Targeting of multiple antigens with multiplex CAR T cells in solid and liquid malignancies

A pharmaceutical composition, comprising a therapeutically effective amount of a first bifunctional compound comprising a first synthetic antigen covalently linked to a first targeting moiety, or a pharmaceutically acceptable salt or stereoisomer thereof, and

• a second bifunctional compound comprising the first synthetic antigen covalently linked to a second targeting moiety, or a pharmaceutically acceptable salt or stereoisomer thereof,
• wherein the first and second targeting moieties specifically bind different tumor associated antigens,
• wherein the synthetic antigen linked to the first or second targeting moiety contains a removable protecting group,
• wherein the synthetic antigen containing the removable protecting group can bind a CAR-T cell after removal of the removable protecting group,
• wherein the synthetic antigen is 4-[(6-methylpyrazin-2-yl) oxy] benzoate (MPOB) anthraquinone-2-carboxylate (AQ), tetraxetan (DOTA), or a peptidic, glysodic or nucleotidic antigen, and
• a pharmaceutically acceptable carrier.

A method of treating cancer, comprising co-administering to a subject in need thereof,

• a) a plurality of subpopulations of bifunctional molecules wherein the plurality comprises
• a1) a therapeutically effective amount of a first subpopulation of bifunctional compounds comprising a synthetic antigen covalently linked to a first targeting moiety that specifically binds a first tumor associated antigen;
• and
• a2) a therapeutically effective amount of a second subpopulation of bifunctional compounds comprising the synthetic antigen covalently linked to a second targeting moiety that specifically binds a second tumor associated antigen, or a pharmaceutically acceptable salt or stereoisomer thereof, wherein the first and second targeting moieties specifically bind different tumor associated antigens;
• wherein the synthetic antigen of the first or second subpopulations comprises a removable protecting group;
• wherein the synthetic antigen comprising the removable protecting group can bind a CAR-T cell after removal of the removable protecting group;
• and
• b) a therapeutically effective number of CAR-T cells, wherein the CAR-T cells comprise an extracellular ligand that specifically binds the synthetic antigen,
• wherein the subject is human.

A kit, comprising:

• a) therapeutically effective amounts of a plurality of subpopulations of bifunctional compounds,
• wherein each bifunctional compound in a first subpopulation comprises a first synthetic antigen covalently linked to a first targeting moiety that specifically binds a first tumor associated antigen, and wherein each bifunctional compound in a second subpopulation comprises the first synthetic antigen covalently linked to a second targeting moiety that specifically binds a second tumor associated antigen, or a pharmaceutically acceptable salt or stereoisomer thereof, wherein the first and second targeting moieties specifically bind different tumor associated antigens, and wherein each subpopulation of bifunctional compounds administered to the patient contains the first synthetic antigen but specifically binds a different tumor associated antigen;
• wherein the plurality of subpopulations of bifunctional compounds is disposed in the same or separate containers,
• wherein the first or second synthetic antigen contains a removable protecting group;
• wherein the synthetic antigen containing the removable protecting group can bind a CAR-T cell after removal of the removable protecting group;
• and
• wherein the synthetic antigen is 4-[(6-methylpyrazin-2-yl) oxy] benzoate (MPOB), anthraquinone-2-carboxylate (AQ), tetraxetan (DOTA), or a peptidic, glysodic or nucleotidic antigen;
• and
• b) printed instructions for co-administering to a cancer patient the therapeutically effective amounts of a plurality of subpopulations of bifunctional compounds and a therapeutically effective number of CAR-T cells, wherein the CAR-T cells comprise an extracellular ligand that specifically binds the synthetic antigen.