binding site

[0077] Exemplary antigen-binding sites that specifically bind to CD38 peptides and can be used in the anti-CD38 T-cell conjugates disclosed herein are described below. Any anti-CD38 antigen-binding site and / or trispecific binding protein described in International Application No. WO 2019 / 074973 can be used in the methods, uses, kits, and compositions disclosed herein. In some embodiments, the CD38 peptide is a human CD38 peptide, also known as ADPRC1. Human CD38 peptides are known in the art and include, but are not limited to, peptides represented by NCBI accession number NP_001766.2 or peptides generated by NCBI gene ID 952. In some embodiments, the antigen-binding site binds to a human CD38 peptide, a non-human primate (e.g., cynomolgus monkey) CD38 peptide, or a human CD38 peptide and a non-human primate (e.g., cynomolgus monkey) CD38 peptide.

[0078] In some embodiments, the antigen-binding site of the CD38 peptide comprises: an antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence containing an amino acid sequence of GYTFTSFN (SEQ ID NO: 31) or GYTFTSYA (SEQ ID NO: 37), a CDR-H2 sequence containing an amino acid sequence of IYPGNGGT (SEQ ID NO: 32) or IYPGQGGT (SEQ ID NO: 38), and a CDR-H3 sequence containing an amino acid sequence of ARTGGLRRAYFTY (SEQ ID NO: 33); or an antibody light chain variable (VL) domain comprising a CDR-L1 sequence containing an amino acid sequence of ESVDSYGNGF (SEQ ID NO: 34) or QSVSSYGQGF (SEQ ID NO: 39), a CDR-L2 sequence containing an amino acid sequence of LAS or GAS, and a CDR-L3 sequence containing an amino acid sequence of QQNKEDPWT (SEQ ID NO: 36). In some embodiments, the antigen-binding site of the CD38 peptide comprises: an antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence containing an amino acid sequence of GYTFTSFN (SEQ ID NO: 31) or GYTFTSYA (SEQ ID NO: 37), a CDR-H2 sequence containing an amino acid sequence of IYPGNGGT (SEQ ID NO: 32) or IYPGQGGT (SEQ ID NO: 38), and a CDR-H3 sequence containing an amino acid sequence of ARTGGLRRAYFTY (SEQ ID NO: 33); or an antibody light chain variable (VL) domain comprising a CDR-L1 sequence containing an amino acid sequence of ESVDSYGNGF (SEQ ID NO: 34) or QSVSSYGQG (SEQ ID NO: 132), a CDR-L2 sequence containing an amino acid sequence of LAS or GAS, and a CDR-L3 sequence containing an amino acid sequence of QQNKEDPWT (SEQ ID NO: 36). In some embodiments, the antigen-binding site of the CD38 peptide comprises 1, 2, 3, 4, 5, or 6 CDRs of the antibody VH and / or VL domain sequences from antigen-binding sites shown in Table A or B (e.g., mAb1, mAb2, mAb3, mAb4, mAb5, mAb6, hhy992, hu5739, hu6284, hhy1195, hhy1370, hyb5739, or hyb6284). In some embodiments, the antigen-binding site of the CD38 peptide comprises 1, 2, 3, 4, 5, or 6 CDRs of the antibody VH and / or VL domain sequences from trispecific binding proteins shown in Table E1 or E2.

[0079] In some embodiments, the antigen-binding site of the CD38 peptide comprises: an antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence containing an amino acid sequence of GYTFTSFN (SEQ ID NO: 31) or GYTFTSYA (SEQ ID NO: 37), a CDR-H2 sequence containing an amino acid sequence of IYPGNGGT (SEQ ID NO: 32) or IYPGQGGT (SEQ ID NO: 38), and a CDR-H3 sequence containing an amino acid sequence of ARTGGLRRAYFTY (SEQ ID NO: 33); and an antibody light chain variable (VL) domain comprising a CDR-L1 sequence containing an amino acid sequence of ESVDSYGNGF (SEQ ID NO: 34) or QSVSSYGQGF (SEQ ID NO: 39), a CDR-L2 sequence containing an amino acid sequence of LAS or GAS, and a CDR-L3 sequence containing an amino acid sequence of QQNKEDPWT (SEQ ID NO: 36).

[0080] In some embodiments, the antigen-binding site of the CD38 peptide comprises: an antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence containing the amino acid sequence GYTFTSFN (SEQ ID NO: 31), a CDR-H2 sequence containing the amino acid sequence IYPGNGGT (SEQ ID NO: 32), and a CDR-H3 sequence containing the amino acid sequence ARTGGLRRAYFTY (SEQ ID NO: 33); or an antibody light chain variable (VL) domain comprising a CDR-L1 sequence containing the amino acid sequence ESVDSYGNGF (SEQ ID NO: 34), a CDR-L2 sequence containing the amino acid sequence LAS, and a CDR-L3 sequence containing the amino acid sequence QQNKEDPWT (SEQ ID NO: 36). In some embodiments, the antigen-binding site of the CD38 peptide comprises: an antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence containing the amino acid sequence GYTFTSFN (SEQ ID NO: 31), a CDR-H2 sequence containing the amino acid sequence IYPGNGGT (SEQ ID NO: 32), and a CDR-H3 sequence containing the amino acid sequence ARTGGLRRAYFTY (SEQ ID NO: 33); and an antibody light chain variable (VL) domain comprising a CDR-L1 sequence containing the amino acid sequence ESVDSYGNGF (SEQ ID NO: 34), a CDR-L2 sequence containing the amino acid sequence LAS, and a CDR-L3 sequence containing the amino acid sequence QQNKEDPWT (SEQ ID NO: 36).

[0081] In some embodiments, the antigen-binding site of the CD38 peptide comprises: an antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence containing the amino acid sequence GYTFTSYA (SEQ ID NO: 37), a CDR-H2 sequence containing the amino acid sequence IYPGQGGT (SEQ ID NO: 38), and a CDR-H3 sequence containing the amino acid sequence ARTGGLRRAYFTY (SEQ ID NO: 33); or an antibody light chain variable (VL) domain comprising a CDR-L1 sequence containing the amino acid sequence QSVSSYGQGF (SEQ ID NO: 39), a CDR-L2 sequence containing the amino acid sequence GAS, and a CDR-L3 sequence containing the amino acid sequence QQNKEDPWT (SEQ ID NO: 36). In some embodiments, the antigen-binding site of the CD38 peptide comprises: an antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence containing the amino acid sequence GYTFTSYA (SEQ ID NO: 37), a CDR-H2 sequence containing the amino acid sequence IYPGQGGT (SEQ ID NO: 38), and a CDR-H3 sequence containing the amino acid sequence ARTGGLRRAYFTY (SEQ ID NO: 33); and an antibody light chain variable (VL) domain comprising a CDR-L1 sequence containing the amino acid sequence QSVSSYGQGF (SEQ ID NO: 39), a CDR-L2 sequence containing the amino acid sequence GAS, and a CDR-L3 sequence containing the amino acid sequence QQNKEDPWT (SEQ ID NO: 36).

[0082] In some embodiments, the VH domain comprises the sequence from the N-terminus to the C-terminus: FR1—CDR-H1—FR2—CDR-H2—FR3—CDR-H3—FR4; wherein FR1 comprises the sequence QVQLVQSGAEVVKPGASVKVSCKAS (SEQ ID NO: 86), QVQLVQSGAEVVKSGASVKVSCKAS (SEQ ID NO: 87), or QVQLVQSGAEVVKPGASVKMSCKAS (SEQ ID NO: 88); wherein FR2 comprises the sequence MHWVKEAPGQRLEWIGY (SEQ ID NO: 90) or MHWVKEAPGQGLEWIGY (SEQ ID NO: 91); wherein FR3 comprises the sequence NYNQKFQGRATLTADTSASTAYMELSSLRSEDTAVYFC (SEQ ID NO: 93) or NYNQKFQGRATLTADTSASTAYMEISSLRSEDTAVYFC (SEQ ID NO: 93). 94); and wherein FR4 contains the sequence WGQGTLVTVSS (SEQ ID NO: 96). In some embodiments, the VL domain contains the sequence from the N-terminus to the C-terminus: FR1—CDR-L1—FR2—CDR-L2—FR3—CDR-L3—FR4; wherein FR1 contains the sequence DIVLTQSPATLSLSPGERATISCRAS (SEQ ID NO: 97); wherein FR2 contains the sequence MHWYQQKPGQPPRLLIY (SEQ ID NO: 99); wherein FR3 contains the sequence SRATGIPARFSGSGSGTDFTLTISPLEPEDFAVYYC (SEQ ID NO: 101); and wherein FR4 contains the sequence FGGGTKLEIK (SEQ ID NO: 103).

[0083] In some embodiments, the VH domain contains an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of SEQ ID NO: 5; and / or the VL domain contains an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of SEQ ID NO: 6. In some embodiments, the VH domain contains an amino acid sequence that is at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 17; and / or the VL domain contains an amino acid sequence that is at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 18. In some embodiments, the VH domain contains an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of SEQ ID NO: 21; and / or the VL domain contains an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of SEQ ID NO: 18.In some embodiments, the VH domain contains an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of SEQ ID NO: 23; and / or the VL domain contains an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of SEQ ID NO: 18. In some embodiments, the VH domain contains an amino acid sequence that is at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 13; and / or the VL domain contains an amino acid sequence that is at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 14.

[0084] In some embodiments, the VH domain contains an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of SEQ ID NO: 5; and the VL domain contains an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of SEQ ID NO: 6. In some embodiments, the VH domain contains an amino acid sequence that is at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 17; and the VL domain contains an amino acid sequence that is at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 18. In some embodiments, the VH domain contains an amino acid sequence that is at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 21; and the VL domain contains an amino acid sequence that is at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 18.In some embodiments, the VH domain contains an amino acid sequence that is at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 23; and the VL domain contains an amino acid sequence that is at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 18. In some embodiments, the VH domain contains an amino acid sequence that is at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 13; and the VL domain contains an amino acid sequence that is at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 14.

[0085] In some embodiments, the VH domain comprises the amino acid sequence of SEQ ID NO: 5; and the VL domain comprises the amino acid sequence of SEQ ID NO: 6. In some embodiments, the VH domain comprises the amino acid sequence of SEQ ID NO: 17; and the VL domain comprises the amino acid sequence of SEQ ID NO: 18. In some embodiments, the VH domain comprises the amino acid sequence of SEQ ID NO: 21; and the VL domain comprises the amino acid sequence of SEQ ID NO: 18. In some embodiments, the VH domain comprises the amino acid sequence of SEQ ID NO: 23; and the VL domain comprises the amino acid sequence of SEQ ID NO: 18. In some embodiments, the VH domain comprises the amino acid sequence of SEQ ID NO: 13; and the VL domain comprises the amino acid sequence of SEQ ID NO: 14.

[0086] In some embodiments, the antigen-binding site of the CD38 peptide comprises: an antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence containing the amino acid sequence GFTFSSYG (SEQ ID NO: 41), a CDR-H2 sequence containing the amino acid sequence IWYDGSNK (SEQ ID NO: 42), and a CDR-H3 sequence containing the amino acid sequence ARMFRGAFDY (SEQ ID NO: 43); or an antibody light chain variable (VL) domain comprising a CDR-L1 sequence containing the amino acid sequence QGIRND (SEQ ID NO: 44), a CDR-L2 sequence containing the amino acid sequence AAS, and a CDR-L3 sequence containing the amino acid sequence LQDYIYYPT (SEQ ID NO: 46). In some embodiments, the antigen-binding site of the CD38 peptide comprises: an antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence containing the amino acid sequence GFTFSSYG (SEQ ID NO: 41), a CDR-H2 sequence containing the amino acid sequence IWYDGSNK (SEQ ID NO: 42), and a CDR-H3 sequence containing the amino acid sequence ARMFRGAFDY (SEQ ID NO: 43); and an antibody light chain variable (VL) domain comprising a CDR-L1 sequence containing the amino acid sequence QGIRND (SEQ ID NO: 44), a CDR-L2 sequence containing the amino acid sequence AAS, and a CDR-L3 sequence containing the amino acid sequence LQDYIYYPT (SEQ ID NO: 46).

[0087] In some embodiments, the VH domain comprises a sequence from the N-terminus to the C-terminus: FR1—CDR-H1—FR2—CDR-H2—FR3—CDR-H3—FR4; wherein FR1 comprises the sequence QVQLVESGGGVVQPGRSLRLSCAAS (SEQ ID NO: 89); wherein FR2 comprises the sequence MHWVRQAPGKGLEWVAV (SEQ ID NO: 92); wherein FR3 comprises the sequence YYADSVKGRFTISGDNSKNTLYLQMNSLRAEDTAVYYC (SEQ ID NO: 95); and wherein FR4 comprises the sequence WGQGTLVTVSS (SEQ ID NO: 96). In some embodiments, the VL domain comprises a sequence from the N-terminus to the C-terminus: FR1—CDR-L1—FR2—CDR-L2—FR3—CDR-L3—FR4; wherein FR1 comprises the sequence AIQMTQSPSSLSASVGDRVTITCRAS (SEQ ID NO: 98); wherein FR2 comprises the sequence GWYQQKPGKAPKLLIY (SEQ ID NO: 100); wherein FR3 comprises the sequence SLQSGVPSRFSGSGSGTDFTLTISGLQPEDSATYYC (SEQ ID NO: 102); and wherein FR4 comprises the sequence WGQGTLVTVSS (SEQ ID NO: 104).

[0088] In some embodiments, the VH domain contains an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of SEQ ID NO: 9; and / or the VL domain contains an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of SEQ ID NO: 10.

[0089] In some embodiments, the VH domain contains an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of SEQ ID NO: 10; and the VL domain contains an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of SEQ ID NO: 10. In some embodiments, the VH domain contains the amino acid sequence of SEQ ID NO: 9; and the VL domain contains the amino acid sequence of SEQ ID NO: 10.

[0090] In some embodiments, the antigen-binding site of the CD38 peptide comprises: an antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence containing the amino acid sequence GYTLTEFS (SEQ ID NO: 2), a CDR-H2 sequence containing the amino acid sequence FDPEDGET (SEQ ID NO: 3), and a CDR-H3 sequence containing the amino acid sequence TTGRFFDWF (SEQ ID NO: 4); or an antibody light chain variable (VL) domain comprising a CDR-L1 sequence containing the amino acid sequence QSVISRF (SEQ ID NO: 7), a CDR-L2 sequence containing the amino acid sequence GAS, and a CDR-L3 sequence containing the amino acid sequence QQDSNLPIT (SEQ ID NO: 11). In some embodiments, the antigen-binding site of the CD38 peptide comprises: an antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence containing the amino acid sequence GYTLTEFS (SEQ ID NO: 2), a CDR-H2 sequence containing the amino acid sequence FDPEDGET (SEQ ID NO: 3), and a CDR-H3 sequence containing the amino acid sequence TTGRFFDWF (SEQ ID NO: 4); and an antibody light chain variable (VL) domain comprising a CDR-L1 sequence containing the amino acid sequence QSVISRF (SEQ ID NO: 7), a CDR-L2 sequence containing the amino acid sequence GAS, and a CDR-L3 sequence containing the amino acid sequence QQDSNLPIT (SEQ ID NO: 11).

[0091] In some embodiments, the antigen-binding site of the CD38 peptide comprises: an antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence containing the amino acid sequence GYAFTTYL (SEQ ID NO: 12), a CDR-H2 sequence containing the amino acid sequence INPGSGST (SEQ ID NO: 15), and a CDR-H3 sequence containing the amino acid sequence ARYAYGY (SEQ ID NO: 16); or an antibody light chain variable (VL) domain comprising a CDR-L1 sequence containing the amino acid sequence QNVGTA (SEQ ID NO: 19), a CDR-L2 sequence containing the amino acid sequence SAS, and a CDR-L3 sequence containing the amino acid sequence QQYSTYPFT (SEQ ID NO: 22). In some embodiments, the antigen-binding site of the CD38 peptide comprises: an antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence containing the amino acid sequence GYAFTTYL (SEQ ID NO: 12), a CDR-H2 sequence containing the amino acid sequence INPGSGST (SEQ ID NO: 15), and a CDR-H3 sequence containing the amino acid sequence ARYAYGY (SEQ ID NO: 16); and an antibody light chain variable (VL) domain comprising a CDR-L1 sequence containing the amino acid sequence QNVGTA (SEQ ID NO: 19), a CDR-L2 sequence containing the amino acid sequence SAS, and a CDR-L3 sequence containing the amino acid sequence QQYSTYPFT (SEQ ID NO: 22).

[0092] In some embodiments, the antigen-binding site of the CD38 peptide comprises: an antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence containing the amino acid sequence GYSFTNYA (SEQ ID NO: 24), a CDR-H2 sequence containing the amino acid sequence ISPHYYGDT (SEQ ID NO: 25), and a CDR-H3 sequence containing the amino acid sequence ARFEGFYYSMDY (SEQ ID NO: 26); or an antibody light chain variable (VL) domain comprising a CDR-L1 sequence containing the amino acid sequence QSLVHSNGNTY (SEQ ID NO: 27), a CDR-L2 sequence containing the amino acid sequence KVS, and a CDR-L3 sequence containing the amino acid sequence SQSTHVPLT (SEQ ID NO: 29). In some embodiments, the antigen-binding site of the CD38 peptide comprises: an antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence containing the amino acid sequence GYSFTNYA (SEQ ID NO: 24), a CDR-H2 sequence containing the amino acid sequence ISPHYGDT (SEQ ID NO: 25), and a CDR-H3 sequence containing the amino acid sequence ARFEGFYYSMDY (SEQ ID NO: 26); and an antibody light chain variable (VL) domain comprising a CDR-L1 sequence containing the amino acid sequence QSLVHSNGNTY (SEQ ID NO: 27), a CDR-L2 sequence containing the amino acid sequence KVS, and a CDR-L3 sequence containing the amino acid sequence SQSTHVPLT (SEQ ID NO: 29).

[0093] In some embodiments, the antigen-binding site of the CD38 peptide comprises: an antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence containing the amino acid sequence GFTFSSYG (SEQ ID NO: 41), a CDR-H2 sequence containing the amino acid sequence IWYDGSNK (SEQ ID NO: 42), and a CDR-H3 sequence containing the amino acid sequence ARDPGLRYFDGGMDV (SEQ ID NO: 106); or an antibody light chain variable (VL) domain comprising a CDR-L1 sequence containing the amino acid sequence QGISSY (SEQ ID NO: 107), a CDR-L2 sequence containing the amino acid sequence AAS, and a CDR-L3 sequence containing the amino acid sequence QQLNSFPYT (SEQ ID NO: 229). In some embodiments, the antigen-binding site of the CD38 peptide comprises: an antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence containing the amino acid sequence GFTFSSYG (SEQ ID NO: 41), a CDR-H2 sequence containing the amino acid sequence IWYDGSNK (SEQ ID NO: 42), and a CDR-H3 sequence containing the amino acid sequence ARDPGLRYFDGGMDV (SEQ ID NO: 106); and an antibody light chain variable (VL) domain comprising a CDR-L1 sequence containing the amino acid sequence QGISSY (SEQ ID NO: 107), a CDR-L2 sequence containing the amino acid sequence AAS, and a CDR-L3 sequence containing the amino acid sequence QQLNSFPYT (SEQ ID NO: 229).

[0094] In some embodiments, the VH domain comprises a sequence from the N-terminus to the C-terminus: FR1—CDR-H1—FR2—CDR-H2—FR3—CDR-H3—FR4; wherein FR1 comprises the sequence QVQLVESGGGVVQPGRSLRLSCAAS (SEQ ID NO: 89); wherein FR2 comprises the sequence MHWVRQAPGKGLEWVAV (SEQ ID NO: 92); wherein FR3 comprises the sequence YYADSVKGRFTISGDNSKNTLYLQMNSLRAEDTAVYYC (SEQ ID NO: 95); and wherein FR4 comprises the sequence WGQGTLVTVSS (SEQ ID NO: 96). In some embodiments, the VL domain comprises a sequence from the N-terminus to the C-terminus: FR1—CDR-L1—FR2—CDR-L2—FR3—CDR-L3—FR4; wherein FR1 comprises the sequence AIQMTQSPSSLSASVGDRVTITCRAS (SEQ ID NO: 98); wherein FR2 comprises the sequence GWYQQKPGKAPKLLIY (SEQ ID NO: 100); wherein FR3 comprises the sequence SLQSGVPSRFSGSGSGTDFTLTISGLQPEDSATYYC (SEQ ID NO: 102); and wherein FR4 comprises the sequence WGQGTLVTVSS (SEQ ID NO: 104).

[0095] In some embodiments, the VH domain contains an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of SEQ ID NO: 108; and / or the VL domain contains an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of SEQ ID NO: 109.

[0096] In some embodiments, the VH domain contains an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of SEQ ID NO: 109; and the VL domain contains an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of SEQ ID NO: 109. In some embodiments, the VH domain contains the amino acid sequence of SEQ ID NO: 108; and the VL domain contains the amino acid sequence of SEQ ID NO: 109.

[0097] In some embodiments, the VH domain contains an amino acid sequence that is at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 110; and / or the VL domain contains an amino acid sequence that is at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 111.

[0098] In some embodiments, the VH domain contains an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of SEQ ID NO: 111; and the VL domain contains an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of SEQ ID NO: 111. In some embodiments, the VH domain contains the amino acid sequence of SEQ ID NO: 110; and the VL domain contains the amino acid sequence of SEQ ID NO: 111.

[0099] In some embodiments, the VH domain contains an amino acid sequence that is at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 116; and / or the VL domain contains an amino acid sequence that is at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 117.

[0100] In some embodiments, the VH domain contains an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of SEQ ID NO: 117; and the VL domain contains an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of SEQ ID NO: 117. In some embodiments, the VH domain contains the amino acid sequence of SEQ ID NO: 116; and the VL domain contains the amino acid sequence of SEQ ID NO: 117.

[0101] In some embodiments, the VH domain contains an amino acid sequence that is at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 112; and / or the VL domain contains an amino acid sequence that is at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 113.

[0102] In some embodiments, the VH domain contains an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of SEQ ID NO: 112; and the VL domain contains an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of SEQ ID NO: 113. In some embodiments, the VH domain contains the amino acid sequence of SEQ ID NO: 112; and the VL domain contains the amino acid sequence of SEQ ID NO: 113.

[0103] In some embodiments, the VH domain contains an amino acid sequence that is at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 118; and / or the VL domain contains an amino acid sequence that is at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 119.

[0104] In some embodiments, the VH domain contains an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of SEQ ID NO: 118; and the VL domain contains an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of SEQ ID NO: 119. In some embodiments, the VH domain contains the amino acid sequence of SEQ ID NO: 118; and the VL domain contains the amino acid sequence of SEQ ID NO: 119.

[0105] In some embodiments, the VH domain contains an amino acid sequence that is at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 114; and / or the VL domain contains an amino acid sequence that is at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 115.

[0106] In some embodiments, the VH domain contains an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of SEQ ID NO: 114; and the VL domain contains an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of SEQ ID NO: 115. In some embodiments, the VH domain contains the amino acid sequence of SEQ ID NO: 114; and the VL domain contains the amino acid sequence of SEQ ID NO: 115.

[0107] In some embodiments, the binding protein disclosed herein comprises 1, 2, 3, 4, 5, or 6 CDR sequences of the antibody sequences shown in Table A. In some embodiments, the binding protein disclosed herein comprises 1, 2, 3, 4, 5, or 6 CDR sequences of the antibody sequences shown in Table B, a VH domain sequence, and / or a VL domain sequence. In some embodiments, the binding protein disclosed herein comprises 1, 2, 3, 4, 5, or 6 CDR sequences of the antibody sequences shown in Table I1 or I2, a VH domain sequence, and / or a VL domain sequence. In some embodiments, the binding protein disclosed herein comprises 1, 2, 3, or 4 polypeptide sequences shown in Table I1 or I2. Table B. Variable domain sequences of anti-CD38 (mAb1-7) and other binding proteins. Note: Bold and underlined text in the above amino acid sequences represent CDR sequences.

[0108] In some embodiments, the binding protein disclosed herein comprises an antigen-binding site that binds to the extracellular domains of both the human CD38 peptide and the cynomolgus monkey CD38 peptide. Exemplary assays for determining whether an antigen-binding site binds to an antigen are described herein and are known in the art. In some embodiments, binding is determined by an ELISA assay, for example, as described below. In some embodiments, binding is determined by an SPR assay, for example, as described below. In some embodiments, binding is determined by flow cytometry using cells expressing the CD38 peptide on their cell surface, for example, as described below. See, for example, Examples 1, 3, and 4.

[0109] In some embodiments, the binding protein disclosed herein binds a purified polypeptide or fragment thereof containing the amino acid sequence SEQ ID NO: 1 and / or 30 (e.g., as measured by ELISA or SPR). In some embodiments, the binding protein disclosed herein binds the polypeptide when the polypeptide containing the amino acid sequence SEQ ID NO: 1 and / or 30 is expressed on the cell surface (e.g., as measured by flow cytometry).

[0110] In some embodiments, the binding protein disclosed herein binds CD38 isotype A polypeptide (e.g., a polypeptide containing the amino acid sequence of SEQ ID NO: 1). In some embodiments, the binding protein disclosed herein binds CD38 isotype E polypeptide (e.g., a polypeptide containing the amino acid sequence of SEQ ID NO: 105 and not containing the complete amino acid sequence of SEQ ID NO: 1, a polypeptide consisting of the amino acid sequence of SEQ ID NO: 105, or a polypeptide consisting substantially of the amino acid sequence of SEQ ID NO: 105). In some embodiments, the binding protein disclosed herein binds both CD38 isotype A polypeptide (e.g., a polypeptide containing the amino acid sequence of SEQ ID NO: 1) and CD38 isotype E polypeptide (e.g., a polypeptide containing the amino acid sequence of SEQ ID NO: 105 and not containing the complete amino acid sequence of SEQ ID NO: 1, a polypeptide consisting of the amino acid sequence of SEQ ID NO: 105, or a polypeptide consisting substantially of the amino acid sequence of SEQ ID NO: 105). Not wanting to be bound by theory, it is believed that binding to the CD38 isotype E peptide may be advantageous, for example, for targeting one or more cells expressing the CD38 isotype E peptide with the binding protein disclosed herein. Human CD38 isoform A extracellular domain polypeptide sequence RWRQQWSGPGTTKRFPETVLARCVKYTEIHPEMRHVDCQSVWDAFKGAFISKHPCNITEEDYQPLMKLGTQTVPCNKILLWSRIKDLAHQFTQVQRDMFTLEDTLLGYLADDLTWCGEFNTSKINYQSCP DWRKDCSNNPVSVFWKTVSRRFAEAACDVVHVMLNGSRSKIFDKNSTFGSVEVHNLQPEKVQTLEAWVIHGGREDSRDLCQDPTIKELESIISKRNIQFSCKNIYRPDKFLQCVKNPEDSSCTSEI (SEQ ID NO: 1) Human CD38 isoform E polypeptide sequence RWRQQWSGPGTTKRFPETVLARCVKYTEIHPEMRHVDCQSVWDAFKGAFISKHPCNITEEDYQPLMKLGTQTVPCNKILLWSRIKDLAHQFTQVQRDMFTLEDTLLGYLADDLTWCGEFNTSKINYQSCPDWRKDCSNNPVSVFWKTVSRRHFWECGSP (SEQ ID NO: 105)

[0111] In some embodiments, the extracellular domain of the human CD38 peptide comprises the amino acid sequence of SEQ ID NO: 1. In some embodiments, the extracellular domain of the cynomolgus monkey CD38 peptide comprises the amino acid sequence of SEQ ID NO: 30. Cynomolgus CD38 polypeptide sequence RWRQQWSGSGTTSRFPETVLARCVKYTEVHPEMRHVDCQSVWDAFKGAFISKYPCNITEEDYQPLVKLGTQTVPCNKTLLWSRIKDLAHQFTQVQRDMFTLEDMLLGYLADDLTWCGEFNTFEINYQSCP DWRKDCSNNPVSVFWKTVSRRFAETACGVVHVMLNGSRSKIFDKNSTFGSVEVHNLQPEKVQALEAWVIHGGREDSRDLCQDPTIKELESIISKRNIRFFCKNIYRPDKFLQCVKNPEDSSCLSGI (SEQ ID NO: 30) Anti-CD28 antigen binding site

[0112] Exemplary antigen-binding sites that specifically bind to the CD28 peptide and can be used in the anti-CD38 T-cell conjugate disclosed herein are described below. Any anti-CD28 antigen-binding site and / or trispecific binding protein described in International Application Nos. WO 2019 / 074973, WO 2017 / 180913, or WO 2020 / 210392 may be used in the methods, uses, kits, and compositions disclosed herein. In some embodiments, the CD28 peptide is a human CD28 peptide, also known as Tp44. Human CD28 peptides are known in the art and include, but are not limited to, peptides represented by NCBI accession numbers XP_011510499.1, XP_011510497.1, XP_011510496.1, NP_001230007.1, NP_001230006.1, or NP_006130.1, or peptides generated by NCBI gene ID 940.

[0113] In some embodiments, the antigen-binding site of the CD28 peptide comprises: an antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence containing the amino acid sequence GYTFTSYY (SEQ ID NO: 139), a CDR-H2 sequence containing the amino acid sequence IYPGNVNT (SEQ ID NO: 140), and a CDR-H3 sequence containing the amino acid sequence TRSHYGLDWNFDV (SEQ ID NO: 141); and / or an antibody light chain variable (VL) domain comprising a CDR-L1 sequence containing the amino acid sequence QNIYVW (SEQ ID NO: 142), a CDR-L2 sequence containing the amino acid sequence KAS, and a CDR-L3 sequence containing the amino acid sequence QQGQTYPY (SEQ ID NO: 144). In some embodiments, the antigen-binding site of the CD28 peptide comprises: an antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence containing the amino acid sequence GYTFTSYY (SEQ ID NO: 139), a CDR-H2 sequence containing the amino acid sequence IYPGNVNT (SEQ ID NO: 140), and a CDR-H3 sequence containing the amino acid sequence TRSHYGLDWNFDV (SEQ ID NO: 141); and an antibody light chain variable (VL) domain comprising a CDR-L1 sequence containing the amino acid sequence QNIYVW (SEQ ID NO: 142), a CDR-L2 sequence containing the amino acid sequence KAS, and a CDR-L3 sequence containing the amino acid sequence QQGQTYPY (SEQ ID NO: 144).

[0114] In some embodiments, the antigen-binding site of the CD28 peptide comprises: an antibody heavy chain variable (VH) domain containing an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of SEQ ID NO: 49; and / or an antibody light chain variable (VL) domain containing an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of SEQ ID NO: 50. In some embodiments, the antigen-binding site of the CD28 peptide comprises: an antibody heavy chain variable (VH) domain containing the amino acid sequence of SEQ ID NO: 49, and / or an antibody light chain variable (VL) domain containing the amino acid sequence of SEQ ID NO: 50. In some embodiments, the antigen-binding site of the CD28 peptide comprises: an antibody heavy chain variable (VH) domain containing the amino acid sequence of SEQ ID NO: 49, and an antibody light chain variable (VL) domain containing the amino acid sequence of SEQ ID NO: 50.

[0115] In some embodiments, the antigen-binding site of the CD28 peptide comprises: an antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence containing the amino acid sequence GFSLSDYG (SEQ ID NO: 212), a CDR-H2 sequence containing the amino acid sequence IWAGGGT (SEQ ID NO: 213), and a CDR-H3 sequence containing the amino acid sequence ARDKGYSYYYSMDY (SEQ ID NO: 214); and / or an antibody light chain variable (VL) domain comprising a CDR-L1 sequence containing the amino acid sequence ESVEYYVTSL (SEQ ID NO: 215), a CDR-L2 sequence containing the amino acid sequence AAS, and a CDR-L3 sequence containing the amino acid sequence QQSRKVPYT (SEQ ID NO: 217). In some embodiments, the antigen-binding site of the CD28 peptide comprises: an antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence containing the amino acid sequence GFSLSDYG (SEQ ID NO: 212), a CDR-H2 sequence containing the amino acid sequence IWAGGGT (SEQ ID NO: 213), and a CDR-H3 sequence containing the amino acid sequence ARDKGYSYYYSMDY (SEQ ID NO: 214); and an antibody light chain variable (VL) domain comprising a CDR-L1 sequence containing the amino acid sequence ESVEYYVTSL (SEQ ID NO: 215), a CDR-L2 sequence containing the amino acid sequence AAS, and a CDR-L3 sequence containing the amino acid sequence QQSRKVPYT (SEQ ID NO: 217).

[0116] In some embodiments, the antigen-binding site of the CD28 peptide comprises: an antibody heavy chain variable (VH) domain containing an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of SEQ ID NO: 51; and / or an antibody light chain variable (VL) domain containing an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of SEQ ID NO: 52. In some embodiments, the antigen-binding site of the CD28 peptide comprises: an antibody heavy chain variable (VH) domain containing the amino acid sequence of SEQ ID NO: 51, and / or an antibody light chain variable (VL) domain containing the amino acid sequence of SEQ ID NO: 52. In some embodiments, the antigen-binding site of the CD28 peptide comprises: an antibody heavy chain variable (VH) domain containing the amino acid sequence of SEQ ID NO: 51, and an antibody light chain variable (VL) domain containing the amino acid sequence of SEQ ID NO: 52.

[0117] In some embodiments, the binding protein disclosed herein comprises 1, 2, 3, 4, 5, or 6 CDR sequences of the antibody sequences shown in Table C. In some embodiments, the binding protein disclosed herein comprises 1, 2, 3, 4, 5, or 6 CDR sequences of the antibody sequences shown in Table C, a VH domain sequence, and / or a VL domain sequence. In some embodiments, the binding protein disclosed herein comprises 1, 2, 3, or 4 polypeptide sequences shown in Table C. Table C. Anti-CD28 antigen binding site sequence. anti-CD3 antigen binding site

[0118] Exemplary antigen-binding sites that specifically bind CD3 peptides and can be used in the anti-CD38 T-cell conjugates considered herein are described below. Any anti-CD3 antigen-binding site and / or trispecific binding protein described in International Application Nos. WO 2019 / 074973, WO 2017 / 180913, or WO 2020 / 210392 may be used in the methods, uses, kits, and compositions disclosed herein. In some embodiments, the CD3 peptide is a human CD3 peptide, including CD3-δ (also known as T3D, IMD19, and CD3-δ), CD3-ε (also known as T3E, IMD18, and TCRE), and CD3-γ (also known as T3G, IMD17, and CD3-γ). Human CD3 peptides are known in the art and include, but are not limited to, peptides represented by NCBI accession numbers XP_006510029.1 or NP_031674.1, or peptides generated by NCBI gene IDs 915, 916, or 917.

[0119] In some embodiments, the antigen-binding site of the CD3 peptide comprises: an antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence containing the amino acid sequence GFTFTKAW (SEQ ID NO: 120), a CDR-H2 sequence containing the amino acid sequence IKDKSNSYAT (SEQ ID NO: 121), and a CDR-H3 sequence containing the amino acid sequence RGVYYALSPFDY (SEQ ID NO: 122); and / or an antibody light chain variable (VL) domain comprising a CDR-L1 sequence containing the amino acid sequence QSLVHX1NX2X3TY (where X1 is E or Q, X2 is A or L, and X3 is Q, R, or F (SEQ ID NO: 131)), a CDR-L2 sequence containing the amino acid sequence KVS, and a CDR-L3 sequence containing the amino acid sequence GQGTQYPFT (SEQ ID NO: 130). In some embodiments, the antigen-binding site of the CD28 peptide comprises: an antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence containing the amino acid sequence GFTFTKAW (SEQ ID NO: 120), a CDR-H2 sequence containing the amino acid sequence IKDKSNSYAT (SEQ ID NO: 121), and a CDR-H3 sequence containing the amino acid sequence RGVYYALSPFDY (SEQ ID NO: 122); and an antibody light chain variable (VL) domain comprising a CDR-L1 sequence containing the amino acid sequence QSLVHX1NX2X3TY (where X1 is E or Q, X2 is A or L, and X3 is Q, R, or F (SEQ ID NO: 131)), a CDR-L2 sequence containing the amino acid sequence KVS, and a CDR-L3 sequence containing the amino acid sequence GQGTQYPFT (SEQ ID NO: 130). In some embodiments, the CDR-L1 sequence is selected from the group consisting of: QSLVHNNANTY (SEQ ID NO: 123), QSLVHQNAQTY (SEQ ID NO: 124), QSLVHENLQTY (SEQ ID NO: 125), QSLVHENLFTY (SEQ ID NO: 126), QSLVHENLRTY (SEQ ID NO: 127), and QSLVHDNAQTY (SEQ ID NO: 128).

[0120] In some embodiments, the antigen-binding site of the CD3 peptide comprises: an antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence containing the amino acid sequence GFTFTKAW (SEQ ID NO: 120), a CDR-H2 sequence containing the amino acid sequence IKDKSNSYAT (SEQ ID NO: 121), and a CDR-H3 sequence containing the amino acid sequence RGVYYALSPFDY (SEQ ID NO: 122); and / or an antibody light chain variable (VL) domain comprising a CDR-L1 sequence containing the amino acid sequence QSLVHNNGNTY (SEQ ID NO: 218), a CDR-L2 sequence containing the amino acid sequence KVS, and a CDR-L3 sequence containing the amino acid sequence GQGTQYPFT (SEQ ID NO: 130). In some embodiments, the antigen-binding site of the CD3 peptide comprises: an antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence containing the amino acid sequence GFTFTKAW (SEQ ID NO: 120), a CDR-H2 sequence containing the amino acid sequence IKDKSNSYAT (SEQ ID NO: 121), and a CDR-H3 sequence containing the amino acid sequence RGVYYALSPFDY (SEQ ID NO: 122); and an antibody light chain variable (VL) domain comprising a CDR-L1 sequence containing the amino acid sequence QSLVHNNGNTY (SEQ ID NO: 218), a CDR-L2 sequence containing the amino acid sequence KVS, and a CDR-L3 sequence containing the amino acid sequence GQGTQYPFT (SEQ ID NO: 130). In some embodiments, the antigen-binding site of the CD3 peptide comprises: an antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence containing the amino acid sequence GFTFTKAW (SEQ ID NO: 120), a CDR-H2 sequence containing the amino acid sequence IKDKSNSYAT (SEQ ID NO: 121), and a CDR-H3 sequence containing the amino acid sequence RGVYYALSPFDY (SEQ ID NO: 122); and / or an antibody light chain variable (VL) domain comprising a CDR-L1 sequence containing the amino acid sequence QSLVHNNANTY (SEQ ID NO: 123), a CDR-L2 sequence containing the amino acid sequence KVS, and a CDR-L3 sequence containing the amino acid sequence GQGTQYPFT (SEQ ID NO: 130).In some embodiments, the antigen-binding site of the CD3 peptide comprises: an antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence containing the amino acid sequence GFTFTKAW (SEQ ID NO: 120), a CDR-H2 sequence containing the amino acid sequence IKDKSNSYAT (SEQ ID NO: 121), and a CDR-H3 sequence containing the amino acid sequence RGVYYALSPFDY (SEQ ID NO: 122); and an antibody light chain variable (VL) domain comprising a CDR-L1 sequence containing the amino acid sequence QSLVHNNANTY (SEQ ID NO: 123), a CDR-L2 sequence containing the amino acid sequence KVS, and a CDR-L3 sequence containing the amino acid sequence GQGTQYPFT (SEQ ID NO: 130).

[0121] In some embodiments, the antigen-binding site of the CD3 polypeptide comprises: an antibody heavy chain variable (VH) domain containing an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of SEQ ID NO: 53 or 138; and / or an antibody light chain variable (VL) domain containing an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of SEQ ID NO: 54, 133, 134, 135, 136, or 137. In some embodiments, the antigen-binding site of the CD3 peptide comprises: an antibody heavy chain variable (VH) domain containing the amino acid sequence of SEQ ID NO: 53 or 138, and / or an antibody light chain variable (VL) domain containing the amino acid sequence of SEQ ID NO: 54, 133, 134, 135, 136, or 137. In some embodiments, the antigen-binding site of the CD3 peptide comprises: an antibody heavy chain variable (VH) domain containing the amino acid sequence of SEQ ID NO: 53 or 138, and an antibody light chain variable (VL) domain containing the amino acid sequence of SEQ ID NO: 54, 133, 134, 135, 136, or 137.

[0122] In some embodiments, the antigen-binding site of the CD3 polypeptide comprises: an antibody heavy chain variable (VH) domain containing an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of SEQ ID NO: 84; and / or an antibody light chain variable (VL) domain containing an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the amino acid sequence of SEQ ID NO: 85. In some embodiments, the antigen-binding site of the CD3 peptide comprises: an antibody heavy chain variable (VH) domain containing the amino acid sequence of SEQ ID NO: 84, and / or an antibody light chain variable (VL) domain containing the amino acid sequence of SEQ ID NO: 85. In some embodiments, the antigen-binding site of the CD3 peptide comprises: an antibody heavy chain variable (VH) domain containing the amino acid sequence of SEQ ID NO: 84, and an antibody light chain variable (VL) domain containing the amino acid sequence of SEQ ID NO: 85.

[0123] In some embodiments, the binding protein disclosed herein comprises 1, 2, 3, 4, 5, or 6 CDR sequences of the antibody sequences shown in Table D. In some embodiments, the binding protein disclosed herein comprises 1, 2, 3, 4, 5, or 6 CDR sequences of the antibody sequences shown in Table D, a VH domain sequence, and / or a VL domain sequence. In some embodiments, the binding protein disclosed herein comprises 1, 2, 3, or 4 polypeptide sequences shown in Table D. Table D. Sequence of anti-CD3 antigen binding sites. Trispecific binding protein formats

[0124] In some embodiments, the anti-CD38 T-cell conjugate disclosed herein is a trispecific binding protein comprising an antigen-binding site binding one or more CD38 peptides, a second antigen-binding site binding a CD28 peptide, and a third antigen-binding site binding a CD3 peptide. Any of the antigen-binding sites described above may be used in the anti-CD38 T-cell conjugate disclosed herein.

[0125] In some embodiments, the trispecific binding protein comprises four polypeptide chains forming three antigen-binding sites, wherein the first polypeptide chain comprises a structure represented by the following formula: V L2 -L1-V L1 -L2-C L [I] Furthermore, the second polypeptide chain contains a structure represented by the following formula: V H1 -L3-V H2 -L4-C H1 -Hinge-C H2 -C H3 [II] Furthermore, the third polypeptide chain contains a structure represented by the following formula: V H3 -C H1 -Hinge-C H2 -C H3 [III] Furthermore, the fourth polypeptide chain contains a structure represented by the following formula: V L3 -C L [IV] in: V L1 It is the variable domain of the first immunoglobulin light chain; V L2 It is the variable domain of the second immunoglobulin light chain; V L3 It is the variable domain of the third immunoglobulin light chain; V H1 It is the variable domain of the first immunoglobulin heavy chain; V H2 It is the variable domain of the second immunoglobulin heavy chain; V H3 It is the variable domain of the third immunoglobulin heavy chain; C L It is the constant structural domain of the immunoglobulin light chain; C H1 It is immunoglobulin C H1 Heavy chain constant structural domain; C H2It is immunoglobulin C H2 Heavy chain constant structural domain; C H3 It is immunoglobulin C H3 Heavy chain constant structural domain; The hinge is the connection of C H1 and C H2 The immunoglobulin hinge region of the structural domain; and L1, L2, L3, and L4 are amino acid linkers.

[0126] In some embodiments, a polypeptide having Formula I and a polypeptide having Formula II form a cross-linked light chain-heavy chain pair. In some embodiments, V H1 and V L1 Forms a binding pair and forms one of the three antigen-binding sites; V H2 and V L2 Forming a binding pair and forming another of the three antigen-binding sites; and V H3 and V L3 A binding pair is formed, forming another of the three antigen-binding sites. In some embodiments, the first and second polypeptide chains have a cross-orientation that forms two different antigen-binding sites. In some embodiments, VH1 and VL1 form a binding pair and form the first antigen-binding site. In some embodiments, VH2 and VL2 form a binding pair and form the second antigen-binding site. In some embodiments, the first antigen-binding site binds to a CD3 polypeptide (e.g., human CD3), and the second antigen-binding site binds to a CD28 polypeptide (e.g., human CD28). In some embodiments, the second antigen-binding site binds to a CD3 polypeptide (e.g., human CD3), and the first antigen-binding site binds to a CD28 polypeptide (e.g., human CD28). In some embodiments, the third and fourth polypeptides form the third antigen-binding site. In some embodiments, VH3 and VL3 form a binding pair and form the third antigen-binding site. In some embodiments, the third antigen-binding site binds to a CD38 polypeptide (e.g., human and optionally cynomolgus monkey CD38). Exemplary binding protein forms with cross-orientation considered for use herein are also described in U.S. Patent Application Serial No. 15 / 487,243 and International Application No. PCT / US2017 / 027488.

[0127] The three antigen-binding sites can be arranged in any combination, including an antigen-binding site binding to CD38 peptide, an antigen-binding site binding to CD28 peptide, and an antigen-binding site binding to CD3 peptide. In some embodiments, V H1 and V L1 Forms a second antigen-binding site for the CD28 polypeptide, V H2 and V L2Forming a third antigen-binding site for the CD3 polypeptide, and V H3 and V L3 Forming the first antigen-binding site for the CD38 polypeptide. In some embodiments, V H1 and V L1 Forms a third antigen-binding site for the CD3 polypeptide, V H2 and V L2 Forming a second antigen-binding site that binds to the CD28 peptide, and V H3 and V L3 Forming the first antigen-binding site for the CD38 polypeptide. In some embodiments, V H1 and V L1 Forms a second antigen-binding site for the CD28 polypeptide, V H2 and V L2 Forming the first antigen-binding site for the CD38 polypeptide, and V H3 and V L3 This forms a third antigen-binding site for the CD3 polypeptide. In some embodiments, V H1 and V L1 Forms a third antigen-binding site for the CD3 polypeptide, V H2 and V L2 Forming the first antigen-binding site for the CD38 polypeptide, and V H3 and V L3 A second antigen-binding site is formed for binding to the CD28 polypeptide. In some embodiments, V H1 and V L1 Forms the first antigen-binding site for the CD38 polypeptide, V H2 and V L2 Forming a third antigen-binding site for the CD3 polypeptide, and V H3 and V L3 A second antigen-binding site is formed for binding to the CD28 polypeptide. In some embodiments, V H1 and V L1 Forms the first antigen-binding site for the CD38 polypeptide, V H2 and V L2 Forming a second antigen-binding site that binds to the CD28 peptide, and V H3 and V L3 It forms a third antigen-binding site that binds to the CD3 polypeptide. Linkers

[0128] In some embodiments, the lengths of linkers L1, L2, L3, and L4 range from zero amino acids (length = 0) to approximately 100 amino acids, or less than 100, 50, 40, 30, 20, or 15 amino acids. The linker lengths can also be 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid. L1, L2, L3, and L4 in a single binding protein may all have the same amino acid sequence or may all have different amino acid sequences.

[0129] Examples of suitable linkers include monoglycine (Gly) residues; diglycine peptides (Gly-Gly); tripeptides (Gly-Gly-Gly); peptides with four glycine residues; peptides with five glycine residues; peptides with six glycine residues; peptides with seven glycine residues; and peptides with eight glycine residues. Other combinations of amino acid residues may be used, such as peptides GGGGSGGGGS (SEQ ID NO: 55), GGGGSGGGGSGGGGS (SEQ ID NO: 56), TKGPS (SEQ ID NO: 57), GQPKAAP (SEQ ID NO: 58), and GGSGSSGSGG (SEQ ID NO: 59). The examples listed above are not intended to limit the scope of this disclosure in any way, and it has been shown that linkers comprising randomly selected amino acids from the group consisting of: valine, leucine, isoleucine, serine, threonine, lysine, arginine, histidine, aspartic acid, glutamic acid, asparagine, glutamine, glycine, and proline. For further description of linker sequences, see, for example, WO 2012135345 and International Application No. PCT / US 2017 / 027488.

[0130] The identity and sequence of amino acid residues in the linker can vary depending on the type of secondary structural element desired to be achieved within the linker. For example, glycine, serine, and alanine are best suited for linkers offering the greatest flexibility. If a more rigid and extensible linker is required, some combinations of glycine, proline, threonine, and serine are useful. Any amino acid residue can be considered as a linker for combination with other amino acid residues to construct larger peptide linkers according to desired properties.

[0131] In some embodiments, at least one of L1, L2, L3, or L4 has an independent length of 0 amino acids. In some embodiments, each of L1, L2, L3, or L4 has an independent length of at least one amino acid. In some embodiments, the length of L1 is at least twice the length of L3. In some embodiments, the length of L2 is at least twice the length of L4. In some embodiments, the length of L1 is at least twice the length of L3, and the length of L2 is at least twice the length of L4. In some embodiments, the length of L1 is 3 to 12 amino acid residues, the length of L2 is 3 to 14 amino acid residues, the length of L3 is 1 to 8 amino acid residues, and the length of L4 is 1 to 3 amino acid residues. In some embodiments, the length of L1 is 5 to 10 amino acid residues, the length of L2 is 5 to 8 amino acid residues, the length of L3 is 1 to 5 amino acid residues, and the length of L4 is 1 to 2 amino acid residues. In some embodiments, the length of L1 is 7 amino acid residues, the length of L2 is 5 amino acid residues, the length of L3 is 1 amino acid residue, and the length of L4 is 2 amino acid residues. In some embodiments, L1 has a length of 10 amino acid residues, L2 has a length of 10 amino acid residues, L3 has a length of 0 amino acid residues, and L4 has a length of 0 amino acid residues. In some embodiments, L1, L2, L3, and L4 each have an independent length selected from 0 to 15 amino acids (e.g., 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 amino acids), wherein at least two of the linkers have a length of 1 to 15 amino acids (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 amino acids). In some embodiments, L1, L2, L3, and L4 each have a length of 0 amino acids.

[0132] In some embodiments, L1, L2, L3, and / or L4 comprise sequences derived from naturally occurring sequences at the junction between the antibody variable domain and the antibody constant domain (e.g., as described in WO 2012 / 135345). For example, in some embodiments, the adapter comprises endogenous V H With C H1 Between structural domains, or endogenous V L With C L A sequence present at the transition between structural domains (e.g., κ or λ). In some embodiments, the connector is contained in endogenous human V H With C H1 Between structural domains, or endogenous human V L With C L Sequences that exist at the transition between structural domains (e.g., human κ or λ).

[0133] In some embodiments, L1, L2, L3, and L4 each independently comprises a sequence of zero amino acids or comprises a sequence selected from the group consisting of: GGGGSGGGGS (SEQ ID NO: 55), GGGGSGGGGSGGGGS (SEQ ID NO: 56), S, RT, TKGPS (SEQ ID NO: 57), GQPKAAP (SEQ ID NO: 58), and GGSGSSGSGG (SEQ ID NO: 59). In some embodiments, L1, L2, L3, and L4 each independently comprises a sequence selected from the group consisting of: GGGGSGGGGS (SEQ ID NO: 55), GGGGSGGGGSGGGGS (SEQ ID NO: 56), S, RT, TKGPS (SEQ ID NO: 57), GQPKAAP (SEQ ID NO: 58), and GGSGSSGSGG (SEQ ID NO: 59).

[0134] In some embodiments, L1 comprises the sequence GQPKAAP (SEQ ID NO: 58), L2 comprises the sequence TKGPS (SEQ ID NO: 57), L3 comprises the sequence S, and L4 comprises the sequence RT. In some embodiments, L1 comprises the sequence GGGGSGGGGS (SEQ ID NO: 55), L2 comprises the sequence GGGGSGGGGS (SEQ ID NO: 55), L3 is 0 amino acids long, and L4 is 0 amino acids long. In some embodiments, L1 comprises the sequence GGSGSSGSGG (SEQ ID NO: 59), L2 comprises the sequence GGSGSSGSGG (SEQ ID NO: 59), L3 is 0 amino acids long, and L4 is 0 amino acids long. In some embodiments, L1 comprises the sequence GGGGSGGGGSGGGGS (SEQ ID NO: 56), L2 is 0 amino acids long, L3 comprises the sequence GGGGSGGGGSGGGGS (SEQ ID NO: 56), and L4 is 0 amino acids long.

[0135] In some embodiments, at least one of L1, L2, L3, or L4 contains the sequence DKTHT (SEQ ID NO: 147). In some embodiments, L1, L2, L3, and L4 contain the sequence DKTHT (SEQ ID NO: 147). Fc regions and constant domains

[0136] In some embodiments, the anti-CD38 T cell binder or binding protein disclosed herein comprises a full-length antibody heavy chain or a polypeptide chain containing an Fc region. In some embodiments, the Fc region is a human Fc region, such as the human IgG1, IgG2, IgG3, or IgG4 Fc region. In some embodiments, the Fc region includes an antibody hinge, C... H1 C H2 C H3 and optional location C H4 Domain. In some embodiments, the Fc region is the human IgG1 Fc region. In some embodiments, the Fc region is the human IgG4 Fc region. In some embodiments, the Fc region includes one or more of the mutations described below.

[0137] In some embodiments, the anti-CD38 T cell binder or binding protein disclosed herein comprises one or two Fc variants. As used herein, the term "Fc variant" refers to a molecule or sequence modified from a natural Fc but still containing a binding site for the rescue remedy FcRn (neonatal Fc receptor). Exemplary Fc variants and their interactions with the remedy receptor are known in the art. Thus, the term "Fc variant" may comprise a molecule or sequence humanized from a non-human natural Fc. Furthermore, natural Fc contains regions that can be removed because they provide structural features or biological activities not required by the antibody-like binding protein of the present invention. Therefore, the term “Fc variant” includes molecules or sequences that lack one or more native Fc sites or residues, or in which one or more Fc sites or residues have been modified, which affect or participate in: (1) disulfide bond formation, (2) incompatibility with selected host cells, (3) N-terminal heterogeneity when expressed in selected host cells, (4) glycosylation, (5) interaction with complement, (6) binding to Fc receptors other than rescue receptors, or (7) antibody-dependent cytotoxicity (ADCC).

[0138] In some embodiments, the Fc region contains one or more mutations that reduce or eliminate Fc receptor binding and / or effector cell function in the Fc region (e.g., Fc receptor-mediated antibody-dependent phagocytosis (ADCP), complement-dependent cytotoxicity (CDC), and / or antibody-dependent cytotoxicity (ADCC)).

[0139] In some embodiments, the Fc region is a human IgG1 Fc region containing one or more amino acid substitutions at positions 234, 235, and / or 329 of human IgG1 according to the EU index. In some embodiments, the amino acid substitutions are L234A, L235A, and / or P329A. In some embodiments, the Fc region is a human IgG1 Fc region containing amino acid substitutions at positions 298, 299, and / or 300 of human IgG1 according to the EU index. In some embodiments, the amino acid substitutions are S298N, T299A, and / or Y300S.

[0140] In some embodiments, the Fc region is a human IgG4 Fc region containing one or more mutations that reduce or eliminate FcγI and / or FcγII binding. In some embodiments, the Fc region is a human IgG4 Fc region containing one or more mutations that reduce or eliminate FcγI and / or FcγII binding but do not affect FcRn binding. In some embodiments, the Fc region is a human IgG4 Fc region containing amino acid substitutions at positions corresponding to positions 228 and / or 409 of human IgG4 according to the EU index. In some embodiments, the amino acid substitutions are S228P and / or R409K. In some embodiments, the Fc region is a human IgG4 Fc region containing amino acid substitutions at positions corresponding to positions 234 and / or 235 of human IgG4 according to the EU index. In some embodiments, the amino acid substitutions are F234A and / or L235A. In some embodiments, the Fc region is a human IgG4 Fc region containing amino acid substitutions at positions corresponding to positions 228, 234, 235, and / or 409 of human IgG4 according to the EU index. In some embodiments, the amino acid substitutions are S228P, F234A, L235A, and / or R409K. In some embodiments, the Fc region is a human IgG4 Fc region containing amino acid substitutions at positions 233-236 of human IgG4 according to the EU index. In some embodiments, the amino acid substitutions are E233P, F234V, L235A, and a deletion at position 236. In some embodiments, the Fc region is a human IgG4 Fc region containing amino acid substitutions at positions 228, 233-236, and / or 409 of human IgG4 according to the EU index. In some embodiments, the amino acid mutations are S228P; E233P, F234V, L235A, and a deletion at position 236; and / or R409K.

[0141] In some embodiments, the anti-CD38 T cell binder or binding protein disclosed herein contains one or more mutations to improve purification, for example, by modulating affinity for purification reagents. For example, it is known that if one of the two Fc regions of the heterodimeric form contains one or more mutations that reduce or eliminate binding to protein A, the heterodimeric binding protein can be selectively purified from its homodimeric form because the heterodimeric form will have moderate affinity for protein A-based purification compared to either homodimeric form and can be selectively eluted from protein A, for example, by using different pH (see, for example, Smith, EJ et al. (2015) Sci. Rep. [Scientific Reports] 5:17943). In some embodiments, the mutation contains substitutions at positions 435 and 436 corresponding to human IgG1 or IgG4 according to the EU index, wherein these amino acid substitutions are H435R and Y436F. In some embodiments, the binding protein comprises a second polypeptide chain and a third polypeptide chain, the second polypeptide chain further containing a linker to C... H1 The first Fc region, which includes the immunoglobulin hinge region and C H2 and C H3 The immunoglobulin heavy chain constant domain, the third polypeptide chain further contains a linker to C. H1 The second Fc region, which includes the immunoglobulin hinge region and C... H2 and C H3 An immunoglobulin heavy chain constant domain; and wherein only one of the first Fc region and the second Fc region contains an amino acid substitution at positions 435 and 436 corresponding to human IgG1 or IgG4 according to the EU index, wherein these amino acid substitutions are H435R and Y436F. In some embodiments, the binding protein disclosed herein comprises club and mortar mutations and one or more mutations to improve purification. In some embodiments, the first and / or second Fc region is a human IgG1 Fc region. In some embodiments, the first and / or second Fc region is a human IgG4 Fc region.

[0142] To improve the yield of certain binding proteins (e.g., bispecific or trispecific binding proteins), the C-type structure can be modified using the "mortar and pestle" technique. H3 The structural domain, this technique is described in detail with several examples in, for example, International Publication No. WO 96 / 027011, Ridgway et al., 1996, Protein Eng. 9: 617-21; and Merchant et al., 1998, Nat. Biotechnol. 16: 677-81. Specifically, changing the two C... H3 The interaction surface of the structural domains increases the content of these two Cs.H3 Heterodimerization of the two heavy chains of the structural domain. (The two C's of these two heavy chains) H3 Each of the domains can be a "pestle" and the other a "mortar." The introduction of disulfide bonds further stabilizes the heterodimer (Merchant et al., 1998; Atwell et al., 1997, J. Mol. Biol. [Journal of Molecular Biology] 270: 26-35) and improves yield. In a particular embodiment, the pestle is located on the second pair of polypeptides having a single variable domain. In other embodiments, the pestle is located on the first pair of polypeptides having a cross-orientation. In other embodiments, C H3 The structural domain does not include the pestle in the mortar.

[0143] In some embodiments, the anti-CD38 T cell binder or binding protein disclosed herein (e.g., a trispecific binding protein) comprises a "pump" mutation on a second polypeptide chain and a "mortar" mutation on a third polypeptide chain. In some embodiments, the binding protein disclosed herein comprises a "pump" mutation on a third polypeptide chain and a "mortar" mutation on a second polypeptide chain. In some embodiments, the "pump" mutation comprises one or more substitutions at positions corresponding to positions 354 and / or 366 of human IgG1 or IgG4 according to the EU index. In some embodiments, the amino acid substitutions are S354C, T366W, T366Y, S354C and T366W, or S354C and T366Y. In some embodiments, the "pump" mutation comprises substitutions at positions corresponding to positions 354 and 366 of human IgG1 or IgG4 according to the EU index. In some embodiments, the amino acid substitutions are S354C and T366W. In some embodiments, the "mortar" mutation comprises one or more substitutions at positions corresponding to positions 407 of human IgG1 or IgG4 according to the EU index and optionally positions 349, 366, and / or 368. In some embodiments, the amino acid substitution is Y407V or Y407T and optionally Y349C, T366S, and / or L368A. In some embodiments, the "mortis" mutation comprises substitutions at positions 349, 366, 368, and 407 corresponding to human IgG1 or IgG4 according to the EU index. In some embodiments, the amino acid substitution is Y349C, T366S, L368A, and Y407V.

[0144] In some embodiments, the second polypeptide chain further includes a first Fc region linked to CH1, the first Fc region comprising an immunoglobulin hinge region and CH2 and CH3 immunoglobulin heavy chain constant domains, wherein the first Fc region contains one or more amino acid substitutions at positions 366 and optionally 354 corresponding to human IgG1 or IgG4 according to the EU index, wherein these amino acid substitutions are T366W or T366Y and optionally S354C; and wherein the third polypeptide chain further includes a second Fc region linked to CH1, the second Fc region comprising an immunoglobulin hinge region and CH2 and CH3 immunoglobulin heavy chain constant domains, wherein the second Fc region contains one or more amino acid substitutions at positions 407 and optionally 349, 366 and / or 368 corresponding to human IgG1 or IgG4 according to the EU index, wherein these amino acid substitutions are Y407V or Y407T and optionally Y349C, T366S and / or L368A.

[0145] In some embodiments, the second polypeptide chain further includes a first Fc region linked to CH1, the first Fc region comprising an immunoglobulin hinge region and CH2 and CH3 immunoglobulin heavy chain constant domains, wherein the first Fc region contains one or more amino acid substitutions at positions 407 and optionally 349, 366 and / or 368 corresponding to human IgG1 or IgG4 according to the EU index, wherein these amino acid substitutions are Y407V or Y407T and optionally Y349C, T366S and / or L368A; and wherein the third polypeptide chain further includes a second Fc region linked to CH1, the second Fc region comprising an immunoglobulin hinge region and CH2 and CH3 immunoglobulin heavy chain constant domains, wherein the second Fc region contains one or more amino acid substitutions at positions 366 and optionally 354 corresponding to human IgG1 or IgG4 according to the EU index, wherein these amino acid substitutions are T366W or T366Y and optionally S354C.

[0146] In some embodiments, the second polypeptide chain further includes a first Fc region linked to CH1, the first Fc region including an immunoglobulin hinge region and CH2 and CH3 immunoglobulin heavy chain constant structural domains, wherein the first Fc region includes an amino acid substitution at position 366 corresponding to human IgG1 or IgG4 according to EU index, wherein the amino acid substitution is T366W; and wherein the third polypeptide chain further includes a second Fc region linked to CH1, the second Fc region including an immunoglobulin hinge region and CH2 and CH3 immunoglobulin heavy chain constant structural domains, wherein the second Fc region includes one or more amino acid substitutions at positions 366, 368 and / or 407 corresponding to human IgG1 or IgG4 according to EU index, wherein these amino acid substitutions are T366S, L368A and / or Y407V.

[0147] In some embodiments, the second polypeptide chain further includes a first Fc region linked to CH1, the first Fc region including an immunoglobulin hinge region and CH2 and CH3 immunoglobulin heavy chain constant domains, wherein the first Fc region includes one or more amino acid substitutions at positions 366, 368 and / or 407 corresponding to human IgG1 or IgG4 according to the EU index, wherein these amino acid substitutions are T366S, L368A and / or Y407V; and wherein the third polypeptide chain further includes a second Fc region linked to CH1, the second Fc region including an immunoglobulin hinge region and CH2 and CH3 immunoglobulin heavy chain constant domains, wherein the second Fc region includes an amino acid substitution at position 366 corresponding to human IgG1 or IgG4 according to the EU index, wherein the amino acid substitution is T366W.

[0148] In some embodiments, the second polypeptide chain further includes a first Fc region linked to CH1, the first Fc region including an immunoglobulin hinge region and CH2 and CH3 immunoglobulin heavy chain constant domains, wherein the first Fc region includes amino acid substitutions at positions 354 and 366 corresponding to human IgG1 or IgG4 according to the EU index, wherein these amino acid substitutions are S354C and T366W; and wherein the third polypeptide chain further includes a second Fc region linked to CH1, the second Fc region including an immunoglobulin hinge region and CH2 and CH3 immunoglobulin heavy chain constant domains, wherein the second Fc region includes amino acid substitutions at positions 349, 366, 368 and 407 corresponding to human IgG1 or IgG4 according to the EU index, wherein these amino acid substitutions are Y349C, T366S, L368A and Y407V. In some embodiments, the second polypeptide chain further includes a first Fc region linked to CH1, the first Fc region comprising an immunoglobulin hinge region and CH2 and CH3 immunoglobulin heavy chain constant domains, wherein the first Fc region contains amino acid substitutions at positions 349, 366, 368, and 407 corresponding to human IgG1 or IgG4 according to the EU index, wherein these amino acid substitutions are Y349C, T366S, L368A, and Y407V; and wherein the third polypeptide chain further includes a second Fc region linked to CH1, the second Fc region comprising an immunoglobulin hinge region and CH2 and CH3 immunoglobulin heavy chain constant domains, wherein the second Fc region contains amino acid substitutions at positions 354 and 366 corresponding to human IgG1 or IgG4 according to the EU index, wherein these amino acid substitutions are S354C and T366W. In some embodiments, the first and / or second Fc regions are human IgG1 Fc regions. In some embodiments, the first and / or second Fc regions are human IgG4 Fc regions.

[0149] In some embodiments, the second polypeptide chain further includes a first Fc region linked to CH1, wherein the first Fc region is a human IgG4 Fc region comprising an immunoglobulin hinge region and CH2 and CH3 immunoglobulin heavy chain constant domains, wherein the first Fc region comprises amino acid substitutions at positions 228, 354, 366, and 409 corresponding to human IgG4 according to the EU index, wherein these amino acid substitutions are S228P, S354C, T366W, and R409K; and wherein the third polypeptide chain further includes a second Fc region linked to CH1, wherein the second Fc region is a human IgG4 Fc region comprising an immunoglobulin hinge region and CH2 and CH3 immunoglobulin heavy chain constant domains. The second Fc region contains amino acid substitutions at positions 228, 349, 366, 368, 407, and 409 corresponding to human IgG4 according to the EU index, wherein these amino acid substitutions are S228P, Y349C, T366S, L368A, Y407V, and R409K. In some embodiments, the second polypeptide chain further includes a first Fc region linked to CH1, wherein the first Fc region is a human IgG4 Fc region comprising an immunoglobulin hinge region and CH2 and CH3 immunoglobulin heavy chain constant domains, wherein the first Fc region contains amino acid substitutions at positions 228, 349, 366, 368, 407, and 409 corresponding to human IgG4 according to the EU index, wherein these amino acid substitutions are S228P, Y349C, T366S, L368A, Y407V, and R409K; and wherein the third polypeptide chain further includes a second Fc region linked to CH1, wherein the second Fc region is a human IgG4 Fc region comprising an immunoglobulin hinge region and CH2 and CH3 immunoglobulin heavy chain constant domains. The second Fc region contains amino acid substitutions at positions 228, 354, 366, and 409 corresponding to human IgG4 according to the EU index, wherein these amino acid substitutions are S228P, S354C, T366W, and R409K.

[0150] In some embodiments, the second polypeptide chain further includes a first Fc region linked to CH1, wherein the first Fc region is a human IgG4 Fc region comprising an immunoglobulin hinge region and CH2 and CH3 immunoglobulin heavy chain constant domains, wherein the first Fc region comprises amino acid substitutions at positions 234, 235, 354, and 366 corresponding to human IgG4 according to the EU index, wherein these amino acid substitutions are F234A, L235A, S354C, and T366W; and wherein the third polypeptide chain further includes a second Fc region linked to CH1, wherein the second Fc region is a human IgG4 Fc region comprising an immunoglobulin hinge region and CH2 and CH3 immunoglobulin heavy chain constant domains. The second Fc region contains amino acid substitutions at positions 234, 235, 349, 366, 368, and 407 corresponding to human IgG4 according to the EU index, wherein these amino acid substitutions are F234A, L235A, Y349C, T366S, L368A, and Y407V. In some embodiments, the second polypeptide chain further includes a first Fc region linked to CH1, wherein the first Fc region is a human IgG4 Fc region comprising an immunoglobulin hinge region and CH2 and CH3 immunoglobulin heavy chain constant domains, wherein the first Fc region comprises amino acid substitutions at positions 234, 235, 349, 366, 368, and 407 of human IgG4 according to the EU index, wherein these amino acid substitutions are F234A, L235A, Y349C, T366S, L368A, and Y407V; and wherein the third polypeptide chain further includes a second Fc region linked to CH1, wherein the second Fc region is a human IgG4 Fc region comprising an immunoglobulin hinge region and CH2 and CH3 immunoglobulin heavy chain constant domains. The second Fc region contains amino acid substitutions at positions 234, 235, 354, and 366 corresponding to human IgG4 according to the EU index, wherein these amino acid substitutions are F234A, L235A, S354C, and T366W.

[0151] In some embodiments, the second polypeptide chain further includes a first Fc region linked to CH1, wherein the first Fc region is a human IgG4 Fc region comprising an immunoglobulin hinge region and CH2 and CH3 immunoglobulin heavy chain constant domains, wherein the first Fc region comprises amino acid substitutions at positions 228, 234, 235, 354, 366, and 409 corresponding to human IgG4 according to the EU index, wherein these amino acid substitutions are S228P, F234A, L235A, S354C, T366W, and R409K; and wherein the third polypeptide chain further includes a second Fc region linked to CH1, wherein the second Fc region is a human IgG4 Fc region comprising an immunoglobulin hinge region and CH2 and CH3 immunoglobulin heavy chain constant domains. The second Fc region contains amino acid substitutions at positions 228, 234, 235, 349, 366, 368, 407, and 409 corresponding to human IgG4 according to the EU index, wherein these amino acid substitutions are S228P, F234A, L235A, Y349C, T366S, L368A, Y407V, and R409K. In some embodiments, the second polypeptide chain further includes a first Fc region linked to CH1, wherein the first Fc region is a human IgG4 Fc region comprising an immunoglobulin hinge region and CH2 and CH3 immunoglobulin heavy chain constant domains, wherein the first Fc region contains amino acid substitutions at positions 228, 234, 235, 349, 366, 368, 407, and 409 corresponding to human IgG4 according to the EU index, wherein these amino acid substitutions are S228P, F234A, L235A, Y349C, T366S, L368A, Y407V, and R409K; and wherein the third polypeptide chain further includes a second Fc region linked to CH1, wherein the second Fc region is a human IgG4 Fc region comprising an immunoglobulin hinge region and CH2 and CH3 immunoglobulin heavy chain constant domains. The second Fc region contains amino acid substitutions at positions 228, 234, 235, 354, 366, and 409 corresponding to human IgG4 according to the EU index, wherein these amino acid substitutions are S228P, F234A, L235A, S354C, T366W, and R409K.

[0152] In some embodiments, the anti-CD38 T cell binder or binding protein disclosed herein contains one or more mutations to improve serum half-life (see, for example, Hinton, PR et al. (2006) J. Immunol. [Journal of Immunology] 176(1):346-56). In some embodiments, the mutation contains substitutions at positions 428 and 434 corresponding to human IgG1 or IgG4 according to the EU index, wherein these amino acid substitutions are M428L and N434S. In some embodiments, the binding protein comprises a second polypeptide chain and a third polypeptide chain, the second polypeptide chain further comprising a first Fc region linked to CH1, the first Fc region comprising an immunoglobulin hinge region and CH2 and CH3 immunoglobulin heavy chain constant domains, and the third polypeptide chain further comprising a second Fc region linked to CH1, the second Fc region comprising an immunoglobulin hinge region and CH2 and CH3 immunoglobulin heavy chain constant domains, wherein the first and / or second Fc regions contain amino acid substitutions at positions 428 and 434 corresponding to human IgG1 or IgG4 according to the EU index, wherein these amino acid substitutions are M428L and N434S. In some embodiments, the binding protein disclosed herein comprises club and mortar mutations and one or more mutations that improve serum half-life. In some embodiments, the first and / or second Fc regions are human IgG1 Fc regions. In some embodiments, the first and / or second Fc regions are human IgG4 Fc regions.

[0153] In some embodiments, the anti-CD38 T cell binder or binding protein disclosed herein comprises one or more mutations to improve the stability of the hinge region and / or dimer interface of, for example, IgG4 (see, for example, Spiess, C. et al. (2013) J. Biol. Chem. 288:26583-26593). In some embodiments, the mutation comprises substitutions at positions 228 and 409 corresponding to human IgG4 according to the EU index, wherein the amino acid substitutions are S228P and R409K. In some embodiments, the binding protein comprises a second polypeptide chain and a third polypeptide chain, the second polypeptide chain further comprising a linker to C H1 The first Fc region, which includes the immunoglobulin hinge region and C H2 and C H3 The immunoglobulin heavy chain constant domain, the third polypeptide chain further contains a linker to C. H1 The second Fc region, which includes the immunoglobulin hinge region and C... H2 and C H3An immunoglobulin heavy chain constant domain; wherein the first Fc region and the second Fc region are human IgG4 Fc regions; and wherein each of the first Fc region and the second Fc region contains amino acid substitutions at positions 228 and 409 corresponding to human IgG4 according to the EU index, wherein these amino acid substitutions are S228P and R409K. In some embodiments, the binding protein disclosed herein comprises club and mortar mutations and one or more mutations that improve stability. In some embodiments, the first and / or second Fc regions are human IgG4 Fc regions.

[0154] In some embodiments, the anti-CD38 T cell binder or binding protein disclosed herein contains one or more mutations to improve purification, for example, by modulating affinity for purification reagents. For example, it is known that if one of the two Fc regions of the heterodimeric form contains one or more mutations that reduce or eliminate binding to protein A, the heterodimeric binding protein can be selectively purified from its homodimeric form because the heterodimeric form will have moderate affinity for protein A-based purification compared to either homodimeric form and can be selectively eluted from protein A, for example, by using different pH (see, for example, Smith, EJ et al. (2015) Sci. Rep. [Scientific Reports] 5:17943). In some embodiments, the mutation contains substitutions at positions 435 and 436 corresponding to human IgG1 or IgG4 according to the EU index, wherein these amino acid substitutions are H435R and Y436F. In some embodiments, the binding protein comprises a second polypeptide chain and a third polypeptide chain, the second polypeptide chain further containing a linker to C... H1 The first Fc region, which includes the immunoglobulin hinge region and C H2 and C H3 The immunoglobulin heavy chain constant domain, the third polypeptide chain further contains a linker to C. H1 The second Fc region, which includes the immunoglobulin hinge region and C... H2 and C H3 An immunoglobulin heavy chain constant domain; and wherein only one of the first Fc region and the second Fc region contains an amino acid substitution at positions 435 and 436 corresponding to human IgG1 or IgG4 according to the EU index, wherein these amino acid substitutions are H435R and Y436F. In some embodiments, the binding protein disclosed herein comprises club and mortar mutations and one or more mutations to improve purification. In some embodiments, the first and / or second Fc region is a human IgG1 Fc region. In some embodiments, the first and / or second Fc region is a human IgG4 Fc region.

[0155] In some embodiments, the anti-CD38 T cell binder or binding protein disclosed herein contains one or more mutations to improve serum half-life (see, for example, Hinton, PR et al. (2006) J. Immunol. [Journal of Immunology] 176(1):346-56). In some embodiments, the mutation contains substitutions at positions 428 and 434 corresponding to human IgG1 or IgG4 according to the EU index, wherein these amino acid substitutions are M428L and N434S. In some embodiments, the binding protein comprises a second polypeptide chain and a third polypeptide chain, the second polypeptide chain further comprising a first Fc region linked to CH1, the first Fc region comprising an immunoglobulin hinge region and CH2 and CH3 immunoglobulin heavy chain constant domains, and the third polypeptide chain further comprising a second Fc region linked to CH1, the second Fc region comprising an immunoglobulin hinge region and CH2 and CH3 immunoglobulin heavy chain constant domains, wherein the first and / or second Fc regions contain amino acid substitutions at positions 428 and 434 corresponding to human IgG1 or IgG4 according to the EU index, wherein these amino acid substitutions are M428L and N434S. In some embodiments, the binding protein disclosed herein comprises club and mortar mutations and one or more mutations that improve serum half-life. In some embodiments, the first and / or second Fc regions are human IgG1 Fc regions. In some embodiments, the first and / or second Fc regions are human IgG4 Fc regions.

[0156] In some embodiments, the anti-CD38 T cell binder or binding protein disclosed herein contains one or more mutations to reduce effector cell function, such as Fc receptor-mediated antibody-dependent phagocytosis (ADCP), complement-dependent cytotoxicity (CDC), and / or antibody-dependent cytotoxicity (ADCC). In some embodiments, the second polypeptide chain further comprises a linker to C H1 The first Fc region, which includes the immunoglobulin hinge region and C H2 and C H3 Immunoglobulin heavy chain constant domain; wherein the third polypeptide chain further comprises a linker to C H1 The second Fc region, which includes the immunoglobulin hinge region and C... H2 and C H3An immunoglobulin heavy chain constant domain; wherein the first Fc region and the second Fc region are human IgG1 Fc regions; and wherein each of the first Fc region and the second Fc region contains an amino acid substitution at positions 234 and 235 corresponding to human IgG1 according to the EU index, wherein these amino acid substitutions are L234A and L235A. In some embodiments, the Fc regions of the second polypeptide chain and the third polypeptide chain are human IgG1 Fc regions, and wherein each of these Fc regions contains an amino acid substitution at positions 234 and 235 corresponding to human IgG1 according to the EU index, wherein these amino acid substitutions are L234A and L235A. In some embodiments, the second polypeptide chain further comprises a linker to C H1 The first Fc region, which includes the immunoglobulin hinge region and C H2 and C H3 Immunoglobulin heavy chain constant domain; wherein the third polypeptide chain further comprises a linker to C H1 The second Fc region, which includes the immunoglobulin hinge region and C... H2 and C H3 An immunoglobulin heavy chain constant domain; wherein the first Fc region and the second Fc region are human IgG1 Fc regions; and wherein each of the first Fc region and the second Fc region contains amino acid substitutions at positions 234, 235, and 329 corresponding to human IgG1 according to the EU index, wherein these amino acid substitutions are L234A, L235A, and P329A. In some embodiments, the Fc regions of the second polypeptide chain and the third polypeptide chain are human IgG1 Fc regions, and wherein each of these Fc regions contains amino acid substitutions at positions 234, 235, and 329 corresponding to human IgG1 according to the EU index, wherein these amino acid substitutions are L234A, L235A, and P329A. In some embodiments, the Fc regions of the second polypeptide chain and the third polypeptide chain are human IgG4 Fc regions, and each of these Fc regions contains amino acid substitutions at positions 234 and 235 corresponding to human IgG4 according to the EU index, wherein these amino acid substitutions are F234A and L235A. In some embodiments, the binding protein comprises a second polypeptide chain and a third polypeptide chain, the second polypeptide chain further containing a linker to C. H1 The first Fc region, which includes the immunoglobulin hinge region and C H2 and C H3 The immunoglobulin heavy chain constant domain, the third polypeptide chain further contains a linker to C. H1 The second Fc region, which includes the immunoglobulin hinge region and C... H2 and C H3An immunoglobulin heavy chain constant domain; and wherein the first Fc region and the second Fc region each contain amino acid substitutions at positions 234 and 235 corresponding to human IgG4 according to the EU index, wherein these amino acid substitutions are F234A and L235A.

[0157] In some embodiments, the anti-CD38 T cell binder or binding protein disclosed herein comprises a club-and-mortar mutation and one or more mutations that reduce effector cell function. In some embodiments, the first and / or second Fc region is a human IgG1 Fc region. In some embodiments, the first and / or second Fc region is a human IgG4 Fc region. For a further description of the Fc mutation at position 329, see, for example, Shields, RL et al. (2001) J. Biol. Chem. 276:6591-6604 and WO 1999051642.

[0158] In some embodiments, the mutation types described above may be combined in any order or combination. For example, the binding protein disclosed herein may comprise two or more "pestle" and "mortar" mutations, one or more mutations that improve serum half-life, one or more mutations that improve IgG4 stability, one or more mutations that improve purification, and / or one or more mutations that reduce the effector cell function described above.

[0159] In some embodiments, the anti-CD38 T cell binder or binding protein disclosed herein comprises antibody fragments, including but not limited to antibody F(ab), F(ab')2, Fab'-SH, Fv, or scFv fragments.

[0160] This document provides exemplary full-chain polypeptide sequences of trispecific binding proteins and is intended for use in any methods, uses, kits, and compositions disclosed herein. In some embodiments, a first polypeptide chain comprises a polypeptide sequence having at least 95% identity with the amino acid sequence of SEQ ID NO: 61, a second polypeptide chain comprises a polypeptide sequence having at least 95% identity with the amino acid sequence of SEQ ID NO: 60, a third polypeptide chain comprises a polypeptide sequence having at least 95% identity with the amino acid sequence of SEQ ID NO: 62, and a fourth polypeptide chain comprises a polypeptide sequence having at least 95% identity with the amino acid sequence of SEQ ID NO: 63. In some embodiments, a first polypeptide chain comprises a polypeptide sequence having at least 95% identity with the amino acid sequence of SEQ ID NO: 61, a second polypeptide chain comprises a polypeptide sequence having at least 95% identity with the amino acid sequence of SEQ ID NO: 64, a third polypeptide chain comprises a polypeptide sequence having at least 95% identity with the amino acid sequence of SEQ ID NO: 65, and a fourth polypeptide chain comprises a polypeptide sequence having at least 95% identity with the amino acid sequence of SEQ ID NO: 63. In some embodiments, the first polypeptide chain comprises a polypeptide sequence having at least 95% identity with the amino acid sequence of SEQ ID NO: 61, the second polypeptide chain comprises a polypeptide sequence having at least 95% identity with the amino acid sequence of SEQ ID NO: 66, the third polypeptide chain comprises a polypeptide sequence having at least 95% identity with the amino acid sequence of SEQ ID NO: 67, and the fourth polypeptide chain comprises a polypeptide sequence having at least 95% identity with the amino acid sequence of SEQ ID NO: 63. In some embodiments, the first polypeptide chain comprises a polypeptide sequence having at least 95% identity with the amino acid sequence of SEQ ID NO: 61, the second polypeptide chain comprises a polypeptide sequence having at least 95% identity with the amino acid sequence of SEQ ID NO: 60, the third polypeptide chain comprises a polypeptide sequence having at least 95% identity with the amino acid sequence of SEQ ID NO: 68, and the fourth polypeptide chain comprises a polypeptide sequence having at least 95% identity with the amino acid sequence of SEQ ID NO: 69. In some embodiments, the first polypeptide chain comprises a polypeptide sequence having at least 95% identity with the amino acid sequence of SEQ ID NO: 61, the second polypeptide chain comprises a polypeptide sequence having at least 95% identity with the amino acid sequence of SEQ ID NO: 64, the third polypeptide chain comprises a polypeptide sequence having at least 95% identity with the amino acid sequence of SEQ ID NO: 70, and the fourth polypeptide chain comprises a polypeptide sequence having at least 95% identity with the amino acid sequence of SEQ ID NO: 69.In some embodiments, the first polypeptide chain comprises a polypeptide sequence having at least 95% identity with the amino acid sequence of SEQ ID NO: 61, the second polypeptide chain comprises a polypeptide sequence having at least 95% identity with the amino acid sequence of SEQ ID NO: 66, the third polypeptide chain comprises a polypeptide sequence having at least 95% identity with the amino acid sequence of SEQ ID NO: 71, and the fourth polypeptide chain comprises a polypeptide sequence having at least 95% identity with the amino acid sequence of SEQ ID NO: 69. In some embodiments, the first polypeptide chain comprises a polypeptide sequence having at least 95% identity with the amino acid sequence of SEQ ID NO: 148, the second polypeptide chain comprises a polypeptide sequence having at least 95% identity with the amino acid sequence of SEQ ID NO: 149, the third polypeptide chain comprises a polypeptide sequence having at least 95% identity with the amino acid sequence of SEQ ID NO: 150, and the fourth polypeptide chain comprises a polypeptide sequence having at least 95% identity with the amino acid sequence of SEQ ID NO: 151. In some embodiments, the first polypeptide chain comprises a polypeptide sequence having at least 95% identity with the amino acid sequence of SEQ ID NO: 152, the second polypeptide chain comprises a polypeptide sequence having at least 95% identity with the amino acid sequence of SEQ ID NO: 153, the third polypeptide chain comprises a polypeptide sequence having at least 95% identity with the amino acid sequence of SEQ ID NO: 154, and the fourth polypeptide chain comprises a polypeptide sequence having at least 95% identity with the amino acid sequence of SEQ ID NO: 155. In some embodiments, the first polypeptide chain comprises a polypeptide sequence having at least 95% identity with the amino acid sequence of SEQ ID NO: 156, the second polypeptide chain comprises a polypeptide sequence having at least 95% identity with the amino acid sequence of SEQ ID NO: 157, the third polypeptide chain comprises a polypeptide sequence having at least 95% identity with the amino acid sequence of SEQ ID NO: 158, and the fourth polypeptide chain comprises a polypeptide sequence having at least 95% identity with the amino acid sequence of SEQ ID NO: 159. In some embodiments, the first polypeptide chain comprises a polypeptide sequence having at least 95% identity with the amino acid sequence of SEQ ID NO: 160, the second polypeptide chain comprises a polypeptide sequence having at least 95% identity with the amino acid sequence of SEQ ID NO: 161, the third polypeptide chain comprises a polypeptide sequence having at least 95% identity with the amino acid sequence of SEQ ID NO: 162, and the fourth polypeptide chain comprises a polypeptide sequence having at least 95% identity with the amino acid sequence of SEQ ID NO: 163.In some embodiments, the first polypeptide chain comprises a polypeptide sequence having at least 95% identity with the amino acid sequence of SEQ ID NO: 164, the second polypeptide chain comprises a polypeptide sequence having at least 95% identity with the amino acid sequence of SEQ ID NO: 165, the third polypeptide chain comprises a polypeptide sequence having at least 95% identity with the amino acid sequence of SEQ ID NO: 166, and the fourth polypeptide chain comprises a polypeptide sequence having at least 95% identity with the amino acid sequence of SEQ ID NO: 167. In some embodiments, the first polypeptide chain comprises a polypeptide sequence having at least 95% identity with the amino acid sequence of SEQ ID NO: 168, the second polypeptide chain comprises a polypeptide sequence having at least 95% identity with the amino acid sequence of SEQ ID NO: 169, the third polypeptide chain comprises a polypeptide sequence having at least 95% identity with the amino acid sequence of SEQ ID NO: 170, and the fourth polypeptide chain comprises a polypeptide sequence having at least 95% identity with the amino acid sequence of SEQ ID NO: 171. In some embodiments, the first polypeptide chain comprises a polypeptide sequence having at least 95% identity with the amino acid sequence of SEQ ID NO: 172, the second polypeptide chain comprises a polypeptide sequence having at least 95% identity with the amino acid sequence of SEQ ID NO: 173, the third polypeptide chain comprises a polypeptide sequence having at least 95% identity with the amino acid sequence of SEQ ID NO: 174, and the fourth polypeptide chain comprises a polypeptide sequence having at least 95% identity with the amino acid sequence of SEQ ID NO: 175. In some embodiments, the first polypeptide chain comprises a polypeptide sequence having at least 95% identity with the amino acid sequence of SEQ ID NO: 176, the second polypeptide chain comprises a polypeptide sequence having at least 95% identity with the amino acid sequence of SEQ ID NO: 177, the third polypeptide chain comprises a polypeptide sequence having at least 95% identity with the amino acid sequence of SEQ ID NO: 178, and the fourth polypeptide chain comprises a polypeptide sequence having at least 95% identity with the amino acid sequence of SEQ ID NO: 179.

[0161] In some embodiments, the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 61, the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 60, the third polypeptide chain comprises the amino acid sequence of SEQ ID NO: 62, and the fourth polypeptide chain comprises the amino acid sequence of SEQ ID NO: 63. In some embodiments, the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 61, the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 64, the third polypeptide chain comprises the amino acid sequence of SEQ ID NO: 65, and the fourth polypeptide chain comprises the amino acid sequence of SEQ ID NO: 63. In some embodiments, the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 61, the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 66, the third polypeptide chain comprises the amino acid sequence of SEQ ID NO: 67, and the fourth polypeptide chain comprises the amino acid sequence of SEQ ID NO: 63. In some embodiments, the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 61, the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 60, the third polypeptide chain comprises the amino acid sequence of SEQ ID NO: 68, and the fourth polypeptide chain comprises the amino acid sequence of SEQ ID NO: 69. In some embodiments, the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 61, the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 64, the third polypeptide chain comprises the amino acid sequence of SEQ ID NO: 70, and the fourth polypeptide chain comprises the amino acid sequence of SEQ ID NO: 69. In some embodiments, the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 61, the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 66, the third polypeptide chain comprises the amino acid sequence of SEQ ID NO: 71, and the fourth polypeptide chain comprises the amino acid sequence of SEQ ID NO: 69. In some embodiments, the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 148, the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 149, the third polypeptide chain comprises the amino acid sequence of SEQ ID NO: 150, and the fourth polypeptide chain comprises the amino acid sequence of SEQ ID NO: 151. In some embodiments, the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 152, the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 153, the third polypeptide chain comprises the amino acid sequence of SEQ ID NO: 154, and the fourth polypeptide chain comprises the amino acid sequence of SEQ ID NO: 155.In some embodiments, the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 156, the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 157, the third polypeptide chain comprises the amino acid sequence of SEQ ID NO: 158, and the fourth polypeptide chain comprises the amino acid sequence of SEQ ID NO: 159. In some embodiments, the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 160, the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 161, the third polypeptide chain comprises the amino acid sequence of SEQ ID NO: 162, and the fourth polypeptide chain comprises the amino acid sequence of SEQ ID NO: 163. In some embodiments, the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 164, the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 165, the third polypeptide chain comprises the amino acid sequence of SEQ ID NO: 166, and the fourth polypeptide chain comprises the amino acid sequence of SEQ ID NO: 167. In some embodiments, the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 168, the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 169, the third polypeptide chain comprises the amino acid sequence of SEQ ID NO: 170, and the fourth polypeptide chain comprises the amino acid sequence of SEQ ID NO: 171. In some embodiments, the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 172, the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 173, the third polypeptide chain comprises the amino acid sequence of SEQ ID NO: 174, and the fourth polypeptide chain comprises the amino acid sequence of SEQ ID NO: 175. In some embodiments, the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 176, the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 177, the third polypeptide chain comprises the amino acid sequence of SEQ ID NO: 178, and the fourth polypeptide chain comprises the amino acid sequence of SEQ ID NO: 179.

[0162] In some embodiments, a trispecific binding protein comprises 1, 2, 3, or 4 polypeptide chains as shown in Table E1 or E2. In some embodiments, a trispecific binding protein comprises 1, 2, 3, or 4 polypeptide chains of a single trispecific binding protein as shown in Table E1 or E2. In some embodiments, a trispecific binding protein comprises 1, 2, 3, or 4 polypeptide chains encoded by a polynucleotide sequence as shown in Table F1 or F2. In some embodiments, a trispecific binding protein comprises 1, 2, 3, or 4 polypeptide chains encoded by one or more polynucleotide sequences of a single trispecific binding protein as shown in Table F1 or F2. Table E1. Full-length sequence of the three specific binding proteins. Table F1. Full-length polynucleotide sequence of the binding protein.

[0163] Standard recombinant DNA methods are used to construct polynucleotides encoding polypeptides that form anti-CD38 T cell binders, these polynucleotides are incorporated into recombinant expression vectors, and such vectors are introduced into host cells. See, for example, Sambrook et al., 2001, *Molecular Cloning: A Laboratory Manual* (Cold Spring Harbor Laboratory Press, 3rd edition). Enzymatic reactions and purification techniques can be performed according to the manufacturer's instructions as commonly practiced in the art or as described herein. Unless specifically defined, the nomenclature, laboratory procedures, and techniques used in relation to analytical chemistry, synthetic organic chemistry, and medical and medicinal chemistry as described herein are those well-known and commonly used in the art. Similarly, conventional techniques are used in chemical synthesis, chemical analysis, drug preparation, formulation, delivery, and patient treatment.

[0164] Certain aspects of this disclosure relate to polynucleotide kits. In some embodiments, one or more polynucleotides are vectors (e.g., expression vectors). These kits are particularly useful for generating one or more of the anti-CD38 T cell binders described herein. For example, kits disclosed herein may contain one or more polynucleotides encoding 1, 2, 3, or 4 polypeptides of an anti-CD38 T cell binder. In some embodiments, kits disclosed herein contain one or more polynucleotides encoding 1, 2, 3, or 4 polypeptide chains of a single trispecific binding protein (e.g., as shown in Tables F1 or F2).

[0165] In some embodiments, the isolated nucleic acid is operatively linked to a heterologous promoter to direct transcription of the nucleic acid sequence encoding the binding protein. The promoter can refer to a nucleic acid control sequence that directs nucleic acid transcription. The first nucleic acid sequence is operatively linked to the second nucleic acid sequence when the first nucleic acid sequence is positioned to have a functional relationship with the second nucleic acid sequence. For example, if the promoter affects the transcription or expression of a coding sequence, the promoter is operatively linked to the coding sequence of the binding protein. Examples of promoters may include, but are not limited to, promoters derived from the genomes of viruses (such as polyomavirus, fowlpox virus, adenovirus (such as adenovirus 2), bovine papillomavirus, avian sarcoma virus, cytomegalovirus, retrovirus, hepatitis B virus, simian virus 40 (SV40), etc.), heterologous eukaryotic promoters (such as actin promoter, immunoglobulin promoter, heat shock promoter, etc.), CAG promoter (Niwa et al., Gene [Gene] 108(2):193-9, 1991), phosphoglycerate kinase (PGK) promoter, tetracycline-inducible promoter (Masui et al., Nucleic Acids Res. [Nucleic Acid Research] 33:e43, 2005), lac system, trp system, tac system, trc system, major operon and promoter regions of bacteriophage λ, promoter of 3-phosphoglycerate kinase, promoter of yeast acid phosphatase, and promoter of yeast α-mating factor. The polynucleotide encoding the binding protein disclosed herein may be under the control of a constitutive promoter, an inducible promoter, or any other suitable promoter described herein or other suitable promoters that will be readily identified by a person skilled in the art.

[0166] In some embodiments, isolated nucleic acids are incorporated into a vector. In some embodiments, the vector is an expression vector. The expression vector may include one or more regulatory sequences operatively linked to the polynucleotide to be expressed. The term "regulatory sequence" includes promoters, enhancers, and other expression control elements (e.g., polyadenylation signals). Examples of suitable enhancers may include, but are not limited to, enhancer sequences from mammalian genes (such as globulins, elastases, albumins, alpha-fetoproteins, insulin, etc.) and enhancer sequences from eukaryotic viruses (such as the SV40 enhancer (bp 100-270) post-OMI, the cytomegalovirus early promoter enhancer, the polytumor enhancer post-OMI, the adenovirus enhancer, etc.). Examples of suitable vectors may include, for example, plasmids, sticky plasmids, episomes, transposons, and viral vectors (e.g., adenovirus, vaccinia virus, Sinderby virus, measles, herpesvirus, lentivirus, retrovirus, adeno-associated virus vector, etc.). The expression vector can be used to transfect host cells, such as bacterial cells, yeast cells, insect cells, and mammalian cells. Biologically functional viruses and plasmid DNA vectors capable of being expressed and replicated in a host are known in the art and can be used to transfect any target cell.

[0167] Other aspects of this disclosure relate to a vector system comprising one or more vectors encoding first, second, third, and fourth polypeptide chains of any anti-CD38 T cell binder described herein. In some embodiments, the vector system comprises a first vector encoding a first polypeptide chain of a binding protein, a second vector encoding a second polypeptide chain of a binding protein, a third vector encoding a third polypeptide chain of a binding protein, and a fourth vector encoding a fourth polypeptide chain of a binding protein. In some embodiments, the vector system comprises a first vector encoding first and second polypeptide chains of a binding protein, and a second vector encoding third and fourth polypeptide chains of a binding protein. In some embodiments, the vector system comprises a first vector encoding first and third polypeptide chains of a binding protein, and a second vector encoding second and fourth polypeptide chains of a binding protein. In some embodiments, the vector system comprises a first vector encoding first and fourth polypeptide chains of a binding protein, and a second vector encoding second and third polypeptide chains of a binding protein. In some embodiments, the vector system comprises a first vector encoding a first, second, third, and fourth polypeptide chain of a binding protein. The one or more vectors of the vector system may be any vector described herein. In some embodiments, the one or more vectors are expression vectors.

[0168] Other aspects of this disclosure relate to isolated host cells comprising one or more isolated polynucleotides, polynucleotide kits, vectors, and / or vector systems described herein. In some embodiments, the host cell is a bacterial cell (e.g., *Escherichia coli* (E. coli) cells). In some embodiments, the host cell is a yeast cell (e.g., *Saccharomyces cerevisiae* cells). In some embodiments, the host cell is an insect cell. Examples of insect host cells may include, for example, fruit fly cells (e.g., S2 cells), fall armyworm cells (e.g., High Five™ cells), and fall armyworm cells (e.g., Sf21 or Sf9 cells). In some embodiments, the host cell is a mammalian cell. Examples of mammalian host cells may include, for example, human embryonic kidney cells (e.g., 293 cells or subclones used for growth in suspension cultures), Expi293TM cells, CHO cells, juvenile hamster kidney cells (e.g., BHK, ATCC CCL 10), mouse supporting cells (e.g., TM4 cells), monkey kidney cells (e.g., CV1 ATCC CCL 70), African green monkey kidney cells (e.g., VERO-76, ATCC CRL-1587), human cervical cancer cells (e.g., HELA, ATCC CCL 2), canine kidney cells (e.g., MDCK, ATCC CCL34), Buffalo rat hepatocytes (e.g., BRL 3A, ATCC CRL 1442), human lung cells (e.g., W138, ATCC CCL 75), human hepatocytes (e.g., Hep G2, HB 8065), mouse mammary tumor cells (e.g., MMT 060562, ATCC CCL51), TRI cells, MRC cells, etc. 5 cells, FS4 cells, human hepatocellular carcinoma lineages (e.g., Hep G2) and myeloma cells (e.g., NS0 and Sp2 / 0 cells).

[0169] Other aspects of this disclosure relate to methods for generating any of the anti-CD38 T cell conjugates described herein. In some embodiments, the method includes a) culturing host cells (e.g., any host cells described herein) under conditions that cause host cells to express the anti-CD38 T cell conjugate, the host cells comprising isolated nucleic acids, vectors, and / or vector systems (e.g., any isolated nucleic acids, vectors, and / or vector systems described herein); and b) isolating the anti-CD38 T cell conjugate from the host cells. Methods of culturing host cells under conditions of protein expression are well known to those skilled in the art. Methods of isolating proteins from cultured host cells are well known to those skilled in the art, including, for example, by affinity chromatography (e.g., two-step affinity chromatography, including protein A affinity chromatography followed by size exclusion chromatography).

[0170] In some embodiments, the anti-CD38 T cell binder disclosed herein is purified by protein A affinity chromatography, κ light chain affinity chromatography (e.g., using Kappa Select resin according to the manufacturer's instructions; GE Healthcare), and optionally λ light chain affinity chromatography (e.g., using Lambda Fab Select resin according to the manufacturer's instructions; GE Healthcare). In some embodiments, the binding protein disclosed herein is purified by protein A affinity chromatography, λ light chain affinity chromatography (e.g., using Lambda Fab Select resin according to the manufacturer's instructions; GE Healthcare), and optionally κ light chain affinity chromatography (e.g., using Kappa Select resin according to the manufacturer's instructions; GE Healthcare). In some embodiments, the binding protein comprises two Fc regions, each containing C H3 A structural domain, and only one C H3 The domain contains amino acid substitutions at positions 435 and 436 corresponding to human IgG1 or IgG4 according to the EU index, wherein these amino acid substitutions are H435R and Y436F. In some embodiments, the binding protein disclosed herein is purified sequentially by protein A affinity chromatography, then by κ light chain affinity chromatography (e.g., using Kappa Select resin according to the manufacturer's instructions; General Health Medical Group), and then optionally by λ light chain affinity chromatography (e.g., using Lambda Fab Select resin according to the manufacturer's instructions; General Health Medical Group). In some embodiments, the binding protein disclosed herein is purified sequentially by protein A affinity chromatography, then by λ light chain affinity chromatography (e.g., using Lambda Fab Select resin according to the manufacturer's instructions; General Health Medical Group), and then optionally by κ light chain affinity chromatography (e.g., using Kappa Select resin according to the manufacturer's instructions; General Health Medical Group). For example, in some embodiments, the binding protein is contacted with protein A, and the binding protein is then subjected to a process suitable for reacting the binding protein with 0 or 2 C-terminals containing amino acid substitutions H435R and Y436F. H3 The binding protein was eluted from protein A under conditions suitable for domain-binding protein separation, contacted with a κ light chain affinity medium (e.g., as used in KappaSelect resin; GE Healthcare), and then subjected to a process suitable for binding the protein to only λC. LElution of the domain-binding protein from the κ light chain affinity medium is performed under conditions suitable for protein A elution (e.g., according to the manufacturer's instructions). Suitable conditions for protein A elution are known in the art, including but not limited to stepwise elution gradients from pH 4.5–2.8. In some embodiments, protein A or a variant of protein A suitable for protein purification is used. In some embodiments, protein A is attached to a substrate or resin, for example, as part of the chromatographic medium. In some embodiments, after elution from the κ light chain affinity medium, the binding protein is contacted with a λ light chain affinity medium (e.g., as used in LambdaFabSelect resin; General Health Medical Group), and the binding protein is subjected to conditions suitable for elution with only κ C. L The binding protein is eluted from a λ light chain affinity medium under conditions of separation (e.g., according to the manufacturer's instructions). In some embodiments, the binding protein disclosed herein is detected using HIC chromatography. In some embodiments, the binding protein comprises: containing λ C L The first polypeptide chain of the structural domain; the C of the second polypeptide chain H3 The domain contains amino acid substitutions at positions 354 and 366 corresponding to human IgG1 or IgG4 according to the EU index, wherein these amino acid substitutions are S354C and T366W; the C of the third polypeptide chain H3 The domain contains amino acid substitutions at positions 349, 366, 368, 407, 435, and 436 corresponding to human IgG1 or IgG4 according to the EU index, wherein these amino acid substitutions are Y349C, T366S, L368A, Y407V, H435R, and Y436F; and contains κ C L The fourth polypeptide chain of the domain. In some embodiments, the binding protein is produced by the host cell. In some embodiments, the binding protein is purified from cell culture medium or host cell extract. In some embodiments, the binding protein is secreted by the host cell or produced and extracted from the host cell (e.g., before contact with protein A). In some embodiments, the binding protein is in cell culture medium or host cell extract when contacted with protein A. In some embodiments, the binding protein is purified from other binding proteins, peptides, and / or other cellular components. III. Therapeutic Compositions, Kits, and Their Application

[0171] In some embodiments, this document provides an anti-CD38 T-cell binder of this disclosure for use in any of the methods described herein. In some embodiments, these methods include administering an effective amount of the anti-CD38 T-cell binder to an individual in need (e.g., an individual who has or has been diagnosed with PTCL) to treat PTCL.

[0172] In some embodiments, the anti-CD38 T-cell binder disclosed herein is used to manufacture a medicament for treating an individual with PTCL, for example, according to any of the methods described herein.

[0173] In some embodiments, a kit or article of manufacture is provided. In some embodiments, the kit comprises the anti-CD38 T cell binder disclosed herein, and optionally, instructions for administering the T cell binder to an individual, for example, according to any of the methods described herein.

[0174] In some embodiments, a pharmaceutical composition is provided. In some embodiments, the pharmaceutical composition comprises an effective amount of the anti-CD38 T cell binder disclosed herein and a pharmaceutically acceptable carrier. In some embodiments, the composition is intended for use according to any method disclosed herein.

[0175] Therapeutic or pharmaceutical compositions containing anti-CD38 T-cell binders or binding proteins are within the scope of this disclosure. Such therapeutic or pharmaceutical compositions may comprise a therapeutically effective amount of a T-cell binder or binding protein, or a mixture of a T-cell binder / binding protein-drug conjugate and a pharmaceutically or physiologically acceptable formulation of a choice suitable for the mode of administration.

[0176] The acceptable formulation material is preferably non-toxic to the recipient at the dosage and concentration used.

[0177] Pharmaceutical compositions may contain formulation materials used to alter, maintain, or preserve, for example, the composition's pH, molar osmolar concentration, viscosity, clarity, color, isotonicity, odor, sterility, stability, dissolution or release rate, adsorption, or permeation. Suitable formulation materials include, but are not limited to, amino acids (e.g., glycine, glutamine, asparagine, arginine, or lysine), antimicrobial agents, antioxidants (e.g., ascorbic acid, sodium sulfite, or sodium bisulfite), buffers (e.g., borates, bicarbonates, Tris-HCl, citrates, phosphates, or other organic acids), fillers (e.g., mannitol or glycine), chelating agents (e.g., ethylenediaminetetraacetic acid (EDTA)), complexing agents (e.g., caffeine, polyvinylpyrrolidone, β-cyclodextrin, or hydroxypropyl-β-cyclodextrin), fillers, monosaccharides, disaccharides, and other carbohydrates (e.g., glucose, mannose, or dextrin), proteins (e.g., serum albumin, gelatin, or immunoglobulins), colorants, flavorings and diluents, emulsifiers, and hydrophilic agents. Polymers (e.g., polyvinylpyrrolidone), low molecular weight peptides, counterions that form salts (e.g., sodium), preservatives (e.g., benzalkonium chloride, benzoic acid, salicylic acid, thimerosal, phenethyl alcohol, methylparaben, propylparaben, chlorhexidine, sorbic acid, or hydrogen peroxide), solvents (e.g., glycerol, propylene glycol, or polyethylene glycol), sugar alcohols (e.g., mannitol or sorbitol), suspending agents, surfactants or wetting agents (e.g., pronicotinic acid; PEG; dehydrated sorbitol esters; polysorbates, such as polysorbate 20 or polysorbate 80; triton; tromethamine; lecithin; cholesterol or tyloxapal), stability enhancers (e.g., sucrose or sorbitol), tensile enhancers (e.g., alkali metal halides— Preferably sodium chloride or potassium chloride—or mannitol or sorbitol), delivery medium, diluent, excipient, and / or pharmaceutical adjuvant (see, for example, Remington's Pharmaceutical Sciences (18th edition, edited by AR Gennaro, Mack Publishing Company, 1990) and subsequent editions, which are incorporated herein by reference for any purpose).

[0178] The optimal pharmaceutical composition will be determined by a technician based on factors such as the intended route of administration, delivery method, and desired dosage. Such a composition can affect the physical state, stability, in vivo release rate, and in vivo clearance rate of the binding protein.

[0179] The primary medium or carrier in a pharmaceutical composition can be aqueous or non-aqueous in nature. For example, a suitable medium or carrier for injection may be water, physiological saline solution, or artificial cerebrospinal fluid, possibly supplemented with other materials commonly found in compositions intended for parenteral administration. Neutral buffered saline or saline mixed with serum albumin are other exemplary mediators. Other exemplary pharmaceutical compositions comprise a Tris buffer of about pH 7.0-8.5 or an acetate buffer of about pH 4.0-5.5, which may further comprise sorbitol or a suitable substitute. In one embodiment of this disclosure, the binding protein composition can be prepared for storage by mixing a selected composition having the desired purity with an optional formulation in the form of a lyophilized cake or aqueous solution. Furthermore, the binding protein can be formulated as a lyophilized product using a suitable excipient, such as sucrose.

[0180] The pharmaceutical compositions disclosed herein may be selected for parenteral or subcutaneous delivery. Alternatively, the compositions may be selected for inhalation or delivery via the digestive tract (e.g., oral administration). The preparation of such pharmaceutically acceptable compositions is within the scope of the art.

[0181] The formulation components are present at concentrations acceptable for the application site. For example, buffer solutions are used to maintain the composition at physiological pH or slightly lower, typically in the pH range of about 5 to about 8.

[0182] When considering parenteral administration, the therapeutic composition used can be in the form of a pyrogen-free, parenterally acceptable aqueous solution containing the desired binding protein in a pharmaceutically acceptable mediator. A particularly suitable mediator for parenteral injection is sterile distilled water, in which the binding protein is formulated as a sterile, isotonic solution and properly stored. Another preparation may involve formulating the desired molecule with an agent that provides controlled or sustained release of the product (such as injectable microspheres, biodegradable particles, polymeric compounds such as polylactic acid or polyglycolic acid, beads, or liposomes), which can then be delivered via accumulation injection. Hyaluronic acid may also be used, which can contribute to increased circulation duration. Other suitable means for introducing the desired molecule include implantable drug delivery devices.

[0183] In one embodiment, the pharmaceutical composition may be formulated for inhalation. For example, the binding protein may be formulated as a dry powder for inhalation. A binding protein inhalation solution may also be formulated with a propellant for aerosol delivery. In another embodiment, the solution may be nebulized.

[0184] It is also anticipated that some formulations can be administered orally. In one embodiment of this disclosure, the binding protein administered in this manner may be formulated with or without those carriers typically used in mixtures of solid dosage forms, such as tablets and capsules. For example, capsules may be designed to release the active portion of the formulation at a point in the gastrointestinal tract at which bioavailability is maximized and pre-systemic degradation is minimized. Additional agents may be included to promote the absorption of the binding protein. Diluents, flavoring agents, low-melting-point waxes, vegetable oils, lubricants, suspending agents, tablet disintegrants, and binders may also be used.

[0185] Another pharmaceutical composition may comprise an effective amount of a mixture of a binding protein and a non-toxic excipient suitable for manufacturing tablets. A unit-dose solution can be prepared by dissolving the tablet in sterile water or other suitable medium. Suitable excipients include, but are not limited to, inert diluents such as calcium carbonate, sodium carbonate, or sodium bicarbonate, lactose, or calcium phosphate; or binders such as starch, gelatin, or gum arabic; or lubricants such as magnesium stearate, stearic acid, or talc.

[0186] Other pharmaceutical compositions disclosed herein will be apparent to those skilled in the art, including formulations involving the sustained or controlled delivery of binding proteins in an formulation. Techniques for formulating a variety of other sustained or controlled delivery methods, such as liposome carriers, biodegradable microparticles or porous beads, and accumulation injection, are also known to those skilled in the art. Other examples of sustained-release formulations include semi-permeable polymer matrices in the form of molded articles, such as films or microcapsules. Sustained-release matrices may include polyesters, hydrogels, polylactic acid, copolymers of L-glutamic acid and γ-ethyl-L-glutamic acid, poly(2-hydroxyethyl-methacrylate), ethylene vinyl acetate, or poly-D(-)-3-hydroxybutyric acid. Sustained-release compositions may also include liposomes, which can be prepared by any of several methods known in the art.

[0187] Pharmaceutical compositions intended for internal administration must typically be sterile. This can be achieved through filtration using a sterile filter membrane. When the composition is lyophilized, sterilization using this method can be performed before or after lyophilization and reconstitution. Compositions intended for parenteral administration can be stored in lyophilized or solution form. Furthermore, parenteral compositions are typically placed in containers with sterile inlets, such as intravenous solution bags or vials with stoppers that can be punctured by a hypodermic needle.

[0188] Once a pharmaceutical composition has been formulated, it can be stored in sterile vials as a solution, suspension, gel, emulsion, solid, or as a dehydrated or lyophilized powder. Such formulations can be stored in ready-to-use form or in a form that requires reconstitution before application (e.g., lyophilized form).

[0189] This disclosure also covers kits for producing single-dose administration units. These kits may each contain a first container with dried protein and a second container with an aqueous formulation. Kits containing single-compartment and multi-compartment pre-filled syringes (e.g., liquid syringes and lyophilized syringes) are also included within the scope of this disclosure.

[0190] The effective amount of a pharmaceutical composition used in treatment will depend, for example, on the treatment context and objectives. Those skilled in the art will understand that the appropriate dose level for treatment will therefore depend in part on the delivered molecule, the indication for the binding protein being used, the route of administration, and the patient's size (weight, body surface area, or organ size) and condition (age and general health). Therefore, clinicians may adjust the dose and modify the route of administration to achieve the best therapeutic effect.

[0191] The frequency of administration will depend on the pharmacokinetic parameters of the binding protein in the formulation used. Typically, clinicians will administer the composition until a dose is reached to achieve the desired effect. Therefore, the composition can be administered as a single dose, two or more doses over time (which may or may not contain the same amount of the desired molecule), or via an implantable device or catheter. Further refinement of the appropriate dosage is routinely performed by those skilled in the art and within the scope of their usual duties. An appropriate dosage can be determined using appropriate dose-response data.

[0192] The pharmaceutical composition may be administered via known methods, such as oral administration; intravenous, intraperitoneal, intracerebral (intra-organ parenchyma), intraventricular, intramuscular, intraocular, intra-arterial, portal vein, or intralesional routes; via a sustained-release system; or via an implantable device. When desired, the composition may be administered by bolus injection or continuous infusion or via an implantable device.

[0193] The composition can also be applied topically by implanting a membrane, sponge, or other suitable material on which the desired molecule has been absorbed or encapsulated. When using an implantable device, the device can be implanted into any suitable tissue or organ, and delivery of the desired molecule can be achieved via diffusion, timed release bolus, or continuous administration. IV. Project

[0194] The following exemplary items represent some aspects of the present invention: 1. An anti-CD38 T-cell binder for use in treating peripheral T-cell lymphoma (PTCL) in individuals in need. 2. The anti-CD38 T cell binder for use according to Item 1 is characterized in that it is a trispecific binding protein comprising a first antigen-binding site that specifically binds to a CD38 polypeptide, a second antigen-binding site that specifically binds to a CD28 polypeptide, and a third antigen-binding site that specifically binds to a CD3 polypeptide. 3. The anti-CD38 T cell binder for use according to Item 1 is characterized in that it is a bispecific binding protein comprising a first antigen-binding site that specifically binds to a CD38 polypeptide and a second antigen-binding site that specifically binds to a CD3 polypeptide. 4. An anti-CD38 T cell binder for use as described in Item 2 or Item 3, wherein the first antigen-binding site of the CD38-binding polypeptide comprises: (a) An antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence containing the amino acid sequence GYTFTSFN (SEQ ID NO: 31) or GYTFTSYA (SEQ ID NO: 37), a CDR-H2 sequence containing the amino acid sequence IYPGNGGT (SEQ ID NO: 32) or IYPGQGGT (SEQ ID NO: 38), and a CDR-H3 sequence containing the amino acid sequence ARTGGLRRAYFTY (SEQ ID NO: 33); and (b) The antibody light chain variable (VL) domain comprises a CDR-L1 sequence containing an amino acid sequence of ESVDSYGNGF (SEQ ID NO: 34) or QSVSSYGQGF (SEQ ID NO: 39), a CDR-L2 sequence containing an amino acid sequence of LAS or GAS, and a CDR-L3 sequence containing an amino acid sequence of QQNKEDPWT (SEQ ID NO: 36). 5. The anti-CD38 T cell binder for use as described in Item 4, wherein the first antigen binding site comprises: (a) An antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence containing the amino acid sequence GYTFTSYA (SEQ ID NO: 37), a CDR-H2 sequence containing the amino acid sequence IYPGQGGT (SEQ ID NO: 38), and a CDR-H3 sequence containing the amino acid sequence ARTGGLRRAYFTY (SEQ ID NO: 33); and (b) The antibody light chain variable (VL) domain comprises a CDR-L1 sequence containing the amino acid sequence QSVSSYGQGF (SEQ ID NO: 39), a CDR-L2 sequence containing the amino acid sequence GAS, and a CDR-L3 sequence containing the amino acid sequence QQNKEDPWT (SEQ ID NO: 36). 6. The anti-CD38 T cell binder for use according to item 4 or item 5, wherein the first antigen binding site comprises a VH domain and a VL domain, the VH domain comprising the amino acid sequence of SEQ ID NO: 13 and the VL domain comprising the amino acid sequence of SEQ ID NO: 14. 7. The anti-CD38 T cell binder for use as described in Item 4, wherein the first antigen binding site comprises: (a) An antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence containing the amino acid sequence GYTFTSFN (SEQ ID NO: 31), a CDR-H2 sequence containing the amino acid sequence IYPGNGGT (SEQ ID NO: 32), and a CDR-H3 sequence containing the amino acid sequence ARTGGLRRAYFTY (SEQ ID NO: 33); and (b) The antibody light chain variable (VL) domain comprises a CDR-L1 sequence containing the amino acid sequence of ESVDSYGNGF (SEQ ID NO: 34), a CDR-L2 sequence containing the amino acid sequence of LAS, and a CDR-L3 sequence containing the amino acid sequence of QQNKEDPWT (SEQ ID NO: 36). 8. An anti-CD38 T cell conjugate for use as described in Item 4 or Item 7, wherein the first antigen binding site comprises: (a) A VH domain and a VL domain, wherein the VH domain contains the amino acid sequence of SEQ ID NO: 5 and the VL domain contains the amino acid sequence of SEQ ID NO: 6; (b) A VH domain and a VL domain, wherein the VH domain contains the amino acid sequence of SEQ ID NO: 17 and the VL domain contains the amino acid sequence of SEQ ID NO: 18; (c) A VH domain and a VL domain, wherein the VH domain contains the amino acid sequence of SEQ ID NO: 21 and the VL domain contains the amino acid sequence of SEQ ID NO: 18; or (d) A VH domain and a VL domain, wherein the VH domain contains the amino acid sequence of SEQ ID NO: 23 and the VL domain contains the amino acid sequence of SEQ ID NO: 18. 9. An anti-CD38 T cell binder for use as described in Item 2 or Item 3, wherein the first antigen-binding site of the CD38-binding polypeptide comprises: (a) An antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence containing the amino acid sequence GFTFSSYG (SEQ ID NO: 41), a CDR-H2 sequence containing the amino acid sequence IWYDGSNK (SEQ ID NO: 42), and a CDR-H3 sequence containing the amino acid sequence ARMFRGAFDY (SEQ ID NO: 43); and (b) The antibody light chain variable (VL) domain comprising a CDR-L1 sequence containing the amino acid sequence QGIRND (SEQ ID NO: 44), a CDR-L2 sequence containing the amino acid sequence AAS, and a CDR-L3 sequence containing the amino acid sequence LQDYIYYPT (SEQ ID NO: 46). 10. The anti-CD38 T cell binder for use according to item 9, wherein the first antigen binding site comprises a VH domain and a VL domain, the VH domain comprising the amino acid sequence of SEQ ID NO: 9 and the VL domain comprising the amino acid sequence of SEQ ID NO: 10. 11. An anti-CD38 T cell binder for use as described in Item 2 or Item 3, wherein the first antigen-binding site of the CD38-binding polypeptide comprises: (a) An antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence containing the amino acid sequence of gytltefs (SEQ ID NO: 2), a CDR-H2 sequence containing the amino acid sequence of fdpedget (SEQ ID NO: 3), and a CDR-H3 sequence containing the amino acid sequence of ttgrffdwf (SEQ ID NO: 4); and (b) The antibody light chain variable (VL) domain comprising a CDR-L1 sequence containing the amino acid sequence of QSVISRF (SEQ ID NO: 7), a CDR-L2 sequence containing the amino acid sequence of GAS, and a CDR-L3 sequence containing the amino acid sequence of qqdsnlpit (SEQ ID NO: 11). 12. The anti-CD38 T cell binder for use according to item 11, wherein the first antigen binding site comprises a VH domain and a VL domain, the VH domain comprising the amino acid sequence of SEQ ID NO: 108 and the VL domain comprising the amino acid sequence of SEQ ID NO: 109. 13. The anti-CD38 T cell binder for use as described in Item 2 or Item 3, wherein the first antigen-binding site of the CD38-binding polypeptide comprises: (a) An antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence containing the amino acid sequence GYAFTTYL (SEQ ID NO: 12), a CDR-H2 sequence containing the amino acid sequence INPGSGST (SEQ ID NO: 15), and a CDR-H3 sequence containing the amino acid sequence ARYAYGY (SEQ ID NO: 16); and (b) The antibody light chain variable (VL) domain comprising a CDR-L1 sequence containing the amino acid sequence QNVGTA (SEQ ID NO: 19), a CDR-L2 sequence containing the amino acid sequence SAS, and a CDR-L3 sequence containing the amino acid sequence QQYSTYPFT (SEQ ID NO: 22). 14. The anti-CD38 T cell binder for use as described in Item 13, wherein the first antigen binding site comprises: (a) A VH domain and a VL domain, wherein the VH domain contains the amino acid sequence of SEQ ID NO: 110 and the VL domain contains the amino acid sequence of SEQ ID NO: 111; or (b) A VH domain and a VL domain, wherein the VH domain contains the amino acid sequence of SEQ ID NO: 116 and the VL domain contains the amino acid sequence of SEQ ID NO: 117. 15. An anti-CD38 T cell binder for use as described in Item 2 or Item 3, wherein the first antigen-binding site of the CD38-binding polypeptide comprises: (a) An antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence containing the amino acid sequence GYSFTNYA (SEQ ID NO: 24), a CDR-H2 sequence containing the amino acid sequence ISPHYGDT (SEQ ID NO: 25), and a CDR-H3 sequence containing the amino acid sequence ARRFEGFYYSMDY (SEQ ID NO: 26); and (b) The antibody light chain variable (VL) domain comprises a CDR-L1 sequence containing the amino acid sequence QSLVHSNGNTY (SEQ ID NO: 27), a CDR-L2 sequence containing the amino acid sequence KVS, and a CDR-L3 sequence containing the amino acid sequence SQSTHVPLT (SEQ ID NO: 29). 16. The anti-CD38 T cell binder for use as described in Item 15, wherein the first antigen binding site comprises: (a) A VH domain and a VL domain, wherein the VH domain contains the amino acid sequence of SEQ ID NO: 112 and the VL domain contains the amino acid sequence of SEQ ID NO: 113; or (b) A VH domain and a VL domain, wherein the VH domain contains the amino acid sequence of SEQ ID NO: 118 and the VL domain contains the amino acid sequence of SEQ ID NO: 119. 17. An anti-CD38 T cell binder for use as described in Item 2 or Item 3, wherein the first antigen-binding site of the CD38-binding polypeptide comprises: (a) An antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence containing the amino acid sequence GFTFSSYG (SEQ ID NO: 41), a CDR-H2 sequence containing the amino acid sequence IWYDGSNK (SEQ ID NO: 42), and a CDR-H3 sequence containing the amino acid sequence ARDPGLRYFDGGMDV (SEQ ID NO: 106); and (b) The antibody light chain variable (VL) domain comprises a CDR-L1 sequence containing the amino acid sequence QGISSY (SEQ ID NO: 107), a CDR-L2 sequence containing the amino acid sequence AAS, and a CDR-L3 sequence containing the amino acid sequence QQLNSFPYT (SEQ ID NO: 229). 18. The anti-CD38 T cell binder for use according to item 17, wherein the first antigen binding site comprises a VH domain and a VL domain, the VH domain comprising the amino acid sequence of SEQ ID NO: 114 and the VL domain comprising the amino acid sequence of SEQ ID NO: 115. 19. The anti-CD38 T cell binder according to any one of items 2 to 18 for use, wherein the antigen-binding site for binding the CD3 polypeptide comprises: (a) An antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence containing the amino acid sequence GFTFTKAW (SEQ ID NO: 120), a CDR-H2 sequence containing the amino acid sequence IKDKSNSYAT (SEQ ID NO: 121), and a CDR-H3 sequence containing the amino acid sequence RGVYYALSPFDY (SEQ ID NO: 122); and (b) The antibody light chain variable (VL) domain comprising a CDR-L1 sequence containing the amino acid sequence QSLVHX1NX2X3TY (where X1 is E or Q, X2 is A or L, and X3 is Q, R, or F) (SEQ ID NO: 131), a CDR-L2 sequence containing the amino acid sequence KVS, and a CDR-L3 sequence containing the amino acid sequence GQGTQYPFT (SEQ ID NO: 130). 20. The anti-CD38 T cell conjugate for use as described in Item 19, wherein the CDR-L1 sequence of the VL domain of the antigen-binding site of the CD3 polypeptide is selected from the group consisting of: QSLVHNNANTY (SEQ ID NO: 123), QSLVHQNAQTY (SEQ ID NO: 124), QSLVHENLQTY (SEQ ID NO: 125), QSLVHENLFTY (SEQ ID NO: 126), QSLVHENLRTY (SEQ ID NO: 127), and QSLVHDNAQTY (SEQ ID NO: 128). 21. The anti-CD38 T cell binder for use according to item 19 or item 20, wherein the antigen-binding site of the CD3-binding polypeptide comprises a VH domain and a VL domain, the VH domain comprising the amino acid sequence of SEQ ID NO: 53 or 138, and the VL domain comprising the amino acid sequence of SEQ ID NO: 54, 133, 134, 135, 136, or 137. 22. The anti-CD38 T cell binder for use according to item 21, wherein the antigen-binding site of the CD3-binding polypeptide comprises a VH domain and a VL domain, the VH domain comprising the amino acid sequence of SEQ ID NO: 53 and the VL domain comprising the amino acid sequence of SEQ ID NO: 54. 23. An anti-CD38 T cell binder for use according to any one of items 2-18, wherein the antigen-binding site for binding the CD3 polypeptide comprises: (a) An antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence containing the amino acid sequence GFTFTKAW (SEQ ID NO: 120), a CDR-H2 sequence containing the amino acid sequence IKDKSNSYAT (SEQ ID NO: 121), and a CDR-H3 sequence containing the amino acid sequence RGVYYALSPFDY (SEQ ID NO: 122); and (b) The antibody light chain variable (VL) domain comprises a CDR-L1 sequence containing the amino acid sequence QSLVHNNGNTY (SEQ ID NO: 218), a CDR-L2 sequence containing the amino acid sequence KVS, and a CDR-L3 sequence containing the amino acid sequence GQGTQYPFT (SEQ ID NO: 130). 24. The anti-CD38 T cell binder for use according to item 23, wherein the third antigen binding site comprises a VH domain and a VL domain, the VH domain comprising the amino acid sequence of SEQ ID NO: 84 and the VL domain comprising the amino acid sequence of SEQ ID NO: 85. 25. An anti-CD38 T cell binder for use according to any one of items 2 and 4-24, wherein the antigen-binding site for binding the CD28 polypeptide comprises: (a) An antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence containing the amino acid sequence GYTFTSYY (SEQ ID NO: 139), a CDR-H2 sequence containing the amino acid sequence IYPGNVNT (SEQ ID NO: 140), and a CDR-H3 sequence containing the amino acid sequence TRSHYGLDWNFDV (SEQ ID NO: 141); and (b) The antibody light chain variable (VL) domain comprises a CDR-L1 sequence containing the amino acid sequence QNIYVW (SEQ ID NO: 142), a CDR-L2 sequence containing the amino acid sequence KAS, and a CDR-L3 sequence containing the amino acid sequence QQGQTYPY (SEQ ID NO: 144). 26. The anti-CD38 T cell binder for use according to item 25, wherein the antigen-binding site of the CD28-binding polypeptide comprises a VH domain and a VL domain, the VH domain comprising the amino acid sequence of SEQ ID NO: 49 and the VL domain comprising the amino acid sequence of SEQ ID NO: 50. 27. An anti-CD38 T cell binder for use according to any one of items 2 and 4-24, wherein the antigen-binding site for binding the CD28 polypeptide comprises: (a) An antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence containing the amino acid sequence GFSLSDYG (SEQ ID NO: 212), a CDR-H2 sequence containing the amino acid sequence IWAGGGT (SEQ ID NO: 213), and a CDR-H3 sequence containing the amino acid sequence ARDKGYSYYYSMDY (SEQ ID NO: 214); and (b) The antibody light chain variable (VL) domain comprises a CDR-L1 sequence containing the amino acid sequence ESVEYYVTSL (SEQ ID NO: 215), a CDR-L2 sequence containing the amino acid sequence AAS, and a CDR-L3 sequence containing the amino acid sequence QQSRKVPYT (SEQ ID NO: 217). 28. The anti-CD38 T cell binder for use according to item 27, wherein the second antigen binding site comprises a VH domain and a VL domain, the VH domain comprising the amino acid sequence of SEQ ID NO: 51 and the VL domain comprising the amino acid sequence of SEQ ID NO: 52. 29. An anti-CD38 T cell binder for use according to any one of items 2 and 4-28, wherein the trispecific binding protein comprises four polypeptide chains forming the three antigen-binding sites, wherein the first polypeptide chain comprises a structure represented by the following formula: V L2 -L1-V L1 -L2-C L [I] Furthermore, the second polypeptide chain contains a structure represented by the following formula: V H1 -L3-V H2 -L4-C H1 -Hinge-C H2 -C H3 [II] Furthermore, the third polypeptide chain contains a structure represented by the following formula: V H3 -C H1 -Hinge-C H2 -C H3 [III] Furthermore, the fourth polypeptide chain contains a structure represented by the following formula: V L3 -C L [IV] in: V L1 It is the variable domain of the first immunoglobulin light chain; V L2 It is the variable domain of the second immunoglobulin light chain; V L3 It is the variable domain of the third immunoglobulin light chain; V H1 It is the variable domain of the first immunoglobulin heavy chain; V H2 It is the variable domain of the second immunoglobulin heavy chain; V H3 It is the variable domain of the third immunoglobulin heavy chain; C L It is the constant structural domain of the immunoglobulin light chain; C H1 It is immunoglobulin C H1 Heavy chain constant structural domain; C H2 It is immunoglobulin C H2 Heavy chain constant structural domain; C H3 It is immunoglobulin C H3 Heavy chain constant structural domain; The hinge is the connection of C H1 and C H2 The immunoglobulin hinge region of the structural domain; and L1, L2, L3, and L4 are amino acid linkers; The polypeptide having Formula I and the polypeptide having Formula II form a cross-linked light chain-heavy chain pair; wherein V H1 and V L1 Forming one of these three antigen-binding sites; among which V H2 and V L2 Forming another of these three antigen-binding sites; and where V H3 and V L3 This forms another of the three antigen-binding sites. 30. The anti-CD38 T cell conjugate as described in Item 29, wherein: (a) V H1 and V L1 This forms the second antigen-binding site for the CD28 peptide, V H2 and V L2 This forms the third antigen-binding site for the CD3-binding polypeptide, and V H3 and V L3 Forming the first antigen-binding site for the CD38 polypeptide; or (b) V H1 and VL1 This forms the third antigen-binding site for the CD3-binding polypeptide, V H2 and V L2 This forms the second antigen-binding site for the CD28 peptide, and V H3 and V L3 This forms the first antigen-binding site for the CD38 polypeptide. 31. The anti-CD38 T cell conjugate as described in Item 29, wherein: (a) V H1 and V L1 This forms the second antigen-binding site for the CD28 peptide, V H2 and V L2 This forms the first antigen-binding site for the CD38 polypeptide, and V H3 and V L3 Forming the third antigen-binding site for the CD3-binding polypeptide; or (b) V H1 and V L1 This forms the third antigen-binding site for the CD3-binding polypeptide, V H2 and V L2 This forms the first antigen-binding site for the CD38 polypeptide, and V H3 and V L3 This forms the second antigen-binding site for the CD28 polypeptide. 32. The anti-CD38 T cell binder for use as described in Item 29, wherein: (a) V H1 and V L1 This forms the first antigen-binding site for the CD38 polypeptide, V H2 and V L2 This forms the third antigen-binding site for the CD3-binding polypeptide, and V H3 and V L3 Forming a second antigen-binding site for the CD28 polypeptide; or (b) V H1 and V L1 This forms the first antigen-binding site for the CD38 polypeptide, V H2 and V L2 This forms the second antigen-binding site for the CD28 peptide, and V H3 and V L3 This forms the third antigen-binding site for the CD3-binding polypeptide. 33. The anti-CD38 T cell binder for use as described in Item 29, wherein: V H1 and V L1This forms the second antigen-binding site for the CD28 peptide, V H2 and V L2 This forms the third antigen-binding site for the CD3-binding polypeptide, and V H3 and V L3 This forms the first antigen-binding site for the CD38 polypeptide; V H1 The CDR-H1 sequence contains the amino acid sequence gytftsyy (SEQ ID NO: 139), the CDR-H2 sequence contains the amino acid sequence iypgnvnt (SEQ ID NO: 140), and the CDR-H3 sequence contains the amino acid sequence trshygldwnfdv (SEQ ID NO: 141). L1 The CDR-L1 sequence contains the amino acid sequence QNIYVW (SEQ ID NO: 142), the CDR-L2 sequence contains the amino acid sequence KAS, and the CDR-L3 sequence contains the amino acid sequence QQGQTYPY (SEQ ID NO: 144). V H2 The CDR-H1 sequence contains the amino acid sequence GFTFTKAW (SEQ ID NO: 120), the CDR-H2 sequence contains the amino acid sequence IKDKSNSYAT (SEQ ID NO: 121), and the CDR-H3 sequence contains the amino acid sequence RGVYYALSPFDY (SEQ ID NO: 122). L2 The CDR-L1 sequence contains the amino acid sequence QSLVHNNANTY (SEQ ID NO: 123), the CDR-L2 sequence contains the amino acid sequence KVS, and the CDR-L3 sequence contains the amino acid sequence GQGTQYPFT (SEQ ID NO: 130); and V H3 The CDR-H1 sequence contains the amino acid sequence GYTFTSYA (SEQ ID NO: 37), the CDR-H2 sequence contains the amino acid sequence IYPGQGGT (SEQ ID NO: 38), and the CDR-H3 sequence contains the amino acid sequence ARTGGLRRAYFTY (SEQ ID NO: 33). L3 The CDR-L1 sequence contains the amino acid sequence QSVSSYGQGF (SEQ ID NO: 39), the CDR-L2 sequence contains the amino acid sequence GAS, and the CDR-L3 sequence contains the amino acid sequence QQNKEDPWT (SEQ ID NO: 36). 34. The anti-CD38 T cell conjugate as described in Item 29, wherein V H1 and V L1 This forms the second antigen-binding site for the CD28 peptide, V H2 and V L2 This forms the third antigen-binding site for the CD3-binding polypeptide, and V H3 and V L3 This forms the first antigen-binding site for the CD38 polypeptide; wherein V H1 Contains the amino acid sequence of SEQ ID NO: 49, and V L1 Contains the amino acid sequence of SEQ ID NO: 50; wherein V H2 Contains the amino acid sequence of SEQ ID NO: 53, and V L2 Contains the amino acid sequence of SEQ ID NO: 54; and wherein V H3 Contains the amino acid sequence of SEQ ID NO: 13, and V L3 It contains the amino acid sequence of SEQ ID NO: 14. 35. An anti-CD38 T cell binder for use according to any one of items 29-34, wherein: (a) L1 contains the sequence GQPKAAP (SEQ ID NO: 58), L2 contains the sequence TKGPS (SEQ ID NO: 57), L3 contains amino acid S, and L4 contains the sequence RT; (b) L1 contains the sequence GGGGSGGGGS (SEQ ID NO: 55), L2 contains the sequence GGGGSGGGGS (SEQ ID NO: 55), L3 has a length of 0 amino acids, and L4 has a length of 0 amino acids; (c) L1 contains the sequence GGSGSSGSGG (SEQ ID NO: 59), L2 contains the sequence GGSGSSGSGG (SEQ ID NO: 59), L3 is 0 amino acids long, and L4 is 0 amino acids long; or (d) L1 contains the sequence GGGGSGGGGSGGGGS (SEQ ID NO: 56), L2 has a length of 0 amino acids, L3 contains the sequence GGGGSGGGGSGGGGS (SEQ ID NO: 56), and L4 has a length of 0 amino acids. 36. An anti-CD38 T cell conjugate for use according to any one of items 29-34, wherein at least one of L1, L2, L3 or L4 contains the sequence DKTHT (SEQ ID NO: 147). 37. The anti-CD38 T cell conjugate for use as described in item 36, wherein L1, L2, L3 and L4 contain the sequence DKTHT (SEQ ID NO: 147). 38. An anti-CD38 T cell conjugate for use according to any one of items 29-37, wherein the hinges of the second polypeptide chain and the third polypeptide chain are C H2 -C H3 The structural domain is the human IgG4 hinge-C H2 -C H3 Structural domain, and in which these hinges - C H2 -C H3 Each of the domains contains an amino acid substitution at positions 234 and 235 corresponding to human IgG4 according to the EU index, wherein these amino acid substitutions are F234A and L235A. 39. An anti-CD38 T cell conjugate for use according to any one of items 29-37, wherein the hinges of the second polypeptide chain and the third polypeptide chain are C H2 -C H3 The structural domain is the human IgG4 hinge-C H2 -C H3 Structural domain, and in which these hinges - C H2 -C H3 Each of the domains contains amino acid substitutions at positions 233-236 corresponding to human IgG4 according to the EU index, wherein these amino acid substitutions are E233P, F234V, L235A and a deletion at position 236. 40. An anti-CD38 T cell conjugate for use according to any one of items 29-37, wherein the hinges of the second polypeptide chain and the third polypeptide chain are C H2 -C H3 The structural domain is the human IgG4 hinge-C H2 -C H3 Structural domain, and in which these hinges - C H2 -C H3 Each of the domains contains amino acid substitutions at positions 228 and 409 corresponding to human IgG4 according to the EU index, wherein these amino acid substitutions are S228P and R409K. 41. An anti-CD38 T cell conjugate for use according to any one of items 29-37, wherein the hinges of the second polypeptide chain and the third polypeptide chain are C H2 -C H3 The structural domain is the human IgG1 hinge-C H2 -C H3 Structural domain, and in which these hinges - C H2 -C H3Each of the domains contains an amino acid substitution at positions 234, 235, and 329 corresponding to human IgG1 according to the EU index, wherein these amino acid substitutions are L234A, L235A, and P329A. 42. An anti-CD38 T cell conjugate for use according to any one of items 29-37, wherein the hinges of the second polypeptide chain and the third polypeptide chain are C H2 -C H3 The structural domain is the human IgG1 hinge-C H2 -C H3 Structural domain, and in which these hinges - C H2 -C H3 Each of the domains contains an amino acid substitution at positions 298, 299, and 300 corresponding to human IgG1 according to the EU index, wherein the amino acid substitutions are S298N, T299A, and Y300S. 43. An anti-CD38 T cell binder for use according to any one of items 29-42, wherein the hinge of the second polypeptide chain is C H2 -C H3 The domain contains amino acid substitutions at positions 349, 366, 368, and 407 corresponding to human IgG1 or IgG4 according to the EU index, wherein these amino acid substitutions are Y349C, T366S, L368A, and Y407V; and wherein the hinge of the third polypeptide chain is C H2 -C H3 The domain contains amino acid substitutions at positions 354 and 366 corresponding to human IgG1 or IgG4 according to the EU index, wherein these amino acid substitutions are S354C and T366W. 44. An anti-CD38 T cell binder for use according to any one of items 29-42, wherein the hinge of the second polypeptide chain is C H2 -C H3 The domain contains amino acid substitutions at positions 354 and 366 corresponding to human IgG1 or IgG4 according to the EU index, wherein these amino acid substitutions are S354C and T366W; and wherein the hinge of the third polypeptide chain is C H2 -C H3 The domain contains amino acid substitutions at positions 349, 366, 368, and 407 corresponding to human IgG1 or IgG4 according to the EU index, wherein these amino acid substitutions are Y349C, T366S, L368A, and Y407V. 45. The anti-CD38 T cell binder for use as described in Item 29, wherein: (a) The first polypeptide chain contains the amino acid sequence of SEQ ID NO: 61, the second polypeptide chain contains the amino acid sequence of SEQ ID NO: 60, the third polypeptide chain contains the amino acid sequence of SEQ ID NO: 62, and the fourth polypeptide chain contains the amino acid sequence of SEQ ID NO: 63; (b) The first polypeptide chain contains the amino acid sequence of SEQ ID NO: 61, the second polypeptide chain contains the amino acid sequence of SEQ ID NO: 64, the third polypeptide chain contains the amino acid sequence of SEQ ID NO: 65, and the fourth polypeptide chain contains the amino acid sequence of SEQ ID NO: 63; (c) The first polypeptide chain contains the amino acid sequence of SEQ ID NO: 61, the second polypeptide chain contains the amino acid sequence of SEQ ID NO: 66, the third polypeptide chain contains the amino acid sequence of SEQ ID NO: 67, and the fourth polypeptide chain contains the amino acid sequence of SEQ ID NO: 63; (d) The first polypeptide chain contains the amino acid sequence of SEQ ID NO: 61, the second polypeptide chain contains the amino acid sequence of SEQ ID NO: 60, the third polypeptide chain contains the amino acid sequence of SEQ ID NO: 68, and the fourth polypeptide chain contains the amino acid sequence of SEQ ID NO: 69; (e) The first polypeptide chain contains the amino acid sequence of SEQ ID NO: 61, the second polypeptide chain contains the amino acid sequence of SEQ ID NO: 64, the third polypeptide chain contains the amino acid sequence of SEQ ID NO: 70, and the fourth polypeptide chain contains the amino acid sequence of SEQ ID NO: 69; (f) The first polypeptide chain contains the amino acid sequence of SEQ ID NO: 61, the second polypeptide chain contains the amino acid sequence of SEQ ID NO: 66, the third polypeptide chain contains the amino acid sequence of SEQ ID NO: 71, and the fourth polypeptide chain contains the amino acid sequence of SEQ ID NO: 69; (g) The first polypeptide chain contains the amino acid sequence of SEQ ID NO: 148, the second polypeptide chain contains the amino acid sequence of SEQ ID NO: 149, the third polypeptide chain contains the amino acid sequence of SEQ ID NO: 150, and the fourth polypeptide chain contains the amino acid sequence of SEQ ID NO: 151; (h) The first polypeptide chain contains the amino acid sequence of SEQ ID NO: 152, the second polypeptide chain contains the amino acid sequence of SEQ ID NO: 153, the third polypeptide chain contains the amino acid sequence of SEQ ID NO: 154, and the fourth polypeptide chain contains the amino acid sequence of SEQ ID NO: 155. (i) The first polypeptide chain contains the amino acid sequence of SEQ ID NO: 156, the second polypeptide chain contains the amino acid sequence of SEQ ID NO: 157, the third polypeptide chain contains the amino acid sequence of SEQ ID NO: 158, and the fourth polypeptide chain contains the amino acid sequence of SEQ ID NO: 159; (j) The first polypeptide chain contains the amino acid sequence of SEQ ID NO: 160, the second polypeptide chain contains the amino acid sequence of SEQ ID NO: 161, the third polypeptide chain contains the amino acid sequence of SEQ ID NO: 162, and the fourth polypeptide chain contains the amino acid sequence of SEQ ID NO: 163; (k) The first polypeptide chain contains the amino acid sequence of SEQ ID NO: 164, the second polypeptide chain contains the amino acid sequence of SEQ ID NO: 165, the third polypeptide chain contains the amino acid sequence of SEQ ID NO: 166, and the fourth polypeptide chain contains the amino acid sequence of SEQ ID NO: 167. (l) The first polypeptide chain contains the amino acid sequence of SEQ ID NO: 168, the second polypeptide chain contains the amino acid sequence of SEQ ID NO: 169, the third polypeptide chain contains the amino acid sequence of SEQ ID NO: 170, and the fourth polypeptide chain contains the amino acid sequence of SEQ ID NO: 171; (m) The first polypeptide chain contains the amino acid sequence of SEQ ID NO: 172, the second polypeptide chain contains the amino acid sequence of SEQ ID NO: 173, the third polypeptide chain contains the amino acid sequence of SEQ ID NO: 174, and the fourth polypeptide chain contains the amino acid sequence of SEQ ID NO: 175; or (n) The first polypeptide chain contains the amino acid sequence of SEQ ID NO: 176, the second polypeptide chain contains the amino acid sequence of SEQ ID NO: 177, the third polypeptide chain contains the amino acid sequence of SEQ ID NO: 178, and the fourth polypeptide chain contains the amino acid sequence of SEQ ID NO: 179. 46. ​​An anti-CD38 T cell binder for use according to any one of items 2-45, wherein the CD38 polypeptide is a human CD38 polypeptide. 47. An anti-CD38 T cell binder for use according to any one of items 2-46, wherein the CD3 polypeptide is a human CD3 polypeptide. 48. An anti-CD38 T cell binder for use according to any one of items 2 and 4-47, wherein the CD28 polypeptide is a human CD28 polypeptide. 49. An anti-CD38 T-cell binder for use according to any one of items 1-48, wherein the PTCL is angioimmunoblastic T-cell lymphoma (AITL), hepatosplenic T-cell lymphoma (HSTL), or adult T-cell leukemia / lymphoma (ATLL), extranodal NK / T-cell lymphoma (ENKTCL), mycosis fungoides (MF), Cezari syndrome (SS), anaplastic large cell ALK-lymphoma (ALCL), PTCL-unspecified (NOS), enteropathy-type T-cell lymphoma (EATL), monomorphic epithelial intestinal T-cell lymphoma (MEITL), primary cutaneous CD4+ lymphoproliferative disorder, subcutaneous panniculitis-like T-cell lymphoma (SCTCL), or primary cutaneous γδ T-cell lymphoma. 50. An anti-CD38 T-cell binder for use as described in any one of items 1-48, wherein the PTCL is hepatosplenic T-cell lymphoma (HSTL), Cezari syndrome (SS), or mycosis fungoides (MF). 51. The anti-CD38 T cell binder for use as described in item 50, wherein the mycosis fungoides is transformed mycosis fungoides. 52. An anti-CD38 T cell conjugate for use according to any one of items 1-51, wherein the cells of the PTCL express CD38 and / or CD28. 53. An anti-CD38 T cell conjugate for use as described in any one of items 1-52, wherein the individual is a human. 54. A pharmaceutical composition comprising an anti-CD38 T cell binder for use according to any one of items 1-53. Example

[0195] The following examples illustrate specific embodiments of this disclosure and their various uses. These examples are presented for illustrative purposes only and should not be construed as limiting the scope of the invention in any way. Example 1: Treatment of PTCL with anti-CD38 / CD28xCD3 trispecific binding protein

[0196] Peripheral T-cell lymphoma (PTCL) is a complex group of clinicopathological entities, often associated with an aggressive clinical course. Despite the introduction of new chemotherapy regimens and a wealth of new agents, progression-free survival in PTCL patients remains to be improved. In recent years, immunotherapy has been established as a major advance in the treatment of various cancers. Trispecific binding proteins targeting CD38, CD3, and CD28 have been developed for the treatment of multiple myeloma and chronic viral infections.

[0197] This example describes a study of the anti-CD38 / CD28xCD3 trispecific binding protein containing the peptide of SEQ ID NO: 60-63 in the treatment of PTCL. Materials and methods Cell lines and cultures

[0198] Human cell lines SEAX, H9 (derived from Cezari syndrome), DERL-2 (derived from hepatosplenic T-cell lymphoma), YT (derived from T / NK cell leukemia), JURKAT and MOLT4 (derived from acute T-cell leukemia), Mac2a, and Mac2b were each established from clinical specimens of one patient. All cell lines (except DERL-2) were grown at 37°C and 5% CO2 in RPMI 1640 Glutamax-I medium supplemented with 10% fetal bovine serum and 100 U / ml penicillin-streptomycin (both from Thermo Fisher Scientific). DERL-2 was grown in RPMI 1640 Glutamax-I medium supplemented with 20% fetal bovine serum, 100 U / ml penicillin-streptomycin (both from Thermo Fisher Scientific), and 20 ng / ml human IL-2 (BioLegend). Immunohistochemistry (IHC)

[0199] IHC analysis was performed on PTCL (n = 261) tissue sections. As previously described [References], IHC was performed on FFPE tissue sections using antibodies against CD3 (mouse mAb, clone F7.2.38, 1:50; Agilent Dako), CD38 (mouse mAb, clone SCP32, 1:100; Leica Biosystems), and CD28 (rabbit mAb, clone D2Z4E, 1:50; Cell Signaling Technology). IHC against CD3 and CD38 was performed on a BOND III or BOND-MAX (Leica Biosystems) automated staining platform. After appropriate antigen retrieval using EDTA buffer (pH 9) (DiaPath), dewaxed tissue sections were manually stained against CD28 using a two-step indirect immunoperoxidase assay (ImmPRESS anti-rabbit; Vector Laboratories). CD28 and CD38 expression was assessed by pathologists on tumor cells. Internal controls were evaluated for both markers: reactive T cells against CD28 and plasma cells against CD38. Expression was assessed using a semi-quantitative scoring method, calculated as the ratio (percentage) of intensity (between 1 and 3, where 3 corresponds to the intensity of the internal control (e.g., plasma cells)) to the proportion (percentage) of stained tumor cells, using the formula: Score = Intensity × Proportion of stained tumor cells H score = Intensity × Percentage of stained tumor cells Multiplex immunofluorescence

[0200] In this study, the TSA-based Opal method was used for immunofluorescence (IF) staining (Akoya Biosciences). Following appropriate antigen retrieval using EDTA buffer pH 9 (Zytomed) according to the manufacturer's instructions, multiplex immunofluorescence analysis was performed on FFPE tissue sections for PTCL-TFH (N=6), Cézari syndrome (N=7), ALCL ALK+ (N=2), and ENKTCL (N=2). Low-expression markers were conjugated with higher-intensity Opals to facilitate spectral acquisition, and vice versa. Opal fluorescent dyes were used at a dilution of 1:200. For multiplex staining, antibodies against CD28 (1:200; Cell Signaling Technologies), CD38 (1:100; Leica Biosystems), PD1 (1:200; Abcam), ICOS (1:200; Abcam), and ALK1 (1:200; Agilent Technologies) were used. Flow cytometry

[0201] For flow cytometry of primary cells, eighteen blood samples were prospectively collected in EDTA tubes, transported at room temperature, and analyzed locally within 24 hours at the Immunology Laboratory of Henri Mondor Hospital. At room temperature, fresh whole blood (100 μL) was stained for 10 minutes with a mixture of antibodies against CD45-Krome Orange (Beckman Coulter), CD3-ECD (Beckman Coulter), CD4-PB (Beckman Coulter), CD8-AA700, CD14-FITC (Beckman Coulter), CD38-PC5.5 or isotype control (Beckman Coulter), and CD28-APC-H7 or isotype control (BD Pharmingen). Based on the T-cell lymphoma subtype, the following additional antibodies were added: anti-CD10 PE (Beckman Coulter), anti-CD7 PE (Beckman Coulter), and anti-KIR3DL1 / DL2 (Miltenyi Biotec). Red blood cells were then lysed using VersaLyse reagent (Beckman Coulter) according to the manufacturer's instructions. Samples were then washed in 1 mL phosphate-buffered saline (PBS) and immediately acquired on a Navios flow cytometer (Beckman Coulter). Data were analyzed using Kaluza 2.1 software. Lymphocytes were gated as CD45+CD14- cells with low side scattering. Tumor T cells were defined according to the T-cell lymphoma subtype: for PTCL-TFH, tumor T cells were defined as CD4+CD3+ / -CD10+ / -CD7+ / - cells; for cutaneous T-cell lymphoma, tumor T cells were defined as CD4+CD3+CD7-KIR3DL2+ cells. Then, the expression of CD38 and CD28 was studied in gated tumor cells, and expression thresholds were set according to isotype controls.

[0202] In flow cytometry of cell lines, multi-parameter flow cytometry immunofluorescence analysis was performed. FlowJo software was used, specifically in LSR Fretessa... TMMulticolor analysis was performed using a combination of monoclonal fluorescently labeled antibodies on an X-20 (BD Biosciences). For cell line phenotypes, CD28-PE (clone CD28.2; BD Biosciences), CD38-APC (clone HIT2; BD Biosciences), and CD3-BV510 (clone SK7; BD Biosciences) antibodies were used according to the manufacturer's instructions, along with corresponding isotype controls, and the immobilizable viability dye eFluor™ 780 (1:1000; Thermo Fisher Scientific). Cytotoxicity assay

[0203] For cytotoxicity assays of cell lines, PBMCs isolated from normal human donors using the Ficoll method were co-incubated with CFSE-labeled tumor cell lines (SEAX, H9, DERL 2, MOLT-4, JURKAT, or YT) in the presence of anti-CD38 / CD28xCD3 trispecific antibodies or indicated single mutant (Δ) or double mutant (ΔΔ) negative controls at dose ranges (0, 8, 40, 200, 1000, 5000, 25000 pM) from healthy human volunteers with normal human donors. The cells were incubated with an effector cell to target cell ratio of 10:1, with 20,000 target cells per well (96-well plate). Spontaneous lysis was measured in separate culture media. After 12 hours of incubation, the lysis of target cell lines by PBMCs was monitored by flow cytometry (MACSQuant Analyzer 16; Miltenyi Biotechnology Co., Ltd.), measured by the percentage of cells double-positive for both CFSE and immobilizable viable dye (eFluor™ 780; 1:1000; Thermo Fisher Scientific Co., Ltd.). Specific lysis of target cells was calculated by subtracting the spontaneous lysis value from that in the culture medium alone.

[0204] For cytotoxicity assays using T lymphocytes as effector cells, monocytes were isolated from PBMCs using the Pan T Cell Isolation Kit (Mitengene Biotechnology Co., Ltd.). Based on an effector cell to target cell ratio of 10:1, T cells were cultured at a rate of 2 × 10⁶ cells / cells. 5 The seeding rate of each well was achieved by inoculating it with CFSE-labeled tumor cell lines. Cytotoxicity assays were performed as previously described. Activation of effector cells and target cells

[0205] To address the activation of PBMCs or target cells (cell lines) mediated by anti-CD38 / CD28xCD3 trispecific binding protein or indicated single mutant (Δ) or double mutant (ΔΔ) negative controls, PBMCs were co-incubated with tumor cell lines (JURKAT or SEAX) or individually for 24 h or 96 h in the presence of dose-range anti-CD38 / CD28xCD3 trispecific binding protein or control antibodies, using an effector cell to target cell ratio of 10:1 similar to cytotoxicity assays. After 24 h of incubation, PBMC activation was monitored by flow cytometry using anti-CD25-PE antibody and CD69-BV421 antibody (Baijin Biotechnology Co., Ltd.). After 96 h of incubation, cell activation was monitored using anti-HLA.DR-APC antibody (BD Parkin) and PD1-PECy7 antibody (Baijin Biotechnology Co., Ltd.). Data were acquired on a FACS MACSQuant Analyzer 16 (Milan Biotech) and analyzed using FlowLogic software v7.2.1. result

[0206] CD38 and CD28 were investigated as potential targets in PTCL. A cohort of FFPE tissue samples (n = 250) were analyzed using IHC and / or fluorescence multiplex labeling with tumor cell markers. CD28 and CD38 showed heterogeneous expression in different PTCL entities. Figure 1A Significantly higher CD38 expression was observed in entities from the innate immune system containing more cytotoxic cells (such as natural killer (NK) cells) compared to entities lacking cytotoxic cells, while CD28 expression showed a significantly different pattern in the opposite direction. Figure 1B ). Figure 1C This study summarized the percentages of samples, measured by IHC semi-quantitative scoring, that showed co-expression of CD38 and CD28, expression of only a single marker, or expression of neither marker. The CD38 and CD28 expression patterns in circulating tumor cells were also analyzed using data from PTCL-follicular helper T-cell (TFH) patients. Figure 1D ) and non-PTCL-TFH patients (e.g., Cezari syndrome and mycosis fungoides; Figure 1E Blood samples were analyzed by flow cytometry. Rafer cases of co-expression of the two markers were identified. IHC staining results for CD3, CD28, and CD38 in different PTCL entities are summarized in [the table / section / etc.]. Figures 4A-4E .

[0207] Angioimmunoblastic T-cell lymphoma (AITL) and PTCL-NOS (PTCL-NOS) are the two most common types, accounting for over 50% of PTCL cases. AITL has a rich tumor microenvironment (TME) and typically contains a large number of CD4+ tumor-infiltrating lymphocytes (TILs). Analysis of this PTCL entity is more complex, but due to multiple labeling using tumor markers such as PD-1 or ICOS, several cases with CD28 expression and some cases with co-expression of CD28 and CD38 have been observed. Figures 2A-2D CD28 staining was detected in atypical lymphoid cells and tumor microenvironment cells. In lymph nodes associated with lymphoma ( Figure 2A Intermediate image and Figure 2C (middle image) or skin ( Figure 2B Intermediate image and Figure 2D CD38 staining was detected in the middle image. In AITL, CD38 staining is heterogeneous in tumor cells, with plasma cells showing strong positivity (…). Figure 2A (Intermediate image), while it was negative in SS ( Figure 2B (Middle image). PD1 staining highlights a large number of tumor cells, while the diffuse polymorphic infiltration area remains unstained. Figure 2A (Right image). AITL multiple staining ( Figure 2A The right image shows different distributions: one type consists of negative tumor cells, another type consists of double-positive (CD28 and CD38) cells, and the third type consists of tumor cells expressing only one target. Multiple staining of SS cells shows that the main co-expression is between CD28 and PD1. Figure 2B (Right image). ALCL tumor cells are positive for CD28 staining ( Figure 2C (Left image), while CD38 staining was not positive ( Figure 2C (Middle image). ALK staining highlights a large number of tumor cells. Multiple staining shows that CD28 is co-expressed with ALK in tumor cells, but not with CD38. Figure 2C Right image). CD28 in ENKTCL skin manifestation ( Figure 2D (Left image) and CD38 ( Figure 2D (Middle image) Staining shows a large number of strongly positive skin tumor cells with marked vascularity. Granulase B (GrB) staining highlights the large number of tumor cells. Immunofluorescence multiplex CD38, CD28, and GrB staining shows co-expression of CD38 and CD28 in GrB-positive cells. Figure 2D (Right image).

[0208] Next, functional studies were conducted to examine the cytotoxic effects of the anti-CD38 / CD28xCD3 trispecific binding protein on relevant cells. In vitro administration of the anti-CD38 / CD28xCD3 binding protein induced cytotoxic lysis in several PTCL cell lines, including SEAX (Cezari syndrome cell line). Figure 3A H9 (transformed MF cell line); Figure 3B ),DERL2 (hepatic and splenic T-cell lymphoma or PTCL cell line of the innate immune system; Figure 3C ), and several leukemia cell lines, including MOLT4 (acute lymphoblastic leukemia cell line); Figure 3D Jurkat (acute T-cell leukemia); Figure 3E ) and YT (T / NK cell leukemia; Figure 3F Cell lines. Mutant forms of the trispecific binding protein with null CD38 and / or CD28 binding sites were also tested to demonstrate the specificity of this effect on the presence of CD3, CD28, and CD38 binding sites. Administration of anti-CD38 / CD28xCD3 binding protein also stimulated effector cells PBMC-specific activation (unactivated T cell targets). The cytotoxic effects of anti-CD38 / CD28xCD3 trispecific binding protein were also observed in two different T leukemia cell lines YTLT ( Figure 5A ) and MOLT4 ( Figure 5B This was demonstrated in a study where these cell lines were used as targets and sorted CD3+ T lymphocytes were used as effector cells (E:T = 10:1). The expression of CD3, CD28, and CD38 in each cell line as determined by flow cytometry is shown in Table 1.

[0209] In summary, these data suggest that trispecific anti-CD38 / CD28xCD3 binding protein has potential therapeutic value as a novel immunotherapy strategy for the treatment of PTCL. Table 1. Summary of CD3, CD28, and CD38 phenotypes of cell lines derived from T-lymphoma or T-leukemia (phenotypes were determined by flow cytometry).

[0210] While this disclosure includes various embodiments, it should be understood that variations and modifications will occur to those skilled in the art. Therefore, the appended claims are intended to cover all such equivalent variations falling within the scope of this disclosure. Furthermore, the section headings used herein are for organizational purposes only and should not be construed as limiting the subject matter.

[0211] Unless otherwise clearly indicated, each embodiment described herein may be combined with any other one or more embodiments. Specifically, unless otherwise clearly indicated, any feature or embodiment indicated as preferred or advantageous may be combined with any other one or more features or embodiments indicated as preferred or advantageous.

[0212] All references cited in this application are explicitly incorporated herein by reference.

Claims

1. A method for treating peripheral T-cell lymphoma (PTCL) in an individual in need, the method comprising administering an effective amount of an anti-CD38 T-cell binder to the individual.

2. The method of claim 1, wherein the anti-CD38 T cell binder is a trispecific binding protein comprising a first antigen-binding site that specifically binds to a CD38 polypeptide, a second antigen-binding site that specifically binds to a CD28 polypeptide, and a third antigen-binding site that specifically binds to a CD3 polypeptide.

3. The method of claim 1, wherein the anti-CD38 T cell binder is a bispecific binding protein comprising a first antigen-binding site that specifically binds to a CD38 polypeptide and a second antigen-binding site that specifically binds to a CD3 polypeptide.

4. The method of claim 2 or claim 3, wherein the first antigen-binding site of the CD38 polypeptide comprises: (a) An antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence containing the amino acid sequence GYTFTSFN (SEQ ID NO: 31) or GYTFTSYA (SEQ ID NO: 37), a CDR-H2 sequence containing the amino acid sequence IYPGNGGT (SEQ ID NO: 32) or IYPGQGGT (SEQ ID NO: 38), and a CDR-H3 sequence containing the amino acid sequence ARTGGLRRAYFTY (SEQ ID NO: 33); and (b) The antibody light chain variable (VL) domain comprises a CDR-L1 sequence containing an amino acid sequence of ESVDSYGNGF (SEQ ID NO: 34) or QSVSSYGQGF (SEQ ID NO: 39), a CDR-L2 sequence containing an amino acid sequence of LAS or GAS, and a CDR-L3 sequence containing an amino acid sequence of QQNKEDPWT (SEQ ID NO: 36).

5. The method of claim 4, wherein the first antigen binding site comprises: (a) An antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence containing the amino acid sequence GYTFTSYA (SEQ ID NO: 37), a CDR-H2 sequence containing the amino acid sequence IYPGQGGT (SEQ ID NO: 38), and a CDR-H3 sequence containing the amino acid sequence ARTGGLRRAYFTY (SEQ ID NO: 33); and (b) The antibody light chain variable (VL) domain comprises a CDR-L1 sequence containing the amino acid sequence QSVSSYGQGF (SEQ ID NO: 39), a CDR-L2 sequence containing the amino acid sequence GAS, and a CDR-L3 sequence containing the amino acid sequence QQNKEDPWT (SEQ ID NO: 36).

6. The method of claim 4 or claim 5, wherein the first antigen binding site comprises a VH domain and a VL domain, the VH domain comprising the amino acid sequence of SEQ ID NO: 13, and the VL domain comprising the amino acid sequence of SEQ ID NO:

14.

7. The method of claim 4, wherein the first antigen binding site comprises: (a) An antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence containing the amino acid sequence GYTFTSFN (SEQ ID NO: 31), a CDR-H2 sequence containing the amino acid sequence IYPGNGGT (SEQ ID NO: 32), and a CDR-H3 sequence containing the amino acid sequence ARTGGLRRAYFTY (SEQ ID NO: 33); and (b) The antibody light chain variable (VL) domain comprises a CDR-L1 sequence containing an amino acid sequence of ESVDSYGNGF (SEQ ID NO: 34), a CDR-L2 sequence containing an amino acid sequence of LAS, and a CDR-L3 sequence containing an amino acid sequence of QQNKEDPWT (SEQ ID NO: 36).

8. The method of claim 4 or claim 7, wherein the first antigen binding site comprises: (a) A VH domain and a VL domain, wherein the VH domain contains the amino acid sequence of SEQ ID NO: 5 and the VL domain contains the amino acid sequence of SEQ ID NO: 6; (b) A VH domain and a VL domain, wherein the VH domain contains the amino acid sequence of SEQ ID NO: 17 and the VL domain contains the amino acid sequence of SEQ ID NO: 18; (c) A VH domain and a VL domain, wherein the VH domain contains the amino acid sequence of SEQ ID NO: 21 and the VL domain contains the amino acid sequence of SEQ ID NO: 18; or (d) A VH domain and a VL domain, wherein the VH domain contains the amino acid sequence of SEQ ID NO: 23 and the VL domain contains the amino acid sequence of SEQ ID NO:

18.

9. The method of claim 2 or claim 3, wherein the first antigen-binding site of the CD38 polypeptide comprises: (a) An antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence containing the amino acid sequence GFTFSSYG (SEQ ID NO: 41), a CDR-H2 sequence containing the amino acid sequence IWYDGSNK (SEQ ID NO: 42), and a CDR-H3 sequence containing the amino acid sequence ARMFRGAFDY (SEQ ID NO: 43); and (b) The antibody light chain variable (VL) domain comprises a CDR-L1 sequence containing the amino acid sequence QGIRND (SEQ ID NO: 44), a CDR-L2 sequence containing the amino acid sequence AAS, and a CDR-L3 sequence containing the amino acid sequence LQDYIYYPT (SEQ ID NO: 46).

10. The method of claim 9, wherein the first antigen binding site comprises a VH domain and a VL domain, the VH domain comprising the amino acid sequence of SEQ ID NO: 9, and the VL domain comprising the amino acid sequence of SEQ ID NO:

10.

11. The method of claim 2 or claim 3, wherein the first antigen-binding site of the CD38 polypeptide comprises: (a) An antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence containing the amino acid sequence GYTLTEFS (SEQ ID NO: 2), a CDR-H2 sequence containing the amino acid sequence FDPEDGET (SEQ ID NO: 3), and a CDR-H3 sequence containing the amino acid sequence TTGRFFDWF (SEQ ID NO: 4); and (b) The antibody light chain variable (VL) domain comprises a CDR-L1 sequence containing the amino acid sequence of QSVISRF (SEQ ID NO: 7), a CDR-L2 sequence containing the amino acid sequence of GAS, and a CDR-L3 sequence containing the amino acid sequence of QQDSNLPIT (SEQ ID NO: 11).

12. The method of claim 11, wherein the first antigen binding site comprises a VH domain and a VL domain, the VH domain comprising the amino acid sequence of SEQ ID NO: 108, and the VL domain comprising the amino acid sequence of SEQ ID NO:

109.

13. The method of claim 2 or claim 3, wherein the first antigen-binding site of the CD38 polypeptide comprises: (a) An antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence containing the amino acid sequence GYAFTTYL (SEQ ID NO: 12), a CDR-H2 sequence containing the amino acid sequence INPGSGST (SEQ ID NO: 15), and a CDR-H3 sequence containing the amino acid sequence ARYAYGY (SEQ ID NO: 16); and (b) The antibody light chain variable (VL) domain comprising a CDR-L1 sequence containing the amino acid sequence QNVGTA (SEQ ID NO: 19), a CDR-L2 sequence containing the amino acid sequence SAS, and a CDR-L3 sequence containing the amino acid sequence QQYSTYPFT (SEQ ID NO: 22).

14. The method of claim 13, wherein the first antigen binding site comprises: (a) A VH domain and a VL domain, wherein the VH domain contains the amino acid sequence of SEQ ID NO: 110 and the VL domain contains the amino acid sequence of SEQ ID NO: 111; or (b) A VH domain and a VL domain, wherein the VH domain contains the amino acid sequence of SEQ ID NO: 116 and the VL domain contains the amino acid sequence of SEQ ID NO:

117.

15. The method of claim 2 or claim 3, wherein the first antigen-binding site of the CD38 polypeptide comprises: (a) An antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence containing the amino acid sequence GYSFTNYA (SEQ ID NO: 24), a CDR-H2 sequence containing the amino acid sequence ISPHYGDT (SEQ ID NO: 25), and a CDR-H3 sequence containing the amino acid sequence ARRFEGFYYSMDY (SEQ ID NO: 26); and (b) The antibody light chain variable (VL) domain comprises a CDR-L1 sequence containing the amino acid sequence QSLVHSNGNTY (SEQ ID NO: 27), a CDR-L2 sequence containing the amino acid sequence KVS, and a CDR-L3 sequence containing the amino acid sequence SQSTHVPLT (SEQ ID NO: 29).

16. The method of claim 15, wherein the first antigen binding site comprises: (a) A VH domain and a VL domain, wherein the VH domain contains the amino acid sequence of SEQ ID NO: 112 and the VL domain contains the amino acid sequence of SEQ ID NO: 113; or (b) A VH domain and a VL domain, wherein the VH domain contains the amino acid sequence of SEQ ID NO: 118 and the VL domain contains the amino acid sequence of SEQ ID NO:

119.

17. The method of claim 2 or claim 3, wherein the first antigen-binding site of the CD38 polypeptide comprises: (a) An antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence containing the amino acid sequence GFTFSSYG (SEQ ID NO: 41), a CDR-H2 sequence containing the amino acid sequence IWYDGSNK (SEQ ID NO: 42), and a CDR-H3 sequence containing the amino acid sequence ARDPGLRYFDGGMDV (SEQ ID NO: 106); and (b) The antibody light chain variable (VL) domain comprises a CDR-L1 sequence containing the amino acid sequence QGISSY (SEQ ID NO: 107), a CDR-L2 sequence containing the amino acid sequence AAS, and a CDR-L3 sequence containing the amino acid sequence QQLNSFPYT (SEQ ID NO: 229).

18. The method of claim 17, wherein the first antigen binding site comprises a VH domain and a VL domain, the VH domain comprising the amino acid sequence of SEQ ID NO: 114, and the VL domain comprising the amino acid sequence of SEQ ID NO:

115.

19. The method of any one of claims 2-18, wherein the antigen-binding site of the CD3-binding polypeptide comprises: (a) An antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence containing the amino acid sequence GFTFTKAW (SEQ ID NO: 120), a CDR-H2 sequence containing the amino acid sequence IKDKSNSYAT (SEQ ID NO: 121), and a CDR-H3 sequence containing the amino acid sequence RGVYYALSPFDY (SEQ ID NO: 122); and (b) The antibody light chain variable (VL) domain comprising a CDR-L1 sequence containing the amino acid sequence QSLVHX1NX2X3TY (where X1 is E or Q, X2 is A or L, and X3 is Q, R, or F) (SEQ ID NO: 131), a CDR-L2 sequence containing the amino acid sequence KVS, and a CDR-L3 sequence containing the amino acid sequence GQGTQYPFT (SEQ ID NO: 130).

20. The method of claim 19, wherein the CDR-L1 sequence of the VL domain of the antigen-binding site of the CD3 polypeptide is selected from the group consisting of: QSLVHNNANTY (SEQ ID NO: 123), QSLVHQNAQTY (SEQ ID NO: 124), QSLVHENLQTY (SEQ ID NO: 125), QSLVHENLFTY (SEQ ID NO: 126), QSLVHENLRTY (SEQ ID NO: 127), and QSLVHDNAQTY (SEQ ID NO: 128).

21. The method of claim 19 or claim 20, wherein the antigen-binding site of the CD3-binding polypeptide comprises a VH domain and a VL domain, the VH domain comprising the amino acid sequence of SEQ ID NO: 53 or 138, and the VL domain comprising the amino acid sequence of SEQ ID NO: 54, 133, 134, 135, 136, or 137.

22. The method of claim 21, wherein the antigen-binding site of the CD3-binding polypeptide comprises a VH domain and a VL domain, the VH domain comprising the amino acid sequence of SEQ ID NO: 53, and the VL domain comprising the amino acid sequence of SEQ ID NO:

54.

23. The method of any one of claims 2-18, wherein the antigen-binding site of the CD3-binding polypeptide comprises: (a) An antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence containing the amino acid sequence GFTFTKAW (SEQ ID NO: 120), a CDR-H2 sequence containing the amino acid sequence IKDKSNSYAT (SEQ ID NO: 121), and a CDR-H3 sequence containing the amino acid sequence RGVYYALSPFDY (SEQ ID NO: 122); and (b) The antibody light chain variable (VL) domain comprises a CDR-L1 sequence containing the amino acid sequence QSLVHNNGNTY (SEQ ID NO: 218), a CDR-L2 sequence containing the amino acid sequence KVS, and a CDR-L3 sequence containing the amino acid sequence GQGTQYPFT (SEQ ID NO: 130).

24. The method of claim 23, wherein the third antigen binding site comprises a VH domain and a VL domain, the VH domain comprising the amino acid sequence of SEQ ID NO: 84, and the VL domain comprising the amino acid sequence of SEQ ID NO:

85.

25. The method of any one of claims 2 and 4-24, wherein the antigen-binding site of the CD28 polypeptide comprises: (a) An antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence containing the amino acid sequence GYTFTSYY (SEQ ID NO: 139), a CDR-H2 sequence containing the amino acid sequence IYPGNVNT (SEQ ID NO: 140), and a CDR-H3 sequence containing the amino acid sequence TRSHYGLDWNFDV (SEQ ID NO: 141); and (b) The antibody light chain variable (VL) domain comprises a CDR-L1 sequence containing the amino acid sequence QNIYVW (SEQ ID NO: 142), a CDR-L2 sequence containing the amino acid sequence KAS, and a CDR-L3 sequence containing the amino acid sequence QQGQTYPY (SEQ ID NO: 144).

26. The method of claim 25, wherein the antigen-binding site of the CD28-binding polypeptide comprises a VH domain and a VL domain, the VH domain comprising the amino acid sequence of SEQ ID NO: 49, and the VL domain comprising the amino acid sequence of SEQ ID NO:

50.

27. The method of any one of claims 2 and 4-24, wherein the antigen-binding site of the CD28 polypeptide comprises: (a) An antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence containing the amino acid sequence GFSLSDYG (SEQ ID NO: 212), a CDR-H2 sequence containing the amino acid sequence IWAGGGT (SEQ ID NO: 213), and a CDR-H3 sequence containing the amino acid sequence ARDKGYSYYYSMDY (SEQ ID NO: 214); and (b) The antibody light chain variable (VL) domain comprises a CDR-L1 sequence containing the amino acid sequence ESVEYYVTSL (SEQ ID NO: 215), a CDR-L2 sequence containing the amino acid sequence AAS, and a CDR-L3 sequence containing the amino acid sequence QQSRKVPYT (SEQ ID NO: 217).

28. The method of claim 27, wherein the second antigen binding site comprises a VH domain and a VL domain, the VH domain comprising the amino acid sequence of SEQ ID NO: 51, and the VL domain comprising the amino acid sequence of SEQ ID NO:

52.

29. The method of any one of claims 2 and 4-28, wherein the trispecific binding protein comprises four polypeptide chains forming the three antigen-binding sites, wherein the first polypeptide chain comprises a structure represented by the following formula: V L2 -L1-V L1 -L2-C L [I] Furthermore, the second polypeptide chain contains a structure represented by the following formula: V H1 -L3-V H2 -L4-C H1 -Hinge-C H2 -C H3 [II] Furthermore, the third polypeptide chain contains a structure represented by the following formula: V H3 -C H1 -Hinge-C H2 -C H3 [III] Furthermore, the fourth polypeptide chain contains a structure represented by the following formula: V L3 -C L [IV] in: V L1 It is the variable domain of the first immunoglobulin light chain; V L2 It is the variable domain of the second immunoglobulin light chain; V L3 It is the variable domain of the third immunoglobulin light chain; V H1 It is the variable domain of the first immunoglobulin heavy chain; V H2 It is the variable domain of the second immunoglobulin heavy chain; V H3 It is the variable domain of the third immunoglobulin heavy chain; C L It is the constant structural domain of the immunoglobulin light chain; C H1 It is immunoglobulin C H1 Heavy chain constant structural domain; C H2 It is immunoglobulin C H2 Heavy chain constant structural domain; C H3 It is immunoglobulin C H3 Heavy chain constant structural domain; The hinge is the connection of C H1 and C H2 The immunoglobulin hinge region of the structural domain; and L1, L2, L3, and L4 are amino acid linkers; The polypeptide having Formula I and the polypeptide having Formula II form a cross-linked light chain-heavy chain pair; wherein V H1 and V L1 Forming one of these three antigen-binding sites; among which V H2 and V L2 Forming another of these three antigen-binding sites; and where V H3 and V L3 This forms another of the three antigen-binding sites.

30. The method of claim 29, wherein: (a) V H1 and V L1 This forms the second antigen-binding site for the CD28 peptide, V H2 and V L2 This forms the third antigen-binding site for the CD3-binding polypeptide, and V H3 and V L3 Forming the first antigen-binding site for the CD38 polypeptide; or (b) V H1 and V L1 This forms the third antigen-binding site for the CD3-binding polypeptide, V H2 and V L2 This forms the second antigen-binding site for the CD28 peptide, and V H3 and V L3 This forms the first antigen-binding site for the CD38 polypeptide.

31. The method of claim 29, wherein: (a) V H1 and V L1 This forms the second antigen-binding site for the CD28 peptide, V H2 and V L2 This forms the first antigen-binding site for the CD38 polypeptide, and V H3 and V L3 Forming the third antigen-binding site for the CD3-binding polypeptide; or (b) V H1 and V L1 This forms the third antigen-binding site for the CD3-binding polypeptide, V H2 and V L2 This forms the first antigen-binding site for the CD38 polypeptide, and V H3 and V L3 This forms the second antigen-binding site for the CD28 polypeptide.

32. The method of claim 29, wherein: (a) V H1 and V L1 This forms the first antigen-binding site for the CD38 polypeptide, V H2 and V L2 This forms the third antigen-binding site for the CD3-binding polypeptide, and V H3 and V L3 Forming a second antigen-binding site for the CD28 polypeptide; or (b) V H1 and V L1 This forms the first antigen-binding site for the CD38 polypeptide, V H2 and V L2 This forms the second antigen-binding site for the CD28 peptide, and V H3 and V L3 This forms the third antigen-binding site for the CD3-binding polypeptide.

33. The method of claim 29, wherein: V H1 and V L1 This forms the second antigen-binding site for the CD28 peptide, V H2 and V L2 This forms the third antigen-binding site for the CD3-binding polypeptide, and V H3 and V L3 This forms the first antigen-binding site for the CD38 polypeptide; V H1 The CDR-H1 sequence contains the amino acid sequence GYTFTSYY (SEQ ID NO: 139), the CDR-H2 sequence contains the amino acid sequence IYPGNVNT (SEQ ID NO: 140), and the CDR-H3 sequence contains the amino acid sequence TRSHYGLDWNFDV (SEQ ID NO: 141). L1 The CDR-L1 sequence contains the amino acid sequence QNIYVW (SEQ ID NO: 142), the CDR-L2 sequence contains the amino acid sequence KAS, and the CDR-L3 sequence contains the amino acid sequence QQGQTYPY (SEQ ID NO: 144). V H2 The CDR-H1 sequence contains the amino acid sequence GFTFTKAW (SEQ ID NO: 120), the CDR-H2 sequence contains the amino acid sequence IKDKSNSYAT (SEQ ID NO: 121), and the CDR-H3 sequence contains the amino acid sequence RGVYYALSPFDY (SEQ ID NO: 122). L2 The CDR-L1 sequence contains the amino acid sequence QSLVHNNANTY (SEQ ID NO: 123), the CDR-L2 sequence contains the amino acid sequence KVS, and the CDR-L3 sequence contains the amino acid sequence GQGTQYPFT (SEQ ID NO: 130); and V H3 The CDR-H1 sequence contains the amino acid sequence GYTFTSYA (SEQ ID NO: 37), the CDR-H2 sequence contains the amino acid sequence IYPGQGGT (SEQ ID NO: 38), and the CDR-H3 sequence contains the amino acid sequence ARTGGLRRAYFTY (SEQ ID NO: 33). L3 The CDR-L1 sequence contains the amino acid sequence QSVSSYGQGF (SEQ ID NO: 39), the CDR-L2 sequence contains the amino acid sequence GAS, and the CDR-L3 sequence contains the amino acid sequence QQNKEDPWT (SEQ ID NO: 36).

34. The method of claim 29, wherein V H1 and V L1 This forms the second antigen-binding site for the CD28 peptide, V H2 and V L2 This forms the third antigen-binding site for the CD3-binding polypeptide, and V H3 and V L3 This forms the first antigen-binding site for the CD38 polypeptide; wherein V H1 Contains the amino acid sequence of SEQ ID NO: 49, and V L1 Contains the amino acid sequence of SEQ ID NO: 50; wherein V H2 Contains the amino acid sequence of SEQ ID NO: 53, and V L2 Contains the amino acid sequence of SEQ ID NO: 54; and wherein V H3 Contains the amino acid sequence of SEQ ID NO: 13, and V L3 It contains the amino acid sequence of SEQ ID NO:

14.

35. The method according to any one of claims 29-34, wherein: (a) L1 contains the sequence GQPKAAP (SEQ ID NO: 58), L2 contains the sequence TKGPS (SEQ ID NO: 57), L3 contains amino acid S, and L4 contains the sequence RT; (b) L1 contains the sequence GGGGSGGGGS (SEQ ID NO: 55), L2 contains the sequence GGGGSGGGGS (SEQ ID NO: 55), L3 has a length of 0 amino acids, and L4 has a length of 0 amino acids; (c) L1 contains the sequence GGSGSSGSGG (SEQ ID NO: 59), L2 contains the sequence GGSGSSGSGG (SEQ ID NO: 59), L3 is 0 amino acids long, and L4 is 0 amino acids long; or (d) L1 contains the sequence GGGGSGGGGSGGGGS (SEQ ID NO: 56), L2 has a length of 0 amino acids, L3 contains the sequence GGGGSGGGGSGGGGS (SEQ ID NO: 56), and L4 has a length of 0 amino acids.

36. The method of any one of claims 29-34, wherein at least one of L1, L2, L3 or L4 comprises the sequence DKTHT (SEQ ID NO: 147).

37. The method of claim 36, wherein L1, L2, L3 and L4 comprise the sequence DKTHT (SEQ ID NO: 147).

38. The method of any one of claims 29-37, wherein the hinges of the second polypeptide chain and the third polypeptide chain are C H2 -C H3 The structural domain is the human IgG4 hinge-C H2 -C H3 Structural domain, and in which these hinges - C H2 -C H3 Each of the domains contains an amino acid substitution at positions 234 and 235 corresponding to human IgG4 according to the EU index, wherein these amino acid substitutions are F234A and L235A.

39. The method of any one of claims 29-37, wherein the hinges of the second polypeptide chain and the third polypeptide chain are C H2 -C H3 The structural domain is the human IgG4 hinge-C H2 -C H3 Structural domain, and in which these hinges - C H2 -C H3 Each of the domains contains amino acid substitutions at positions 233-236 corresponding to human IgG4 according to the EU index, wherein these amino acid substitutions are E233P, F234V, L235A and a deletion at position 236.

40. The method of any one of claims 29-37, wherein the hinges of the second polypeptide chain and the third polypeptide chain are C H2 -C H3 The structural domain is the human IgG4 hinge-C H2 -C H3 Structural domain, and in which these hinges - C H2 -C H3 Each of the domains contains amino acid substitutions at positions 228 and 409 corresponding to human IgG4 according to the EU index, wherein these amino acid substitutions are S228P and R409K.

41. The method of any one of claims 29-37, wherein the hinges of the second polypeptide chain and the third polypeptide chain are C H2 -C H3 The structural domain is the human IgG1 hinge-C H2 -C H3 Structural domain, and in which these hinges - C H2 -C H3 Each of the domains contains an amino acid substitution at positions 234, 235, and 329 corresponding to human IgG1 according to the EU index, wherein these amino acid substitutions are L234A, L235A, and P329A.

42. The method of any one of claims 29-37, wherein the hinges of the second polypeptide chain and the third polypeptide chain are C H2 -C H3 The structural domain is the human IgG1 hinge-C H2 -C H3 Structural domain, and in which these hinges - C H2 -C H3 Each of the domains contains an amino acid substitution at positions 298, 299, and 300 corresponding to human IgG1 according to the EU index, wherein the amino acid substitutions are S298N, T299A, and Y300S.

43. The method of any one of claims 29-42, wherein the hinge of the second polypeptide chain is C H2 -C H3 The domain contains amino acid substitutions at positions 349, 366, 368, and 407 corresponding to human IgG1 or IgG4 according to the EU index, wherein these amino acid substitutions are Y349C, T366S, L368A, and Y407V; and wherein the hinge of the third polypeptide chain is C H2 -C H3 The domain contains amino acid substitutions at positions 354 and 366 corresponding to human IgG1 or IgG4 according to the EU index, wherein these amino acid substitutions are S354C and T366W.

44. The method of any one of claims 29-42, wherein the hinge-C of the second polypeptide chain H2 -C H3 The domain contains amino acid substitutions at positions 354 and 366 corresponding to human IgG1 or IgG4 according to the EU index, wherein these amino acid substitutions are S354C and T366W; and wherein the hinge of the third polypeptide chain is C H2 -C H3 The domain contains amino acid substitutions at positions 349, 366, 368, and 407 corresponding to human IgG1 or IgG4 according to the EU index, wherein these amino acid substitutions are Y349C, T366S, L368A, and Y407V.

45. The method of claim 29, wherein: (a) The first polypeptide chain contains the amino acid sequence of SEQ ID NO: 61, the second polypeptide chain contains the amino acid sequence of SEQ ID NO: 60, the third polypeptide chain contains the amino acid sequence of SEQ ID NO: 62, and the fourth polypeptide chain contains the amino acid sequence of SEQ ID NO: 63; (b) The first polypeptide chain contains the amino acid sequence of SEQ ID NO: 61, the second polypeptide chain contains the amino acid sequence of SEQ ID NO: 64, the third polypeptide chain contains the amino acid sequence of SEQ ID NO: 65, and the fourth polypeptide chain contains the amino acid sequence of SEQ ID NO: 63; (c) The first polypeptide chain contains the amino acid sequence of SEQ ID NO: 61, the second polypeptide chain contains the amino acid sequence of SEQ ID NO: 66, the third polypeptide chain contains the amino acid sequence of SEQ ID NO: 67, and the fourth polypeptide chain contains the amino acid sequence of SEQ ID NO: 63; (d) The first polypeptide chain contains the amino acid sequence of SEQ ID NO: 61, the second polypeptide chain contains the amino acid sequence of SEQ ID NO: 60, the third polypeptide chain contains the amino acid sequence of SEQ ID NO: 68, and the fourth polypeptide chain contains the amino acid sequence of SEQ ID NO: 69; (e) The first polypeptide chain contains the amino acid sequence of SEQ ID NO: 61, the second polypeptide chain contains the amino acid sequence of SEQ ID NO: 64, the third polypeptide chain contains the amino acid sequence of SEQ ID NO: 70, and the fourth polypeptide chain contains the amino acid sequence of SEQ ID NO: 69; (f) The first polypeptide chain contains the amino acid sequence of SEQ ID NO: 61, the second polypeptide chain contains the amino acid sequence of SEQ ID NO: 66, the third polypeptide chain contains the amino acid sequence of SEQ ID NO: 71, and the fourth polypeptide chain contains the amino acid sequence of SEQ ID NO: 69; (g) The first polypeptide chain contains the amino acid sequence of SEQ ID NO: 148, the second polypeptide chain contains the amino acid sequence of SEQ ID NO: 149, the third polypeptide chain contains the amino acid sequence of SEQ ID NO: 150, and the fourth polypeptide chain contains the amino acid sequence of SEQ ID NO:

151. (h) The first polypeptide chain contains the amino acid sequence of SEQ ID NO: 152, the second polypeptide chain contains the amino acid sequence of SEQ ID NO: 153, the third polypeptide chain contains the amino acid sequence of SEQ ID NO: 154, and the fourth polypeptide chain contains the amino acid sequence of SEQ ID NO:

155. (i) The first polypeptide chain contains the amino acid sequence of SEQ ID NO: 156, the second polypeptide chain contains the amino acid sequence of SEQ ID NO: 157, the third polypeptide chain contains the amino acid sequence of SEQ ID NO: 158, and the fourth polypeptide chain contains the amino acid sequence of SEQ ID NO:

159. (j) The first polypeptide chain contains the amino acid sequence of SEQ ID NO: 160, the second polypeptide chain contains the amino acid sequence of SEQ ID NO: 161, the third polypeptide chain contains the amino acid sequence of SEQ ID NO: 162, and the fourth polypeptide chain contains the amino acid sequence of SEQ ID NO:

163. (k) The first polypeptide chain contains the amino acid sequence of SEQ ID NO: 164, the second polypeptide chain contains the amino acid sequence of SEQ ID NO: 165, the third polypeptide chain contains the amino acid sequence of SEQ ID NO: 166, and the fourth polypeptide chain contains the amino acid sequence of SEQ ID NO:

167. (l) The first polypeptide chain contains the amino acid sequence of SEQ ID NO: 168, the second polypeptide chain contains the amino acid sequence of SEQ ID NO: 169, the third polypeptide chain contains the amino acid sequence of SEQ ID NO: 170, and the fourth polypeptide chain contains the amino acid sequence of SEQ ID NO: 171; (m) The first polypeptide chain contains the amino acid sequence of SEQ ID NO: 172, the second polypeptide chain contains the amino acid sequence of SEQ ID NO: 173, the third polypeptide chain contains the amino acid sequence of SEQ ID NO: 174, and the fourth polypeptide chain contains the amino acid sequence of SEQ ID NO: 175; or (n) The first polypeptide chain contains the amino acid sequence of SEQ ID NO: 176, the second polypeptide chain contains the amino acid sequence of SEQ ID NO: 177, the third polypeptide chain contains the amino acid sequence of SEQ ID NO: 178, and the fourth polypeptide chain contains the amino acid sequence of SEQ ID NO:

179.

46. ​​The method of any one of claims 2-45, wherein the CD38 polypeptide is a human CD38 polypeptide.

47. The method of any one of claims 2-46, wherein the CD3 polypeptide is a human CD3 polypeptide.

48. The method of any one of claims 2 and 4-47, wherein the CD28 polypeptide is a human CD28 polypeptide.

49. The method of any one of claims 1-48, wherein the PTCL is angioimmunoblastic T-cell lymphoma (AITL), hepatosplenic T-cell lymphoma (HSTL), adult T-cell leukemia / lymphoma (ATLL), extranodal NK / T-cell lymphoma (ENKTCL), mycosis fungoides (MF), Cezari syndrome (SS), anaplastic large cell ALK-lymphoma (ALCL), PTCL-nospecific (NOS), enteropathy-type T-cell lymphoma (EATL), monomorphic epithelial intestinal T-cell lymphoma (MEITL), primary cutaneous CD4+ lymphoproliferative disorder, subcutaneous panniculitis-like T-cell lymphoma (SCTCL), or primary cutaneous γδ T-cell lymphoma.

50. The method of any one of claims 1-48, wherein the PTCL is hepatosplenic T-cell lymphoma (HSTL), Cezari syndrome (SS), or mycosis fungoides (MF).

51. The method of claim 49 or claim 50, wherein the mycosis fungoides (MF) is transformed mycosis fungoides.

52. The method of any one of claims 1-51, wherein the PTCL cells express CD38 and / or CD28.

53. The method of any one of claims 1-52, wherein the individual is a person.

54. An anti-CD38 T-cell binder for use in a method of treating peripheral T-cell lymphoma (PTCL) in an individual in need, the method comprising administering an effective amount of the anti-CD38 T-cell binder to the individual.

55. Use of anti-CD38 T-cell binders in the manufacture of drugs for the treatment of peripheral T-cell lymphoma (PTCL) in individuals in need.

56. A kit comprising an anti-CD38 T-cell binder and instructions for use in administering an effective amount of the anti-CD38 T-cell binder to an individual with peripheral T-cell lymphoma (PTCL) according to the method described in any one of claims 1-53.

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