US 12,433,954 B2
Methods of activating anti-CD19 chimeric antigen receptor (CAR) T cells using amphiphilic ligand conjugates comprising CAR-targeting protein sequence motifs
Darrell J. Irvine, Arlington, MA (US); Karl Dane Wittrup, Boston, MA (US); Naveen Mehta, Somerville, MA (US); Leyuan Ma, Brookline, MA (US); and Benjamin Cossette, Durham, NC (US)
Assigned to Massachusetts Institute of Technology, Cambridge, MA (US)
Filed by Massachusetts Institute of Technology, Cambridge, MA (US)
Filed on Mar. 1, 2021, as Appl. No. 17/188,813.
Claims priority of provisional application 63/019,053, filed on May 1, 2020.
Prior Publication US 2021/0338833 A1, Nov. 4, 2021
Int. Cl. A61K 47/68 (2017.01); A61K 40/11 (2025.01); A61K 40/31 (2025.01); A61K 40/42 (2025.01); A61K 47/54 (2017.01); A61P 37/04 (2006.01); C07K 16/28 (2006.01)
CPC A61K 47/6889 (2017.08) [A61K 40/11 (2025.01); A61K 40/31 (2025.01); A61K 40/4211 (2025.01); A61K 47/543 (2017.08); A61P 37/04 (2018.01); C07K 16/2803 (2013.01); A61K 2239/31 (2023.05); A61K 2239/38 (2023.05); A61K 2239/48 (2023.05); C07K 2317/622 (2013.01); C07K 2317/92 (2013.01); C07K 2319/74 (2013.01)] 23 Claims
OG exemplary drawing
 
1. A method of activating anti-CD19 CAR T cells or increasing proliferation of anti-CD19 CAR T cells in a subject, comprising administering an amphiphilic ligand conjugate, wherein the amphiphilic ligand conjugate comprises:
an anti-CD19 CAR ligand or a multimer thereof; and
a lipid operably linked to the ligand or the multimer,
wherein the anti-CD19 CAR ligand comprises a peptide comprising a sequence motif, wherein the sequence motif binds an anti-CD19 CAR antigen recognition domain, wherein the anti-CD19 CAR antigen recognition domain comprises an antigen recognition domain derived from FMC63, and wherein binding of the sequence motif to the anti-CD19 CAR antigen recognition domain activates a T cell expressing a CAR comprising the anti-CD19 CAR antigen recognition domain, wherein:
(1) the sequence motif comprises the amino acid sequence [Arg-Xaa1-Cys-Pro-Trp-Xaa2-Cys-Xaa3-Xaa4-Xaa5] (SEQ ID NO: 7), wherein the peptide is capable of forming an intra-peptidyl disulfide bridge;
wherein Xaa1 is selected from Ile, Leu, Met, Val, and Arg;
wherein Xaa2 is selected from Ala, Glu, His, Ser, Asp, and Asn;
wherein Xaa3 is selected from Leu, Arg, Ala, Met, Ser, Val, Ile, and Lys;
wherein Xaa4 is selected from Ser, Val, Gln, Ile, Pro, Lys, Glu, and His; and
wherein Xaa5 is selected from Leu, Ile, Arg, His, Gln, and Trp;
(2) the sequence motif comprises [Arg-Xaa1-Cys-Pro-Trp-Xaa2-Cys-Xaa3-Xaa4-Xaa5] (SEQ ID NO: 131), wherein the peptide is capable of forming an intra-peptidyl disulfide bridge;
wherein Xaa1 is selected from Leu, Ile, and Val;
wherein Xaa2 is Ser or Lys;
wherein Xaa3 is selected from Arg, Ile, Val, and Met;
wherein Xaa4 is selected from Glu, Lys, and Pro; and
wherein Xaa5 is selected from Leu, Gln, Phe, and Ile;
(3) the sequence motif comprises [Arg-Xaa1-Cys-Pro-Trp-Xaa2-Cys-Xaa3-Xaa4-Xaa5] (SEQ ID NO: 132), wherein the peptide is capable of forming an intra-peptidyl disulfide bridge;
wherein Xaa1 is selected from Leu, Ile, and Met;
wherein Xaa2 is selected from Ser, Asn, Asp, and Gly;
wherein Xaa3 is selected from Leu, Met, Ser, Arg, and Lys;
wherein Xaa4 is selected from Glu, Gln, and Pro; and
wherein Xaa5 is Leu or Ile;
(4) the sequence motif comprises [Arg-Xaa1-Cys-Pro-Trp-Xaa2-Cys-Xaa3-Glu-Leu] (SEQ ID NO: 133), wherein the peptide is capable of forming an intra-peptidyl disulfide bridge;
wherein Xaa1 is Leu or Ile;
wherein Xaa2 is selected from Ser, Asn, and Asp; and
wherein Xaa3 is selected from Gln, Ile, Val, Lys, and Arg; or
(5) the sequence motif comprises an amino acid sequence selected from: RHCPWNCSLL (SEQ ID NO: 5), RICPWSCRAP (SEQ ID NO: 6), and SEQ ID NOs: 10, 12-15, and 20.