US 11,945,865 B2
Blockade of CD7 expression and chimeric antigen receptors for immunotherapy of T-cell malignancies
Yi Tian Png, Singapore (SG); Natasha Vinanica, Singapore (SG); Takahiro Kamiya, Tokyo (JP); and Dario Campana, Singapore (SG)
Assigned to NATIONAL UNIVERSITY OF SINGAPORE, Singapore (SG)
Filed by National University of Singapore, Singapore (SG)
Filed on Jul. 14, 2022, as Appl. No. 17/865,292.
Application 17/865,292 is a continuation of application No. 15/821,153, filed on Nov. 22, 2017, granted, now 11,440,958.
Claims priority of provisional application 62/425,398, filed on Nov. 22, 2016.
Claims priority of provisional application 62/543,696, filed on Aug. 10, 2017.
Prior Publication US 2022/0348655 A1, Nov. 3, 2022
This patent is subject to a terminal disclaimer.
Int. Cl. A61K 38/00 (2006.01); A61K 35/17 (2015.01); A61K 39/00 (2006.01); A61P 35/02 (2006.01); C07K 14/705 (2006.01); C07K 14/725 (2006.01); C07K 16/28 (2006.01); C07K 16/30 (2006.01)
CPC C07K 16/2803 (2013.01) [A61K 35/17 (2013.01); A61K 39/001129 (2018.08); A61P 35/02 (2018.01); C07K 14/7051 (2013.01); C07K 14/70517 (2013.01); C07K 14/70578 (2013.01); C07K 16/3061 (2013.01); A61K 38/00 (2013.01); A61K 2039/5156 (2013.01); A61K 2039/5158 (2013.01); A61K 2039/804 (2018.08); C07K 2317/56 (2013.01); C07K 2317/622 (2013.01); C07K 2317/76 (2013.01); C07K 2319/02 (2013.01); C07K 2319/03 (2013.01); C07K 2319/04 (2013.01); C07K 2319/05 (2013.01); C07K 2319/33 (2013.01); C07K 2319/74 (2013.01)] 15 Claims
 
1. An expression vector comprising:
a) a first nucleic acid comprising a nucleotide sequence encoding a target-CD7 binding molecule linked to an endoplasmic reticulum (ER) localizing domain, wherein said target-CD7 binding molecule comprises a first scFv antibody that binds to an endogenous CD7 in an engineered immune cell, and wherein the ER localizing domain comprises an amino acid sequence selected from the group consisting of SEQ ID NO:8, SEQ ID NO:9, and SEQ ID NO: 13, and wherein the first nucleic acid comprises a nucleotide sequence encoding a CD8 alpha signal peptide; and
b) a second nucleic acid comprises a nucleotide sequence encoding a CD7 CAR, wherein said CD7 CAR comprises a second scFv antibody that binds to CD7, a transmembrane domain, and an intracellular signaling domain;
wherein the first scFv antibody and the second scFv antibody comprise:
(i) a heavy chain variable domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 1, comprising:
a heavy chain (HC) complementarity determining region (CDR) 1 comprising amino acids 31-35 of SEQ ID NO: 1,
a HC CDR2 comprising amino acids 50-65 of SEQ ID NO: 1, and
a HC CDR3 comprising amino acids 98-106 of SEQ ID NO: 1, and
a light chain variable domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 2, comprising:
a light chain (LC) CDR1 comprising amino acids 28-38 of SEQ ID NO: 2,
a LC CDR2 comprising amino acids 54-60 of SEQ ID NO: 2, and
a LC CDR3 comprising amino acids 93-101 of SEQ ID NO: 2,
(ii) a heavy chain variable domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 14, comprising:
a HC CDR1 comprising amino acids 31-35 of SEQ ID NO: 14,
a HC CDR2 comprising amino acids 50-66 of SEQ ID NO: 14, and
a HC CDR3 comprising amino acids 99-112 of SEQ ID NO: 14, and
a light chain variable domain having at least 95% sequence identity to the amino acid sequence SEQ ID NO: 15, comprising:
a LC CDR1 comprising amino acids 24-34 of SEQ ID NO: 15,
a LC CDR2 comprising amino acids 50-56 of SEQ ID NO: 15, and
a LC CDR3 comprising amino acids 89-97 of SEQ ID NO: 15, or
(iii) a heavy chain variable domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 16, comprising:
a HC CDR1 comprising amino acids 31-35 of SEQ ID NO: 16,
a HC CDR2 comprising amino acids 50-66 of SEQ ID NO: 16, and
a HC CDR3 comprising amino acids 99-109 of SEQ ID NO: 16, and
a light chain variable domain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 17, comprising:
a LC CDR1 comprising amino acids 24-38 of SEQ ID NO: 17,
a LC CDR2 comprising amino acids 54-60 of SEQ ID NO: 17, and
a LC CDR3 comprising amino acids 93-101 of SEQ ID NO: 17,
wherein the binding of endogenous CD7 to the CD7 binding domain reduces surface expression of the endogenous CD7 in the engineered immune cell, thereby increasing the effectiveness of the engineered immune cell against CD7 positive cancer cells.