Chimeric antigen receptors that bind to prostate specific membrane antigen

Patent metadata

Field	Value
Publication number	US-12428492-B2
Application number	US-202017788723-A
Country	US
Kind code	B2
Filing date	Dec 14, 2020
Priority date	Dec 23, 2019
Publication date	Sep 30, 2025
Grant date	Sep 30, 2025

Abstract

The present invention relates to a novel chimeric antigen receptor (CAR) comprising an antigen-binding fragment which binds specifically to PSMA antigen, and a method of manufacturing high-quality CAR T cell products by transfection and/or transduction of T cells therewith, which allows to effectively treat tumors in vivo alone or in combination with pharmaceutical drugs, such chemotherapies, biopharmaceutical drugs, such as antibodies, or small-molecule drugs, such as protein kinase inhibitors.

First claim

The invention claimed is:

1. A humanized chimeric antigen receptor (CAR) comprising:
(a) an antigen-binding fragment which binds specifically to a prostate specific membrane antigen (PSMA), wherein said antigen-binding fragment against said PSMA contains the following 6 QDR sequences: GFTFSDYY (SEQ ID NO: 9) as CDR-H1, ISDGGYYT (SEQ ID NO: 10) as CDR-H2, TRGFPLLRHGAMDYWG (SEQ ID NO: 11) as CDR-H3, QNVDTN (SEQ ID NO: 12) as CDR-L1, SAS (SEQ ID NO: 13) as CDR-L2 and QQYDSYPYT (SEQ ID NO: 14) as CDR-L3,
(b) a transmembrane domain,
(c) an intracellular signaling domain comprising a CD3ζ cytoplasmic domain,
(d) a co-stimulatory domain comprising a CD28 and/or a 4-1BB cytoplasmic domain, and
(e) an Fc IgG1 derived hinge region disposed between the antigen-binding fragment of part (a) and the transmembrane domain of part (b) so as to provide a physical spacer therebetween for optimal target recognition.

2. The humanized chimeric antigen receptor according to claim 1 characterized in that said chimeric antigen receptor contains at least one humanized light chain selected from the group consisting of SEQ ID NO:26, SEQ ID NO:28, SEQ ID NO:30, SEQ ID NO:32, SEQ ID NO:34, and SEQ ID NO:36 or at least one heavy chain having an amino acid sequence selected from the group consisting of SEQ ID NO:16, SEQ ID NO:18, SEQ ID NO:20, SEQ ID NO:22, and SEQ ID NO:24.

3. A nucleic acid coding for the humanized chimeric antigen receptor according to claim 1 .

4. A vector coding for the humanized chimeric antigen receptor according to claim 1 , characterized in that said vector contains a nucleic acid coding for said humanized chimeric antigen receptor.

5. A vector coding for a chimeric antigen receptor characterized in that said vector comprises SEQ ID NO:37, SEQ ID NO:38, or a sequence which is at least 95% identical to the entirety of SEQ ID NO: 37 or SEQ ID NO: 38.

6. An in vitro method of providing immune cells that express the humanized chimeric antigen receptor according to claim 1 , said method comprising the steps of:
(i) isolating immune cells from a donor by leukapheresis;
(ii) transfecting/transducing the immune cells with a vector containing a nucleic acid coding for said humanized chimeric antigen receptor; and
(iii) isolating and amplifying the transfected/transduced immune cells that express said humanized chimeric antigen receptor.

7. An immune cell containing the genetic information coding for the humanized chimeric antigen receptor according to claim 1 .

8. The humanized chimeric antigen receptor according to claim 1 or a nucleic acid coding for said humanized chimeric antigen receptor or a vector containing such a nucleic acid formulated for use in the treatment of prostate cancer and/or prostate-derived tumors.

9. An agent having anti-tumor activity comprising the humanized chimeric antigen receptor according to claim 1 , a nucleic acid coding for said humanized chimeric antigen receptor, and/or a vector containing such a nucleic acid formulated for use in the treatment of solid tumor expressing PSMA, wherein said agent is introduced into the solid tumor.

10. A method of treating prostate cancer, a prostate-derived tumor, or a solid tumor expressing PSMA in a subject in need thereof, said method comprising the steps of administering to said subject transfected/transduced immune cells isolated and amplified by the method of claim 6 simultaneously or contemporaneously with the administration of (a) one or more chemotherapeutically active agents selected from the group consisting of taxol derivatives, 5-fluorouracil, cyclophosphamide, mitoxantrone, docetaxel and capacitaxel, and/or (b) one or more biopharmaceutical agents selected from the group consisting of protein kinase inhibitors, enzyme inhibitors, anti-genomic therapeutics, and T cell checkpoint antagonists.

11. The immune cells of claim 7 , wherein said immune cells are selected from the group consisting of T-cells, natural killer cells (NK), invariant natural killer T-cells (iNKT), diverse natural killer cells (dNKT), cytokine-induced killer cells (CIK) and γ-δ T-cells.

12. A humanized chimeric antigen receptor (CAR) that consists essentially of:
a. an antigen-binding fragment that binds specifically to a prostate specific membrane antigen (PSMA), wherein said antigen-binding fragment against said PSMA contains the following 6 DR sequences: GFTFSDYY (SEQ ID NO: 9) as CDR-H1, ISDGGYYT (SEQ ID NO: 10) as CDR-H2, TRGFPLLRHGAMDYWG (SEQ ID NO: 11) as CDR-H3, QNVDTN (SEQ ID NO: 12) as CDR-L1, SAS (SEQ ID NO: 13) as CDR-L2 and QQYDSYPYT (SEQ ID NO: 14) as CDR-L3,
b. a transmembrane domain,
c. a CD3ζ cytoplasmic domain acting as an intracellular signaling domain,
d. at least one co-stimulatory domain selected from the group consisting of a CD28 and 4-1BB cytoplasmic domain, and
e. an Fc IgG1 derived hinge region disposed between the antigen-binding fragment of part (a) and the transmembrane domain of part (b) so as to provide a physical spacer therebetween for optimal target recognition.

Assignees

• Univ Freiburg Albert Ludwigs

Inventors

• Cathomen Toni
• Wolf Philipp
• Alzubi Jamal
• Schultze-Seemann Susanne
• Kuckuck Irina

Classifications

• A61K40/4276

  Prostate specific membrane antigen [PSMA] · CPC title

• A61K40/31

  Chimeric antigen receptors [CAR] · CPC title

• A61K40/11

  T-cells, e.g. tumour infiltrating lymphocytes [TIL] or regulatory T [Treg] cells; Lymphokine-activated killer [LAK] cells · CPC title

• C07K2319/03

  containing a transmembrane segment · CPC title

• C07K2319/02

  containing a signal sequence · CPC title