CHIMERIC ANTIGEN RECEPTOR T CELL SWITCHES AND USES THEREOF
Disclosed herein are switches for regulating the activity of a chimeric antigen receptor effector cells (CAR-ECs). The switches generally comprise a chimeric antigen receptor-interacting domain (CAR-ID) and a target interacting domain (TID). The switch may further comprise a linker. Further disclosed herein are methods of using the switches for the treatment of one or more conditions or diseases in a subject in need thereof.
1 . A switch for activating a chimeric antigen receptor-effector cell (CAR-EC), the switch comprising:
a. a chimeric antigen receptor-interacting domain (CAR-ID) that interacts with a chimeric antigen receptor on the CAR-EC; and
b. a target interacting domain (TID) comprising an unnatural amino acid, wherein the TID interacts with a surface molecule on a target cell.
2 . The switch of claim 1 , wherein the CAR-ID is attached to the TID.
3 . The switch of claim 1 , wherein the CAR-ID is attached to the TID via the unnatural amino acid.
4 . The switch of claim 1 , wherein the CAR-ID is site-specifically attached to the unnatural amino acid of the TID.
5 . The switch of claim 1 , further comprising a linker.
6 . The switch of claim 5 , wherein the linker attaches the CAR-ID to the TID.
7 . The switch of claim 5 , wherein the linker attaches the CAR-ID to the TID via the unnatural amino acid.
8 . The switch of claim 5 , wherein the linker site-specifically attaches the CAR-ID to the unnatural amino acid of the TID.
9 . The switch of claim 5 , wherein the linker comprises an aminooxy group, azide group cyclooctyne group, or a combination thereof at one or more termini.
10 . The switch of claim 1 , wherein the CAR-ID comprises a small molecule.
11 . The switch of claim 10 , wherein the small molecule is a hapten.
12 . The switch of claim 10 , wherein the small molecule is fluorescein isothiocyanate (FITC).
13 . The switch of claim 12 , wherein the TID is conjugated to an isothiocyanate of FITC.
14 . The switch of claim 12 , further comprising a linker that is conjugated to an isothiocyanate of FITC.
15 . The switch of claim 10 , wherein the small molecule is biotin.
16 . The switch of claim 1 , wherein the TID is based on or derived from at least a portion of an antibody.
17 . The switch of claim 1 , wherein the TID is based on or derived from at least a portion of a single chain variable fragment (scFv).
18 . The switch of claim 1 , wherein the TID is based on or derived from at least a portion of an anti-CD19 antibody.
19 . The switch of claim 1 , wherein the TID is based on or derived from at least a portion of a single chain variable domain (scFv) of an anti-CD19 antibody.
20 . The switch of claim 1 , wherein the TID is based on or derived from at least a portion of an antibody selected from the group consisting of anti-CD20, anti-CD22, anti-CD33, anti-BMSA, anti-CEA, anti-CLL1, anti-CS1, anti-EGFR, and anti-Her2.
21 . The switch of claim 1 , wherein the TID is based on or derived from at least a portion of a single chain variable domain (scFv) of an antibody selected from the group consisting of anti-CD20, anti-CD22, anti-CD33, anti-BMSA, anti-CEA, anti-CLL1, anti-CS1, anti-EGFR, and anti-Her2.
22 . The switch of claim 16 , wherein the unnatural amino acid is inserted in the portion of the antibody from which the TID is based or derived.
23 . The switch of claim 16 , wherein the unnatural amino acid replaces an amino acid of the antibody from which the TID is based or derived.
24 . A composition comprising a plurality of switches for activating a chimeric antigen receptor-effector cell (CAR-EC), wherein a switch of the plurality of switches comprises (a) a chimeric antigen receptor-interacting domain (CAR-ID) that interacts with a chimeric antigen receptor on the CAR-EC; and (b) a target interacting domain (TID) that interacts with a surface molecule on a target cell, wherein at least about 60% of the switches are structurally homologous.
25 . (canceled)
26 . A composition comprising a plurality of switches for activating a chimeric antigen receptor-effector cell (CAR-EC), wherein a switch of the plurality of switches comprises (a) a chimeric antigen receptor-interacting domain (CAR-ID) that interacts with a chimeric antigen receptor on the CAR-EC; and (b) a target interacting domain (TID) comprising a polypeptide, wherein the CAR-ID is attached to the same predetermined site in the TID in at least 60% of the switches.
27 - 33 . (canceled)
34 . The composition of claim 26 , wherein the predetermined site in the TID is an amino acid residue.
35 . The composition of claim 34 , wherein the amino acid residue is an unnatural amino acid.
36 . A switch intermediate for activating a chimeric antigen receptor-effector cell (CAR-EC), the switch intermediate comprising:
a. a chimeric antigen receptor-interacting domain (CAR-ID) comprising a small molecule, wherein the CAR-ID interacts with a chimeric antigen receptor on the CAR-EC; and
b. a linker connected to the CAR-ID, wherein the linker does not comprise a region that interacts with the CAR-EC and the linker does not comprise a region that interacts with a surface molecule on a target cell.
37 .- 40 . (canceled)
41 . The switch intermediate of claim 36 , wherein the CAR-ID comprises fluorescein isothiocyanate (FITC).
42 . A switch intermediate for activating a chimeric antigen receptor-effector cell (CAR-EC), the switch intermediate comprising:
a. a target interacting domain (TID) comprising an unnatural amino acid, wherein the TID interacts with a surface molecule on a target cell; and
b. a linker connected to the TID, wherein the linker does not comprise a region that directly interacts with the CAR-EC and the linker does not comprise a region that directly interacts with the target cell.
43 .- 49 . (canceled)
50 . The switch intermediate of claim 42 , wherein the TID is based on or derived from an antibody or antibody fragment.
51 . The switch intermediate of claim 42 , wherein the TID comprises one or more unnatural amino acids.
52 . A method of producing a switch comprising (a) a chimeric antigen receptor-interacting domain (CAR-ID); and (b) a target interacting domain (TID), the method comprising:
a. coupling a first linker to the TID to produce a first switch intermediate comprising the first linker conjugated to the TID;
b. coupling the first switch intermediate to the CAR-ID, thereby producing the switch.
53 .- 60 . (canceled)