Claudin-18.2-specific immunoreceptors and t cell epitopes

Abstract

The present invention provides Claudin-18.2-specific immunoreceptors (T cell receptors and artificial T cell receptors (chimeric antigen receptors; CARs)) and T cell epitopes which are useful for immunotherapy.

1 . A peptide comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 2, 3, 4, 5, 6 and 7 or a variant of said amino acid sequence. 
     
     
         2 . The peptide of  claim 1  which is 100 or less, 50 or less, 20 or less, or 10 or less amino acids long. 
     
     
         3 . The peptide of  claim 1  or  2  which consists of the amino acid sequence selected from the group consisting of SEQ ID NOs: 2, 3, 4, 5, 6 and 7 or a variant of said amino acid sequence. 
     
     
         4 . A nucleic acid comprising a nucleotide sequence encoding the peptide of any one of  claims 1  to  3 . 
     
     
         5 . The nucleic acid of  claim 4  which is a recombinant nucleic acid. 
     
     
         6 . A cell comprising the nucleic acid of  claim 4  or  5 . 
     
     
         7 . A cell that presents the peptide of any one of  claims 1  to  3  or a procession product thereof. 
     
     
         8 . An immunoreactive cell which is reactive with a peptide of any one of  claims 1  to  3 . 
     
     
         9 . A binding agent which binds to a peptide of  claim 1 , optionally in a complex with an MHC molecule. 
     
     
         10 . A T cell receptor which binds to a peptide of  claim 1 , optionally in a complex with an MHC molecule, and preferably is reactive with said peptide, or a polypeptide chain of said T cell receptor. 
     
     
         11 . A T cell receptor α-chain or a T cell receptor comprising said T cell receptor α-chain, wherein said T cell receptor α-chain is selected from the group consisting of:
 (i) a T cell receptor α-chain comprising at least one, preferably two, more preferably all three of the CDR sequences of a T cell receptor α-chain selected from the group consisting of SEQ ID NOs: 8, 10, 12, 14, 16 and 18 or a variant thereof and 
 (ii) a T cell receptor α-chain comprising a T cell receptor α-chain sequence selected from the group consisting of SEQ ID NOs: 8, 10, 12, 14, 16 and 18 or a fragment thereof, or a variant of said sequence or fragment. 
 
     
     
         12 . A T cell receptor β-chain or a T cell receptor comprising said T cell receptor β-chain, wherein said T cell receptor β-chain is selected from the group consisting of:
 (i) a T cell receptor β-chain comprising at least one, preferably two, more preferably all three of the CDR sequences of a T cell receptor β-chain selected from the group consisting of SEQ ID NOs: 9, 11, 13, 15, 17 and 19 or a variant thereof 
 and 
 (ii) a T cell receptor β-chain comprising a T cell receptor β-chain sequence selected from the group consisting of SEQ ID NOs: 9, 11, 13, 15, 17 and 19 or a fragment thereof, or a variant of said sequence or fragment. 
 
     
     
         13 . A T cell receptor selected from the group consisting of:
 (I) a T cell receptor comprising:
 (i) a T cell receptor α-chain comprising at least one, preferably two, more preferably all three of the CDR sequences of the T cell receptor α-chain of SEQ ID NO: x or a variant thereof, and 
 (ii) a T cell receptor β-chain comprising at least one, preferably two, more preferably all three of the CDR sequences of a T cell receptor β-chain of SEQ ID NO: x+1 or a variant thereof; 
   wherein x selected from the group consisting of 8, 10, 12, 14, 16 and 18   and   (II) a T cell receptor comprising:
 (i) a T cell receptor α-chain comprising the T cell receptor α-chain sequence of SEQ ID NO: x or a fragment thereof, or a variant of said sequence or fragment, and 
 (ii) a T cell receptor β-chain comprising the T cell receptor β-chain sequence of SEQ ID NO: x+1 or a fragment thereof, or a variant of said sequence or fragment; 
   wherein x selected from the group consisting of 8, 10, 12, 14, 16 and 18.   
     
     
         14 . An artificial T cell receptor which binds to claudin-18.2 (CLDN18.2). 
     
     
         15 . The artificial T cell receptor of  claim 14  which binds to an extracellular domain of CLDN18.2. 
     
     
         16 . The artificial T cell receptor of  claim 14  or  15  wherein said binding is a specific binding. 
     
     
         17 . The artificial T cell receptor of any one of  claims 14  to  16  which comprises a binding domain for CLDN18.2. 
     
     
         18 . The artificial T cell receptor of  claim 17  wherein the binding domain for CLDN18.2 is comprised by an exodomain of said artificial T cell receptor. 
     
     
         19 . The artificial T cell receptor of  claim 17  or  18 , wherein the binding domain for CLDN18.2 comprises a single-chain variable fragment (scFv) of a CLDN18.2 antibody. 
     
     
         20 . The artificial T cell receptor of any one of  claims 17  to  19 , wherein the binding domain for CLDN18.2 comprises a variable region of a heavy chain of an immunoglobulin (VH) with a specificity for CLDN18.2 (VH(CLDN18.2)) and a variable region of a light chain of an immunoglobulin (VL) with a specificity for CLDN18.2 (VL(CLDN18.2)). 
     
     
         21 . The artificial T cell receptor of  claim 20 , wherein said heavy chain variable region (VH) and the corresponding light chain variable region (VL) are connected via a peptide linker, preferably a peptide linker comprising the amino acid sequence (GGGGS)3. 
     
     
         22 . The artificial T cell receptor of any one of  claims 17  to  21 , wherein the binding domain for CLDN18.2 comprises a VH(CLDN18.2) comprising an amino acid sequence represented by SEQ ID NO: 23 or a fragment thereof, or a variant of said amino acid sequence or fragment. 
     
     
         23 . The artificial T cell receptor of any one of  claims 17  to  22 , wherein the binding domain for CLDN18.2 comprises a VL(CLDN18.2) comprising an amino acid sequence represented by SEQ ID NO: 30 or a fragment thereof, or a variant of said amino acid sequence or fragment. 
     
     
         24 . The artificial T cell receptor of any one of  claims 17  to  23 , wherein the binding domain for CLDN18.2 comprises a VH(CLDN18.2) comprising an amino acid sequence represented by SEQ ID NO: 23 or a fragment thereof, or a variant of said amino acid sequence or fragment and a VL(CLDN18.2) comprising an amino acid sequence represented by SEQ ID NO:
 30 or a fragment thereof, or a variant of said amino acid sequence or fragment. 
 
     
     
         25 . The artificial T cell receptor of any one of  claims 17  to  24 , wherein the binding domain for CLDN18.2 comprises an amino acid sequence represented by SEQ ID NO: 35 or a fragment thereof, or a variant of said amino acid sequence or fragment. 
     
     
         26 . The artificial T cell receptor of any one of  claims 14  to  25  which comprises a transmembrane domain. 
     
     
         27 . The artificial T cell receptor of any one of  claims 14  to  26  which comprises a T cell signaling domain. 
     
     
         28 . The artificial T cell receptor of  claim 27 , wherein the T cell signaling domain comprises CD3-zeta, preferably the endodomain of CD3-zeta, optionally in combination with CD28. 
     
     
         29 . The artificial T cell receptor of any one of  claims 14  to  28  which comprises a signal peptide which directs the nascent protein into the endoplasmic reticulum. 
     
     
         30 . The artificial T cell receptor of any one of  claims 14  to  29  which comprises a spacer region which links the binding domain for CLDN18.2 to the transmembrane domain. 
     
     
         31 . The artificial T cell receptor of any one of  claims 14  to  30  which comprises the structure:
 NH2—signal peptide—binding domain for CLDN18.2—spacer region—transmembrane domain—T cell signaling domain—COOH. 
 
     
     
         32 . The artificial T cell receptor of any one of  claims 14  to  31  which comprises the amino acid sequence according to SEQ ID NO: 41 or a fragment thereof, or a variant of said amino acid sequence or fragment. 
     
     
         33 . A nucleic acid comprising a nucleotide sequence encoding the T cell receptor chain or T cell receptor of any one of  claims 10  to  13  or encoding the artificial T cell receptor of any one of  claims 14  to  32 . 
     
     
         34 . A cell comprising the T cell receptor chain or T cell receptor of any one of  claims 10  to  13  or the artificial T cell receptor of any one of  claims 14  to  32  and/or comprising a nucleic acid comprising a nucleotide sequence encoding the T cell receptor chain or T cell receptor or encoding the artificial T cell receptor. 
     
     
         35 . A method of producing an immunoreactive cell comprising the step of transducing a T cell with a nucleic acid of  claim 33 . 
     
     
         36 . A pharmaceutical composition comprising one or more of:
 (i) the peptide of any one of  claims 1  to  3 ;   (ii) the nucleic acid of any one of  claims 4 ,  5  and  33 ;   (iii) the cell of any one of  claims 6 ,  7  and  34 ;   (iv) the immunoreactive cell of  claim 8 ;   (v) the binding agent of  claim 9 ;   (vi) the T cell receptor of any one of  claims 10  to  13 ; and   (vii) the artificial T cell receptor of any one of  claims 14  to  32 .   
     
     
         37 . The pharmaceutical composition of  claim 36  which further comprises a pharmaceutically acceptable carrier. 
     
     
         38 . A method of treating or preventing a cancer disease comprising administering to a patient the pharmaceutical composition of  claim 36  or  37 . 
     
     
         39 . The peptide of any one of  claims 1  to  3 , the nucleic acid of any one of  claims 4 ,  5  and  33 , the cell of any one of  claims 6 ,  7  and  34 , the immunoreactive cell of  claim 8 , the binding agent of  claim 9 , the T cell receptor of any one of  claims 10  to  13 , or the artificial T cell receptor of any one of  claims 14  to  32  for use in therapy, in particular for use in treating or preventing cancer. 
     
     
         40 . A method for inducing an immune response in a subject, comprising administering to the subject a pharmaceutical composition of  claim 36 . 
     
     
         41 . A method for stimulating, priming and/or expanding T cells, comprising contacting T cells with one or more of: the peptide of any one of  claims 1  to  3 , the nucleic acid of  claim 4  or  5  and/or the cell of  claim 6  or  7 . 
     
     
         42 . A method of killing cancer cells in a subject, comprising the step of providing to the subject a therapeutically effective amount of the peptide of any one of  claims 1  to  3 , the nucleic acid of any one of  claims 4 ,  5  and  33 , the cell of any one of  claims 6 ,  7  and  34 , the immunoreactive cell of  claim 8 , the binding agent of  claim 9 , the T cell receptor of any one of  claims 10  to  13 , or the artificial T cell receptor of any one of  claims 14  to  32 . 
     
     
         43 . A method for determining an immune response in a subject, comprising determining T cells reactive with a peptide of any one of  claims 1  to  3  in a biological sample isolated from the subject.