Granted Patent B2

US 12,622,973 · App. 18/392,939 · Granted May 12, 2026

Inventors: Philip Stewart Low (West Lafayette, IN); Boning Zhang (West Lafayette, IN); John V. Napoleon (West Lafayette, IN)

Assignee: Purdue Research Foundation

A61K47/55A61K31/437A61K31/4535A61K31/4545A61K31/7076A61K40/11A61K40/31A61K40/4211A61K47/62A61K47/6835A61K49/0043A61K49/0052A61K2239/48

Abstract

Claims (38)

1 . A system to rejuvenate an exhausted classical CAR T cell comprising at least two components:

a first component that is a conjugate comprising a targeting ligand covalently linked to a payload drug; and

a second component that is a targeting ligand binding module linked to a membrane-anchoring module,

wherein:

the payload drug is a Toll Like Receptor 7 (TLR7) agonist,

the targeting ligand and the targeting ligand binding molecule are fluorescein isothiocyanate (FITC) and an anti-FITC antibody or fragment thereof, respectively; or tacrolimus (FK506) and FK506-binding protein (FKBP), respectively,

the membrane-anchoring module is a folate receptor,

the targeting ligand binding module of the second component recognizes the targeting ligand in the first component with high affinity to form a complex,

the payload drug re-activates the CAR T cell through an antigen-independent pathway, and

the membrane-anchoring module mediates internalization of the at least two components complex into the exhausted classical CAR T cell.

2 . The system according to claim 1 , wherein the membrane-anchoring module is a folate receptor alpha (FRα).

3 . The system according to claim 1 , wherein the first component comprises a releasable linker between the targeting ligand and the payload drug.

4 . The system according to claim 1 , wherein the first component comprises a non-releasable linker between the targeting ligand and the payload drug.

5 . The system according to claim 1 , wherein the binding affinity between the targeting ligand and the ligand-binding module is in a sub-nanomolar range.

6 . The system according to claim 1 , wherein the TLR7 agonist has the structure of

7 . The system according to claim 1 , wherein targeting ligand and the targeting ligand binding molecule are fluorescein isothiocyanate (FITC) and an anti-FITC antibody or fragment thereof, respectively, and the first component is a fluorescein isothiocyanate-TLR7 agonist having the structure:

8 . The system according to claim 1 , wherein the targeting ligand and the targeting ligand binding molecule are tacrolimus (FK506) and FK506-binding protein (FKBP), respectively, and the first component is a FK506-TLR7 agonist having the structure:

9 . The system according to claim 1 , wherein the targeting ligand and the targeting ligand binding molecule are fluorescein isothiocyanate (FITC) and an anti-FITC scFv, respectively, and wherein the first component is one of the following:

10 . The system according to claim 1 , wherein the first component comprises a spacer between the targeting ligand and the payload drug, wherein the spacer is selected from the group consisting of the following structures:

wherein n is 1-12.

11 . A method to rejuvenate an exhausted CAR T cell, comprising:

a. providing to the exhausted CAR T cell a first component comprising a conjugate comprising, a targeting ligand covalently linked to a payload of drug through a releasable linker or a non-releasable linker;

b. providing said exhausted CAR T cell a second component comprising a fusion receptor, wherein the fusion receptor comprises a targeting ligand binding module linked to a membrane-anchoring module;

c. wherein the targeting ligand binding module of the second component binds to the targeting ligand in the first component to form a complex,

d. the membrane-bound receptor module mediates internalization of the complex into the exhausted CAR T cell; and

e. the payload drug re-activates the CAR T cell through an antigen-independent pathway;

wherein:

the payload drug is a Toll Like Receptor 7 (TLR7) agonist,

the targeting ligand and the targeting ligand binding molecule are fluorescein isothiocyanate (FITC) and an anti-FITC antibody or fragment thereof, respectively, or tacrolimus (FK506) and FK506-binding protein (FKBP), respectively, and

the membrane-anchoring module is a folate receptor.

12 . The method according to claim 11 , wherein the payload drug executes its function within the endosome of the exhausted CAR T cell, and the targeting ligand and the payload drug are linked by a non-releasable linker.

13 . The method according to claim 11 , wherein the payload drug executes its function as a free drug in the cytosol of the exhausted CAR T cell, and the targeting ligand and the payload drug are linked by a releasable linker.

14 . The method according to claim 11 , wherein the TLR7 agonist has the structure:

15 . The method according to claim 11 , wherein the first component is a targeting ligand and the targeting ligand binding molecule are fluorescein isothiocyanate (FITC) and an anti-FITC antibody or fragment thereof, respectively, and the first component is a fluorescein isothiocyanate-TLR7 agonist having the structure:

16 . The method according to claim 11 , wherein the targeting ligand and the targeting ligand binding molecule are tacrolimus (FK506) and FK506-binding protein (FKBP), respectively, and the first component is a FK506-TLR7 agonist having the structure:

17 . The method according to claim 11 , wherein the targeting ligand and the targeting ligand binding molecule are fluorescein isothiocyanate (FITC) and an anti-FITC scFv, respectively, and wherein the first component is one of the following;

18 . The method according to claim 11 , wherein the linker between the targeting ligand and the payload drug is selected from the group consisting of the following structures:

wherein n is 1-12.

Assignments (1)

Continuity (3)

References Cited (166)