Patent application EP19166421A (EP), filed 2019-03-29
Publication number: EP3714944A1
Title: CAR FOR USE IN THE TREATMENT OF HVG DISEASE
Abstract: The invention provides an optimized and far potent chimeric antigen receptor for its use in the treatment of HvG disease in a patient having received a transplant, for use in suppressing the host's immune response directed against the transplant. The fusion protein is adapted for use in suppressing the immune rejection of a transplant which contains or expresses HLA-A*02 in a recipient patient who is negative for HLA-A*02, i.e. the patient prior to transplantation does not express HLA-A*02. The fusion protein is a chimeric antigen receptor (CAR), which upon expression in regulatory T-cells (T<sub>reg</sub>) causes a specific suppressor activity of the regulatory T-cells in the presence of HLA-A*02.
IPC codes: A61P  37/    06            A I
C07K  16/    28            A I
Claims: number: 1
text: Fusion protein comprising a single-chain variable fragment antibody domain (scFv), a hinge, a transmembrane domain, an intracellular hCD28 signalling domain and an intracellular hCD3ζ (hCD3 zeta) signalling domain forming a chimeric antigen receptor having specificity for HLA-A*02 (CAR-A*02) or having specificity for SLA-01*0401 (CAR-SLA-01*0401) for use in the treatment of HvG disease in a patient, characterized by the single-chain variable fragment antibody domain (scFv) having an amino acid sequence which is SEQ ID NO: 26 or SEQ ID NO: 28.
number: 2
text: Fusion protein according to claim 1, characterized in that the hinge and the transmembrane domain have an amino acid sequence of SEQ ID NO: 13, the hCD28 signalling domain has an amino acid sequence of SEQ ID NO: 14, and the hCD3ζ (hCD3 zeta) signalling domain has an amino acid sequence of SEQ ID NO: 15.
number: 3
text: Fusion protein according to claim 1, characterized in that the hinge is a hΔFc IgG domain having an amino acid sequence of SEQ ID NO: 20.
number: 4
text: Fusion protein according to one of the preceding claims, characterized in that the hinge and the transmembrane domain, which is a CD8 transmembrane domain, have an amino acid sequence of SEQ ID NO: 13, the hCD28 signalling domain has an amino acid sequence of SEQ ID NO: 14, and the hCD3 ζ domain has an amino acid sequence of SEQ ID NO: 15, or the hCD28 signalling domain including the hCD3ζ signalling domain have an amino acid sequence of SEQ ID NO: 21.
number: 5
text: Fusion protein according to one of the preceding claims, characterized in that it is expressed in a CD4<+>CD25<+>CD127<low> HLA-A*02 negative human regulatory T (Treg) cell.
number: 6
text: Fusion protein according to one of the preceding claims, characterized in that the patient is HLA-A*02 negative and in that the patient contains or is intended to contain a solid tissue transplant which is HLA-A*02 positive.
number: 7
text: Fusion protein according to one of the preceding claims, characterized in that a signal peptide of SEQ ID NO: 27 is arranged at the N-terminus of SEQ ID NO: 26 or SEQ ID NO: 28.
number: 8
text: Fusion protein according to one of the preceding claims, characterized in that it comprises or consists of, from N-terminus to C-terminus, one scFv domain having an amino acid sequence of SEQ ID NO: 26 or SEQ ID NO: 28, a hinge and a transmembrane domain having an amino acid sequence of SEQ ID NO: 13, a hCD28 signalling domain having an amino acid sequence of SEQ ID NO: 14, and a hCD3ζ (hCD3 zeta) signalling domain having an amino acid sequence of SEQ ID NO: 15, optionally additionally a signal peptide of SEQ ID NO: 27 at the N-terminus.
number: 9
text: Fusion protein according to one of the preceding claims, characterized in that it comprises or consists of, from N-terminal to C-terminal, one scFv domain having an amino acid sequence of SEQ ID NO: 26 or SEQ ID NO: 28, a hΔFc IgG domain as a hinge having an amino acid sequence of SEQ ID NO: 20, a hCD28 transmembrane domain and a hCD28/hCD3 signalling domain having an amino acid sequence of SEQ ID NO: 21, optionally additionally a signal peptide of SEQ ID NO: 27 at the N-terminus.
number: 10
text: Fusion protein according to one of the preceding claims, characterized in that it is expressed from a nucleic acid sequence encoding the fusion protein with optionally an additional N-terminal secretory leader peptide.
number: 11
text: Fusion protein according to one of the preceding claims, characterized in that it is expressed from a nucleic acid sequence encoding the fusion protein with an additional N-terminal secretory leader peptide and an additional C-terminal P2A-hFOXP3 having an amino acid sequence of SEQ ID NO: 22.
number: 12
text: Fusion protein according to one of claims 10 to 11, characterized in that the leader peptide has an amino acid sequence of SEQ ID NO: 24.
number: 13
text: Fusion protein according to one of the preceding claims, characterized by providing suppressor activity to a CD4<+>CD25<+>CD127<low>HLA-A*02 negative human regulatory T (Treg) cell in the presence of HLA-A*02 positive solid tissue or in the presence of SLA-01*0401 positive solid tissue.
number: 14
text: Fusion protein according to claim 12, characterized by providing for homing capability to secondary lymphoid organs in a CD4<+>CD25<+>CD127<low>HLA-A*02 negative human regulatory T (Treg) cell.
number: 15
text: Process for providing a human regulatory T (Treg) cell having suppressor activity in the presence of HLA-A*02 positive solid tissue, comprising the steps of
number: 16
text: Process according to claim 15, characterized in that isolating the human regulatory T cells is isolating HLA-A*02 negative human regulatory T cells.
number: 17
text: Process according to one of claims 15 to 16, characterized in that the nucleic acid sequence is comprised in a retroviral vector that is packaged in a retroviral particle and is introduced into the isolated Treg cells by transduction.
number: 18
text: Process according to one of claims 15 to 17, characterized in that following step b., the Treg cells are kept in culture for 24 h, followed by isolating Treg cells expressing the fusion protein.
number: 19
text: Process according to claim 18, characterized in that the Treg cells are kept in culture in a medium containing low dose IL-2, which medium does not contain an agent stimulating expansion of Treg cells.
number: 20
text: Treg cell containing a nucleic acid construct encoding a fusion protein according to one of claims 1 to 14.
Office action: search_report, dated 2020-09-30
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Final disposition: pending (2020-09-30)