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WO2025027003A1 - Méthodes et utilisations pour des activateurs de lymphocytes t anti-cd38 dans le traitement de lymphomes t périphériques - Google Patents

Méthodes et utilisations pour des activateurs de lymphocytes t anti-cd38 dans le traitement de lymphomes t périphériques Download PDF

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Publication number
WO2025027003A1
WO2025027003A1 PCT/EP2024/071517 EP2024071517W WO2025027003A1 WO 2025027003 A1 WO2025027003 A1 WO 2025027003A1 EP 2024071517 W EP2024071517 W EP 2024071517W WO 2025027003 A1 WO2025027003 A1 WO 2025027003A1
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amino acid
seq
acid sequence
sequence
polypeptide
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Helgi Van De Velde
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Sanofi SA
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Sanofi SA
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Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
    • C07K16/2896—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against molecules with a "CD"-designation, not provided for elsewhere
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A61P35/02—Antineoplastic agents specific for leukemia
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
    • C07K16/2803—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the immunoglobulin superfamily
    • C07K16/2809—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the immunoglobulin superfamily against the T-cell receptor (TcR)-CD3 complex
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
    • C07K16/2803—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the immunoglobulin superfamily
    • C07K16/2818—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the immunoglobulin superfamily against CD28 or CD152
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/30—Immunoglobulins specific features characterized by aspects of specificity or valency
    • C07K2317/31—Immunoglobulins specific features characterized by aspects of specificity or valency multispecific

Definitions

  • PTCL peripheral T-cell lymphoma
  • 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.
  • 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.
  • the first antigen binding site that binds 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
  • 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).
  • VH antibody heavy chain variable
  • 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.
  • 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 GYTFTSFN (SEQ ID NO:31), a CDR-H2 sequence comprising the amino acid sequence of IYPGNGGT (SEQ ID NO:32), and a CDR-H3 sequence comprising the amino acid sequence of 2 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), a CDR-L2 sequence comprising the amino acid sequence of LAS, and a CDR-L3 sequence comprising the amino acid sequence of QQNKEDPWT (VH) domain comprising
  • the first antigen binding site comprises a VH domain comprising the amino acid sequence of SEQ ID NO:5, and a VL domain comprising the amino acid sequence of SEQ ID NO:6. In some embodiments, the first antigen binding site comprises a VH domain comprising the amino acid sequence of SEQ ID NO:17, and a VL domain comprising the amino acid sequence of SEQ ID NO:18. In some embodiments, the first antigen binding site comprises a VH domain comprising the amino acid sequence of SEQ ID NO:21, and a VL domain comprising the amino acid sequence of SEQ ID NO:18.
  • the first antigen binding site comprises a VH domain comprising the amino acid sequence of SEQ ID NO:23, and a VL domain comprising the amino acid sequence of SEQ ID NO:18.
  • the first antigen binding site that binds a CD38 polypeptide comprises: (a) an antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence comprising the amino acid sequence of GFTFSSYG (SEQ ID NO:41), a CDR-H2 sequence comprising the amino acid sequence of IWYDGSNK (SEQ ID NO:42), and a CDR-H3 sequence comprising the amino acid sequence of ARMFRGAFDY (SEQ ID NO:43); and (b) an antibody light chain variable (VL) domain comprising a CDR-L1 sequence comprising the amino acid sequence of QGIRND (SEQ ID NO:44), a CDR-L2 sequence comprising the amino acid sequence of AAS, and a CDR-L3 sequence comprising the amino acid sequence of
  • the first antigen binding site comprises a VH domain comprising the amino acid sequence of SEQ ID NO:9, and a VL domain comprising the amino acid sequence of SEQ ID NO:10.
  • the first antigen binding site that binds a CD38 polypeptide comprises: (a) an antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence comprising the amino acid sequence of GYTLTEFS (SEQ ID NO:2), a CDR-H2 sequence comprising the amino acid sequence of FDPEDGET (SEQ ID NO:3), and a CDR-H3 sequence comprising the amino acid sequence of TTGRFFDWF (SEQ ID NO:4); and (b) an antibody light chain variable (VL) domain comprising a CDR-L1 sequence comprising the amino acid sequence of QSVISRF (SEQ ID NO:7), a CDR-L2 sequence comprising the amino acid sequence of GAS, and a CDR-L3 sequence comprising the amino acid sequence of QQDSN
  • the first antigen binding site that binds a CD38 polypeptide comprises: (a) an antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence comprising the amino acid sequence of GYAFTTYL (SEQ ID NO:12), a CDR-H2 sequence comprising the amino acid sequence of INPGSGST (SEQ ID NO:15), and a CDR-H3 sequence comprising the amino acid sequence of ARYAYGY (SEQ ID NO:16); and (b) an antibody light chain variable (VL) domain comprising a CDR- L1 sequence comprising the amino acid sequence of QNVGTA (SEQ ID NO:19), a CDR- L2 sequence comprising the amino acid sequence of SAS, and a CDR-L3 sequence comprising the amino acid sequence of QQYSTYPFT (SEQ ID NO:22).
  • VH antibody heavy chain variable
  • the first antigen binding site comprises a VH domain comprising the amino acid sequence of SEQ ID NO:110, and a VL domain comprising the amino acid sequence of SEQ ID NO:111. In some embodiments, the first antigen binding site comprises a VH domain comprising the amino acid sequence of SEQ ID NO:116, and a VL domain comprising the amino acid sequence of SEQ ID NO:117.
  • the first antigen binding site that binds a CD38 polypeptide comprises: (a) an antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence comprising the amino acid sequence of GYSFTNYA (SEQ ID NO:24), a CDR-H2 sequence comprising the amino acid sequence of ISPYYGDT (SEQ ID NO:25), and a CDR-H3 sequence comprising the amino acid sequence of ARRFEGFYYSMDY (SEQ ID NO:26); and (b) an antibody light chain variable (VL) domain comprising a CDR-L1 sequence comprising the amino acid sequence of QSLVHSNGNTY (SEQ ID NO:27), a CDR-L2 sequence comprising the amino acid sequence of KVS, and a CDR-L3 sequence comprising the amino acid sequence of SQSTHVPLT (SEQ ID NO:29).
  • VH antibody heavy chain variable
  • the first antigen binding site comprises a VH domain comprising the amino acid sequence of SEQ ID NO:112, and a VL domain comprising the amino acid sequence of SEQ ID NO:113. In some embodiments, the first antigen binding site comprises a VH domain comprising the amino acid sequence of SEQ ID NO:118, and a VL domain comprising the amino acid sequence of SEQ ID NO:119.
  • the first antigen binding site that binds a CD38 polypeptide comprises: (a) an antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence comprising the amino acid sequence of GFTFSSYG (SEQ ID NO:41), a CDR-H2 sequence comprising the amino acid sequence of IWYDGSNK (SEQ ID NO:42), and a CDR-H3 sequence comprising the amino acid sequence of ARDPGLRYFDGGMDV (SEQ ID NO:106); and (b) an antibody light chain variable (VL) domain comprising a 4
  • the first antigen binding site comprises a VH domain comprising the amino acid sequence of SEQ ID NO:114, and a VL domain comprising the amino acid sequence of SEQ ID NO:115.
  • the antigen binding site that binds a CD3 polypeptide comprises: (a) an antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence comprising the amino acid sequence of GFTFTKAW (SEQ ID NO:120), a CDR-H2 sequence comprising the amino acid sequence of IKDKSNSYAT (SEQ ID NO:121), and a CDR-H3 sequence comprising the amino acid sequence of RGVYYALSPFDY (SEQ ID NO:122); and (b) an antibody light chain variable (VL) domain comprising a CDR-L1 sequence comprising the amino acid sequence of QSLVHNNGNTY (SEQ ID NO:218), a CDR-L2 sequence comprising the amino acid sequence of KVS, and a CDR-L3 sequence comprising the amino acid sequence of GQGTQYPFT (SEQ ID NO:130).
  • VH antibody heavy chain variable
  • the third antigen binding site comprises a VH domain comprising the amino acid sequence of SEQ ID NO:84, and a VL domain comprising the amino acid sequence of SEQ ID NO:85.
  • the antigen binding site that binds a CD3 polypeptide comprises: (a) an antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence comprising the amino acid sequence of GFTFTKAW (SEQ ID NO:120), a CDR-H2 sequence comprising the amino acid sequence of IKDKSNSYAT (SEQ ID NO:121), and a CDR-H3 sequence comprising the amino acid sequence of RGVYYALSPFDY (SEQ ID NO:122); and (b) an antibody light chain variable (VL) domain comprising a CDR-L1 sequence comprising the amino acid sequence of QSLVHX1NX2X3TY, wherein X1 is E or Q, X2 is A or L, and X3 is Q, R, or F (SEQ ID NO:
  • the CDR-L1 sequence of the VL domain of the antigen binding site that binds a 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).
  • the VH domain of the antigen binding site that binds a CD3 polypeptide comprises the amino acid sequence of SEQ ID NO:53 or 138, and the VL domain of the 5
  • the antigen binding site that binds a CD3 polypeptide comprises the amino acid sequence of SEQ ID NO:54, 133, 134, 135, 136, or 137.
  • the antigen binding site that binds a CD28 polypeptide comprises: (a) an antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence comprising the amino acid sequence of GYTFTSYY (SEQ ID NO:139), a CDR-H2 sequence comprising the amino acid sequence of IYPGNVNT (SEQ ID NO:140), and a CDR-H3 sequence comprising the amino acid sequence of TRSHYGLDWNFDV (SEQ ID NO:141); and (b) an antibody light chain variable (VL) domain comprising a CDR-L1 sequence comprising the amino acid sequence of QNIYVW (SEQ ID NO:142), a CDR-L2 sequence comprising the amino acid sequence of KAS, and a CDR-L3 sequence compris
  • the antigen binding site that binds a CD28 polypeptide comprises a VH domain comprising the amino acid sequence of SEQ ID NO:49, and a VL domain comprising the amino acid sequence of SEQ ID NO:50.
  • the antigen binding site that binds a CD28 polypeptide comprises: (a) an antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence comprising the amino acid sequence of GFSLSDYG (SEQ ID NO:212), a CDR-H2 sequence comprising the amino acid sequence of IWAGGGT (SEQ ID NO:213), and a CDR-H3 sequence comprising the amino acid sequence of ARDKGYSYYYSMDY (SEQ ID NO:214); and (b) an antibody light chain variable (VL) domain comprising a CDR-L1 sequence comprising the amino acid sequence of ESVEYYVTSL (SEQ ID NO:215), a CDR-L2 sequence comprising the amino acid sequence of AAS, and
  • the antigen binding site that binds a CD28 polypeptide comprises a VH domain comprising the amino acid sequence of SEQ ID NO:51, and a VL domain comprising the amino acid sequence of SEQ ID NO:52.
  • the antigen binding site that binds a CD28 polypeptide comprises a VH domain comprising the amino acid sequence of SEQ ID NO:49, and a VL domain comprising the amino acid sequence of SEQ ID NO:50
  • the antigen binding site that binds a CD3 polypeptide comprises a VH domain comprising the amino acid sequence of SEQ ID NO:53 and a VL domain comprising the amino acid sequence of SEQ ID NO:54.
  • the trispecific binding protein comprises four polypeptide chains that form the three antigen 6
  • a first polypeptide chain comprises a structure represented by the formula: VL2-L1-VL1-L2-CL [I] and a second polypeptide chain comprises a structure represented by the formula: VH1-L3-VH2-L4-CH1-hinge-CH2-CH3 [II] and a third polypeptide chain comprises a structure represented by the formula: V H3 -C H1 -hinge-C H2 -C H3 [III] and a fourth polypeptide chain comprises a structure represented by the formula: VL3-CL [IV] wherein: V L1 is a first immunoglobulin light chain variable domain; VL2 is a second immunoglobulin light chain variable domain; VL3 is a third immunoglobulin light chain variable domain; V H1 is a first immunoglobulin heavy chain variable domain; VH2 is a second immunoglobulin heavy chain variable domain; VH3 is a third immunoglobulin heavy chain variable domain; C L is an immunoglobulin
  • VH1 and V L1 form the second antigen binding site that binds a CD28 polypeptide
  • V H2 and V L2 form the third antigen binding site that binds a CD3 polypeptide
  • V H3 and V L3 form the first antigen binding site that binds a CD38 polypeptide
  • VH1 and VL1 form the third antigen binding site that binds a CD3 polypeptide
  • VH2 and VL2 form the second antigen binding site that binds a CD28 polypeptide
  • V H3 and V L3 form the first antigen binding site that binds a CD38 polypeptide.
  • VH1 and VL1 form the second antigen binding site that binds a CD28 polypeptide
  • VH2 and VL2 form the first antigen binding site that binds a CD38 polypeptide
  • V H3 and V L3 form the third 7
  • V H1 and V L1 form the third antigen binding site that binds a CD3 polypeptide
  • VH2 and VL2 form the first antigen binding site that binds a CD38 polypeptide
  • VH3 and VL3 form the second antigen binding site that binds a CD28 polypeptide
  • V H1 and V L1 form the first antigen binding site that binds a CD38 polypeptide
  • VH2 and VL2 form the third antigen binding site that binds a CD3 polypeptide
  • VH3 and VL3 form the second antigen binding site that binds a CD28 polypeptide.
  • V H1 and V L1 form the first antigen binding site that binds a CD38 polypeptide
  • VH2 and VL2 form the second antigen binding site that binds a CD28 polypeptide
  • VH3 and VL3 form the third antigen binding site that binds a CD3 polypeptide.
  • L 1 comprises the sequence GQPKAAP (SEQ ID NO:58)
  • L 2 comprises the sequence TKGPS (SEQ ID NO:57)
  • L3 comprises the amino acid S
  • L4 comprises the sequence RT.
  • L1 comprises the sequence GGGGSGGGGS (SEQ ID NO:55)
  • L2 comprises the sequence GGGGSGGGGS (SEQ ID NO:55)
  • L 3 is 0 amino acids in length
  • L 4 is 0 amino acids in length.
  • L1 comprises the sequence GGSGSSGSGG (SEQ ID NO:59)
  • L2 comprises the sequence GGSGSSGSGG (SEQ ID NO:59)
  • L3 is 0 amino acids in length
  • L 4 is 0 amino acids in length.
  • L 1 comprises the sequence GGGGSGGGGSGGGGS (SEQ ID NO:56), L 2 is 0 amino acids in length, L3 comprises the sequence GGGGSGGGGSGGGGS (SEQ ID NO:56), and L4 is 0 amino acids in length.
  • at least one of L 1 , L 2 , L 3 or L 4 comprises the sequence DKTHT (SEQ ID NO:147).
  • L 1 , L 2 , L 3 and L 4 comprise the sequence DKTHT (SEQ ID NO:147).
  • the hinge-CH2-CH3 domains of the second and the third polypeptide chains are human IgG4 hinge-CH2-CH3 domains, and wherein the hinge-C H2 -C H3 domains each comprise amino acid substitutions at positions corresponding to positions 234 and 235 of human IgG4 according to EU Index, wherein the amino acid substitutions are F234A and L235A.
  • the hinge-C H2 -C H3 domains of the second and the third polypeptide chains are human IgG4 hinge-C H2 -C H3 domains, and wherein the hinge-C H2 -C H3 domains each comprise amino acid substitutions at positions corresponding to positions 233-236 of human IgG4 according to EU Index, wherein the amino acid substitutions are E233P, F234V, L235A, and a deletion at 236.
  • the hinge-C H2 -C H3 domains of the second and the third polypeptide chains are human IgG4 hinge-CH2-CH3 domains, and wherein the hinge-CH2- CH3 domains each comprise amino acid substitutions at positions corresponding to positions 228 and 409 of human IgG4 according to EU Index, wherein the amino acid substitutions 8
  • the hinge-C H2 -C H3 domains of the second and the third polypeptide chains are human IgG1 hinge-CH2-CH3 domains, and wherein the hinge-CH2-CH3 domains each comprise amino acid substitutions at positions corresponding to positions 234, 235, and 329 of human IgG1 according to EU Index, wherein the amino acid substitutions are L234A, L235A, and P329A.
  • the hinge-CH2- CH3 domains of the second and the third polypeptide chains are human IgG1 hinge-CH2-CH3 domains, and wherein the hinge-C H2 -C H3 domains each comprise amino acid substitutions at positions corresponding to positions 298, 299, and 300 of human IgG1 according to EU Index, wherein the amino acid substitutions are S298N, T299A, and Y300S.
  • the hinge-C H2 -C H3 domain of the second polypeptide chain comprises amino acid substitutions at positions corresponding to positions 349, 366, 368, and 407 of human IgG1 or IgG4 according to EU Index, wherein the amino acid substitutions are Y349C, T366S, L368A, and Y407V; and wherein the hinge-CH2-CH3 domain of the third polypeptide chain comprises amino acid substitutions at positions corresponding to positions 354 and 366 of human IgG1 or IgG4 according to EU Index, wherein the amino acid substitutions are S354C and T366W.
  • the hinge-CH2-CH3 domain of the second polypeptide chain comprises amino acid substitutions at positions corresponding to positions 354 and 366 of human IgG1 or IgG4 according to EU Index, wherein the amino acid substitutions are S354C and T366W; and wherein the hinge-CH2- C H3 domain of the third polypeptide chain comprises amino acid substitutions at positions corresponding to positions 349, 366, 368, and 407 of human IgG1 or IgG4 according to EU Index, wherein the amino acid substitutions are Y349C, T366S, L368A, and Y407V.
  • VH1 and VL1 form the second antigen binding site that binds a CD28 polypeptide
  • V H2 and V L2 form the third antigen binding site that binds a CD3 polypeptide
  • VH3 and VL3 form the first antigen binding site that binds a CD38 polypeptide
  • VH1 comprises a CDR-H1 sequence comprising the amino acid sequence of GYTFTSYY (SEQ ID NO:139), a CDR-H2 sequence comprising the amino acid sequence of IYPGNVNT (SEQ ID NO:140), and a CDR-H3 sequence comprising the amino acid sequence of TRSHYGLDWNFDV (SEQ ID NO:141)
  • VL1 comprises a CDR-L1 sequence comprising the amino acid sequence of QNIYVW (SEQ ID NO:142), a CDR-L2 sequence comprising the amino acid sequence of KAS, and a CDR-L3 sequence comprising the amino acid sequence of QQGQTYPY (SEQ ID NO
  • VL2 comprises a CDR-L1 sequence comprising the amino acid sequence of QSLVHNNANTY (SEQ ID NO:123), a CDR-L2 sequence comprising the amino acid sequence of KVS, and a CDR-L3 sequence comprising the amino acid sequence of GQGTQYPFT (SEQ ID NO:130); and VH3 comprises 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 VL3 comprises a CDR-L1 sequence comprising the amino acid sequence of QSVSSYGQGF (SEQ ID NO:39), a CDR-L2 sequence comprising the
  • VH1 and VL1 form the second antigen binding site that binds a CD28 polypeptide
  • VH2 and VL2 form the third antigen binding site that binds a CD3 polypeptide
  • V H3 and V L3 form the first antigen binding site that binds a CD38 polypeptide
  • VH1 comprises the amino acid sequence of SEQ ID NO:49 and VL1 comprises the amino acid sequence of SEQ ID NO:50
  • VH2 comprises the amino acid sequence of SEQ ID NO:53 and V L2 comprises the amino acid sequence of SEQ ID NO:54
  • V H3 comprises the amino acid sequence of SEQ ID NO:13 and V L3 comprises the amino acid sequence of SEQ ID NO:14.
  • 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
  • the fourth polypeptide chain comprises the amino acid sequence of SEQ ID NO:63.
  • 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
  • the fourth polypeptide chain comprises the amino acid sequence of SEQ ID NO:63.
  • 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
  • the fourth polypeptide chain comprises the amino acid sequence of SEQ ID NO:63.
  • 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, 10
  • 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
  • the fourth polypeptide chain comprises the amino acid sequence of SEQ ID NO:151.
  • 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
  • the fourth polypeptide chain comprises the amino acid sequence of SEQ ID NO:155.
  • 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
  • the fourth polypeptide chain comprises the amino acid sequence of SEQ ID NO:159.
  • 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
  • the fourth polypeptide chain comprises the amino acid sequence of SEQ ID NO:163.
  • 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
  • the fourth polypeptide chain comprises the amino acid sequence of SEQ ID NO:167.
  • 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
  • the fourth polypeptide chain comprises the amino acid sequence of SEQ ID 11
  • 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
  • the fourth polypeptide chain comprises the amino acid sequence of SEQ ID NO:175.
  • 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
  • the fourth polypeptide chain comprises the amino acid sequence of SEQ ID NO:179.
  • the CD38 polypeptide is a human CD38 polypeptide.
  • the CD3 polypeptide is a human CD3 polypeptide.
  • the CD28 polypeptide is a human CD28 polypeptide.
  • 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), Sezary syndrome (SS), anaplastic large cell ALK- lymphoma (ALCL), PTCL-not otherwise specified (NOS), enteropathy-type T-cell lymphoma (EATL), monomorphic epitheliotropic intestinal T-cell lymphoma (MEITL), primary cutaneous CD4+ lymphoproliferative disorder, subcutaneous T-cell lymphoma panniculitis-like (SCTCL), or primary cutaneous ⁇ ⁇ T-cell lymphoma.
  • AITL angioimmunoblastic T- cell lymphoma
  • HSTL hepatosplenic T-cell lymphoma
  • ATLL adult T-cell
  • the PTCL is HSTL, SS, or MF.
  • the mycosis fungoides is transformed mycosis fungoides (tMF or MFt).
  • cells of the PTCL express CD38 and/or CD28.
  • the individual is a human.
  • an anti-CD38 T-cell engager for use in a method for treating peripheral T-cell lymphoma (PTCL) in an individual in need thereof, said method comprising administering to the individual an effective amount of the anti- CD38 T-cell engager according to any one of the above embodiments.
  • an anti-CD38 T-cell engager according to any one of the above embodiments in the manufacture of a medicament for treating peripheral T-cell lymphoma (PTCL) in an individual in need thereof.
  • a kit comprising an anti-CD38 T-cell engager (e.g., an effective amount thereof, optionally in combination with a pharmaceutically acceptable carrier) and instructions for administering to an individual with 12
  • FIGS.1A-1E show expression and distribution of CD38 (left in each subtype) and CD28 (right in each subtype) through PTCL subtypes by immunochemistry (IHC). Data per entity are expressed according to IHC score.
  • FIG.1A Distribution of CD38 (left) and CD28 (right) expression with significant difference between PTCLs with a cytotoxic or non-cytotoxic profile (FIG.1B). Distribution of target expression pattern of CD38 and CD28 in tumour cells (FIG.1C).
  • the letters A to J at the top of the figure show the following patterns of CD38 and CD28 expression for each tumor type (to be read from the bottom of the figure to the top):
  • E CD38+CD28+, CD38+CD28- and CD38-CD28+;
  • F CD38+CD28+, CD38-CD28- and CD38+CD28-;
  • a to C at the left of the figure show the following patterns of CD38 and CD28 expression for each patient (to be read from the left of the figure to the right): A: CD38-CD28+, CD38+CD28-, CD38+CD28+, CD38-CD28-; B: CD38-CD28+, CD38+CD28+, CD38-CD28-; C: CD38- CD28+, CD38-CD28-.
  • the letters A to D at the left of the figure show the following patterns of CD38 and CD28 expression for each patient (to be read from the left of the figure to the 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-. 13
  • FIGS.2A-2D show IHC analyses for CD38 and CD28 in four T-cell lymphoma subtypes and multiplex Immunofluorescence for CD38, CD28, and neoplastic cell markers such as PD-1.
  • AITL angioimmunoblastic T-cell lymphoma
  • SS Sezary Syndrome
  • ALK+ Lymphoma ALK+ Lymphoma
  • ENKTCL extranodal NK/T-cell lymphoma
  • FIGS.3A-3F show the cytotoxic effect of anti-CD38/CD28xCD3 trispecific binding protein in different T-lymphoma cell lines (FIGS.3A-3C) or T-leukemia cell lines (FIGS.3D-3F).
  • Cases are grouped by diagnosis; shown are cutaneous anaplastic large cell lymphoma (cALCL; FIG.4A top panel), lymphomatoid papulosis (FIG.4A top panel), mycosis fungoides (FIG.4A middle panel), transformed mycosis fungoides (MFt; FIG.4A bottom panel), Sezary syndrome (FIG.4B), primary cutaneous CD4+ TCL (FIG.4B), SCTCL (Subcutaneous T-cell lymphoma, panniculitis-like) (FIG.4B), Primary cutaneous ⁇ T-cell lymphoma (FIG.4B), extranodal NK/T-cell lymphoma (ENKTCL; FIG.4C top panel), enteropathy associated T-cell lymphoma (FIG.4C middle panel), MEITL (Monomorphic epitheliotropic intestinal T-cell lymphoma) (FIG.4C middle panel), hepatosplenic T-cell lymphoma (HSTL;
  • the disclosure provides, inter alia, methods for treating PTCLs with an anti- CD38 T-cell engager, as well as uses, compositions, and kits related thereto. These are based at least in part on the demonstration herein that anti-CD38 T-cell engagers (e.g., an anti-CD38/CD28xCD3 trispecific binding protein) are able to induce cytotoxic lysis of various PTCL cell lines, including those from Sezary syndrome, MF (e.g., transformed MF), and hepatosplenic T-cell lymphoma in the presence of human T cells.
  • anti-CD38 T-cell engagers e.g., an anti-CD38/CD28xCD3 trispecific binding protein
  • MF e.g., transformed MF
  • hepatosplenic T-cell lymphoma in the presence of human T cells.
  • polynucleotide refers to single-stranded or double- stranded nucleic acid polymers of at least 10 nucleotides in length.
  • the nucleotides comprising the polynucleotide can be ribonucleotides or deoxyribonucleotides or a modified form of either type of nucleotide. Such modifications 15
  • polynucleotide specifically includes single-stranded and double-stranded forms of DNA.
  • An "isolated polynucleotide” is a polynucleotide of genomic, cDNA, or synthetic origin or some combination thereof, which: (1) is not associated with all or a portion of a polynucleotide in which the isolated polynucleotide is found in nature, (2) is linked to a polynucleotide to which it is not linked in nature, or (3) does not occur in nature as part of a larger sequence.
  • An "isolated polypeptide” is one that: (1) is free of at least some other polypeptides with which it would normally be found, (2) is essentially free of other polypeptides from the same source, e.g., from the same species, (3) is expressed by a cell from a different species, (4) has been separated from at least about 50 percent of polynucleotides, lipids, carbohydrates, or other materials with which it is associated in nature, (5) is not associated (by covalent or noncovalent interaction) with portions of a polypeptide with which the "isolated polypeptide" is associated in nature, (6) is operably associated (by covalent or noncovalent interaction) with a polypeptide with which it is not associated in nature, or (7) does not occur in nature.
  • Such an isolated polypeptide can be encoded by genomic DNA, cDNA, mRNA or other RNA, of synthetic origin, or any combination thereof.
  • the isolated polypeptide is substantially free from polypeptides or other contaminants that are found in its natural environment that would interfere with its use (therapeutic, diagnostic, prophylactic, research or otherwise).
  • Naturally occurring antibodies typically comprise a tetramer. Each such tetramer is typically composed of two identical pairs of polypeptide chains, each pair having one full-length "light" chain (typically having a molecular weight of about 25 kDa) and one full-length "heavy” chain (typically having a molecular weight of about 50-70 kDa).
  • heavy chain and light chain refer to any immunoglobulin polypeptide having sufficient variable domain sequence to confer specificity for a target antigen.
  • the amino-terminal portion of each light and heavy chain typically includes a variable domain of about 100 to 110 or more amino acids that typically is responsible for antigen recognition.
  • the carboxy-terminal portion of each chain typically defines a constant domain responsible for effector function.
  • a full-length heavy chain immunoglobulin polypeptide includes a variable domain 16
  • V H and three constant domains (C H1 , C H2 , and C H3 ), wherein the V H domain is at the amino-terminus of the polypeptide and the CH3 domain is at the carboxyl-terminus, and a full-length light chain immunoglobulin polypeptide includes a variable domain (VL) and a constant domain (C L ), wherein the V L domain is at the amino-terminus of the polypeptide and the CL domain is at the carboxyl-terminus.
  • VL variable domain
  • C L constant domain
  • Human light chains are typically classified as kappa and lambda light chains, and human heavy chains are typically classified as mu, delta, gamma, alpha, or epsilon, and define the antibody's isotype as IgM, IgD, IgG, IgA, and IgE, respectively.
  • IgG has several subclasses, including, but not limited to, IgG1, IgG2, IgG3, and IgG4.
  • IgM has subclasses including, but not limited to, IgM1 and IgM2.
  • IgA is similarly subdivided into subclasses including, but not limited to, IgA1 and IgA2.
  • variable and constant domains typically are joined by a "J" region of about 12 or more amino acids, with the heavy chain also including a "D” region of about 10 more amino acids.
  • the variable regions of each light/heavy chain pair typically form an antigen binding site.
  • the variable domains of naturally occurring antibodies typically exhibit the same general structure of relatively conserved framework regions (FR) joined by three hypervariable regions, also called complementarity determining regions or CDRs.
  • CDR set refers to a group of three CDRs that occur in a single variable region capable of binding the antigen. The exact boundaries of these CDRs have been defined differently according to different systems. The system described by Kabat (Kabat et al., SEQUENCES OF PROTEINS OF IMMUNOLOGICAL INTEREST (National Institutes of Health, Bethesda, Md.
  • These regions may be referred to as Chothia CDRs, which have boundaries that overlap with Kabat CDRs.
  • Other boundaries defining CDRs overlapping with the Kabat CDRs have been described by Padlan, 1995, FASEB J.9: 133- 39; MacCallum, 1996, J. Mol. Biol.262(5): 732-45; and Lefranc, 2003, Dev. Comp. Immunol.27: 55-77.
  • CDR boundary definitions may not strictly follow one of the herein systems, but will nonetheless overlap with the Kabat CDRs, although they may be shortened or lengthened in light of prediction or experimental findings that particular residues or groups of residues or even entire CDRs do not significantly impact antigen binding.
  • the methods used herein may utilize CDRs defined according to any of these systems, although certain embodiments use Kabat or Chothia defined CDRs. Identification of predicted CDRs using the amino acid sequence is well known in the field, such as in Martin, A.C. "Protein sequence and structure analysis of antibody variable domains," In Antibody Engineering, Vol.2. Kontermann R., Dübel S., eds. Springer-Verlag, Berlin, p. 33–51 (2010).
  • the amino acid sequence of the heavy and/or light chain variable domain may be also inspected to identify the sequences of the CDRs by other conventional methods, e.g., by comparison to known amino acid sequences of other heavy and light chain variable regions to determine the regions of sequence hypervariability.
  • the numbered sequences may be aligned by eye, or by employing an alignment program such as one of the CLUSTAL suite of programs, as described in Thompson, 1994, Nucleic Acids Res.22: 4673-80.
  • Molecular models are conventionally used to correctly delineate framework and CDR regions and thus correct the sequence-based assignments.
  • CDR/FR definition in an immunoglobulin light or heavy chain is to be determined based on IMGT definition (Lefranc et al. Dev.
  • Fc refers to a molecule comprising the sequence of a non-antigen-binding fragment resulting from digestion of an antibody or produced by other means, whether in monomeric or multimeric form, and can contain the hinge region.
  • the original immunoglobulin source of the native Fc is preferably of human origin and can be any of the immunoglobulins.
  • Fc molecules are made up of monomeric polypeptides that can be linked into dimeric or multimeric forms by covalent (i.e., disulfide bonds) and non- covalent association.
  • the number of intermolecular disulfide bonds between monomeric subunits of native Fc molecules ranges from 1 to 4 depending on class (e.g., IgG, IgA, and IgE) or subclass (e.g., IgG1, IgG2, IgG3, IgA1, IgGA2, and IgG4).
  • class e.g., IgG, IgA, and IgE
  • subclass e.g., IgG1, IgG2, IgG3, IgA1, IgGA2, and IgG4
  • a F(ab) fragment typically includes one light chain and the VH and CH1 domains of one heavy chain, wherein the V H -C H1 heavy chain portion of the F(ab) fragment cannot form a disulfide bond with another heavy chain polypeptide.
  • a F(ab) fragment can also include one light chain containing two variable domains separated by an amino acid linker and one heavy chain containing two variable domains separated by an amino acid linker and a CH1 domain.
  • a F(ab') fragment typically includes one light chain and a portion of one heavy chain that contains more of the constant region (between the C H1 and C H2 domains), such that an interchain disulfide bond can be formed between two heavy chains to form a F(ab') 2 molecule.
  • a "recombinant" molecule is one that has been prepared, expressed, created, or isolated by recombinant means.
  • anti-CD38 T-cell engager refers to a multispecific (e.g., bispecific or trispecific) binding molecule that is able to specifically bind CD38 (e.g., expressed on the surface of a target cell) as well as one or more T cell-expressed surface proteins, including without limitation CD3 and/or CD28.
  • binding protein refers to a non-naturally occurring (or recombinant or engineered) molecule that specifically binds to at least one target antigen.
  • binding protein refers to a binding protein that specifically binds to two different antigen targets.
  • a bispecific binding protein binds to two different antigens.
  • the bispecific binding protein can comprise two antigen binding sites, each antigen binding site specifically binding to a different target antigen (e.g., CD38 and CD3).
  • the term "trispecific binding protein” refers to a binding protein that specifically binds to three different antigen targets.
  • a trispecific binding protein binds to three different antigens.
  • the trispecific binding protein can comprise three antigen binding sites, each antigen binding site specifically binding to a different target antigen (e.g., CD38, CD28, and CD3).
  • the term "trivalent binding protein” refers to a binding protein that has three antigen binding sites.
  • the trivalent binding protein can bind to three antigen targets.
  • the trivalent binding protein can comprise three antigen binding sites, each antigen binding site specifically binding to a different target antigen 19
  • a binding protein can be trispecific and trivalent.
  • One embodiment of the disclosure provides binding proteins having biological and immunological specificity to two or three target antigens. Another embodiment of the disclosure provides nucleic acid molecules comprising nucleotide sequences encoding polypeptide chains that form such binding proteins. Another embodiment of the disclosure provides expression vectors comprising nucleic acid molecules comprising nucleotide sequences encoding polypeptide chains that form such binding proteins. Yet another embodiment of the disclosure provides host cells that express such binding proteins (i.e., comprising nucleic acid molecules or vectors encoding polypeptide chains that form such binding proteins).
  • the term "swapability" as used herein refers to the interchangeability of variable domains within the binding protein format and with retention of folding and ultimate binding affinity. "Full swapability” refers to the ability to swap the order of both V H1 and VH2 domains, and therefore the order of VL1 and VL2 domains, in the polypeptide chain of formula I or the polypeptide chain of formula II (i.e., to reverse the order) while maintaining full functionality of the binding protein as evidenced by the retention of binding affinity. Furthermore, it should be noted that the designations V H and V L refer only to the domain's location on a particular protein chain in the final format.
  • VH1 and V H2 could be derived from V L1 and V L2 domains in parent antibodies and placed into the V H1 and V H2 positions in the binding protein.
  • V L1 and V L2 could be derived from VH1 and VH2 domains in parent antibodies and placed in the VH1 and VH2 positions in the binding protein.
  • the VH and VL designations refer to the present location and not the original location in a parent antibody.
  • V H and V L domains are therefore "swappable.”
  • the term "antigen” or “target antigen” or “antigen target” as used herein refers to a molecule or a portion of a molecule that is capable of being bound by a binding protein, and additionally is capable of being used in an animal to produce antibodies capable of binding to an epitope of that antigen.
  • a target antigen may have one or more epitopes. With respect to each target antigen recognized by a binding protein, the binding protein is capable of competing with an intact antibody that recognizes the target antigen.
  • "CD38” is a cluster of differentiation 38 polypeptide and is a glycoprotein found on the surface of many immune cells. In some embodiments, an anti-CD38 T-cell engager of the present disclosure binds the extracellular domain of one or more CD38 polypeptide(s). Exemplary CD38 extracellular domain polypeptide sequences include, but 20
  • the CD38 polypeptide is a human CD38 polypeptide.
  • "CD28" is cluster of differentiation 28 polypeptide and is 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, e.g., NCBI Gene ID No.940 and NP_001230006. In some embodiments, the CD28 polypeptide is a human CD28 polypeptide.
  • CD3 is cluster of differentiation factor 3 polypeptide and is a T-cell surface protein that is typically part of the T cell receptor (TCR) complex formed by TCR alpha/beta or gamma/delta heterodimers in complex with CD3-epsilon, -gamma, -delta, and -zeta.
  • TCR T cell receptor
  • CD3 polynucleotide and polypeptide sequences are known in the art; see, e.g., NCBI Gene ID No.915 and NP_000723 for CD3-delta, NCBI Gene ID No.916 and NP_000724 for CD3-epsilon, NCBI Gene ID No.917 and NP_000064 for CD3-gamma, and NCBI Gene ID No.919 and NP_000725 for CD3-zeta.
  • the CD3 polypeptide is a human CD3 polypeptide.
  • An "isolated" binding protein is one that has been identified and separated and/or recovered from a component of its natural environment.
  • the binding protein will be purified: (1) to greater than 95% by weight of antibody as determined by the Lowry method, and most preferably more than 99% by weight, (2) to a degree sufficient to obtain at least 15 residues of N-terminal or internal amino acid sequence by use of a spinning cup sequenator, or (3) to homogeneity by SDS-PAGE under reducing or nonreducing conditions using Coomassie blue or, preferably, silver stain.
  • Isolated binding proteins include the binding protein in situ within recombinant cells since at least one component of the binding protein's natural environment will not be present.
  • the terms "substantially pure” or “substantially purified” as used herein refer to a compound or species that is the predominant species present (i.e., on a molar basis it is more abundant than any other individual species in the composition).
  • a substantially purified fraction is a composition wherein the species comprises at least about 50% (on a molar basis) of all macromolecular species present.
  • a substantially pure composition will comprise more than about 80%, 85%, 90%, 95%, or 99% of all macromolar species present in the composition.
  • the species is purified to essential homogeneity (contaminant species cannot be detected in the composition by conventional detection methods) wherein the composition consists essentially of a single macromolecular species.
  • epitopope includes any determinant, preferably a polypeptide determinant, capable of specifically binding to an immunoglobulin or T-cell receptor.
  • epitope determinants include chemically active surface groupings of molecules such as amino acids, sugar side chains, phosphoryl groups, or sulfonyl groups, and, in certain embodiments, may have specific three-dimensional structural characteristics and/or specific charge characteristics.
  • An epitope is a region of an antigen that is bound by an antibody or binding protein.
  • a binding protein is said to specifically bind an antigen when it preferentially recognizes its target antigen in a complex mixture of proteins and/or macromolecules.
  • a binding protein is said to specifically bind an antigen when the equilibrium dissociation constant is ⁇ 10 -8 M, more preferably when the equilibrium dissociation constant is ⁇ 10 -9 M, and most preferably when the dissociation constant is ⁇ 10 -10 M.
  • the dissociation constant (K D ) of a binding protein can be determined, for example, by surface plasmon resonance. Generally, surface plasmon resonance analysis measures real-time binding interactions between ligand (a target antigen on a biosensor matrix) and analyte (a binding protein in solution) by surface plasmon resonance (SPR) using the BIAcore system (Pharmacia Biosensor; Piscataway, NJ).
  • Surface plasmon analysis can also be performed by immobilizing the analyte (binding protein on a biosensor matrix) and presenting the ligand (target antigen).
  • K D refers to the dissociation constant of the interaction between a particular binding protein and a target antigen.
  • binding protein refers to the ability of a binding protein or an antigen-binding fragment thereof to bind to an antigen containing an epitope with an Kd of at least about 1 x 10 -6 M, 1 x 10 -7 M, 1 x 10 -8 M, 1 x 10 -9 M, 1 x 10 -10 M, 1 x 10 -11 M, 1 x 10 -12 M, or more, and/or to bind to an epitope with an affinity that is at least two-fold greater than its affinity for a nonspecific antigen.
  • a binding protein of the present disclosure binds to two or more antigens, e.g., a human and a cynomologus monkey CD38 polypeptide. 22
  • an antigen binding domain and/or binding protein of the present disclosure “cross reacts” with human and cynomolgus monkey CD38 polypeptides, e.g., CD38 extracellular domains, such as SEQ ID NO:1 (human CD38 isoform A), SEQ ID NO:105 (human CD38 isoform E) and SEQ ID NO:30 (cynomolgus monkey CD38).
  • a binding protein binding to antigen 1 (Ag1) is “cross-reactive” to antigen 2 (Ag2) when the EC 50 s are in a similar range for both antigens.
  • a binding protein binding to Ag1 is cross-reactive to Ag2 when the ratio of affinity of Ag2 to affinity of Ag1 is equal or less than 10 (for instance 5, 2, 1 or 0.5), affinities being measured with the same method for both antigens.
  • a binding protein binding to Ag1 is “not significantly cross-reactive” to Ag2 when the affinities are very different for the two antigens. Affinity for Ag2 may not be measurable if the binding response is too low.
  • a binding protein binding to Ag1 is not significantly cross-reactive to Ag2, when the binding response of the binding protein to Ag2 is less than 5% of the binding response of the same binding protein to Ag1 in the same experimental setting and at the same antibody concentration.
  • L 1 which is located on the light chain between the C-terminus of the V L2 and the N- terminus of the VL1 domain
  • L2 which is located on the light chain between the C- terminus of the VL1 and the N-terminus of the CL domain.
  • the heavy chain linkers are known as L 3 , which is located between the C-terminus of the V H1 and the N-terminus of the VH2 domain; and L4, which is located between the C-terminus of the VH2 and the N- terminus of the CH1 domain.
  • vector refers to any molecule (e.g., nucleic acid, plasmid, or virus) that is used to transfer coding information to a host cell.
  • vector includes a nucleic acid molecule that is capable of transporting another nucleic acid to which it has been linked.
  • plasmid refers to a circular double-stranded DNA molecule into which additional DNA segments may be inserted.
  • viral vector Another type of vector, wherein additional DNA segments may be inserted into the viral genome.
  • vectors are capable of autonomous replication in a host cell into which they are introduced (e.g., bacterial vectors having a bacterial origin of replication and episomal mammalian vectors).
  • Other vectors e.g., non-episomal mammalian vectors
  • certain 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").
  • expression vectors of utility in recombinant DNA techniques are often in the form of plasmids.
  • plasmid and "vector” may be used interchangeably herein, as a plasmid is the most commonly used form of vector. However, the disclosure is intended to include other forms of expression vectors, such as viral vectors (e.g., replication defective retroviruses, adenoviruses, and adeno-associated viruses), which serve equivalent functions.
  • viral vectors e.g., replication defective retroviruses, adenoviruses, and adeno-associated viruses
  • recombinant host cell refers to a cell into which a recombinant expression vector has been introduced. A recombinant host cell or host cell is intended to refer not only to the particular subject cell, but also to the progeny of such a cell.
  • host cell A wide variety of host cell expression systems can be used to express the binding proteins, including bacterial, yeast, baculoviral, and mammalian expression systems (as well as phage display expression systems).
  • An example of a suitable bacterial expression vector is pUC19.
  • a host cell is transformed or transfected with one or more recombinant expression vectors carrying DNA fragments encoding the polypeptide chains of the binding protein such that the polypeptide chains are expressed in the host cell and, preferably, secreted into the medium in which the host cells are cultured, from which medium the binding protein can be recovered.
  • transformation refers to a change in a cell's genetic characteristics, and a cell has been transformed when it has been modified to contain a new DNA.
  • a cell is transformed where it is genetically modified from its native state.
  • the transforming DNA may recombine with that of the cell by physically integrating into a chromosome of the cell, or may be maintained transiently as an episomal element without being replicated, or may replicate independently as a plasmid.
  • a cell is considered to have been stably transformed when the DNA is replicated with the division of the cell.
  • non-naturally occurring refers to an object that is not found in nature or that has been structurally modified or synthesized by man.
  • the twenty conventional amino acids and their abbreviations follow conventional usage. Stereoisomers (e.g., D-amino acids) of the twenty conventional amino acids; unnatural amino acids and analogs such as ⁇ -, ⁇ -disubstituted amino acids, N- alkyl amino acids, lactic acid, and other unconventional amino acids may also be suitable components for the polypeptide chains of the binding proteins.
  • Examples of unconventional amino acids include: 4-hydroxyproline, ⁇ -carboxyglutamate, ⁇ -N,N,N- trimethyllysine, ⁇ -N-acetyllysine, O-phosphoserine, N-acetylserine, N-formylmethionine, 3-methylhistidine, 5-hydroxylysine, ⁇ -N-methylarginine, and other similar amino acids and imino acids (e.g., 4-hydroxyproline).
  • the left-hand direction is the amino terminal direction and the right-hand direction is the carboxyl- terminal direction, in accordance with standard usage and convention.
  • Naturally occurring residues may be divided into classes based on common side chain properties: (1) hydrophobic: Met, Ala, Val, Leu, Ile, Phe, Trp, Tyr, Pro; 25
  • Conservative amino acid substitutions may involve exchange of a member of one of these classes with another member of the same class.
  • Non-conservative substitutions may involve the exchange of a member of one of these classes for a member from another class.
  • a skilled artisan will be able to determine suitable variants of the polypeptide chains of the binding proteins using well-known techniques. For example, one skilled in the art may identify suitable areas of a polypeptide chain that may be changed without destroying activity by targeting regions not believed to be important for activity. Alternatively, one skilled in the art can identify residues and portions of the molecules that are conserved among similar polypeptides.
  • the terms "patient,” “individual,” or “subject” as used herein include humans and animals (e.g., mammals, such as dogs, pigs, horses, cats, cows, etc.).
  • treatment or “treat” as used herein refer to both therapeutic treatment and prophylactic or preventative measures. Those in need of treatment include those having a disorder as well as those prone to have the disorder or those in which the disorder is to be prevented.
  • binding proteins can be used to treat humans with cancer, or humans susceptible to cancer, or ameliorate cancer in a human subject.
  • the binding proteins can also be used to prevent cancer in a human patient.
  • the cancer is a PTCL.
  • pharmaceutical composition or “therapeutic composition” as used herein refer to a compound or composition capable of inducing a desired therapeutic effect when properly administered to a patient. 26
  • the term "pharmaceutically acceptable carrier” or “physiologically acceptable carrier” as used herein refers to one or more formulation materials suitable for accomplishing or enhancing the delivery of a binding protein.
  • the terms "effective amount” and “therapeutically effective amount” when used in reference to a pharmaceutical composition comprising one or more binding proteins refer to an amount or dosage sufficient to produce a desired therapeutic result. More specifically, a therapeutically effective amount is an amount of a binding protein sufficient to inhibit, for some period of time, one or more of the clinically defined pathological processes associated with the condition being treated. The effective amount may vary depending on the specific binding protein that is being used, and also depends on a variety of factors and conditions related to the patient being treated and the severity of the disorder.
  • binding protein is to be administered in vivo, factors such as the age, weight, and health of the patient as well as dose response curves and toxicity data obtained in preclinical animal work would be among those factors considered.
  • the determination of an effective amount or therapeutically effective amount of a given pharmaceutical composition is well within the ability of those skilled in the art.
  • One embodiment of the disclosure provides a pharmaceutical composition comprising a pharmaceutically acceptable carrier and a therapeutically effective amount of a binding protein. II. Methods and Uses [0070] Certain aspects of the present disclosure relate to methods of treating peripheral T-cell lymphoma (PTCL) in an individual, e.g., in need thereof.
  • PTCL peripheral T-cell lymphoma
  • the methods comprise administering to the individual an effective amount of an anti-CD38 T- cell engager of the present disclosure.
  • the individual is a human.
  • the individual has or has been diagnosed with PTCL.
  • 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), Sezary syndrome (SS), anaplastic large cell ALK- lymphoma (ALCL), PTCL-not otherwise specified (NOS), enteropathy-type T-cell lymphoma (EATL), monomorphic epitheliotropic intestinal T-cell lymphoma (MEITL), primary cutaneous CD4+ lymphoproliferative disorder, subcutaneous T-cell lymphoma panniculitis-like (SCTCL), or primary cutaneous ⁇ ⁇ T-cell lymphoma.
  • AITL angioimmunoblastic T-cell lymphoma
  • HSTL hepatosplenic T-cell lymphoma
  • ATLL adult T-cell
  • the MF is transformed MF.
  • the PTCL is HSTL, Sezary syndrome, or MF (e.g., transformed MF).
  • cells of the PTCL express CD38 and/or CD28.
  • cells of the PTCL express CD38 and CD28.
  • cells of the PTCL express CD38.
  • cells of the PTCL express CD28.
  • expression of CD38 and/or CD28 by cells of the PTCL can be measured in a sample from the PTCL, e.g., a sample comprising PTCL cells.
  • a sample obtained from the PTCL of the individual comprises cells (e.g., PTCL cells) that express CD38 and/or CD28.
  • the methods of the present disclosure further comprise measuring expression of CD38 and/or CD28 in a sample obtained from the individual, e.g., a sample comprising PTCL cells.
  • the methods of the present disclosure further comprise obtaining a sample from the individual, e.g., a sample comprising PTCL cells.
  • the sample is from a tumor biopsy.
  • anti-CD38 T-cell engagers Any of the anti-CD38 T-cell engagers, bispecific binding proteins, or trispecific binding proteins described herein may find use in the methods, uses, kits, and compositions of the present disclosure.
  • Anti-CD38 T-cell engagers TCEs
  • Certain aspects of the present disclosure relate to anti-CD38 T-cell engagers.
  • the anti-CD38 T-cell engager binds to human CD38, e.g., expressed on the surface of a cell (such as a PTCL cell).
  • the anti-CD38 T-cell engager binds to human CD28, e.g., expressed on the surface of a cell (such as a PTCL cell). In some embodiments, the anti-CD38 T-cell engager binds to human CD3, e.g., expressed on the surface of a cell (such as a PTCL cell). In some embodiments, the anti- CD38 T-cell engager binds to human CD38 and human CD3. In some embodiments, the anti-CD38 T-cell engager binds to human CD38, human CD28, and human CD3. Exemplary human CD38, CD28, and CD3 polypeptides are known in the art and disclosed herein.
  • the anti-CD38 T-cell engager is a bispecific binding protein (e.g., a bispecific antibody) comprising a first antigen binding site that specifically binds a CD38 polypeptide and a second antigen binding site that specifically binds a CD3 28
  • a bispecific binding protein e.g., a bispecific antibody
  • the anti-CD38 T-cell engager can be a bispecific antibody comprising at least a first antigen binding domain that specifically binds a CD38 polypeptide and at least a second antigen binding domain that specifically binds a CD3 polypeptide.
  • the anti-CD38 T-cell engager is a trispecific binding protein.
  • the binding proteins comprise an antigen binding site that specifically binds a CD38 polypeptide, an antigen binding site that specifically binds a CD28 polypeptide, and an antigen binding site that specifically binds a CD3 polypeptide.
  • a binding protein induces apoptosis of a CD38+ cell.
  • a binding protein recruits a T cell to a CD38+ cell and optionally activates the T cell (e.g., though TCR stimulation and/or costimulation).
  • Any of the anti-CD38 T- cell engagers described herein may find use in the methods, uses, kits, and compositions of the present disclosure.
  • Anti-CD38 antigen binding sites [0077] Exemplary antigen binding sites that specifically bind CD38 polypeptides and may find use in the anti-CD38 T-cell engagers of the present disclosure are described infra. Any of the anti-CD38 antigen binding sites and/or trispecific binding proteins described in International Appl. No. WO2019/074973 may find use in the methods, uses, kits, and compositions of the present disclosure.
  • the CD38 polypeptide is a human CD38 polypeptide, also known as ADPRC1.
  • Human CD38 polypeptides are known in the art and include, without limitation, the polypeptide represented by NCBI Accession Number NP_001766.2, or a polypeptide produced from NCBI Gene ID Number 952.
  • the antigen binding site binds a human CD38 polypeptide, a non- human primate (e.g., cynomolgus monkey) CD38 polypeptide, or a human CD38 polypeptide and a non-human primate (e.g., cynomolgus monkey) CD38 polypeptide.
  • the antigen binding site that binds a CD38 polypeptide comprises: 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); or 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 29
  • the antigen binding site that binds a CD38 polypeptide comprises: 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); or an antibody light chain variable (VL) domain comprising a CDR-L1 sequence comprising the amino acid sequence of ESVDSYGNGF (SEQ ID NO:34) or QSVSSYGQG (SEQ ID NO:36).
  • VH antibody heavy chain variable
  • the antigen binding site that binds a CD38 polypeptide comprises: 1, 2, 3, 4, 5, or 6 CDRs from an antibody VH and/or VL domain sequence of an antigen binding site (e.g., mAb1, mAb2, mAb3, mAb4, mAb5, mAb6, hhy992, hu5739, hu6284, hhy1195, hhy1370, hyb5739, or hyb6284) as shown in Table A or B.
  • an antigen binding site e.g., mAb1, mAb2, mAb3, mAb4, mAb5, mAb6, hhy992, hu5739, hu6284, hhy1195, hhy1370, hyb5739, or hyb6284
  • the antigen binding site that binds a CD38 polypeptide comprises: 1, 2, 3, 4, 5, or 6 CDRs from an antibody VH and/or VL domain sequence of a trispecific binding protein shown in Table E1 or E2.
  • the antigen binding site that binds a CD38 polypeptide comprises: 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 an antibody light chain variable (VL) domain comprising a CDR-L1 sequence comprising the amino acid sequence of ESVDSYGNGF (SEQ ID NO:31) or GYTFTSYA (S
  • the antigen binding site that binds a CD38 polypeptide comprises: an antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence comprising the amino acid sequence of GYTFTSFN (SEQ ID NO:31), a CDR-H2 sequence comprising the amino acid sequence of IYPGNGGT (SEQ ID NO:32), and a CDR-H3 sequence comprising the amino acid sequence of ARTGGLRRAYFTY (SEQ ID NO:33); 30
  • VH antibody heavy chain variable domain comprising a CDR-H1 sequence comprising the amino acid sequence of GYTFTSFN (SEQ ID NO:31), a CDR-H2 sequence comprising the amino acid sequence of IYPGNGGT (SEQ ID NO:32), and a CDR-H3 sequence comprising the amino acid sequence of ARTGGLRRAYFTY (SEQ ID NO:33);
  • VL antibody light chain variable domain
  • a CDR-L1 sequence comprising the amino acid sequence of ESVDSYGNGF (SEQ ID NO:34)
  • CDR-L2 sequence comprising the amino acid sequence of LAS
  • CDR-L3 sequence comprising the amino acid sequence of QQNKEDPWT (SEQ ID NO:36).
  • the antigen binding site that binds a CD38 polypeptide comprises: an antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence comprising the amino acid sequence of GYTFTSFN (SEQ ID NO:31), a CDR-H2 sequence comprising the amino acid sequence of IYPGNGGT (SEQ ID NO:32), and a CDR-H3 sequence comprising the amino acid sequence of ARTGGLRRAYFTY (SEQ ID NO:33); and an antibody light chain variable (VL) domain comprising a CDR-L1 sequence comprising the amino acid sequence of ESVDSYGNGF (SEQ ID NO:34), a CDR-L2 sequence comprising the amino acid sequence of LAS, and a CDR-L3 sequence comprising the amino acid sequence of QQNKEDPWT (SEQ ID NO:36).
  • VH antibody heavy chain variable
  • the antigen binding site that binds a CD38 polypeptide comprises: 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); or 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).
  • VH antibody heavy chain variable
  • the antigen binding site that binds a CD38 polypeptide comprises: 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 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).
  • VH antibody heavy chain variable
  • the VH domain comprises the sequence, from N- terminus to C-terminus, FR1—CDR-H1—FR2—CDR-H2—FR3—CDR-H3—FR4; where FR1 comprises the sequence QVQLVQSGAEVVKPGASVKVSCKAS (SEQ ID NO:86), 31
  • QVQLVQSGAEVVKSGASVKVSCKAS (SEQ ID NO:87), or QVQLVQSGAEVVKPGASVKMSCKAS (SEQ ID NO:88); where FR2 comprises the sequence MHWVKEAPGQRLEWIGY (SEQ ID NO:90) or MHWVKEAPGQGLEWIGY (SEQ ID NO:91); where FR3 comprises the sequence NYNQKFQGRATLTADTSASTAYMELSSLRSEDTAVYFC (SEQ ID NO:93) or NYNQKFQGRATLTADTSASTAYMEISSLRSEDTAVYFC (SEQ ID NO:94); and where FR4 comprises the sequence WGQGTLVTVSS (SEQ ID NO:96).
  • the VL domain comprises the sequence, from N-terminus to C-terminus, FR1—CDR-L1— FR2—CDR-L2—FR3—CDR-L3—FR4; where FR1 comprises the sequence DIVLTQSPATLSLSPGERATISCRAS (SEQ ID NO:97); where FR2 comprises the sequence MHWYQQKPGQPPRLLIY (SEQ ID NO:99); where FR3 comprises the sequence SRATGIPARFSGSGSGTDFTLTISPLEPEDFAVYYC (SEQ ID NO:101); and where FR4 comprises the sequence FGGGTKLEIK (SEQ ID NO:103).
  • the VH domain comprises 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:5; and/or the VL domain comprises 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:6.
  • the VH domain comprises 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 comprises 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.
  • the VH domain comprises 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/or the VL domain comprises 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 32
  • the VH domain comprises 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/or the VL domain comprises 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/or the VL domain comprises 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
  • the VH domain comprises 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 comprises 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.
  • the VH domain comprises 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:5; and the VL domain comprises 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:6.
  • the VH domain comprises 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 comprises 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 comprises 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
  • VL domain comprises 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.
  • the VH domain comprises 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 comprises 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.
  • the VH domain comprises 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 comprises 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.
  • 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.
  • 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.
  • the antigen binding site that binds a CD38 polypeptide comprises: an antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence comprising the amino acid sequence of GFTFSSYG (SEQ ID NO:41), a CDR-H2 sequence 34
  • VL antibody light chain variable domain comprising a CDR-L1 sequence comprising the amino acid sequence of QGIRND (SEQ ID NO:44), a CDR-L2 sequence comprising the amino acid sequence of AAS, and a CDR-L3 sequence comprising the amino acid sequence of LQDYIYYPT (SEQ ID NO:46).
  • the antigen binding site that binds a CD38 polypeptide comprises: an antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence comprising the amino acid sequence of GFTFSSYG (SEQ ID NO:41), a CDR-H2 sequence comprising the amino acid sequence of IWYDGSNK (SEQ ID NO:42), and a CDR-H3 sequence comprising the amino acid sequence of ARMFRGAFDY (SEQ ID NO:43); and an antibody light chain variable (VL) domain comprising a CDR-L1 sequence comprising the amino acid sequence of QGIRND (SEQ ID NO:44), a CDR-L2 sequence comprising the amino acid sequence of AAS, and a CDR-L3 sequence comprising the amino acid sequence of LQDYIYYPT (SEQ ID NO:46).
  • VH antibody heavy chain variable
  • the VH domain comprises the sequence, from N- terminus to C-terminus, FR1—CDR-H1—FR2—CDR-H2—FR3—CDR-H3—FR4; where FR1 comprises the sequence QVQLVESGGGVVQPGRSLRLSCAAS (SEQ ID NO:89); where FR2 comprises the sequence MHWVRQAPGKGLEWVAV (SEQ ID NO:92); where FR3 comprises the sequence YYADSVKGRFTISGDNSKNTLYLQMNSLRAEDTAVYYC (SEQ ID NO:95); and where FR4 comprises the sequence WGQGTLVTVSS (SEQ ID NO:96).
  • the VL domain comprises the sequence, from N-terminus to C-terminus, FR1—CDR-L1—FR2—CDR-L2—FR3—CDR-L3—FR4; where FR1 comprises the sequence AIQMTQSPSSLSASVGDRVTITCRAS (SEQ ID NO:98); where FR2 comprises the sequence GWYQQKPGKAPKLLIY (SEQ ID NO:100); where FR3 comprises the sequence SLQSGVPSRFSGSGSGTDFTLTISGLQPEDSATYYC (SEQ ID NO:102); and where FR4 comprises the sequence WGQGTLVTVSS (SEQ ID NO:104).
  • the VH domain comprises 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:9; and/or the VL domain comprises 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 35
  • the VH domain comprises 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:9; and the VL domain comprises 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:10.
  • the VH domain comprises the amino acid sequence of SEQ ID NO:9; and the VL domain comprises the amino acid sequence of SEQ ID NO:10.
  • the antigen binding site that binds a CD38 polypeptide comprises: an antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence comprising the amino acid sequence of GYTLTEFS (SEQ ID NO:2), a CDR-H2 sequence comprising the amino acid sequence of FDPEDGET (SEQ ID NO:3), and a CDR-H3 sequence comprising the amino acid sequence of TTGRFFDWF (SEQ ID NO:4); or an antibody light chain variable (VL) domain comprising a CDR-L1 sequence comprising the amino acid sequence of QSVISRF (SEQ ID NO:7), a CDR-L2 sequence comprising the amino acid sequence of GAS, and a CDR-L3 sequence comprising the amino acid sequence of QQDSNLPIT (SEQ ID NO:11).
  • the antigen binding site that binds a CD38 polypeptide comprises: an antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence comprising the amino acid sequence of GYTLTEFS (SEQ ID NO:2), a CDR-H2 sequence comprising the amino acid sequence of FDPEDGET (SEQ ID NO:3), and a CDR-H3 sequence comprising the amino acid sequence of TTGRFFDWF (SEQ ID NO:4); and an antibody light chain variable (VL) domain comprising a CDR-L1 sequence comprising the amino acid sequence of QSVISRF (SEQ ID NO:7), a CDR-L2 sequence comprising the amino acid sequence of GAS, and a CDR-L3 sequence comprising the amino acid sequence of QQDSNLPIT (SEQ ID NO:11).
  • VH antibody heavy chain variable
  • the antigen binding site that binds a CD38 polypeptide comprises: an antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence comprising the amino acid sequence of GYAFTTYL (SEQ ID NO:12), a CDR-H2 sequence comprising the amino acid sequence of INPGSGST (SEQ ID NO:15), and a CDR-H3 sequence comprising the amino acid sequence of ARYAYGY (SEQ ID NO:16); 36
  • VH antibody heavy chain variable
  • VL antibody light chain variable domain
  • a CDR-L1 sequence comprising the amino acid sequence of QNVGTA (SEQ ID NO:19)
  • a CDR-L2 sequence comprising the amino acid sequence of SAS
  • a CDR-L3 sequence comprising the amino acid sequence of QQYSTYPFT (SEQ ID NO:22).
  • the antigen binding site that binds a CD38 polypeptide comprises: an antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence comprising the amino acid sequence of GYAFTTYL (SEQ ID NO:12), a CDR-H2 sequence comprising the amino acid sequence of INPGSGST (SEQ ID NO:15), and a CDR-H3 sequence comprising the amino acid sequence of ARYAYGY (SEQ ID NO:16); and an antibody light chain variable (VL) domain comprising a CDR-L1 sequence comprising the amino acid sequence of QNVGTA (SEQ ID NO:19), a CDR-L2 sequence comprising the amino acid sequence of SAS, and a CDR-L3 sequence comprising the amino acid sequence of QQYSTYPFT (SEQ ID NO:22).
  • VH antibody heavy chain variable
  • the antigen binding site that binds a CD38 polypeptide comprises: an antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence comprising the amino acid sequence of GYSFTNYA (SEQ ID NO:24), a CDR-H2 sequence comprising the amino acid sequence of ISPYYGDT (SEQ ID NO:25), and a CDR-H3 sequence comprising the amino acid sequence of ARRFEGFYYSMDY (SEQ ID NO:26); or an antibody light chain variable (VL) domain comprising a CDR-L1 sequence comprising the amino acid sequence of QSLVHSNGNTY (SEQ ID NO:27), a CDR-L2 sequence comprising the amino acid sequence of KVS, and a CDR-L3 sequence comprising the amino acid sequence of SQSTHVPLT (SEQ ID NO:29).
  • VH antibody heavy chain variable
  • the antigen binding site that binds a CD38 polypeptide comprises: an antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence comprising the amino acid sequence of GYSFTNYA (SEQ ID NO:24), a CDR-H2 sequence comprising the amino acid sequence of ISPYYGDT (SEQ ID NO:25), and a CDR-H3 sequence comprising the amino acid sequence of ARRFEGFYYSMDY (SEQ ID NO:26); and an antibody light chain variable (VL) domain comprising a CDR-L1 sequence comprising the amino acid sequence of QSLVHSNGNTY (SEQ ID NO:27), a CDR-L2 sequence comprising the amino acid sequence of KVS, and a CDR-L3 sequence comprising the amino acid sequence of SQSTHVPLT (SEQ ID NO:29).
  • VH antibody heavy chain variable
  • the antigen binding site that binds a CD38 polypeptide comprises: an antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence comprising the amino acid sequence of GFTFSSYG (SEQ ID NO:41), a CDR-H2 sequence comprising the amino acid sequence of IWYDGSNK (SEQ ID NO:42), and a CDR-H3 37
  • VH antibody heavy chain variable
  • ARDPGLRYFDGGMDV SEQ ID NO:106
  • VL antibody light chain variable domain comprising a CDR-L1 sequence comprising the amino acid sequence of QGISSY (SEQ ID NO:107), a CDR-L2 sequence comprising the amino acid sequence of AAS, and a CDR-L3 sequence comprising the amino acid sequence of QQLNSFPYT (SEQ ID NO:229).
  • the antigen binding site that binds a CD38 polypeptide comprises: an antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence comprising the amino acid sequence of GFTFSSYG (SEQ ID NO:41), a CDR-H2 sequence comprising the amino acid sequence of IWYDGSNK (SEQ ID NO:42), and a CDR-H3 sequence comprising the amino acid sequence of ARDPGLRYFDGGMDV (SEQ ID NO:106); and an antibody light chain variable (VL) domain comprising a CDR-L1 sequence comprising the amino acid sequence of QGISSY (SEQ ID NO:107), a CDR-L2 sequence comprising the amino acid sequence of AAS, and a CDR-L3 sequence comprising the amino acid sequence of QQLNSFPYT (SEQ ID NO:229).
  • VH antibody heavy chain variable
  • the VH domain comprises the sequence, from N- terminus to C-terminus, FR1—CDR-H1—FR2—CDR-H2—FR3—CDR-H3—FR4; where FR1 comprises the sequence QVQLVESGGGVVQPGRSLRLSCAAS (SEQ ID NO:89); where FR2 comprises the sequence MHWVRQAPGKGLEWVAV (SEQ ID NO:92); where FR3 comprises the sequence YYADSVKGRFTISGDNSKNTLYLQMNSLRAEDTAVYYC (SEQ ID NO:95); and where FR4 comprises the sequence WGQGTLVTVSS (SEQ ID NO:96).
  • the VL domain comprises the sequence, from N-terminus to C-terminus, FR1—CDR-L1—FR2—CDR-L2—FR3—CDR-L3—FR4; where FR1 comprises the sequence AIQMTQSPSSLSASVGDRVTITCRAS (SEQ ID NO:98); where FR2 comprises the sequence GWYQQKPGKAPKLLIY (SEQ ID NO:100); where FR3 comprises the sequence SLQSGVPSRFSGSGSGTDFTLTISGLQPEDSATYYC (SEQ ID NO:102); and where FR4 comprises the sequence WGQGTLVTVSS (SEQ ID NO:104).
  • the VH domain comprises 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:108; and/or the VL domain comprises 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 38
  • the VH domain comprises 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:108; and the VL domain comprises 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:109; and the VL domain comprises 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%,
  • the VH domain comprises the amino acid sequence of SEQ ID NO:108; and the VL domain comprises the amino acid sequence of SEQ ID NO:109. [0097] In some embodiments, the VH domain comprises 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 comprises 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.
  • the VH domain comprises 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 the VL domain comprises 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.
  • 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. [0099] In some embodiments, the VH domain comprises 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; 39
  • VL domain comprises 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.
  • the VH domain comprises 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 the VL domain comprises 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.
  • 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.
  • the VH domain comprises 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 comprises 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.
  • the VH domain comprises 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 the VL domain comprises 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.
  • 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. [0103] In some embodiments, the VH domain comprises 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%, 40
  • the VL domain comprises 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.
  • the VH domain comprises 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 the VL domain comprises 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.
  • 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.
  • the VH domain comprises 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 comprises 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.
  • the VH domain comprises 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 the VL domain comprises 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.
  • the VH domain comprises the amino acid sequence of SEQ ID NO:114; and the VL domain comprises the amino acid sequence of SEQ ID NO:115. 41
  • a binding protein of the present disclosure comprises 1, 2, 3, 4, 5, or 6 CDR sequences of an antibody sequence shown in Table A. In some embodiments, a binding protein of the present disclosure comprises 1, 2, 3, 4, 5, or 6 CDR sequences, a VH domain sequence, and/or a VL domain sequence of an antibody sequence shown in Table B. In some embodiments, a binding protein of the present disclosure comprises 1, 2, 3, 4, 5, or 6 CDR sequences, a VH domain sequence, and/or a VL domain sequence of an antibody sequence shown in Table I1 or I2. In some embodiments, a binding protein of the present disclosure comprises 1, 2, 3, or 4 polypeptide sequences shown in Table I1 or I2. 42 43
  • Table B Variable domain sequences of anti-CD38 (mAb1-7) and other binding proteins. 44
  • a binding protein of the present disclosure binds a purified polypeptide or fragment thereof comprising the amino acid sequence of SEQ ID NO:1 and/or 30 (e.g., as measured by ELISA or SPR). In some embodiments, a binding protein of the present disclosure binds a polypeptide or comprising the amino acid sequence of SEQ ID NO:1 and/or 30 when expressed on the surface of a cell (e.g., as measured by flow cytometry). [0110] In some embodiments, a binding protein of the present disclosure binds to a CD38 isoform A polypeptide (e.g., comprising the amino acid sequence of SEQ ID NO:1).
  • a binding protein of the present disclosure binds to a CD38 isoform E polypeptide (e.g., comprising the amino acid sequence of SEQ ID NO:105 and not comprising the full amino acid sequence of SEQ ID NO:1, consisting of the amino acid sequence of SEQ ID NO:105, or consisting essentially of the amino acid sequence of SEQ ID NO:105).
  • a CD38 isoform E polypeptide e.g., comprising the amino acid sequence of SEQ ID NO:105 and not comprising the full amino acid sequence of SEQ ID NO:1, consisting of the amino acid sequence of SEQ ID NO:105, or consisting essentially of the amino acid sequence of SEQ ID NO:105.
  • a binding protein of the present disclosure binds to a CD38 isoform A polypeptide (e.g., comprising the amino acid sequence of SEQ ID NO:1) and a CD38 isoform E polypeptide (e.g., comprising the amino acid sequence of SEQ ID NO:105 and not comprising the full amino acid sequence of SEQ ID NO:1, consisting of the amino acid sequence of SEQ ID NO:105, or consisting essentially of the amino acid sequence of SEQ ID NO:105).
  • a CD38 isoform E polypeptide can be advantageous, e.g., in targeting a binding protein of the present disclosure to cell(s) expressing a CD38 isoform E polypeptide.
  • Human CD38 isoform A extracellular domain polypeptide sequence RWRQQWSGPGTTKRFPETVLARCVKYTEIHPEMRHVDCQSVWDAFKGAFISKHPCN ITEEDYQPLMKLGTQTVPCNKILLWSRIKDLAHQFTQVQRDMFTLEDTLLGYLADDL TWCGEFNTSKINYQSCPDWRKDCSNNPVSVFWKTVSRRFAEAACDVVHVMLNGSR SKIFDKNSTFGSVEVHNLQPEKVQTLEAWVIHGGREDSRDLCQDPTIKELESIISKRNI QFSCKNIYRPDKFLQCVKNPEDSSCTSEI (SEQ ID NO:1) Human CD38 isoform E polypeptide sequence RWRQQWSGPGTTKRFPETVLARCVKYTEIHPEMRHVDCQSVWDAFKGAFISKHPCN ITEEDYQPLMKLGTQTVPCNKILLWSRIKDLAHQFTQVQRDMFTLEDTLLGYLADDL
  • the extracellular domain of a human CD38 polypeptide comprises the amino acid sequence of SEQ ID NO:1. In some embodiments, the extracellular domain of a cynomolgus monkey CD38 polypeptide comprises the amino acid sequence of SEQ ID NO:30.
  • Human CD28 polypeptides are known in the art and include, without limitation, the polypeptides 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 a polypeptide produced from NCBI Gene ID Number 940.
  • the antigen binding site that binds a CD28 polypeptide comprises: an antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence comprising the amino acid sequence of GYTFTSYY (SEQ ID NO:139), a CDR-H2 sequence comprising the amino acid sequence of IYPGNVNT (SEQ ID NO:140), and a CDR-H3 sequence comprising the amino acid sequence of TRSHYGLDWNFDV (SEQ ID NO:141) and/or an antibody light chain variable (VL) domain comprising a CDR-L1 sequence comprising the amino acid sequence of QNIYVW (SEQ ID NO:142), a CDR-L2 sequence comprising the amino acid sequence of KAS, and a CDR-L3 sequence comprising the amino acid sequence of QQGQTYPY (SEQ ID NO:144).
  • the antigen binding site that binds a CD28 polypeptide comprises: an antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence comprising the amino
  • SEQ ID NO:139 a CDR-H2 sequence comprising the amino acid sequence of IYPGNVNT (SEQ ID NO:140), and a CDR-H3 sequence comprising the amino acid sequence of TRSHYGLDWNFDV (SEQ ID NO:141) and an antibody light chain variable (VL) domain comprising a CDR-L1 sequence comprising the amino acid sequence of QNIYVW (SEQ ID NO:142), a CDR-L2 sequence comprising the amino acid sequence of KAS, and a CDR-L3 sequence comprising the amino acid sequence of QQGQTYPY (SEQ ID NO:144).
  • the antigen binding site that binds a CD28 polypeptide comprises: an antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence comprising the amino acid sequence of GFSLSDYG (SEQ ID NO:212), a CDR-H2 sequence comprising the amino acid sequence of IWAGGGT (SEQ ID NO:213), and a CDR-H3 sequence comprising the amino acid sequence of ARDKGYSYYYSMDY (SEQ ID NO:214) and/or an antibody light chain variable (VL) domain comprising a CDR-L1 sequence comprising the amino acid sequence of ESVEYYVTSL (SEQ ID NO:215), a CDR-L2 sequence comprising the amino acid sequence of AAS, and a CDR-L3 sequence comprising the amino acid sequence of QQSRKVPYT (SEQ ID NO:217).
  • VH antibody heavy chain variable
  • IWAGGGT SEQ ID NO:2173
  • CDR-H3 sequence comprising the amino acid sequence of ARDKGYSYYYSMDY (SEQ ID NO:214) and an antibody light chain variable (VL) domain comprising a CDR-L1 sequence comprising the amino acid sequence of ESVEYYVTSL (SEQ ID NO:215), a CDR-L2 sequence comprising the amino acid sequence of AAS, and a CDR-L3 sequence comprising the amino acid sequence of QQSRKVPYT (SEQ ID NO:217).
  • the antigen binding site that binds a CD28 polypeptide comprises: an antibody heavy chain variable (VH) domain comprising 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:51, and/or an antibody light chain variable (VL) domain comprising 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:52.
  • VH antibody heavy chain variable
  • Human CD3 polypeptides are known in the art and include, without limitation, the polypeptides represented by NCBI Accession Numbers XP_006510029.1 or NP_031674.1, or a polypeptide produced from NCBI Gene ID Numbers 915, 916, or 917.
  • the antigen binding site that binds a CD28 polypeptide comprises: an antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence comprising the amino acid sequence of GFTFTKAW (SEQ ID NO:120), a CDR-H2 sequence comprising the amino acid sequence of IKDKSNSYAT (SEQ ID NO:121), and a CDR-H3 sequence comprising the amino acid sequence of RGVYYALSPFDY (SEQ ID NO:122) and an antibody light chain variable (VL) domain comprising a CDR-L1 sequence comprising the amino acid sequence of QSLVHX1NX2X3TY, wherein 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 comprising the amino acid sequence of KVS, and a CDR-L3 sequence comprising the amino acid sequence of GQGTQYPFT (SEQ ID NO:130).
  • VH antibody heavy chain variable
  • 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).
  • the antigen binding site that binds a CD3 polypeptide comprises: an antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence 51
  • GFTFTKAW SEQ ID NO:120
  • CDR-H2 sequence comprising the amino acid sequence of IKDKSNSYAT (SEQ ID NO:121)
  • CDR-H3 sequence comprising the amino acid sequence of RGVYYALSPFDY (SEQ ID NO:122) and/or an antibody light chain variable (VL) domain comprising a CDR-L1 sequence comprising the amino acid sequence of QSLVHNNGNTY (SEQ ID NO:218), a CDR-L2 sequence comprising the amino acid sequence of KVS, and a CDR-L3 sequence comprising the amino acid sequence of GQGTQYPFT (SEQ ID NO:130).
  • the antigen binding site that binds a CD3 polypeptide comprises: an antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence comprising the amino acid sequence of GFTFTKAW (SEQ ID NO:120), a CDR-H2 sequence comprising the amino acid sequence of IKDKSNSYAT (SEQ ID NO:121), and a CDR-H3 sequence comprising the amino acid sequence of RGVYYALSPFDY (SEQ ID NO:122) and/or an antibody light chain variable (VL) domain comprising a CDR-L1 sequence comprising the amino acid sequence of QSLVHNNANTY (SEQ ID NO:123), a CDR-L2 sequence comprising the amino acid sequence of KVS, and a CDR-L3 sequence comprising the amino acid sequence of GQGTQYPFT (SEQ ID NO:130).
  • VH antibody heavy chain variable
  • the antigen binding site that binds a CD3 polypeptide comprises: an antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence comprising the amino acid sequence of GFTFTKAW (SEQ ID NO:120), a CDR-H2 sequence comprising the amino acid sequence of IKDKSNSYAT (SEQ ID NO:121), and a CDR-H3 sequence comprising the amino acid sequence of RGVYYALSPFDY (SEQ ID NO:122) and an antibody light chain variable (VL) domain comprising a CDR-L1 sequence comprising the amino acid sequence of QSLVHNNANTY (SEQ ID NO:123), a CDR-L2 sequence comprising the amino acid sequence of KVS, and a CDR-L3 sequence comprising the amino acid sequence of GQGTQYPFT (SEQ ID NO:130).
  • VH antibody heavy chain variable
  • the antigen binding site that binds a CD3 polypeptide comprises: an antibody heavy chain variable (VH) domain comprising 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:53 or 138, and/or an antibody light chain variable (VL) domain comprising 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: 54, 133, 134,
  • the antigen binding site that binds a CD3 polypeptide comprises: an antibody heavy chain variable (VH) domain comprising the amino acid sequence of SEQ ID NO: 53 or 138, and/or an antibody light chain variable (VL) domain comprising the amino acid sequence of SEQ ID NO: 54, 133, 134, 135, 136, or 137.
  • the antigen binding site that binds a CD3 polypeptide comprises: an antibody heavy chain variable (VH) domain comprising the amino acid sequence of SEQ ID NO: 53 or 138, and an antibody light chain variable (VL) domain comprising the amino acid sequence of SEQ ID NO: 54, 133, 134, 135, 136, or 137.
  • the antigen binding site that binds a CD3 polypeptide comprises: an antibody heavy chain variable (VH) domain comprising 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:84, and/or an antibody light chain variable (VL) domain comprising 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:85.
  • VH antibody heavy chain variable
  • the antigen binding site that binds a CD3 polypeptide comprises: an antibody heavy chain variable (VH) domain comprising the amino acid sequence of SEQ ID NO:84, and/or an antibody light chain variable (VL) domain comprising the amino acid sequence of SEQ ID NO:85.
  • the antigen binding site that binds a CD3 polypeptide comprises: an antibody heavy chain variable (VH) domain comprising the amino acid sequence of SEQ ID NO:84, and an antibody light chain variable (VL) domain comprising the amino acid sequence of SEQ ID NO:85.
  • a binding protein of the present disclosure comprises 1, 2, 3, 4, 5, or 6 CDR sequences of an antibody sequence shown in Table D. In some embodiments, a binding protein of the present disclosure comprises 1, 2, 3, 4, 5, or 6 CDR sequences, a VH domain sequence, and/or a VL domain sequence of an antibody sequence shown in Table D. In some embodiments, a binding protein of the present disclosure comprises 1, 2, 3, 4, 5, or 6 CDR sequences, a VH domain sequence, and/or a VL domain sequence of an antibody sequence shown in Table D. In some embodiments, a binding protein of the present disclosure comprises 1, 2, 3, or 4 polypeptide sequences shown in Table D. Table D. Anti-CD3 antigen binding site sequences. 54
  • an anti-CD38 T-cell engager of the present disclosure is a trispecific binding protein comprising an antigen binding site that binds one or more CD38 polypeptides, a second antigen binding site that binds a CD28 polypeptide, and a third antigen binding site that binds a CD3 polypeptide. Any of the antigen binding sites described supra may find use in the anti-CD38 T-cell engagers of the present disclosure.
  • the trispecific binding protein comprises four polypeptide chains that form the three antigen binding sites, wherein a first polypeptide chain comprises a structure represented by the formula: V L2 -L 1 -V L1 -L 2 -C L [I] and a second polypeptide chain comprises a structure represented by the formula: VH1-L3-VH2-L4-CH1-hinge-CH2-CH3 [II] and a third polypeptide chain comprises a structure represented by the formula: V H3 -C H1 -hinge-C H2 -C H3 [III] and a fourth polypeptide chain comprises a structure represented by the formula: VL3-CL [IV] wherein: V L1 is a first immunoglobulin light chain variable domain; VL2 is a second immunoglobulin light chain variable domain; V L3 is a third immunoglobulin light chain variable domain; V H1 is a first immunoglobulin heavy chain variable domain; VH2 is a
  • the polypeptide of formula I and the polypeptide of formula II form a cross-over light chain-heavy chain pair.
  • V H1 and V L1 form a binding pair and one of the three antigen binding sites
  • V H2 and V L2 form a binding pair and another of the three antigen binding sites
  • VH3 and VL3 form a binding 56
  • the first polypeptide chain and the second polypeptide chain have a cross-over orientation that forms two distinct antigen binding sites.
  • the VH1 and VL1 form a binding pair and form the first antigen binding site.
  • the VH2 and VL2 form a binding pair and form the second antigen binding site.
  • the first antigen binding site binds a CD3 polypeptide (e.g., human CD3)
  • the second antigen binding site binds a CD28 polypeptide (e.g., human CD28).
  • the second antigen binding site binds a CD3 polypeptide (e.g., human CD3), and the first antigen binding site binds a CD28 polypeptide (e.g., human CD28).
  • the third polypeptide and the fourth polypeptide form a third antigen binding site.
  • the VH3 and VL3 form a binding pair and form the third antigen binding site.
  • the third antigen binding site binds a CD38 polypeptide (e.g., human and optionally cynomolgus monkey CD38). Exemplary binding protein formats with cross-over orientations contemplated for use herein are also described in U.S. Pat. Appl. Ser.
  • the three antigen binding sites may be arranged in any combination that includes an antigen binding site that binds a CD38 polypeptide, an antigen binding site that binds a CD28 polypeptide, and an antigen binding site that binds a CD3 polypeptide.
  • VH1 and VL1 form the second antigen binding site that binds a CD28 polypeptide
  • V H2 and V L2 form the third antigen binding site that binds a CD3 polypeptide
  • V H3 and V L3 form the first antigen binding site that binds a CD38 polypeptide.
  • VH1 and VL1 form the third antigen binding site that binds a CD3 polypeptide
  • VH2 and VL2 form the second antigen binding site that binds a CD28 polypeptide
  • VH3 and V L3 form the first antigen binding site that binds a CD38 polypeptide
  • VH1 and VL1 form the second antigen binding site that binds a CD28 polypeptide
  • VH2 and VL2 form the first antigen binding site that binds a CD38 polypeptide
  • V H3 and V L3 form the third antigen binding site that binds a CD3 polypeptide.
  • V H1 and V L1 form the third antigen binding site that binds a CD3 polypeptide
  • VH2 and VL2 form the first antigen binding site that binds a CD38 polypeptide
  • VH3 and VL3 form the second antigen binding site that binds a CD28 polypeptide
  • V H1 and V L1 form the first antigen binding site that binds a CD38 polypeptide
  • VH2 and VL2 form the third antigen binding site that binds a CD3 polypeptide
  • VH3 and VL3 form the second antigen binding site that binds a CD28 polypeptide.
  • V H1 and V L1 form the first antigen binding site that binds a CD38 polypeptide, 57
  • the linkers can also be 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acids long.
  • L1, L2, L3 and L 4 in one binding protein may all have the same amino acid sequence or may all have different amino acid sequences.
  • linkers include a single glycine (Gly) residue; a diglycine peptide (Gly-Gly); a tripeptide (Gly-Gly-Gly); a peptide with four glycine residues; a peptide with five glycine residues; a peptide with six glycine residues; a peptide with seven glycine residues; and a peptide with eight glycine residues.
  • amino acid residues may be used such as the peptide GGGGSGGGGS (SEQ ID NO: 55), the peptide GGGGSGGGGSGGGGS (SEQ ID NO: 56), the peptide TKGPS (SEQ ID NO: 57), the peptide GQPKAAP (SEQ ID NO:58), and the peptide GGSGSSGSGG (SEQ ID NO:59).
  • linkers comprising randomly selected amino acids selected from the group consisting of valine, leucine, isoleucine, serine, threonine, lysine, arginine, histidine, aspartate, glutamate, asparagine, glutamine, glycine, and proline have been shown to be suitable in the binding proteins.
  • linker sequences see, e.g., WO2012135345 and International Application No. PCT/US2017/027488.
  • the identity and sequence of amino acid residues in the linker may vary depending on the type of secondary structural element necessary to achieve in the linker.
  • L1, L2, L3 or L4 is independently 0 amino acids in length.
  • L1, L2, L3 or L4 are each independently at least one amino acid in length.
  • the length of L 1 is at least twice the length of L 3 .
  • the length of L 2 is at least twice the length of L 4 .
  • the length of L1 is at least twice the length of L3, and the length of L2 is at least 58
  • L 1 is 3 to 12 amino acid residues in length
  • L 2 is 3 to 14 amino acid residues in length
  • L3 is 1 to 8 amino acid residues in length
  • L4 is 1 to 3 amino acid residues in length.
  • L1 is 5 to 10 amino acid residues in length
  • L 2 is 5 to 8 amino acid residues in length
  • L 3 is 1 to 5 amino acid residues in length
  • L4 is 1 to 2 amino acid residues in length.
  • L1 is 7 amino acid residues in length
  • L2 is 5 amino acid residues in length
  • L3 is 1 amino acid residue in length
  • L 4 is 2 amino acid residues in length.
  • L 1 is 10 amino acid residues in length
  • L2 is 10 amino acid residues in length
  • L3 is 0 amino acid residue in length
  • L4 is 0 amino acid residues in length.
  • L1, L2, L3, and L4 each have an independently selected length 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).
  • L1, L2, L3, and L4 are each 0 amino acids in length.
  • L 1 , L 2 , L 3 , and/or L 4 comprise a sequence derived from a naturally occurring sequence at the junction between an antibody variable domain and an antibody constant domain (e.g., as described in WO2012/135345).
  • the linker comprises a sequence found at the transition between an endogenous V H and C H1 domain, or between an endogenous V L and C L domain (e.g., kappa or lambda).
  • the linker comprises a sequence found at the transition between an endogenous human V H and C H1 domain, or between an endogenous human V L and C L domain (e.g., human kappa or lambda).
  • L1, L2, L3 and L4 each independently are zero amino acids in length or comprise 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).
  • L1, L2, L3 and L4 each independently comprise 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).
  • L1 comprises the sequence GQPKAAP (SEQ ID NO: 58)
  • L 2 comprises the sequence TKGPS (SEQ ID NO:57)
  • L 3 comprises the sequence S
  • L 4 comprises the sequence RT.
  • L1 comprises the sequence GGGGSGGGGS (SEQ ID NO:55)
  • L2 comprises the sequence GGGGSGGGGS (SEQ ID NO:55)
  • L 3 is 0 amino acids in length
  • L 4 is 0 amino acids in length.
  • L 1 comprises the sequence GGSGSSGSGG (SEQ ID NO:59)
  • L 2 comprises the sequence GGSGSSGSGG (SEQ ID NO:59)
  • L3 is 0 amino acids in length
  • L4 is 0 amino acids in length
  • L1 comprises the sequence GGGGSGGGGSGGGGS (SEQ ID NO:56)
  • L 2 is 0 amino acids in length
  • L 3 comprises the sequence GGGGSGGGGSGGGGS (SEQ ID NO:56)
  • L4 is 0 amino acids in length.
  • at least one of L1, L2, L3 or L4 comprises the sequence DKTHT (SEQ ID NO:147).
  • an anti-CD38 T-cell engager or binding protein of the present disclosure comprises a full-length antibody heavy chain or a polypeptide chain comprising an Fc region.
  • the Fc region is a human Fc region, e.g., a human IgG1, IgG2, IgG3, or IgG4 Fc region.
  • the Fc region includes an antibody hinge, C H1 , C H2 , C H3 , and optionally C H4 domains.
  • the Fc region is a human IgG1 Fc region. In some embodiments, the Fc region is a human IgG4 Fc region. In some embodiments, the Fc region includes one or more of the mutations described infra.
  • an anti-CD38 T-cell engager or binding protein of the present disclosure includes one or two Fc variants.
  • the term "Fc variant” as used herein refers to a molecule or sequence that is modified from a native Fc but still comprises a binding site for the salvage receptor, FcRn (neonatal Fc receptor). Exemplary Fc variants, and their interaction with the salvage receptor, are known in the art.
  • Fc variant can comprise a molecule or sequence that is humanized from a non-human native Fc.
  • a native Fc comprises regions that can be removed because they provide structural features or biological activity that are not required for the antibody-like binding proteins of the invention.
  • the term “Fc variant” comprises a molecule or sequence that lacks one or more native Fc sites or residues, or in which one or more Fc sites or residues has be modified, that affect or are involved in: (1) disulfide bond formation, (2) incompatibility with a selected host cell, (3) N-terminal heterogeneity upon expression in a selected host cell, (4) glycosylation, (5) interaction with complement, (6) binding to an Fc receptor other than a salvage receptor, or (7) antibody-dependent cellular cytotoxicity (ADCC).
  • the Fc region comprises one or more mutations that reduce or eliminate Fc receptor binding and/or effector function of the Fc region (e.g., Fc receptor- 60
  • the Fc region is a human IgG1 Fc region comprising one or more amino acid substitutions at positions corresponding to positions 234, 235, and/or 329 of human IgG1 according to EU Index.
  • the amino acid substitutions are L234A, L235A, and/or P329A.
  • the Fc region is a human IgG1 Fc region comprising amino acid substitutions at positions corresponding to positions 298, 299, and/or 300 of human IgG1 according to EU Index.
  • the amino acid substitutions are S298N, T299A, and/or Y300S.
  • the Fc region is a human IgG4 Fc region comprising one or more mutations that reduce or eliminate Fc ⁇ I and/or Fc ⁇ II binding.
  • the Fc region is a human IgG4 Fc region comprising one or more mutations that reduce or eliminate Fc ⁇ I and/or Fc ⁇ II binding but do not affect FcRn binding.
  • the Fc region is a human IgG4 Fc region comprising amino acid substitutions at positions corresponding to positions 228 and/or 409 of human IgG4 according to EU Index.
  • the amino acid substitutions are S228P and /or R409K.
  • the Fc region is a human IgG4 Fc region comprising amino acid substitutions at positions corresponding to positions 234 and/or 235 of human IgG4 according to EU Index.
  • the amino acid substitutions are F234A and/or L235A.
  • the Fc region is a human IgG4 Fc region comprising amino acid substitutions at positions corresponding to positions 228, 234, 235, and/or 409 of human IgG4 according to EU Index.
  • the amino acid substitutions are S228P, F234A, L235A, and /or R409K.
  • the Fc region is a human IgG4 Fc region comprising amino acid substitutions at positions corresponding to positions 233-236 of human IgG4 according to EU Index.
  • the amino acid substitutions are E233P, F234V, L235A, and a deletion at 236.
  • the Fc region is a human IgG4 Fc region comprising amino acid mutations at substitutions corresponding to positions 228, 233-236, and/or 409 of human IgG4 according to EU Index.
  • the amino acid mutations are S228P; E233P, F234V, L235A, and a deletion at 236; and/or R409K.
  • an anti-CD38 T-cell engager or binding protein of the present disclosure comprises one or more mutations to improve purification, e.g., by modulating the affinity for a purification reagent.
  • a purification reagent for example, it is known that heterodimeric binding proteins can be selectively purified away from their homodimeric 61
  • the mutation comprises substitutions at positions corresponding to positions 435 and 436 of human IgG1 or IgG4 according to EU Index, wherein the amino acid substitutions are H435R and Y436F.
  • the binding protein comprises a second polypeptide chain further comprising a first Fc region linked to CH1, the first Fc region comprising an immunoglobulin hinge region and CH2 and C H3 immunoglobulin heavy chain constant domains, and a third polypeptide chain further comprising a second Fc region linked to C H1 , the second Fc region comprising an immunoglobulin hinge region and CH2 and CH3 immunoglobulin heavy chain constant domains; and wherein only one of the first and the second Fc regions comprises amino acid substitutions at positions corresponding to positions 435 and 436 of human IgG1 or IgG4 according to EU Index, wherein the amino acid substitutions are H435R and Y436F.
  • a binding protein of the present disclosure comprises knob and hole mutations and one or more mutations to improve purification.
  • the first and/or second Fc regions are human IgG1 Fc regions.
  • the first and/or second Fc regions are human IgG4 Fc regions.
  • the C H3 domains can be altered by the "knob-into-holes" technology which 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.
  • the interaction surfaces of the two C H3 domains are altered to increase the heterodimerisation of both heavy chains containing these two CH3 domains.
  • Each of the two CH3 domains (of the two heavy chains) can be the "knob," while the other is the "hole.”
  • the introduction of a disulfide bridge further stabilizes the heterodimers (Merchant et al., 1998; Atwell et al., 1997, J. Mol. Biol.270: 26-35) and increases the yield.
  • the knob is on the second pair of polypeptides with a single variable domain. In other embodiments, the knob is on the first pair of polypeptides having the cross-over orientation.
  • an anti-CD38 T-cell engager or binding protein of the present disclosure comprises a “knob” mutation on the 62
  • a binding protein of the present disclosure comprises a “knob” mutation on the third polypeptide chain and a “hole” mutation on the second polypeptide chain.
  • the “knob” mutation comprises substitution(s) at positions corresponding to positions 354 and/or 366 of human IgG1 or IgG4 according to EU Index.
  • the amino acid substitutions are S354C, T366W, T366Y, S354C and T366W, or S354C and T366Y.
  • the “knob” mutation comprises substitutions at positions corresponding to positions 354 and 366 of human IgG1 or IgG4 according to EU Index.
  • the amino acid substitutions are S354C and T366W.
  • the “hole” mutation comprises substitution(s) at positions corresponding to positions 407 and, optionally, 349, 366, and/or 368 and of human IgG1 or IgG4 according to EU Index.
  • the amino acid substitutions are Y407V or Y407T and optionally Y349C, T366S, and/or L368A.
  • the “hole” mutation comprises substitutions at positions corresponding to positions 349, 366, 368, and 407 of human IgG1 or IgG4 according to EU Index.
  • the amino acid substitutions are Y349C, T366S, L368A, and Y407V.
  • the second polypeptide chain further comprises 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 comprises amino acid substitution(s) at positions corresponding to positions 366 and optionally 354 of human IgG1 or IgG4 according to EU Index, wherein the amino acid substitutions are T366W or T366Y and optionally S354C; and wherein the third polypeptide chain further comprises 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 comprises amino acid substitution(s) at positions corresponding to positions 407 and optionally 349, 366, and/or 368 and of human IgG1 or IgG4 according to EU Index, wherein the amino acid substitutions are Y407V or Y407T and optionally Y349
  • the second polypeptide chain further comprises 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 comprises amino acid substitution(s) at positions corresponding to positions 407 and optionally 349, 366, and/or 368 and of human IgG1 or IgG4 according to EU Index, wherein the amino acid substitutions are Y407V or Y407T and optionally Y349C, T366S, and/or 63
  • the third polypeptide chain further comprises 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 comprises amino acid substitution(s) at positions corresponding to positions 366 and optionally 354 of human IgG1 or IgG4 according to EU Index, wherein the amino acid substitutions are T366W or T366Y and optionally S354C.
  • the second polypeptide chain further comprises 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 comprises amino acid substitution at position corresponding to position 366 of human IgG1 or IgG4 according to EU Index, wherein the amino acid substitution is T366W; and wherein the third polypeptide chain further comprises 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 comprises amino acid substitution(s) at positions corresponding to positions 366, 368, and/or 407 and of human IgG1 or IgG4 according to EU Index, wherein the amino acid substitutions are T366S, L368A, and/or Y407V.
  • the second polypeptide chain further comprises 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 comprises amino acid substitution(s) at positions corresponding to positions 366, 368, and/or 407 and of human IgG1 or IgG4 according to EU Index, wherein the amino acid substitutions are T366S, L368A, and/or Y407V; and wherein the third polypeptide chain further comprises 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 comprises amino acid substitution at position corresponding to position 366 of human IgG1 or IgG4 according to EU Index, wherein the amino acid substitution is T366W.
  • the second polypeptide chain further comprises 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 comprises amino acid substitutions at positions corresponding to positions 354 and 366 of human IgG1 or IgG4 according to EU Index, wherein the amino acid substitutions are S354C and T366W; and wherein the third polypeptide chain further comprises a second Fc region 64
  • the second Fc region comprising an immunoglobulin hinge region and CH2 and CH3 immunoglobulin heavy chain constant domains, wherein the second Fc region comprises amino acid substitutions at positions corresponding to positions 349, 366, 368, and 407 of human IgG1 or IgG4 according to EU Index, wherein the amino acid substitutions are Y349C, T366S, L368A, and Y407V.
  • the second polypeptide chain further comprises 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 comprises amino acid substitutions at positions corresponding to positions 349, 366, 368, and 407 of human IgG1 or IgG4 according to EU Index, wherein the amino acid substitutions are Y349C, T366S, L368A, and Y407V; and wherein the third polypeptide chain further comprises 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 comprises amino acid substitutions at positions corresponding to positions 354 and 366 of human IgG1 or IgG4 according to EU Index, wherein the amino acid substitutions are S354C and T366W.
  • 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.
  • the second polypeptide chain further comprises 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 corresponding to positions 228, 354, 366, and 409 of human IgG4 according to EU Index, wherein the amino acid substitutions are S228P, S354C, T366W, and R409K; and wherein the third polypeptide chain further comprises 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, wherein the second F
  • the second polypeptide chain further comprises 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 corresponding to positions 228, 349, 366, 368, 407, and 409 of human IgG4 according to EU Index, wherein the amino acid 65
  • substitutions are S228P, Y349C, T366S, L368A, Y407V, and R409K; and wherein the third polypeptide chain further comprises 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, wherein the second Fc region comprises amino acid substitutions at positions corresponding to positions 228, 354, 366, and 409 of human IgG4 according to EU Index, wherein the amino acid substitutions are S228P, S354C, T366W, and R409K.
  • the second polypeptide chain further comprises 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 corresponding to positions 234, 235, 354, and 366 of human IgG4 according to EU Index, wherein the amino acid substitutions are F234A, L235A, S354C, and T366W; and wherein the third polypeptide chain further comprises 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, wherein the second Fc region comprises amino acid substitutions at positions corresponding to positions 234, 235, 349, 366, 368, and 407 of human IgG4 according to EU Index, wherein the amino acid substitutions are
  • the second polypeptide chain further comprises 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 corresponding to positions 234, 235, 349, 366, 368, and 407 of human IgG4 according to EU Index, wherein the amino acid substitutions are F234A, L235A, Y349C, T366S, L368A, and Y407V; and wherein the third polypeptide chain further comprises 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, wherein the second Fc region comprises amino acid substitutions at positions corresponding to positions 234, 235, 354, and 366 of human IgG4 according to EU Index,
  • the second polypeptide chain further comprises 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 66
  • the first Fc region comprises amino acid substitutions at positions corresponding to positions 228, 234, 235, 354, 366, and 409 of human IgG4 according to EU Index, wherein the amino acid substitutions are S228P, F234A, L235A, S354C, T366W, and R409K; and wherein the third polypeptide chain further comprises 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, wherein the second Fc region comprises amino acid substitutions at positions corresponding to positions 228, 234, 235, 349, 366, 368, 407, and 409 of human IgG4 according to EU Index, wherein the amino acid substitutions are S228P, F234A, L235A, Y349C, T366S, L368A, Y407V, and R409K.
  • the second polypeptide chain further comprises 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 corresponding to positions 228, 234, 235, 349, 366, 368, 407, and 409 of human IgG4 according to EU Index, wherein the amino acid substitutions are S228P, F234A, L235A, Y349C, T366S, L368A, Y407V, and R409K; and wherein the third polypeptide chain further comprises 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, wherein the second Fc region comprises amino acid substitutions at positions corresponding to positions 228, 234, 235
  • an anti-CD38 T-cell engager or binding protein of the present disclosure comprises one or more mutations to improve serum half-life (See e.g., Hinton, P.R. et al. (2006) J. Immunol.176(1):346-56).
  • the mutation comprises substitutions at positions corresponding to positions 428 and 434 of human IgG1 or IgG4 according to EU Index, wherein the amino acid substitutions are M428L and N434S.
  • a binding protein of the present disclosure comprises knob and hole mutations and one or more mutations to improve serum half-life.
  • the first and/or second Fc regions are human IgG1 Fc regions.
  • the first and/or second Fc regions are human IgG4 Fc regions.
  • an anti-CD38 T-cell engager or binding protein of the present disclosure comprises one or more mutations to improve stability, e.g., of the hinge region and/or dimer interface of IgG4 (See e.g., Spiess, C. et al. (2013) J. Biol. Chem.
  • the mutation comprises substitutions at positions corresponding to positions 228 and 409 of human IgG4 according to EU Index, wherein the amino acid substitutions are S228P and R409K.
  • the binding protein comprises a 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 a 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 second Fc regions are human IgG4 Fc regions; and wherein the first and the second Fc regions each comprise amino acid substitutions at positions corresponding to positions 228 and 409 of human IgG4 according to EU Index, wherein the amino acid substitutions are S228P and R409K.
  • a binding protein of the present disclosure comprises knob and hole mutations and one or more mutations to improve stability.
  • the first and/or second Fc regions are human IgG4 Fc regions.
  • an anti-CD38 T-cell engager or binding protein of the present disclosure comprises one or more mutations to improve purification, e.g., by modulating the affinity for a purification reagent.
  • a binding protein of the present disclosure comprises knob and hole mutations and one or more mutations to improve purification.
  • an anti-CD38 T-cell engager or binding protein of the present disclosure comprises one or more mutations to improve serum half-life (See e.g., Hinton, P.R. et al. (2006) J. Immunol.176(1):346-56).
  • the mutation comprises substitutions at positions corresponding to positions 428 and 434 of human IgG1 or IgG4 according to EU Index, wherein the amino acid substitutions are M428L and N434S.
  • a binding protein of the present disclosure comprises knob and hole mutations and one or more mutations to improve serum half-life.
  • the first and/or second Fc regions are human IgG1 Fc regions.
  • the first and/or second Fc regions are human IgG4 Fc regions.
  • an anti-CD38 T-cell engager or binding protein of the present disclosure comprises one or more mutations to reduce effector function, e.g., Fc receptor-mediated antibody-dependent cellular phagocytosis (ADCP), complement- dependent cytotoxicity (CDC), and/or antibody-dependent cellular cytotoxicity (ADCC).
  • the second polypeptide chain further comprises a first Fc region linked to C H1 , the first Fc region comprising an immunoglobulin hinge region and C H2 and C H3 69
  • the Fc regions of the second and the third polypeptide chains are human IgG1 Fc regions, and wherein the Fc regions each comprise amino acid substitutions at positions corresponding to positions 234 and 235 of human IgG1 according to EU Index, wherein the amino acid substitutions are L234A and L235A.
  • the second polypeptide chain further comprises 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 third polypeptide chain further comprises a second Fc region linked to C H1 , the second Fc region comprising an immunoglobulin hinge region and CH2 and CH3 immunoglobulin heavy chain constant domains; wherein the first and second Fc regions are human IgG1 Fc regions; and wherein the first and the second Fc regions each comprise amino acid substitutions at positions corresponding to positions 234, 235, and 329 of human IgG1 according to EU Index, wherein the amino acid substitutions are L234A, L235A, and P329A.
  • the Fc regions of the second and the third polypeptide chains are human IgG1 Fc regions, and wherein the Fc regions each comprise amino acid substitutions at positions corresponding to positions 234, 235, and 329 of human IgG1 according to EU Index, wherein the amino acid substitutions are L234A, L235A, and P329A.
  • the Fc regions of the second and the third polypeptide chains are human IgG4 Fc regions, and the Fc regions each comprise amino acid substitutions at positions corresponding to positions 234 and 235 of human IgG4 according to EU Index, wherein the amino acid substitutions are F234A and L235A.
  • the binding protein comprises a second polypeptide chain further comprising a first Fc region linked to C H1 , the first Fc region comprising an immunoglobulin hinge region and CH2 and CH3 immunoglobulin heavy chain constant domains, and a third polypeptide chain further comprising a second Fc region linked to C H1 , the second Fc region comprising an immunoglobulin hinge region and C H2 and C H3 immunoglobulin heavy chain constant domains; and wherein the first and the second Fc regions each comprise amino acid substitutions at positions corresponding to positions 234 70
  • an anti-CD38 T-cell engager or binding protein of the present disclosure comprises knob and hole mutations and one or more mutations to reduce effector function.
  • the first and/or second Fc regions are human IgG1 Fc regions.
  • the first and/or second Fc regions are human IgG4 Fc regions.
  • a binding protein of the present disclosure can comprise two or more of the “knob” and “hole” mutations, the one or more mutations to improve serum half-life, the one or more mutations to improve IgG4 stability, the one or more mutations to improve purification, and/or the one or more mutations to reduce effector function described supra.
  • an anti-CD38 T-cell engager or binding protein of the present disclosure comprises an antibody fragment, including but not limited to antibody F(ab), F(ab’)2, Fab’-SH, Fv, or scFv fragments.
  • the first polypeptide chain comprises a polypeptide sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:61
  • the second polypeptide chain comprises a polypeptide sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:60
  • the third polypeptide chain comprises a polypeptide sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:62
  • the fourth polypeptide chain comprises a polypeptide sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:63.
  • the first polypeptide chain comprises a polypeptide sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:61
  • the second polypeptide chain comprises a polypeptide sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:64
  • the third polypeptide chain comprises a polypeptide sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:65
  • the fourth polypeptide chain comprises a polypeptide sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:63.
  • the first polypeptide chain comprises a polypeptide sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:61, the 71
  • the second polypeptide chain comprises a polypeptide sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:66
  • the third polypeptide chain comprises a polypeptide sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:67
  • the fourth polypeptide chain comprises a polypeptide sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:63.
  • the first polypeptide chain comprises a polypeptide sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:61
  • the second polypeptide chain comprises a polypeptide sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:60
  • the third polypeptide chain comprises a polypeptide sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:68
  • the fourth polypeptide chain comprises a polypeptide sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:69.
  • the first polypeptide chain comprises a polypeptide sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:61
  • the second polypeptide chain comprises a polypeptide sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:64
  • the third polypeptide chain comprises a polypeptide sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:70
  • the fourth polypeptide chain comprises a polypeptide sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:69.
  • the first polypeptide chain comprises a polypeptide sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:61
  • the second polypeptide chain comprises a polypeptide sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:66
  • the third polypeptide chain comprises a polypeptide sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:71
  • the fourth polypeptide chain comprises a polypeptide sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:69.
  • the first polypeptide chain comprises a polypeptide sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:148
  • the second polypeptide chain comprises a polypeptide sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:149
  • the third polypeptide chain comprises a polypeptide sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:150
  • the fourth polypeptide chain comprises a polypeptide sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:151.
  • the first polypeptide chain comprises a polypeptide sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:152
  • the second polypeptide chain comprises a polypeptide sequence that is at least 95% identical to the amino acid sequence of SEQ ID 72
  • the third polypeptide chain comprises a polypeptide sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:154
  • the fourth polypeptide chain comprises a polypeptide sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:155.
  • the first polypeptide chain comprises a polypeptide sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:156
  • the second polypeptide chain comprises a polypeptide sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:157
  • the third polypeptide chain comprises a polypeptide sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:158
  • the fourth polypeptide chain comprises a polypeptide sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:159.
  • the first polypeptide chain comprises a polypeptide sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:160
  • the second polypeptide chain comprises a polypeptide sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:161
  • the third polypeptide chain comprises a polypeptide sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:162
  • the fourth polypeptide chain comprises a polypeptide sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:163.
  • the first polypeptide chain comprises a polypeptide sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:164
  • the second polypeptide chain comprises a polypeptide sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:165
  • the third polypeptide chain comprises a polypeptide sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:166
  • the fourth polypeptide chain comprises a polypeptide sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:167.
  • the first polypeptide chain comprises a polypeptide sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:168
  • the second polypeptide chain comprises a polypeptide sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:169
  • the third polypeptide chain comprises a polypeptide sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:170
  • the fourth polypeptide chain comprises a polypeptide sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:171.
  • the first polypeptide chain comprises a polypeptide sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:172
  • the second polypeptide chain comprises a polypeptide sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:173
  • the third polypeptide chain comprises a polypeptide sequence that is at least 95% identical to the amino acid sequence of 73
  • the fourth polypeptide chain comprises a polypeptide sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:175.
  • the first polypeptide chain comprises a polypeptide sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:176
  • the second polypeptide chain comprises a polypeptide sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:177
  • the third polypeptide chain comprises a polypeptide sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:178
  • the fourth polypeptide chain comprises a polypeptide sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:179.
  • 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
  • the fourth polypeptide chain comprises the amino acid sequence of SEQ ID NO:63.
  • 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
  • the fourth polypeptide chain comprises the amino acid sequence of SEQ ID NO:63.
  • 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
  • the fourth polypeptide chain comprises the amino acid sequence of SEQ ID NO:63.
  • 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
  • the fourth polypeptide chain comprises the amino acid sequence of SEQ ID NO:69.
  • 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
  • the fourth polypeptide chain comprises the amino acid sequence of SEQ ID NO:69.
  • 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
  • the fourth polypeptide chain comprises the amino acid sequence of SEQ ID NO:69.
  • the first polypeptide chain comprises the amino acid 74
  • 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
  • the fourth polypeptide chain comprises the amino acid sequence of SEQ ID NO:155.
  • 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
  • the fourth polypeptide chain comprises the amino acid sequence of SEQ ID NO:159.
  • 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
  • the fourth polypeptide chain comprises the amino acid sequence of SEQ ID NO:163.
  • 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
  • the fourth polypeptide chain comprises the amino acid sequence of SEQ ID NO:167.
  • 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
  • the fourth polypeptide chain comprises the amino acid sequence of SEQ ID NO:171.
  • 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
  • the fourth polypeptide chain comprises the amino acid sequence of SEQ ID NO:175.
  • 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
  • the fourth polypeptide chain comprises the amino acid sequence of SEQ ID NO:179.
  • a trispecific binding protein comprises 1, 2, 3, or 4 polypeptide chains shown in Table E1 or E2.
  • a trispecific binding protein comprises 1, 2, 3, or 4 polypeptide chains of a single trispecific binding protein shown in Table E1 or E2.
  • a trispecific binding protein comprises 1, 2, 3, or 4 polypeptide chains encoded by a polynucleotide sequence shown in Table F1 or F2.
  • a trispecific binding protein comprises 1, 2, 3, or 4 polypeptide chains encoded by the polynucleotide sequence(s) of a single trispecific binding protein shown in Table F1 or F2.
  • Table E1 Full-length sequences of trispecific binding proteins.
  • Standard recombinant DNA methodologies are used to construct the polynucleotides that encode the polypeptides which form the anti-CD38 T-cell engagers, incorporate these polynucleotides into recombinant expression vectors, and introduce such vectors into host cells. See e.g., Sambrook et al., 2001, MOLECULAR CLONING: A LABORATORY MANUAL (Cold Spring Harbor Laboratory Press, 3rd ed.). Enzymatic reactions and purification techniques may be performed according to manufacturer's specifications, as commonly accomplished in the art, or as described herein.
  • kits of polynucleotides relate to kits of polynucleotides.
  • one or more of the polynucleotides is a vector (e.g., an expression vector).
  • the kits may find use, inter alia, in producing one or more of the anti-CD38 T-cell engagers described herein.
  • kits of the present disclosure may include one or more polynucleotides encoding 1, 2, 3, or 4 polypeptides of an anti-CD38 T-cell engager.
  • a kit of the present disclosure comprises one or more polynucleotides encoding 1, 2, 3, or 4 polypeptide chains of a single trispecific binding protein, e.g., as shown in Table F1 or F2.
  • the isolated nucleic acid is operably linked to a heterologous promoter to direct transcription of the binding protein-coding nucleic acid sequence.
  • a promoter may refer to nucleic acid control sequences which direct transcription of a nucleic acid.
  • a first nucleic acid sequence is operably linked to a second nucleic acid sequence when the first nucleic acid sequence is placed in a functional relationship with the second nucleic acid sequence.
  • a promoter is operably linked to a coding sequence of a binding protein if the promoter affects the transcription or expression of the coding sequence.
  • promoters may include, but are not limited to, promoters obtained from the genomes of viruses (such as polyoma virus, fowlpox virus, adenovirus (such as Adenovirus 2), bovine papilloma virus, avian sarcoma virus, cytomegalovirus, a retrovirus, hepatitis-B virus, Simian Virus 40 (SV40), and the like), from heterologous eukaryotic promoters (such as the actin promoter, an immunoglobulin promoter, from heat- shock promoters, and the like), the CAG-promoter (Niwa et al., Gene 108(2):193-9, 1991), the phosphoglycerate kinase (PGK)-promoter, a tetracycline-inducible promoter (Masui et al., 124
  • viruses such as polyoma virus, fowlpox virus, adenovirus (such as
  • the isolated nucleic acid is incorporated into a vector.
  • the vector is an expression vector.
  • Expression vectors 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).
  • suitable enhancers may include, but are not limited to, enhancer sequences from mammalian genes (such as globin, elastase, albumin, ⁇ -fetoprotein, insulin and the like), and enhancer sequences from a eukaryotic cell virus (such as SV40 enhancer on the late side of the replication origin (bp 100-270), the cytomegalovirus early promoter enhancer, the polyoma enhancer on the late side of the replication origin, adenovirus enhancers, and the like).
  • mammalian genes such as globin, elastase, albumin, ⁇ -fetoprotein, insulin and the like
  • enhancer sequences from a eukaryotic cell virus (such as
  • suitable vectors may include, for example, plasmids, cosmids, episomes, transposons, and viral vectors (e.g., adenoviral, vaccinia viral, Sindbis-viral, measles, herpes viral, lentiviral, retroviral, adeno-associated viral vectors, etc.).
  • Expression vectors can be used to transfect host cells, such as, for example, bacterial cells, yeast cells, insect cells, and mammalian cells.
  • Biologically functional viral and plasmid DNA vectors capable of expression and replication in a host are known in the art, and can be used to transfect any cell of interest.
  • the vector system comprises one or more vectors encoding a first, second, third, and fourth polypeptide chain of any of the anti- CD38 T-cell engagers described herein.
  • the vector system comprises a first vector encoding the first polypeptide chain of the binding protein, a second vector encoding the second polypeptide chain of the binding protein, a third vector encoding the third polypeptide chain of the binding protein, and a fourth vector encoding the fourth polypeptide chain of the binding protein.
  • the vector system comprises a first vector encoding the first and second polypeptide chains of the binding protein, and a second vector encoding the third and fourth polypeptide chains of the binding protein.
  • the vector system comprises a first vector encoding the first and third polypeptide chains of the binding protein, and a second vector encoding the second and 125
  • the vector system comprises a first vector encoding the first and fourth polypeptide chains of the binding protein, and a second vector encoding the second and third polypeptide chains of the binding protein. In some embodiments, the vector system comprises a first vector encoding the first, second, third, and fourth polypeptide chains of the binding protein.
  • the one or more vectors of the vector system may be any of the vectors described herein. In some embodiments, the one or more vectors are expression vectors. [0168] Other aspects of the present disclosure relate to an isolated host cell comprising one or more isolated polynucleotides, polynucleotide kits, vectors, and/or vector systems described herein.
  • the host cell is a bacterial cell (e.g., an E. coli cell). In some embodiments, the host cell is a yeast cell (e.g., an S. cerevisiae cell). In some embodiments, the host cell is an insect cell. Examples of insect host cells may include, for example, Drosophila cells (e.g., S2 cells), Trichoplusia ni cells (e.g., High FiveTM cells), and Spodoptera frugiperda cells (e.g., Sf21 or Sf9 cells). In some embodiments, the host cell is a mammalian cell.
  • Drosophila cells e.g., S2 cells
  • Trichoplusia ni cells e.g., High FiveTM cells
  • Spodoptera frugiperda cells e.g., Sf21 or Sf9 cells.
  • the host cell is a mammalian cell.
  • mammalian host cells may include, for example, human embryonic kidney cells (e.g., 293 or 293 cells subcloned for growth in suspension culture), Expi293TM cells, CHO cells, baby hamster kidney cells (e.g., BHK, ATCC CCL 10), mouse sertoli 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 carcinoma cells (e.g., HELA, ATCC CCL 2), canine kidney cells (e.g., MDCK, ATCC CCL 34), buffalo rat liver cells (e.g., BRL 3A, ATCC CRL 1442), human lung cells (e.g., W138, ATCC CCL 75), human liver cells (e.g., Hep G2, HB 8065), mouse mammary tumor cells (e.g., MMT 060562, ATCC CCL51), a
  • the method includes a) culturing a host cell (e.g., any of the host cells described herein) comprising an isolated nucleic acid, vector, and/or vector system (e.g., any of the isolated nucleic acids, vectors, and/or vector systems described herein) under conditions such that the host cell expresses the anti-CD38 T-cell engager; and b) isolating the anti-CD38 T-cell engager from the host cell.
  • a host cell e.g., any of the host cells described herein
  • an isolated nucleic acid, vector, and/or vector system e.g., any of the isolated nucleic acids, vectors, and/or vector systems described herein
  • an anti-CD38 T-cell engager of the present disclosure is purified by protein A affinity chromatography, kappa light chain affinity chromatography (e.g., using a KappaSelect resin according to manufacturer’s instructions; GE Healthcare), and optionally lambda light chain affinity chromatography (e.g., using a LambdaFabSelect resin according to manufacturer’s instructions; GE Healthcare).
  • a binding protein of the present disclosure is purified by Protein A affinity chromatography, lambda light chain affinity chromatography (e.g., using a LambdaFabSelect resin according to manufacturer’s instructions; GE Healthcare), and optionally kappa light chain affinity chromatography (e.g., using a KappaSelect resin according to manufacturer’s instructions; GE Healthcare).
  • the binding protein comprises two Fc regions, each comprising a CH3 domain, and only one of the CH3 domains comprises amino acid substitutions at positions corresponding to positions 435 and 436 of human IgG1 or IgG4 according to EU Index, wherein the amino acid substitutions are H435R and Y436F.
  • a binding protein of the present disclosure is purified by protein A affinity chromatography, then kappa light chain affinity chromatography (e.g., using a KappaSelect resin according to manufacturer’s instructions; GE Healthcare), then optionally lambda light chain affinity chromatography (e.g., using a LambdaFabSelect resin according to manufacturer’s instructions; GE Healthcare) in sequence.
  • a binding protein of the present disclosure is purified by Protein A affinity chromatography, then lambda light chain affinity chromatography (e.g., using a LambdaFabSelect resin according to manufacturer’s instructions; GE Healthcare), then optionally kappa light chain affinity chromatography (e.g., using a KappaSelect resin according to manufacturer’s instructions; GE Healthcare) in sequence.
  • the binding protein is contacted with Protein A, eluted from Protein A under conditions suitable for isolating the binding protein away from binding proteins comprising either 0 or 2 C H3 domains comprising the amino acid substitutions are H435R and Y436F, contacted with a kappa light chain affinity medium (e.g., as used in the KappaSelect resin; GE Healthcare), and eluted from the kappa light chain affinity medium under conditions suitable for isolating the binding protein away from binding proteins comprising only lambda C L domains (e.g., according to manufacturer’s instructions).
  • Conditions suitable for the Protein A elution are known in the art, including without limitation a stepwise elution gradient from pH4.5-2.8.
  • Protein A or a Protein A variant useful for protein purification is employed.
  • the Protein A is attached to a substrate or resin, e.g., as part of a chromatography medium.
  • the binding protein is contacted with a lambda light chain affinity medium (e.g., as used in the LambdaFabSelect resin; GE Healthcare), and eluted from the lambda light chain affinity medium under conditions suitable for isolating the binding protein away from binding proteins comprising only kappa CL domains (e.g., according to manufacturer’s instructions).
  • a binding protein of the present disclosure is detected using HIC chromatography.
  • the binding protein comprises: a first polypeptide chain that comprises a lambda CL domain; a CH3 domain of a second polypeptide chain that comprises amino acid substitutions at positions corresponding to positions 354 and 366 of human IgG1 or IgG4according to EU Index, wherein the amino acid substitutions are S354C and T366W; a CH3 domain of a third polypeptide chain that comprises amino acid substitutions at positions corresponding to positions 349, 366, 368, 407, 435, and 436 of human IgG1 or IgG4according to EU Index, wherein the amino acid substitutions are Y349C, T366S, L368A, Y407V, H435R, and Y436F; and a fourth polypeptide chain that comprises a kappa CL domain.
  • the binding protein is produced by a host cell. In some embodiments, the binding protein is purified from a cell culture medium or host cell extract. In some embodiments, the binding proteins are secreted by a host cell or produced and extracted from a host cell (e.g., before being contacted with Protein A). In some embodiments, the binding protein is in a cell culture medium or host cell extract when contacted with Protein A. In some embodiments, the binding protein is purified away from other binding proteins, polypeptides, and/or other cellular components. III. Therapeutic compositions, kits, and administration thereof [0171] In some embodiments, provided herein is the anti-CD38 T-cell engager of the present disclosure for use in any of the methods described herein.
  • the methods comprise administering an effective amount of the anti-CD38 T-cell engager to an individual in need thereof, e.g., to an individual that has or has been diagnosed with PTCL for treatment of PTCL.
  • an anti-CD38 T-cell engager of the present disclosure is used in the manufacture of a medicament for treating PTCL in an individual, e.g., according to any of the methods described herein.
  • a kit or article of manufacture is provided.
  • the kit comprises an anti-CD38 T-cell engager of the present disclosure and, 128
  • a pharmaceutical composition is provided.
  • the pharmaceutical composition comprises an effective amount of an anti- CD38 T-cell engager of the present disclosure and a pharmaceutically acceptable carrier.
  • the composition is for use according to any of the methods of the present disclosure.
  • Therapeutic or pharmaceutical compositions comprising anti-CD38 T-cell engagers or binding proteins are within the scope of the disclosure.
  • Such therapeutic or pharmaceutical compositions can comprise a therapeutically effective amount of a T-cell engager or binding protein, or T-cell engager/binding protein-drug conjugate, in admixture with a pharmaceutically or physiologically acceptable formulation agent selected for suitability with the mode of administration.
  • Acceptable formulation materials preferably are nontoxic to recipients at the dosages and concentrations employed.
  • the pharmaceutical composition can contain formulation materials for modifying, maintaining, or preserving, for example, the pH, osmolarity, viscosity, clarity, color, isotonicity, odor, sterility, stability, rate of dissolution or release, adsorption, or penetration of the composition.
  • Suitable formulation materials include, but are not limited to, amino acids (such as glycine, glutamine, asparagine, arginine, or lysine), antimicrobials, antioxidants (such as ascorbic acid, sodium sulfite, or sodium hydrogen-sulfite), buffers (such as borate, bicarbonate, Tris-HCl, citrates, phosphates, or other organic acids), bulking agents (such as mannitol or glycine), chelating agents (such as ethylenediamine tetraacetic acid (EDTA)), complexing agents (such as caffeine, polyvinylpyrrolidone, beta-cyclodextrin, or hydroxypropyl-beta-cyclodextrin), fillers, monosaccharides, disaccharides, and other carbohydrates (such as glucose, mannose, or dextrins), proteins (such as serum albumin, gelatin, or immunoglobulins), coloring, flavoring and diluting agents, emuls
  • tyloxapal tyloxapal
  • stability enhancing agents such as sucrose or sorbitol
  • tonicity enhancing agents such as alkali metal halides – preferably sodium or potassium chloride – or mannitol sorbitol
  • delivery vehicles diluents, excipients and/or pharmaceutical adjuvants (see, e.g., REMINGTON'S PHARMACEUTICAL SCIENCES (18th Ed., A.R. Gennaro, ed., Mack Publishing Company 1990), and subsequent editions of the same, incorporated herein by reference for any purpose).
  • the optimal pharmaceutical composition will be determined by a skilled artisan depending upon, for example, the intended route of administration, delivery format, and desired dosage.
  • the primary vehicle or carrier in a pharmaceutical composition can be either aqueous or non-aqueous in nature.
  • a suitable vehicle or carrier for injection can be water, physiological saline solution, or artificial cerebrospinal fluid, possibly supplemented with other materials common in compositions for parenteral administration.
  • Neutral buffered saline or saline mixed with serum albumin are further exemplary vehicles.
  • Other exemplary pharmaceutical compositions comprise Tris buffer of about pH 7.0-8.5, or acetate buffer of about pH 4.0-5.5, which can further include sorbitol or a suitable substitute.
  • binding protein compositions can be prepared for storage by mixing the selected composition having the desired degree of purity with optional formulation agents in the form of a lyophilized cake or an aqueous solution. Further, the binding protein can be formulated as a lyophilizate using appropriate excipients such as sucrose. [0180]
  • the pharmaceutical compositions of the disclosure can be selected for parenteral delivery or subcutaneous. Alternatively, the compositions can be selected for inhalation or for delivery through the digestive tract, such as orally. The preparation of such pharmaceutically acceptable compositions is within the skill of the art.
  • the formulation components are present in concentrations that are acceptable to the site of administration.
  • the therapeutic compositions for use can be in the form of a pyrogen-free, parenterally acceptable, aqueous solution comprising the desired binding protein in a pharmaceutically acceptable vehicle.
  • a particularly suitable vehicle for parenteral injection is sterile distilled water in which a 130
  • binding protein is formulated as a sterile, isotonic solution, properly preserved.
  • Yet another preparation can involve the formulation of the desired molecule with an agent, such as injectable microspheres, bio-erodible particles, polymeric compounds (such as polylactic acid or polyglycolic acid), beads, or liposomes, that provides for the controlled or sustained release of the product which can then be delivered via a depot injection.
  • Hyaluronic acid can also be used, and this can have the effect of promoting sustained duration in the circulation.
  • Other suitable means for the introduction of the desired molecule include implantable drug delivery devices.
  • a pharmaceutical composition can be formulated for inhalation.
  • a binding protein can be formulated as a dry powder for inhalation.
  • Binding protein inhalation solutions can also be formulated with a propellant for aerosol delivery. In yet another embodiment, solutions can be nebulized. [0184] It is also contemplated that certain formulations can be administered orally. In one embodiment of the disclosure, binding proteins that are administered in this fashion can be formulated with or without those carriers customarily used in the compounding of solid dosage forms such as tablets and capsules. For example, a capsule can be designed to release the active portion of the formulation at the point in the gastrointestinal tract when bioavailability is maximized and pre-systemic degradation is minimized. Additional agents can be included to facilitate absorption of the binding protein.
  • Diluents, flavorings, low melting point waxes, vegetable oils, lubricants, suspending agents, tablet disintegrating agents, and binders can also be employed.
  • Another pharmaceutical composition can involve an effective quantity of binding proteins in a mixture with non-toxic excipients that are suitable for the manufacture of tablets. By dissolving the tablets in sterile water, or another appropriate vehicle, solutions can be prepared in unit-dose form.
  • Suitable excipients include, but are not limited to, inert diluents, such as calcium carbonate, sodium carbonate or bicarbonate, lactose, or calcium phosphate; or binding agents, such as starch, gelatin, or acacia; or lubricating agents such as magnesium stearate, stearic acid, or talc.
  • inert diluents such as calcium carbonate, sodium carbonate or bicarbonate, lactose, or calcium phosphate
  • binding agents such as starch, gelatin, or acacia
  • lubricating agents such as magnesium stearate, stearic acid, or talc.
  • Additional pharmaceutical compositions of the disclosure will be evident to those skilled in the art, including formulations involving binding proteins in sustained- or controlled-delivery formulations. Techniques for formulating a variety of other sustained- or controlled-delivery means, such as liposome carriers, bio-erodible microparticles or porous beads and depot injections, are also known
  • Sustained release matrices can include polyesters, hydrogels, polylactides, copolymers of L-glutamic acid and gamma ethyl-L-glutamate, poly(2-hydroxyethyl-methacrylate), ethylene vinyl acetate, or poly-D(-)-3-hydroxybutyric acid.
  • Sustained-release compositions can also include liposomes, which can be prepared by any of several methods known in the art. [0187]
  • Pharmaceutical compositions to be used for in vivo administration typically must be sterile. This can be accomplished by filtration through sterile filtration membranes.
  • compositions for parenteral administration can be stored in lyophilized form or in a solution.
  • parenteral compositions generally are placed into a container having a sterile access port, for example, an intravenous solution bag or vial having a stopper pierceable by a hypodermic injection needle.
  • sterile access port for example, an intravenous solution bag or vial having a stopper pierceable by a hypodermic injection needle.
  • kits for producing a single-dose administration unit can each contain both a first container having a dried protein and a second container having an aqueous formulation. Also included within the scope of this disclosure are kits containing single and multi-chambered pre-filled syringes (e.g., liquid syringes and lyosyringes).
  • the effective amount of a pharmaceutical composition to be employed therapeutically will depend, for example, upon the therapeutic context and objectives.
  • the route of administration of the pharmaceutical composition is in accord with known methods, e.g., orally; through injection by intravenous, intraperitoneal, intracerebral (intraparenchymal), intracerebroventricular, intramuscular, intraocular, intraarterial, intraportal, or intralesional routes; by sustained release systems; or by implantation devices.
  • compositions can be administered by bolus injection or continuously by infusion, or by implantation device.
  • the composition can also be administered locally via implantation of a membrane, sponge, or other appropriate material onto which the desired molecule has been absorbed or encapsulated.
  • an implantation device is used, the device can be implanted into any suitable tissue or organ, and delivery of the desired molecule can be via diffusion, timed- release bolus, or continuous administration.
  • Items [0194] are representative of some aspects of the invention: 1. An anti-CD38 T-cell engager for use in the treatment of peripheral T-cell lymphoma (PTCL) in an individual in need thereof. 2.
  • the anti-CD38 T-cell engager for use according to Item 1 characterized in that it 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.
  • the anti-CD38 T-cell engager for use according to Item 1 characterized in that it 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.
  • the anti-CD38 T-cell engager for use according to Item 2 or Item 3, wherein the first antigen binding site that binds 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 G
  • VH antibody heavy chain variable domain comprising a CDR-H1 sequence comprising the amino acid sequence of GYTF
  • VL antibody light chain variable domain comprising a CDR-L1 sequence comprising the amino acid sequence of ESVDSYGNGF (SEQ ID NO:34), a CDR- L2 sequence comprising the amino acid sequence of LAS, and a CDR-L3 sequence comprising the amino acid sequence of QQNKEDPWT (SEQ ID NO:36).
  • the anti-CD38 T-cell engager for use according to Item 2 or Item 3, wherein the first antigen binding site that binds a CD38 polypeptide comprises: (a) an antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence comprising the amino acid sequence of GFTFSSYG (SEQ ID NO:41), a CDR-H2 sequence comprising the amino acid sequence of IWYDGSNK (SEQ ID NO:42), and a CDR-H3 sequence comprising the amino acid sequence of ARMFRGAFDY (SEQ ID NO:43); and (b) an antibody light chain variable (VL) domain comprising a CDR-L1 sequence comprising the amino acid sequence of QGIRND (SEQ ID NO:44), a CDR-L2 sequence comprising the amino acid sequence of AAS, and a CDR-L3 sequence comprising the amino acid sequence of LQDYIYYPT (SEQ ID NO:46).
  • VH antibody heavy chain variable
  • the anti-CD38 T-cell engager for use according to Item 2 or Item 3, wherein the first antigen binding site that binds a CD38 polypeptide comprises: (a) an antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence comprising the amino acid sequence of gytltefs (SEQ ID NO:2), a CDR-H2 sequence comprising the amino acid sequence of fdpedget (SEQ ID NO:3), and a CDR-H3 sequence comprising the amino acid sequence of ttgrffdwf (SEQ ID NO:4); and (b) an antibody light chain variable (VL) domain comprising a CDR-L1 sequence comprising the amino acid sequence of QSVISRF (SEQ ID NO:7), a CDR-L2 sequence comprising the amino acid sequence of GAS, and a CDR-L3 sequence comprising the amino acid sequence of qqdsnlpit (SEQ ID NO:11).
  • VH antibody heavy chain variable
  • VH antibody heavy chain variable
  • VH antibody heavy chain variable domain comprising a CDR
  • the anti-CD38 T-cell engager for use according to Item 2 or Item 3, wherein the first antigen binding site that binds a CD38 polypeptide comprises: (a) an antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence comprising the amino acid sequence of GYSFTNYA (SEQ ID NO:24), a CDR-H2 sequence comprising the amino acid sequence of ISPYYGDT (SEQ ID NO:25), and a CDR-H3 sequence comprising the amino acid sequence of ARRFEGFYYSMDY (SEQ ID NO:26); and (b) an antibody light chain variable (VL) domain comprising a CDR-L1 sequence comprising the amino acid sequence of QSLVHSNGNTY (SEQ ID NO:27), a CDR-L2 sequence comprising the amino acid sequence of KVS, and a CDR-L3 sequence comprising the amino acid sequence of SQSTHVPLT (SEQ ID NO:29).
  • VH antibody heavy chain variable domain comprising a C
  • VH antibody heavy chain variable domain comprising a CDR-H1 sequence comprising the amino acid sequence of GFTFSSYG (SEQ ID NO:41), a CDR-H2 sequence comprising the amino acid sequence of IWYDGSNK (SEQ ID NO:42), and a CDR-H3 sequence comprising the amino acid sequence of ARDPGLRYFDGGMDV (SEQ ID NO:106); and
  • VL antibody light chain variable domain
  • a CDR-L1 sequence comprising the amino acid sequence of QGISSY (SEQ ID NO:107), a CDR-L2 sequence comprising the amino acid sequence of AAS, and a CDR-L3 sequence comprising the amino acid sequence of QQLNSFPYT (SEQ ID NO:229).
  • the anti-CD38 T-cell engager of any one for use according to any one of Items 2 to 18, wherein the antigen binding site that binds a CD3 polypeptide comprises: (a) an antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence comprising the amino acid sequence of GFTFTKAW (SEQ ID NO:120), a CDR-H2 sequence comprising the amino acid sequence of IKDKSNSYAT (SEQ ID NO:121), and a CDR-H3 sequence comprising the amino acid sequence of RGVYYALSPFDY (SEQ ID NO:122); and (b) an antibody light chain variable (VL) domain comprising a CDR-L1 sequence comprising the amino acid sequence of QSLVHX1NX2X3TY, wherein X1 is E or Q, X 2 is A or L, and X 3 is Q, R, or F (SEQ ID NO:131), a CDR-L2 sequence comprising the amino acid sequence of KVS, and a CDR-L
  • the anti-CD38 T-cell engager for use according to any one of Items 2-18, wherein the antigen binding site that binds a CD3 polypeptide comprises: (a) an antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence comprising the amino acid sequence of GFTFTKAW (SEQ ID NO:120), a CDR-H2 sequence comprising the amino acid sequence of IKDKSNSYAT (SEQ ID NO:121), and a CDR-H3 sequence comprising the amino acid sequence of RGVYYALSPFDY (SEQ ID NO:122); and (b) an antibody light chain variable (VL) domain comprising a CDR-L1 sequence comprising the amino acid sequence of QSLVHNNGNTY (SEQ ID NO:218), a CDR-L2 sequence comprising the amino acid sequence of KVS, and a CDR-L3 sequence comprising the amino acid sequence of GQGTQYPFT (SEQ ID NO:130).
  • VH antibody heavy chain variable
  • VL antibody
  • the anti-CD38 T-cell engager for use according to any one of Item 2 and 4- 24, wherein the antigen binding site that binds a CD28 polypeptide comprises: (a) an antibody heavy chain variable (VH) domain comprising a CDR-H1 sequence comprising the amino acid sequence of GYTFTSYY (SEQ ID NO:139), a CDR-H2 sequence comprising the amino acid sequence of IYPGNVNT (SEQ ID NO:140), and a CDR-H3 sequence comprising the amino acid sequence of TRSHYGLDWNFDV (SEQ ID NO:141); and (b) an antibody light chain variable (VL) domain comprising a CDR-L1 sequence comprising the amino acid sequence of QNIYVW (SEQ ID NO:142), a CDR-L2 sequence comprising the amino acid sequence of KAS, and a CDR-L3 sequence comprising the amino acid sequence of QQGQTYPY (SEQ ID NO:144).
  • VH antibody heavy chain variable
  • VH antibody heavy chain variable
  • the anti-CD38 T-cell engager for use according to any one of Items 2 and 4- 28, wherein the trispecific binding protein comprises four polypeptide chains that form the three antigen binding sites, wherein a first polypeptide chain comprises a structure represented by the formula: VL2-L1-VL1-L2-CL [I] and a second polypeptide chain comprises a structure represented by the formula: V H1 -L 3 -V H2 -L 4 -C H1 -hinge-C H2 -C H3 [II] and a third polypeptide chain comprises a structure represented by the formula: VH3-CH1-hinge-CH2-CH3 [III] and a fourth polypeptide chain comprises a structure represented by the formula: V L3 -C L [IV] 140
  • VL1 is a first immunoglobulin light chain variable domain
  • VL2 is a second immunoglobulin light chain variable domain
  • VL3 is a third immunoglobulin light chain variable domain
  • V H1 is a first immunoglobulin heavy chain variable domain
  • V H2 is a second immunoglobulin heavy chain variable domain
  • VH3 is a third immunoglobulin heavy chain variable domain
  • CL is an immunoglobulin light chain constant domain
  • CH1 is an immunoglobulin CH1 heavy chain constant domain
  • C H2 is an immunoglobulin C H2 heavy chain constant domain
  • C H3 is an immunoglobulin C H3 heavy chain constant domain
  • hinge is an immunoglobulin hinge region connecting the CH1 and CH2 domains
  • L1, L2, L3 and L4 are amino acid linkers; wherein the polypeptide of formula I and the polypeptide of formula II form a cross-over light chain-heavy chain pair; wherein V H1 and V L1 form one of the three antigen binding sites; wherein
  • VH1 and VL1 form the second antigen binding site that binds a CD28 polypeptide
  • VH2 and VL2 form the third antigen binding site that binds a CD3 polypeptide
  • VH3 and VL3 form the first antigen binding site that binds a CD38 polypeptide
  • V H1 and V L1 form the third antigen binding site that binds a CD3 polypeptide
  • VH2 and VL2 form the second antigen binding site that binds a CD28 polypeptide
  • V H3 and V L3 form the first antigen binding site that binds a CD38 polypeptide.
  • V H1 and V L1 form the second antigen binding site that binds a CD28 polypeptide
  • V H2 and VL2 form the first antigen binding site that binds a CD38 polypeptide
  • VH3 and VL3 form the third antigen binding site that binds a CD3 polypeptide
  • V H1 and V L1 form the third antigen binding site that binds a CD3 polypeptide
  • VH2 and VL2 form the first antigen binding site that binds a CD38 polypeptide
  • VH3 and VL3 form the second antigen binding site that binds a CD28 polypeptide.
  • V H1 and V L1 form the first antigen binding site that binds a CD38 polypeptide
  • V H2 and VL2 form the third antigen binding site that binds a CD3 polypeptide
  • VH3 and VL3 form the second antigen binding site that binds a CD28 polypeptide
  • V H1 and V L1 form the first antigen binding site that binds a CD38 polypeptide
  • VH2 and VL2 form the second antigen binding site that binds a CD28 polypeptide
  • VH3 and VL3 form the third antigen binding site that binds a CD3 polypeptide.
  • VH1 and VL1 form the second antigen binding site that binds a CD28 polypeptide
  • V H2 and V L2 form the third antigen binding site that binds a CD3 polypeptide
  • V H3 and V L3 form the first antigen binding site that binds a CD38 polypeptide
  • VH1 comprises a CDR-H1 sequence comprising the amino acid sequence of gytftsyy (SEQ ID NO:139), a CDR-H2 sequence comprising the amino acid sequence of iypgnvnt (SEQ ID NO:140), and a CDR-H3 sequence comprising the amino acid sequence of trshygldwnfdv (SEQ ID NO:141)
  • VL1 comprises a CDR-L1 sequence comprising the amino acid sequence of QNIYVW (SEQ ID NO:142), a CDR-L2 sequence comprising the amino acid sequence of KAS
  • V H3 comprises 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 V L3 comprises 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).
  • V H1 and VL1 form the second antigen binding site that binds a CD28 polypeptide
  • VH2 and VL2 form the third antigen binding site that binds a CD3 polypeptide
  • VH3 and VL3 form the first antigen binding site that binds a CD38 polypeptide
  • V H1 comprises the amino acid sequence of SEQ ID NO:49 and VL1 comprises the amino acid sequence of SEQ ID NO:50
  • VH2 comprises the amino acid sequence of SEQ ID NO:53 and VL2 comprises the amino acid sequence of SEQ ID NO:54
  • V H3 comprises the amino acid sequence of SEQ ID NO:13 and V L3 comprises the amino acid sequence of SEQ ID NO:14.
  • L 1 comprises the sequence GQPKAAP (SEQ ID NO:58), L 2 comprises the sequence TKGPS (SEQ ID NO:57), L3 comprises the amino acid S, and L4 comprises the sequence RT;
  • L1 comprises the sequence GGGGSGGGGS (SEQ ID NO:55), L2 comprises the sequence GGGGSGGGGS (SEQ ID NO:55), L 3 is 0 amino acids in length, and L 4 is 0 amino acids in length;
  • L1 comprises the sequence GGSGSSGSGG (SEQ ID NO:59), L2 comprises the sequence GGSGSSGSGG (SEQ ID NO:59), L 3 is 0 amino acids in length, and L 4 is 0 amino acids in length; or 143
  • L 1 comprises the sequence GGGGSGGGGSGGGGS (SEQ ID NO:56), L 2 is 0 amino acids in length, L3 comprises the sequence GGGGSGGGGSGGGGS (SEQ ID NO:56), and L4 is 0 amino acids in length.
  • L1, L2, L3 or L4 comprises the sequence DKTHT (SEQ ID NO:147).
  • L1, L2, L3 and L 4 comprise the sequence DKTHT (SEQ ID NO:147).
  • the anti-CD38 T-cell engager for use according to any one of Items 29-37, wherein the hinge-CH2-CH3 domains of the second and the third polypeptide chains are human IgG4 hinge-C H2 -C H3 domains, and wherein the hinge-C H2 -C H3 domains each comprise amino acid substitutions at positions corresponding to positions 234 and 235 of human IgG4 according to EU Index, wherein the amino acid substitutions are F234A and L235A. 39.
  • the anti-CD38 T-cell engager for use according to any one of Items 29-37, wherein the hinge-C H2 -C H3 domains of the second and the third polypeptide chains are human IgG4 hinge-CH2-CH3 domains, and wherein the hinge-CH2-CH3 domains each comprise amino acid substitutions at positions corresponding to positions 233-236 of human IgG4 according to EU Index, wherein the amino acid substitutions are E233P, F234V, L235A, and a deletion at 236. 40.
  • the anti-CD38 T-cell engager for use according to any one of Items 29-37, wherein the hinge-C H2 -C H3 domains of the second and the third polypeptide chains are human IgG4 hinge-C H2 -C H3 domains, and wherein the hinge-C H2 -C H3 domains each comprise amino acid substitutions at positions corresponding to positions 228 and 409 of human IgG4 according to EU Index, wherein the amino acid substitutions are S228P and R409K. 41.
  • the anti-CD38 T-cell engager for use according to any one of Items 29-37, wherein the hinge-C H2 -C H3 domains of the second and the third polypeptide chains are human IgG1 hinge-CH2-CH3 domains, and wherein the hinge-CH2-CH3 domains each comprise amino acid substitutions at positions corresponding to positions 234, 235, and 329 of human IgG1 according to EU Index, wherein the amino acid substitutions are L234A, L235A, and P329A. 144
  • the anti-CD38 T-cell engager for use according to any one of Items 29-37, wherein the hinge-CH2-CH3 domains of the second and the third polypeptide chains are human IgG1 hinge-CH2-CH3 domains, and wherein the hinge-CH2-CH3 domains each comprise amino acid substitutions at positions corresponding to positions 298, 299, and 300 of human IgG1 according to EU Index, wherein the amino acid substitutions are S298N, T299A, and Y300S. 43.
  • the anti-CD38 T-cell engager for use according to any one of Items 29-42, wherein the hinge-C H2 -C H3 domain of the second polypeptide chain comprises amino acid substitutions at positions corresponding to positions 349, 366, 368, and 407 of human IgG1 or IgG4 according to EU Index, wherein the amino acid substitutions are Y349C, T366S, L368A, and Y407V; and wherein the hinge-C H2 -C H3 domain of the third polypeptide chain comprises amino acid substitutions at positions corresponding to positions 354 and 366 of human IgG1 or IgG4 according to EU Index, wherein the amino acid substitutions are S354C and T366W. 44.
  • the anti-CD38 T-cell engager for use according to any one of Items 29-42, wherein the hinge-CH2-CH3 domain of the second polypeptide chain comprises amino acid substitutions at positions corresponding to positions 354 and 366 of human IgG1 or IgG4 according to EU Index, wherein the amino acid substitutions are S354C and T366W; and wherein the hinge-C H2 -C H3 domain of the third polypeptide chain comprises amino acid substitutions at positions corresponding to positions 349, 366, 368, and 407 of human IgG1 or IgG4 according to EU Index, wherein the amino acid substitutions are Y349C, T366S, L368A, and Y407V. 45.
  • the anti-CD38 T-cell engager for use according to Item 29, wherein: (a) 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; (b) 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; 145
  • 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;
  • 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;
  • 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;
  • the first polypeptide chain comprises the amino acid sequence of
  • 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;
  • 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;
  • 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;
  • the first polypeptide chain comprises the amino acid sequence of SEQ
  • the anti-CD38 T-cell engager 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 147
  • AITL angioimmunoblastic T-cell lymphoma
  • HSTL hepatosplenic T-cell lymphoma
  • ATLL adult T-cell leukemia/lymphoma
  • ENKTCL mycosis fungoides
  • SS Sezary syndrome
  • ALK- lymphoma ALCL
  • NOS anaplastic large cell ALK- lymphoma
  • NOS enteropathy-type T-cell lymphoma
  • MEITL monomorphic epitheliotropic intestinal T-cell lymphoma
  • SCTCL subcutaneous T-cell lymphoma panniculitis-like
  • SCTCL primary cutaneous ⁇ ⁇ T-cell lymphoma. 50.
  • the anti-CD38 T-cell engager for use according to any one of Items 1-48, wherein the PTCL is hepatosplenic T-cell lymphoma (HSTL), Sezary syndrome (SS), or mycosis fungoides (MF).
  • HSTL hepatosplenic T-cell lymphoma
  • SS Sezary syndrome
  • MF mycosis fungoides
  • 51. The anti-CD38 T-cell engager for use according to Item 50, wherein the mycosis fungoides is transformed mycosis fungoides.
  • 52. The anti-CD38 T-cell engager for use according to any one of Items 1-51, wherein cells of the PTCL express CD38 and/or CD28.
  • 53. The anti-CD38 T-cell engager for use according to any one of Items 1-52, wherein the individual is a human. 54.
  • Example 1 Treatment of PTCL with anti-CD38/CD28xCD3 trispecific binding proteins
  • PTCL Peripheral T-cell lymphoma
  • Immunotherapy has established itself in recent years as a major advance in the treatment of several cancers. Trispecific binding proteins that 148
  • This Example describes the investigation of anti-CD38/CD28xCD3 trispecific binding proteins comprising the polypeptides of SEQ ID Nos:60-63 in the treatment of PTCL.
  • Materials and Methods Cell lines and culture [0198] The human cell lines SEAX, H9 (derived from Sezary 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 established from clinical specimens of one patient each.
  • IHC was carried out on FFPE tissue sections, as previously described [ref], using antibodies against CD3 (mouse mAb, clone F7.2.38, 1: 50; Agilent Dako), CD38 (mouse mAb, clone SCP32, 1: 100; Leica Biosystems), CD28 (rabbit mAb, clone D2Z4E, 1: 50; Cell Signaling Technology).
  • IHC was performed on a BOND III or a BOND-MAX (Leica Biosystems) automated stainer platform for CD3 and CD38.
  • H Score intensity x percentage of tumor cells stained Multiplex immunofluorescence [0200]
  • CD28 (1:200; Cell Signaling Technology), CD38 (1:100, Leica Biosystems), PD1 (1:200; Abcam), ICOS (1:200; Abcam), ALK1 (1:200; Agilent Dako) antibodies were used.
  • Flow cytometry For flow cytometry on primary cells, eighteen blood samples were prospectively collected in EDTA tubes, shipped at room temperature and analyzed locally in the Immunology laboratory of Henri Mondor's hospital within 24 hours.
  • the following additional antibodies were added depending on the T-cell lymphoma sub-type: anti-CD10 PE (Beckman Coulter), anti-CD7 PE (Beckman Coulter) and anti-KIR3DL1/DL2 (Miltenyi Biotec).
  • Tumor T cells were defined according to the T-cell lymphoma sub-type: for PTCL-TFH tumor T cells were defined as CD4+CD3+/-CD10+/- CD7+/- cells, and for Cutanous T cell lymphoma tumor T cells were defined as CD4+CD3+CD7-KIR3DL2+ cells. The expression of CD38 and CD28 was then studied on gated tumor cells and threshold of expression was set according to isotype controls. 150
  • Cytotoxicity assays For cytotoxicity assays on cell lines, human healthy volunteers’ PBMCs, isolated from normal human donors by Ficoll separation, were incubated with CFSE (1:50000; Biolegend) labelled tumor cell lines (SEAX, H9, DERL 2, MOLT-4, JURKAT or YT) in the presence of a dose range (0, 8, 40, 200, 1000, 5000, 25000pM) of anti-CD38/CD28xCD3 trispecific antibody or indicated single mutants ( ⁇ ) or double-mutant ( ⁇ ) negative controls using an effector-to-target ratio of 10:1 with 20000 target cells per well (96-well plate). Spontaneous lysis was measured, in culture medium alone.
  • Target cell line lysis by PBMCs was monitored with flow cytometry (MACSQuant Analyzer 16; Miltenyi Biotec) by measuring the percentage of CFSE and Fixable Viability Dye (eFluorTM 780; 1:1000; ThermoFisher Scientific) double positive cells. Specific lysis of target cells was calculating by by subtracting the value of spontaneous lysis in medium alone.
  • flow cytometry MMSQuant Analyzer 16; Miltenyi Biotec
  • Fixable Viability Dye eFluorTM 780; 1:1000; ThermoFisher Scientific
  • T-cells were inoculated in at a rate of 2 ⁇ 10 5 /well according to effector-to-target ratio of 10:1 with CFSE labelled tumor cell lines. Cytotoxicity assay was performed as previously described. Effector and target activation [0205] To address the PBMC or target (cell lines) activation mediated by either anti-CD38/CD28xCD3 trispecific binding protein or indicated single mutants ( ⁇ ) or double- mutant ( ⁇ ) negative controls.
  • PBMCs peripheral blood mononuclear cells
  • tumor cell lines JURKAT or SEAX
  • SEAX tumor cell lines
  • CD28 and CD38 showed heterogenous expression among the different entities of PTCL (FIG.1A). Significantly higher CD38 expression was observed in entities from the innate immune system containing more cytotoxic cells like natural killer (NK) cells than those entities with no cytotoxic cells, whereas CD28 expression showed significant differences in the opposite pattern (FIG.1B).
  • FIG.1C summarizes the percentage of samples showing co-expression of CD38 and CD28, expression of only a single marker, or expression of neither marker, as measured by IHC semi-quantitative scoring.
  • CD38 and CD28 expression pattern on circulating tumor cells was also analyzed by flow cytometry with blood samples from PTCL-T follicular helper (TFH) (FIG.1D) and non- PTCL-TFH patients (e.g., Sezary syndrome and mycosis fungoides; FIG.1E). Rarer cases with co-expression of both markers were identified. IHC staining results for CD3, CD28, and CD38 in various PTCL entities are summarized in FIGS.4A-4E. [0207] Angioimmunoblastic T-cell lymphoma (AITL) and PTCL-not otherwise specified (PTCL-NOS) are the 2 most frequent categories, accounting for >50% of PTCLs.
  • AITL Angioimmunoblastic T-cell lymphoma
  • PTCL-NOS PTCL-not otherwise specified
  • AITL has a rich tumor microenvironment (TME) that typically harbors plenty of CD4+tumor infiltrating lymphocytes, (TIL)-T-cells.
  • TEM tumor microenvironment
  • TIL CD4+tumor infiltrating lymphocytes
  • Analysis was more complicated for this PTCL entity but, thanks to multiplex labelling with tumor markers like PD-1 or ICOS, several cases with CD28 expression and some cases with co-expression of CD28 and CD38 were observed (FIGS.2A- 2D).
  • CD28 staining was detected in atypical lymphoid cells and in tumor microenvironment cells.
  • CD38 staining was detected in lymph node (FIG.2A middle panel and FIG.2C middle panel) or skin (FIG.2B middle panel and FIG.2D middle panel) with lymphoma.
  • CD38 staining was heterogeneous in neoplastic cells and strongly positive in plasma cells for AITL (FIG.2A middle panel) and negative for SS (FIG.2B middle panel).
  • PD1 staining highlighted numerous neoplastic cells sparing the diffuse polymorphic infiltrate (FIG.2A 152
  • CD28 (FIG.2D left panel) and CD38 (FIG.2D middle panel) staining in skin presentation of ENKTCL showed numerous and strongly positive skin neoplastic cells with obvious angiotropism.
  • Granzyme B (GrB) staining highlighted numerous neoplastic cells.
  • Immunofluoresecence multiplex CD38, CD28 and GrB staining showed the co-expression of CD38 and CD28 in the GrB positive cells (FIG. 2D right panel).
  • SEAX a Sezary syndrome cell line
  • H9 MF transformed cell line
  • DERL2 hepatosplenic T-cell lymphoma or PTCL of the innate immune system cell line
  • MOLT4 acute lymphoblastic leukemia cell line
  • Jurkat acute T-cell leukemia
  • YT T/NK cell leukemia

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Abstract

L'invention concerne des méthodes de traitement d'un lymphome T périphérique (PTCL) à l'aide d'un activateur de lymphocytes T anti-CD38, ainsi que des utilisations, des compositions et des kits associés à ceux-ci.
PCT/EP2024/071517 2023-07-31 2024-07-30 Méthodes et utilisations pour des activateurs de lymphocytes t anti-cd38 dans le traitement de lymphomes t périphériques Pending WO2025027003A1 (fr)

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Citations (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1996027011A1 (fr) 1995-03-01 1996-09-06 Genentech, Inc. Procede d'obtention de polypeptides heteromultimeriques
WO1999051642A1 (fr) 1998-04-02 1999-10-14 Genentech, Inc. Variants d'anticorps et fragments de ceux-ci
WO2012135345A1 (fr) 2011-03-28 2012-10-04 Sanofi Protéines de liaison à des anticorps-like à double région variable ayant une orientation croisée de région de liaison
EP2567976A2 (fr) * 2005-03-23 2013-03-13 Genmab A/S Anticorps diriges contre CD38 pour le traitement du myelome multiple
WO2017180913A2 (fr) 2016-04-13 2017-10-19 Sanofi Protéines de liaison trispécifiques et/ou trivalentes
WO2019074973A2 (fr) 2017-10-10 2019-04-18 Sanofi Anticorps anti-cd38 et procédés d'utilisation
WO2020076853A1 (fr) 2018-10-09 2020-04-16 Sanofi Protéines de liaison anti-cd38, anti-cd28 et anti-cd3 trispécifiques et procédés d'utilisation pour traiter une infection virale
WO2020210392A1 (fr) 2019-04-09 2020-10-15 Sanofi Protéines de liaison trispécifiques, procédés et utilisations connexes
WO2022061098A1 (fr) * 2020-09-18 2022-03-24 Regeneron Pharmaceuticals, Inc. Molécules de liaison à l'antigène se liant à cd38 et/ou cd28 et leurs utilisations

Patent Citations (11)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1996027011A1 (fr) 1995-03-01 1996-09-06 Genentech, Inc. Procede d'obtention de polypeptides heteromultimeriques
WO1999051642A1 (fr) 1998-04-02 1999-10-14 Genentech, Inc. Variants d'anticorps et fragments de ceux-ci
EP2567976A2 (fr) * 2005-03-23 2013-03-13 Genmab A/S Anticorps diriges contre CD38 pour le traitement du myelome multiple
WO2012135345A1 (fr) 2011-03-28 2012-10-04 Sanofi Protéines de liaison à des anticorps-like à double région variable ayant une orientation croisée de région de liaison
WO2017180913A2 (fr) 2016-04-13 2017-10-19 Sanofi Protéines de liaison trispécifiques et/ou trivalentes
WO2019074973A2 (fr) 2017-10-10 2019-04-18 Sanofi Anticorps anti-cd38 et procédés d'utilisation
US11186649B2 (en) 2017-10-10 2021-11-30 Sanofi Anti-CD38 antibodies and methods of use
WO2020076853A1 (fr) 2018-10-09 2020-04-16 Sanofi Protéines de liaison anti-cd38, anti-cd28 et anti-cd3 trispécifiques et procédés d'utilisation pour traiter une infection virale
US11530268B2 (en) 2018-10-09 2022-12-20 Sanofi Trispecific anti-CD38, anti-CD28, and anti-CD3 binding proteins and methods of use for treating viral infection
WO2020210392A1 (fr) 2019-04-09 2020-10-15 Sanofi Protéines de liaison trispécifiques, procédés et utilisations connexes
WO2022061098A1 (fr) * 2020-09-18 2022-03-24 Regeneron Pharmaceuticals, Inc. Molécules de liaison à l'antigène se liant à cd38 et/ou cd28 et leurs utilisations

Non-Patent Citations (24)

* Cited by examiner, † Cited by third party
Title
"NCBI", Database accession no. XP_ 011510499.1
"REMINGTON'S PHARMACEUTICAL SCIENCES", 1990, MACK PUBLISHING COMPANY
CALABRETTA ELEONORA ET AL: "The Many Facets of CD38 in Lymphoma: From Tumor-Microenvironment Cell Interactions to Acquired Resistance to Immunotherapy", CELLS, vol. 9, no. 4, 26 March 2020 (2020-03-26), pages 802, XP055913527, DOI: 10.3390/cells9040802 *
CHOTHIA ET AL., NATURE, vol. 342, 1989, pages 877 - 83
CHOTHIALESK, J. MOL. BIOL., vol. 196, 1987, pages 901 - 17
DUPUY AURÉLIE ET AL: "The CD38/CD3xCD28 Trispecic Antibody (SAR442257) Potentially Represents a Novel Therapeutic Strategy for Peripheral T-Cell Lymphoma", BLOOD, 2 November 2023 (2023-11-02), pages 1 - 3, XP093217772, Retrieved from the Internet <URL:https://watermark.silverchair.com/blood-985-main.pdf?token=AQECAHi208BE49Ooan9kkhW_Ercy7Dm3ZL_9Cf3qfKAc485ysgAABHQwggRwBgkqhkiG9w0BBwagggRhMIIEXQIBADCCBFYGCSqGSIb3DQEHATAeBglghkgBZQMEAS4wEQQMwBnXUz5X-mC7lIRAAgEQgIIEJw1s9c2jLzXORbGKrwMeOJI2HKbeKGaDDMz7LwlPvNwNjzrv0qCv8b7fBZ07GfQpLc0pcSzUVQ3GBlEI5iqiZ> *
HINTON, P.R. ET AL., J. IMMUNOL., vol. 176, no. 1, 2006, pages 346 - 56
HUANG HUI-QIANG ET AL: "Daratumumab Monotherapy for Patients with Relapsed or Refractory (R/R) Natural Killer/T-Cell Lymphoma (NKTCL), Nasal Type: Updated Results from an Open-Label, Single-Arm, Multicenter Phase 2 Study", BLOOD, AMERICAN SOCIETY OF HEMATOLOGY, US, vol. 134, 13 November 2019 (2019-11-13), pages 1568, XP086673233, ISSN: 0006-4971, DOI: 10.1182/BLOOD-2019-123446 *
J. MOL. BIOL., vol. 270, pages 26 - 35
LAN WU ET AL: "Trispecific antibodies enhance the therapeutic efficacy of tumor-directed T cells through T cell receptor co-stimulation", NATURE CANCER, vol. 1, no. 1, 18 November 2019 (2019-11-18), pages 86 - 98, XP055716458, DOI: 10.1038/s43018-019-0004-z *
LEFRANC ET AL., DEV. COMP. IMMUNOL., vol. 27, no. 1, 2003, pages 55 - 77
MACCALLUM, J. MOL. BIOL., vol. 262, no. 5, 1996, pages 732 - 45
MARTIN, A.C.: "Antibody Engineering,", vol. 2, 2010, SPRINGER-VERLAG, article "Protein sequence and structure analysis of antibody variable domains", pages: 33 - 51
MASUI ET AL., NUCLEIC ACIDS RES., vol. 33, 2005, pages 43
MERCHANT ET AL., NAT. BIOTECHNOL., vol. 16, 1998, pages 677 - 81
NIWA ET AL., GENE, vol. 108, no. 2, 1991, pages 193 - 9
PADLAN, FASEB J., vol. 9, 1995, pages 133 - 39
RIDGWAY ET AL., PROTEIN ENG., vol. 9, 1996, pages 617 - 21
SAMBROOK ET AL.: "MOLECULAR CLONING: A LABORATORY MANUAL", 2001, COLD SPRING HARBOR LABORATORY PRESS
SHIELDS, R.L., J. BIOL. CHEM., vol. 276, 2001, pages 6591 - 6604
SMITH, E.J. ET AL., SCI. REP., vol. 5, 2015, pages 17943
SPIESS, C. ET AL., J. BIOL. CHEM., vol. 288, 2013, pages 26583 - 26593
TAPIA-GALISTEO ANTONIO ET AL: "Bi- and trispecific immune cell engagers for immunotherapy of hematological malignancies", JOURNAL OF HEMATOLOGY & ONCOLOGY, vol. 16, no. 1, 27 July 2023 (2023-07-27), London UK, XP093200437, ISSN: 1756-8722, DOI: 10.1186/s13045-023-01482-w *
THOMPSON, NUCLEIC ACIDS RES., vol. 22, 1994, pages 4673 - 80

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