CHIMERIC ANTIGEN RECEPTOR-EXPRESSING T CELLS AS ANTI-CANCER THERAPEUTICS
Cytotoxic lymphocytes expressing chimeric antigen receptors (CAR) that target and bind small conjugate molecules (SCM) are disclosed, as well as methods of using the cells and the SCMs in the treatment of cancer.
1 . A two component cancer therapeutic comprising:
(a) a small conjugate molecule (SCM) comprising a targeted moiety conjugated to a tumor receptor ligand, wherein the tumor receptor ligand is selected from the group consisting of folate, DUPA, and CCK2R ligand; and
(b) chimeric antigen receptor (CAR)-expressing cytotoxic lymphocytes, wherein the CAR is a fusion protein comprising a recognition region, a co-stimulation domain and an activation signaling domain, and wherein the CAR has binding specificity for the targeted moiety or can be bound by the targeted moiety.
2 . The two component cancer therapeutic of claim 1 , wherein the targeted moiety and the ligand are conjugated via a linker domain.
3 . The two component cancer therapeutic of claim 1 , wherein the targeted moiety is a molecule selected from the group consisting of 2,4-dinitrophenol (DNP), 2,4,6-trinitrophenol (TNP), biotin, digoxigenin, fluorescein, fluorescein isothiocyanate (FITC), NHS-fluorescein, pentafluorophenyl ester (PFP), tetrafluorophenyl ester (TFP), a knottin, a centyrin, and a DARPin.
4 . The two component cancer therapeutic of claim 2 , wherein the linker domain is selected from the group consisting of polyethylene glycol (PEG), polyproline, a hydrophilic amino acid, a sugar, an unnatural peptideoglycan, polyvinylpyrrolidone, and pluronic F-127.
5 . The two component cancer therapeutic of claim 1 , wherein the targeted moiety is FITC.
6 . The two component cancer therapeutic of claim 4 , wherein the targeted moiety is FITC and the linker is (PEG) 12 .
7 . The two component cancer therapeutic of claim 1 , wherein the recognition region of the CAR is a single chain fragment variable (scFv) region of an antibody with binding specificity for the targeted moiety.
8 . The two component cancer therapeutic of claim 5 , wherein the recognition region of the CAR is a single chain fragment variable (scFv) region of an anti-FITC antibody.
9 . The two component cancer therapeutic of claim 1 , wherein the co-stimulation domain of the CAR is selected from the group consisting of CD28, CD137 (4-1BB), CD134 (OX40), and CD278 (ICOS).
10 . The two component cancer therapeutic of claim 1 , wherein the activation signaling domain of the CAR is the T cell CD3ζ chain or Fc receptor γ.
11 . The two component cancer therapeutic of claim 5 , wherein the recognition region is a single chain fragment variable (scFv) region of an anti-FITC antibody, wherein the co-stimulation domain is CD137 (4-1BB), and wherein the activation signaling domain is the T cell CD3ζ chain.
13 - 19 . (canceled)
20 . CAR-expressing lymphocytes, wherein the CAR is a fusion protein comprising a recognition region, a co-stimulation domain and an activation signaling domain, wherein the recognition region of the CAR is a single chain fragment variable (scFv) region of an anti-FITC antibody and wherein the CAR has binding specificity for a selected targeted moiety or can be bound by the targeted moiety.
21 . The CAR-expressing cytotoxic lymphocytes of claim 20 , wherein the recognition region of the CAR is a single chain fragment variable (scFv) region of an antibody with binding specificity for the targeted moiety.
22 . (canceled)
23 . The CAR-expressing cytotoxic lymphocytes of claim 20 , wherein the co-stimulation domain of the CAR is selected from the group consisting of CD28, CD137 (4-1BB), CD134 (OX40), and CD278 (ICOS).
24 . The CAR-expressing cytotoxic lymphocytes of claim 20 , wherein the activation signaling domain of the CAR is the T cell CD3ζ chain or Fc receptor γ.
25 . The CAR-expressing cytotoxic lymphocytes of claim 20 , wherein the recognition region is a single chain fragment variable (scFv) region of an anti-FITC antibody, wherein the co-stimulation domain is CD137 (4-1BB), and wherein the activation signaling domain is the T cell CD3ζ chain.
26 . The CAR-expressing cytotoxic lymphocytes of claim 20 , wherein the targeted moiety is a molecule selected from the group consisting of 2,4-dinitrophenol (DNP), 2,4,6-trinitrophenol (TNP), biotin, digoxigenin, fluorescein, fluorescein isothiocyanate (FITC), NHS-fluorescein, pentafluorophenyl ester (PFP), tetrafluorophenyl ester (TFP), a knottin, a centyrin, and a DARPin.
27 . The CAR-expressing cytotoxic lymphocytes of claim 20 , wherein the cytotoxic lymphocyte are one or more cell types selected from the group consisting of cytotoxic T cells, natural killer (NK) cells, and lymphokine-activated killer (LAK) cells.
28 - 48 . (canceled)
49 . The two component cancer therapeutic of claim 6 , wherein the recognition region of the CAR is a single chain fragment variable (scFv) region of an anti-FITC antibody.
50 . The two component cancer therapeutic of claim 6 , wherein the recognition region is a single chain fragment variable (scFv) region of an anti-FITC antibody, wherein the co-stimulation domain is CD137 (4-1BB), and wherein the activation signaling domain is the T cell CD3ζ chain.