SALVAGE CHIMERIC ANTIGEN RECEPTOR SYSTEMS
The invention provides improved compositions for adoptive cell therapies for cancers.
1 . A salvage chimeric antigen receptor (CAR) comprising:
a) an extracellular antigen binding domain;
b) a multimerization domain;
c) a transmembrane domain;
d) one or more intracellular co-stimulatory signaling domains; and/or
e) a primary signaling domain.
2 . (canceled)
3 . The salvage CAR of claim 1 , wherein the extracellular antigen binding domain comprises an antibody or antigen binding fragment thereof, wherein the antigen binding fragment thereof is selected from the group consisting of: a Camel Ig, Ig NAR, Fab fragments, Fab′ fragments, F(ab)′2 fragments, F(ab)′3 fragments, Fv, single chain Fv antibody (“scFv”), bis-scFv, (scFv)2, minibody, diabody, triabody, tetrabody, disulfide stabilized Fv protein (“dsFv”), and single-domain antibody (sdAb, Nanobody).
4 .- 5 . (canceled)
6 . The salvage CAR of claim 1 , wherein:
(a) the extracellular antigen binding domain binds an antigen expressed on a cancer cell;
(b) the extracellular antigen binding domain binds an antigen expressed on a solid cancer cell;
(c) the extracellular antigen binding domain binds an antigen expressed on a liquid cancer cell;
(d) the extracellular antigen binding domain binds an antigen expressed on a malignant B cell;
(e) the extracellular antigen binding domain binds an antigen expressed on a malignant plasma cell; and/or
(f) the extracellular antigen binding domain binds an antigen selected from the group consisting of: alpha folate receptor, 5T4, αvβ6 integrin, BCMA, B7-H3, B7-H6, CAIX, CD16, CD19, CD20, CD22, CD30, CD33, CD37, CD44, CD44v6, CD44v7/8, CD70, CD79a, CD79b, CD123, CD138, CD171, CEA, CSPG4, EGFR, EGFR family including ErbB2 (HER2), EGFRVIII, EGP2, EGP40, EPCAM, EphA2, EpCAM, FAP, fetal AchR, FRα, GD2, GD3, Glypican-3 (GPC3), HLA-A1+MAGE1, HLA-A2+MAGE1, HLA-A3+MAGE1, HLA-A1+NY-ESO-1, HLA-A2+NY-ESO-1, HLA-A3+NY-ESO-1, IL-11Rα, IL-13Rα2, Lambda, Lewis-Y, Kappa, Mesothelin, Muc1, Muc16, NCAM, NKG2D Ligands, NY-ESO-1, PRAME, PSCA, PSMA, ROR1, SSX, Survivin, TAG72, TEMs, VEGFR2, and WT-1.
7 .- 8 . (canceled)
9 . The salvage CAR of claim 1 , wherein the extracellular antigen binding domain binds BCMA or CD19.
10 .- 14 . (canceled)
15 . The salvage CAR of claim 1 , wherein the multimerization domain is selected from the group consisting of: an FKBP polypeptide, an FRB polypeptide, a calcineurin polypeptide, a cyclophilin polypeptide, a bacterial DHFR polypeptide, a PYL1 polypeptide, an ABI1 polypeptide, a GIB1 polypeptide, a GAI polypeptide, and variants thereof.
16 . (canceled)
17 . The salvage CAR of claim 1 , wherein the multimerization domain is selected from the group consisting of: an FKBP12 polypeptide and an FRB T2098L polypeptide.
18 . The salvage CAR of claim 1 , wherein the transmembrane domain is isolated from a polypeptide selected from the group consisting of: alpha or beta chain of the T-cell receptor, CDδ, CD3ε, CDγ, CD3ζ, CD4, CD5, CD8α, CD9, CD 16, CD22, CD27, CD28, CD33, CD37, CD45, CD64, CD71, CD80, CD86, CD 134, CD137, CD152, CD154, AMN, and PD1.
19 . (canceled)
20 . The salvage CAR of claim 1 , wherein the transmembrane domain is isolated from CD8α.
21 . The salvage CAR of claim 1 , wherein:
(a) the one or more co-stimulatory signaling domains and/or primary signaling domains comprise an immunoreceptor tyrosine activation motif (ITAM);
(b) the one or more co-stimulatory signaling domains are isolated from a co-stimulatory molecule selected from the group consisting of: TLR1, TLR2, TLR3, TLR4, TLR5, TLR6, TLR7, TLR8, TLR9, TLR10, CARD11, CD2, CD7, CD27, CD28, CD30, CD40, CD54 (ICAM), CD83, CD134 (OX40), CD137 (4-1BB), CD278 (ICOS), DAP10, LAT, NKD2C, SLP76, TRIM, and ZAP70; and/or
(c) the primary signaling domain isolated from a polypeptide selected from the group consisting of: FcRγ, FcRβ, CD3γ, CD3δ, CD3ε, CD3ζ, CD22, CD79a, CD79b, and CD66d.
22 .- 23 . (canceled)
24 . The salvage CAR of claim 1 , wherein the one or more co-stimulatory signaling domains is isolated from CD137.
25 . (canceled)
26 . The salvage CAR of claim 1 , wherein the primary signaling domain isolated from a CD3.
27 . The salvage CAR of claim 1 , further comprising:
(a) a CD8α hinge region polypeptide; and/or
(b) a spacer region.
28 .- 29 . (canceled)
30 . The salvage CAR of claim 1 , further comprising a signal peptide selected from the group consisting of: an IgG1 heavy chain signal polypeptide, a CD8α signal polypeptide, or a human GM-CSF receptor alpha signal polypeptide.
31 . (canceled)
32 . The salvage CAR of claim 1 , comprising a signal peptide, an anti-BCMA scFv, a linker, an FRB (T82L) multimerization domain, a CD8α hinge and transmembrane domain, a 4-1BB co-stimulatory domain, and a CD3 ζ primary signaling domain.
33 . The salvage CAR of claim 1 , comprising an amino acid sequence having at least 90% identity to the sequence set forth in SEQ ID NO: 2.
34 . (canceled)
35 . A dimerizable salvage receptor comprising:
a) an antigen binding domain;
b) a linker;
c) a multimerization domain;
d) a hinge domain; and
e) an anchor domain.
36 .- 39 . (canceled)
40 . The dimerizable salvage receptor of claim 35 , wherein the antigen binding domain comprises an antibody or antigen binding fragment thereof selected from the group consisting of: a Camel Ig, Ig NAR, Fab fragments, Fab′ fragments, F(ab)′2 fragments, F(ab)′3 fragments, Fv, single chain Fv antibody (“scFv”), bis-scFv, (scFv)2, minibody, diabody, triabody, tetrabody, disulfide stabilized Fv protein (“dsFv”), and single-domain antibody (sdAb, Nanobody).
41 .- 42 . (canceled)
43 . The dimerizable salvage receptor of claim 35 , wherein:
(a) the antigen binding domain binds an antigen expressed on a cancer cell;
(b) the antigen binding domain binds an antigen expressed on a solid cancer cell;
(c) the antigen binding domain binds an antigen expressed on a liquid cancer cell;
(d) the antigen binding domain binds an antigen expressed on a malignant B cell;
(e) the antigen binding domain binds an antigen expressed on a malignant plasma cell; and/or
(f) the antigen binding domain binds an antigen selected from the group consisting of: alpha folate receptor, 5T4, αvβ6 integrin, BCMA, B7-H3, B7-H6, CAIX, CD16, CD19, CD20, CD22, CD30, CD33, CD37, CD44, CD44v6, CD44v7/8, CD70, CD79a, CD79b, CD123, CD138, CD171, CEA, CSPG4, EGFR, EGFR family including ErbB2 (HER2), EGFRvIII, EGP2, EGP40, EPCAM, EphA2, EpCAM, FAP, fetal AchR, FRα, GD2, GD3, Glypican-3 (GPC3), HLA-A1+MAGE1, HLA-A2+MAGE1, HLA-A3+MAGE1, HLA-A1+NY-ESO-1, HLA-A2+NY-ESO-1, HLA-A3+NY-ESO-1, IL-11Rα, IL-13Rα2, Lambda, Lewis-Y, Kappa, Mesothelin, Muc1, Muc16, NCAM, NKG2D Ligands, NY-ESO-1, PRAME, PSCA, PSMA, ROR1, SSX, Survivin, TAG72, TEMs, VEGFR2, and WT-1.
44 .- 45 . (canceled)
46 . The dimerizable salvage receptor of claim 35 , wherein the antigen binding domain binds BCMA or CD19.
47 .- 51 . (canceled)
52 . The dimerizable salvage receptor of claim 35 , wherein the multimization domain is selected from the group consisting of: an FKBP polypeptide, an FRB polypeptide, a calcineurin polypeptide, a cyclophilin polypeptide, a bacterial DHFR polypeptide, a PYL1 polypeptide, an ABI1 polypeptide, a GIB1 polypeptide, a GAI polypeptide, and variants thereof.
53 . (canceled)
54 . The dimerizable salvage receptor of claim 35 , wherein the multimerization domain is selected from the group consisting of: an FKBP12 polypeptide and an FRB T2098L polypeptide.
55 . The dimerizable salvage receptor of claim 35 , wherein the hinge domain is selected from the group consisting essentially of: a CD4 hinge, a CD8α hinge, a PD-1 hinge, and a CD152 hinge.
56 . The dimerizable salvage receptor of claim 35 , wherein the anchor domain is selected from the group consisting of: a GPI molecule and a transmembrane domain; and/or
wherein the anchor domain comprises a transmembrane region of a polypeptide selected from the group consisting of: the alpha or beta chain of the T-cell receptor, CDδ, CD3ε, CDγ, CD3L, CD4, CD5, CD8α, CD9, CD 16, CD22, CD27, CD28, CD33, CD37, CD45, CD64, CD71, CD80, CD86, CD 134, CD137, CD152, CD154, AMN, and PD1.
57 . (canceled)
58 . The dimerizable salvage receptor of claim 35 , comprising a signal peptide, an anti-CD19 scFv, a linker, and an FKBP12 multimerization domain.
59 . (canceled)
60 . The dimerizable salvage receptor of claim 35 , comprising a polypeptide sequence having at least 90% identity to a sequence set forth in any one of SEQ ID NOs: 3-5.
61 . (canceled)
62 . A polynucleotide encoding a salvage CAR according to claim 1 .
63 . A polynucleotide encoding a dimerizable salvage receptor according to claim 35 .
64 . A vector encoding the polynucleotide according to claim 62 and the polynucleotide according to claim 63 .
65 .- 74 . (canceled)
75 . A cell comprising the vector according to claim 64 .
76 .- 78 . (canceled)
79 . The cell of claim 75 , wherein the cell is:
a) a hematopoietic cell;
b) an immune effector cell;
c) CD3+, CD4+, CD8+, or a combination thereof;
d) a T cell; and/or
e) a cytotoxic T lymphocyte (CTL), a tumor infiltrating lymphocyte (TIL), or a helper T cell.
80 .- 84 . (canceled)
85 . A composition comprising the cell according to claim 79 .
86 . A composition comprising a physiologically acceptable carrier and the cell according to claim 79 .
87 .- 88 . (canceled)
89 . A salvage CAR system comprising:
a) a CAR T cell comprising a salvage CAR according to claim 1 that binds a first antigen;
b) a dimerizable salvage receptor according to claim 35 that binds a second antigen; and
c) a bridging factor.
90 .- 107 . (canceled)
108 . A method for decreasing the number of relapsed/refractory cancer cells in a subject, comprising administering to the subject:
a) an effective amount of a dimerizable salvage receptor according to claim 35 , wherein the subject has previously been administered immune effector cells comprising a salvage CAR according to claim 1 ; and
b) an effective amount of a bridging factor that binds the multimerization domains of the dimerizable salvage receptor and the salvage CAR.
109 .- 110 . (canceled)
111 . A method of treating a cancer in a subject in need thereof, comprising administering to the subject:
a) an effective amount of a dimerizable salvage receptor according to claim 35 , wherein the subject has previously been administered immune effector cells comprising a salvage CAR according to claim 1 ; and
b) an effective amount of a bridging factor that binds the multimerization domains of the dimerizable salvage receptor and the salvage CAR.
112 .- 125 . (canceled)