	US 11,981,731 B2
	Targeting cytotoxic cells with chimeric receptors for adoptive immunotherapy
Gregory Beatty, Philadelphia, PA (US); Boris Engels, Arlington, MA (US); Neeraja Idamakanti, Burlington, MA (US); Carl H. June, Merion Station, PA (US); Andreas Loew, Boston, MA (US); Michael C. Milone, Cherry Hill, NJ (US); Huijuan Song, Shanghai (CN); Enxiu Wang, Upper Darby, PA (US); and Qilong Wu, Brighton, MA (US)
Assigned to The Trustees of the University of Pennsylvania, Philadelphia, PA (US); and Novartis AG, Basel (CH)
Filed by Novartis AG, Basel (CH); and The Trustees of the University of Pennsylvania, Philadelphia, PA (US)
Filed on Jan. 15, 2020, as Appl. No. 16/743,772.
Application 16/743,772 is a continuation of application No. 15/511,973, granted, now 10,577,417, previously published as PCT/US2015/050715, filed on Sep. 17, 2015.
Claims priority of application No. PCT/CN2014/086694 (WO), filed on Sep. 17, 2014; and application No. PCT/CN2014/090578 (WO), filed on Nov. 7, 2014.
Prior Publication US 2020/0377589 A1, Dec. 3, 2020
This patent is subject to a terminal disclaimer.
Int. Cl. A61K 35/17 (2015.01); C07K 14/705 (2006.01); C07K 14/725 (2006.01); C07K 14/735 (2006.01); C07K 14/74 (2006.01); C07K 16/28 (2006.01); C07K 16/30 (2006.01); C12N 5/0783 (2010.01)

CPC C07K 16/28 (2013.01) [A61K 35/17 (2013.01); C07K 14/705 (2013.01); C07K 14/70503 (2013.01); C07K 14/7051 (2013.01); C07K 14/70535 (2013.01); C07K 14/70539 (2013.01); C07K 14/7056 (2013.01); C07K 14/70596 (2013.01); C07K 16/30 (2013.01); C12N 5/0636 (2013.01); C07K 2317/21 (2013.01); C07K 2317/56 (2013.01); C07K 2317/565 (2013.01); C07K 2317/622 (2013.01); C07K 2317/74 (2013.01); C07K 2319/02 (2013.01); C07K 2319/03 (2013.01); C07K 2319/70 (2013.01); C07K 2319/74 (2013.01); C12N 2510/00 (2013.01)]

17 Claims

1. An isolated nucleic acid molecule encoding a natural killer cell immune-function receptor-chimeric antigen receptor (NKR-CAR), wherein the NKR-CAR comprises an antigen binding domain that binds to mesothelin, and:

(i) a NKR transmembrane domain and a cytoplasmic domain; or

(ii) a transmembrane domain and a NKR cytoplasmic domain,

wherein the antigen binding domain that binds mesothelin comprises: a heavy chain complementarity determining region 1 (HC CDR1), a heavy chain complementary determining region 2 (HC CDR2), and a heavy chain complementary determining region 3 (HC CDR3), a light chain complementarity determining region 1 (LC CDR1), a light chain complementary determining region 2 (LC CDR2), and a light chain complementary determining region 3 (LC CDR3), and wherein:

(a) the HC CDR1, HC CDR2, and HC CDR3 are of the anti-mesothelin heavy chain amino acid sequence of SEQ ID NO:229, and the LC CDR1, LC CDR2, and LC CDR3 are of the anti-mesothelin light chain amino acid sequence of SEQ ID NO:229;

(b) the HC CDR1, HC CDR2, and HC CDR3 are of the anti-mesothelin heavy chain amino acid sequence of SEQ ID NO:230, and the LC CDR1, LC CDR2, and LC CDR3 are of the anti-mesothelin light chain amino acid sequence of SEQ ID NO:230;

(c) the HC CDR1, HC CDR2, and HC CDR3 are of the anti-mesothelin heavy chain amino acid sequence of SEQ ID NO:231, and the LC CDR1, LC CDR2, and LC CDR3 are of the anti-mesothelin light chain amino acid sequence of SEQ ID NO:231;

(d) the HC CDR1, HC CDR2, and HC CDR3 are of the anti-mesothelin heavy chain amino acid sequence of SEQ ID NO:232, and the LC CDR1, LC CDR2, and LC CDR3 are of the anti-mesothelin light chain amino acid sequence of SEQ ID NO:232;

(e) the HC CDR1, HC CDR2, and HC CDR3 are of the anti-mesothelin heavy chain amino acid sequence of SEQ ID NO:233, and the LC CDR1, LC CDR2, and LC CDR3 are of the anti-mesothelin light chain amino acid sequence of SEQ ID NO:233;

(f) the HC CDR1, HC CDR2, and HC CDR3 are of the anti-mesothelin heavy chain amino acid sequence of SEQ ID NO:235, and the LC CDR1, LC CDR2, and LC CDR3 are of the anti-mesothelin light chain amino acid sequence of SEQ ID NO:235;

(g) the HC CDR1, HC CDR2, and HC CDR3 are of the anti-mesothelin heavy chain amino acid sequence of SEQ ID NO:236, and the LC CDR1, LC CDR2, and LC CDR3 are of the anti-mesothelin light chain amino acid sequence of SEQ ID NO:236;

(h) the HC CDR1, HC CDR2, and HC CDR3 are of the anti-mesothelin heavy chain amino acid sequence of SEQ ID NO:237, and the LC CDR1, LC CDR2, and LC CDR3 are of the anti-mesothelin light chain amino acid sequence of SEQ ID NO:237;

(i) the HC CDR1, HC CDR2, and HC CDR3 are of the anti-mesothelin heavy chain amino acid sequence of SEQ ID NO:238, and the LC CDR1, LC CDR2, and LC CDR3 are of the anti-mesothelin light chain amino acid sequence of SEQ ID NO:238;

(j) the HC CDR1, HC CDR2, and HC CDR3 are of the anti-mesothelin heavy chain amino acid sequence of SEQ ID NO:239, and the LC CDR1, LC CDR2, and LC CDR3 are of the anti-mesothelin light chain amino acid sequence of SEQ ID NO:239;

(kl) the HC CDR1, HC CDR2, and HC CDR3 are of the anti-mesothelin heavy chain amino acid sequence of SEQ ID NO:241, and the LC CDR1, LC CDR2, and LC CDR3 are of the anti-mesothelin light chain amino acid sequence of SEQ ID NO:241;

(l) the HC CDR1, HC CDR2, and HC CDR3 are of the anti-mesothelin heavy chain amino acid sequence of SEQ ID NO:242, and the LC CDR1, LC CDR2, and LC CDR3 are of the anti-mesothelin light chain amino acid sequence of SEQ ID NO:242;

(m) the HC CDR1, HC CDR2, and HC CDR3 are of the anti-mesothelin heavy chain amino acid sequence of SEQ ID NO:243, and the LC CDR1, LC CDR2, and LC CDR3 are of the anti-mesothelin light chain amino acid sequence of SEQ ID NO:243;

(n) the HC CDR1, HC CDR2, and HC CDR3 are of the anti-mesothelin heavy chain amino acid sequence of SEQ ID NO:244, and the LC CDR1, LC CDR2, and LC CDR3 are of the anti-mesothelin light chain amino acid sequence of SEQ ID NO:244;

(o) the HC CDR1, HC CDR2, and HC CDR3 are of the anti-mesothelin heavy chain amino acid sequence of SEQ ID NO:245, and the LC CDR1, LC CDR2, and LC CDR3 are of the anti-mesothelin light chain amino acid sequence of SEQ ID NO:245;

(p) the HC CDR1, HC CDR2, and HC CDR3 are of the anti-mesothelin heavy chain amino acid sequence of SEQ ID NO:246, and the LC CDR1, LC CDR2, and LC CDR3 are of the anti-mesothelin light chain amino acid sequence of SEQ ID NO:246;

(q) the HC CDR1, HC CDR2, and HC CDR3 are of the anti-mesothelin heavy chain amino acid sequence of SEQ ID NO:247, and the LC CDR1, LC CDR2, and LC CDR3 are of the anti-mesothelin light chain amino acid sequence of SEQ ID NO:247;

(r) the HC CDR1, HC CDR2, and HC CDR3 are of the anti-mesothelin heavy chain amino acid sequence of SEQ ID NO:248, and the LC CDR1, LC CDR2, and LC CDR3 are of the anti-mesothelin light chain amino acid sequence of SEQ ID NO:248;

(s) the HC CDR1, HC CDR2, and HC CDR3 are of the anti-mesothelin heavy chain amino acid sequence of SEQ ID NO:249, and the LC CDR1, LC CDR2, and LC CDR3 are of the anti-mesothelin light chain amino acid sequence of SEQ ID NO:249;

(t) the HC CDR1, HC CDR2, and HC CDR3 are of the anti-mesothelin heavy chain amino acid sequence of SEQ ID NO:250, and the LC CDR1, LC CDR2, and LC CDR3 are of the anti-mesothelin light chain amino acid sequence of SEQ ID NO:250;

(u) the HC CDR1, HC CDR2, and HC CDR3 are of the anti-mesothelin heavy chain amino acid sequence of SEQ ID NO:251, and the LC CDR1, LC CDR2, and LC CDR3 are of the anti-mesothelin light chain amino acid sequence of SEQ ID NO:251;

(v) the HC CDR1, HC CDR2, and HC CDR3 are of the anti-mesothelin heavy chain amino acid sequence of SEQ ID NO:252, and the LC CDR1, LC CDR2, and LC CDR3 are of the anti-mesothelin light chain amino acid sequence of SEQ ID NO:252; or

(w) the HC CDR1, HC CDR2, and HC CDR3 are of the anti-mesothelin heavy chain amino acid sequence of SEQ ID NO:253, and the LC CDR1, LC CDR2, and LC CDR3 are of the anti-mesothelin light chain amino acid sequence of SEQ ID NO:253; and

wherein the NKR-CAR further comprises:

a killer cell immunoglobulin-like receptor chimeric antigen receptor (KIR-CAR), wherein the KIR-CAR comprises one or both of a transmembrane domain from a KIR (a KIR transmembrane domain) or a cytoplasmic domain comprising a functional signaling domain from a KIR (a KIR cytoplasmic domain), wherein the KIR transmembrane or cytoplasmic domain comprises a transmembrane or cytoplasmic domain of a protein selected from the group consisting of: KIR2DL1, KIR2DL2, KIR2DL4, KIR2DL5A, KIR2DL5B, KIR2DS1, KIR2DS3, KIR2DS4, KIR2DS5, KIR3DL1, KIR3DS1, KIR3DL2, KIR3DL3, KIR2DP1 and KIR3DP1;

a natural cytotoxicity receptor chimeric antigen receptor (NCR-CAR), wherein the NCR-CAR comprises one or both of a transmembrane domain from a NCR (a NCR transmembrane domain) or a cytoplasmic domain comprising a functional signaling domain from a NCR (a NCR cytoplasmic domain), wherein the NCR transmembrane or cytoplasmic domain comprises a transmembrane or cytoplasmic domain of a protein selected from the group consisting of: NKp30 and NKp44;

a signaling lymphocyte activation molecule family chimeric antigen receptor (SLAMF-CAR), wherein the SLAMF-CAR comprises one or both of a transmembrane domain from a SLAMF (a SLAMF transmembrane domain) or a cytoplasmic domain comprising a functional signaling domain from a SLAMF (a SLAMF cytoplasmic domain);

a Fc receptor chimeric antigen receptor (FcR-CAR), wherein the FcR-CAR comprises one or both of a transmembrane domain from a FcR selected from CD16 or CD64, or a cytoplasmic domain comprising a functional signaling domain from a FcR selected from CD16 or CD64; or

a Ly49 receptor chimeric antigen receptor (Ly49-CAR), wherein the Ly49-CAR comprises one or both of a transmembrane domain from Ly49 (a Ly49 transmembrane domain) or a cytoplasmic domain comprising a functional signaling domain from Ly49 (a Ly49 cytoplasmic domain).