WO2025093596A2

WO2025093596A2 - Antigen recognizing construct capable of binding to a preferentially expressed antigen in melanoma (prame) peptide and a t cell receptor (tcr) having antigenic specificity for prame as well as corresponding nucleic acid sequence, vector, host cell, pharmaceutical composition and kit

Info

Publication number: WO2025093596A2
Application number: PCT/EP2024/080675
Authority: WO
Inventors: Lia WALCHER; Lorenz KNACKSTEDT; Lucia Poncette; Elisa Kieback
Original assignee: T Knife GmbH
Current assignee: T Knife GmbH
Priority date: 2023-10-30
Filing date: 2024-10-30
Publication date: 2025-05-08
Anticipated expiration: 2026-04-30
Also published as: WO2025093596A3

Abstract

The present invention inter alia relates to an antigen recognizing construct capable of binding to a Preferentially Expressed Antigen in Melanoma (FRAME) peptide, a T cell receptor having antigenic specificity for FRAME, a nucleic acid sequence encoding said antigen recognizing construct or T cell receptor, a vector comprising said nucleic acid sequence, and a host cell comprising said antigen recognizing construct or T cell receptor. The invention also relates to the antigen recognizing construct or T cell receptor, the nucleic acid sequence, vector or host cell for use in medicine or for use in detection, diagnosis, prognosis, prevention and/or treatment of a disease. Additionally, the present invention relates to a method of treating a disease as well as a pharmaceutical composition and a kit.

Description

ANTIGEN RECOGNIZING CONSTRUCT CAPABLE OF BINDING TO A PREFERENTIALLY EXPRESSED ANTIGEN IN MELANOMA (PRAME) PEPTIDE AND A T CELL RECEPTOR (TCR) HAVING ANTIGENIC SPECIFICITY FOR PRAME AS WELL AS CORRESPONDING NUCLEIC ACID SEQUENCE, VECTOR, HOST CELL, PHARMACEUTICAL COMPOSITION AND KIT

FIELD OF THE INVENTION

[001] The present application claims priority of the European Patent application No. 23206799.1 , filed October 30, 2023, the disclosure of which is incorporated herein by reference in its entirety.

[002] The present invention relates to an antigen recognizing construct capable of binding to a Preferentially Expressed Antigen in Melanoma (PRAME) peptide, a T cell receptor (TCR) having antigenic specificity for PRAME, a nucleic acid sequence encoding said antigen recognizing construct or said T cell receptor, a respective vector comprising the nucleic acid sequence and a host cell comprising the antigen recognizing construct or the T cell receptor. The invention also relates to the antigen recognizing construct or the T cell receptor, the nucleic acid sequence, the vector, or the host cell, for use in medicine or for use in detection, diagnosis, prognosis, prevention and/or treatment of a disease. The present invention further relates to a use of the antigen recognizing construct or the T cell receptor, the nucleic acid sequence, the vector, or the host cell, for the manufacture of a medicament for treating a disease. The present invention also relates to a method of treating a disease, comprising the step of administering a therapeutically effective amount of the antigen recognizing construct or the T cell receptor, the nucleic acid sequence, the vector, or the host cell. Additionally, the present invention also relates to a pharmaceutical composition comprising the antigen recognizing construct or the T cell receptor, the nucleic acid sequence, the vector, or the host cell of the invention and also to a respective kit for use in medicine.

BACKGROUND OF THE INVENTION

[003] Cancer testis antigens (CTAs) have been considered promising targets for adoptive T cell therapy thanks to their restricted expression in somatic normal tissues, re- expression in many cancer types, and immunogenic nature (Fratta E. et al., Mol. Oncol. 2011 ;5: 164-182). CTA re-expression in tumors has been linked with worse clinical outcome in various cancers (Yao J. et al.; Cancer Immunol. Res. 2014; 2:371-379). Among all candidate CTAs, MAGE-A3, NY-ESO-1 , and PRAME have shown great potential as prognostic biomarkers and immunotherapeutic targets.

[004] PRAME is a tumor-associated antigen expressed in a wide variety of tumors. Specifically, PRAME is re-expressed in a wide range of cancers, including melanoma, renal cell cancer, non-small cell lung cancer (NSCLC), neuroblastoma, breast cancer, multiple myeloma, acute leukemia, chronic myeloid leukemia, multiple sarcoma subtypes, and primary and metastatic uveal melanoma (Al-Khadairi G. and Decock J.; Cancers (Basel). 2019 Jul; 11 (7): 984). It is not expressed in normal tissues, except testis.

[005] The encoded protein acts as a repressor of retinoic acid receptor, and likely confers a growth advantage to cancer cells via this function. PRAME overexpression in triple negative breast cancer has also been found to promote cancer cell motility through induction of the epithelial-to-mesenchymal transition (Al-Khadairi et al., Journal of Translational Medicine 2019; 17: 9).

[006] PRAME may be expressed in any of a variety of human cancers such as, for example, pancreatic cancers (e.g., pancreatic carcinoma), colorectal cancers, lung cancers (e.g., lung adenocarcinoma), endometrial cancers, ovarian cancers (e.g., epithelial ovarian cancer), prostate cancers, melanomas, thyroid cancers, and breast cancers, as well as some instances of myeloid leukemias such as AML.

[007] There is thus a need for novel therapies targeting PRAME peptides. Accordingly, it is an object of the invention to provide such therapies.

SUMMARY OF THE INVENTION

[008] This object is inter alia accomplished by the antigen recognizing constructs, the TCRs, nucleic acids, vectors, host cells, methods, compositions, and kits having the features of the respective independent claims.

[009] In a first aspect, the invention provides an antigen recognizing construct capable of binding to a Preferentially Expressed Antigen in Melanoma (PRAME) peptide, wherein the antigen recognizing construct comprises a complementary determining region 3 (CDR3) of the a-chain having at least 90% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 3, 9, 15, 21 , 27, 33, 39, 45, 51 , 57, 63, 69, and 75, and/or wherein the antigen recognizing construct comprises a complementary determining region 3 (CDR3) of the p-chain having at least 90% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 6, 12, 18, 24, 30, 36, 42, 48, 54, 60, 66, 72 and 78.

[0010] It is further provided an antigen recognizing construct, wherein the antigen recognizing construct comprises a complementary determining region 3 (CDR3) of the a- chain having at least 90% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 3, 9, 15, 21 , 27, 33, 39, 45, 51 , 57, 63, 69, and 75, and/or wherein the antigen recognizing construct comprises a complementary determining region 3 (CDR3) of the p-chain having at least 90% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 6, 12, 18, 24, 30, 36, 42, 48, 54, 60, 66, 72 and 78.

[0011] In a second aspect, the invention provides a T cell receptor (TCR) having antigenic specificity for Preferentially Expressed Antigen in Melanoma (PRAME), wherein the TCR comprises:

(a) an a-chain CDR1 comprising or consisting of the amino acid sequence of SEQ ID NO: 1 , an a-chain CDR2 comprising or consisting of the amino acid sequence of SEQ ID NO: 2, and an a-chain CDR3 comprising or consisting of the amino acid sequence of SEQ ID NO: 3; or

(b) an a-chain CDR1 comprising or consisting of the amino acid sequence of SEQ ID NO: 7, an a-chain CDR2 comprising or consisting of the amino acid sequence of SEQ ID NO: 8, and an a-chain CDR3 comprising or consisting of the amino acid sequence of SEQ ID NO: 9; or

(c) an a-chain CDR1 comprising or consisting of the amino acid sequence of SEQ ID NO: 13, an a-chain CDR2 comprising or consisting of the amino acid sequence of SEQ ID NO: 14, and an a-chain CDR3 comprising or consisting of the amino acid sequence of SEQ ID NO: 15; or

(d) an a-chain CDR1 comprising or consisting of the amino acid sequence of SEQ ID NO: 19, an a-chain CDR2 comprising or consisting of the amino acid sequence of SEQ ID NO: 20, and an a-chain CDR3 comprising or consisting of the amino acid sequence of SEQ ID NO: 21 ; or

(e) an a-chain CDR1 comprising or consisting of the amino acid sequence of SEQ ID NO: 25, an a-chain CDR2 comprising or consisting of the amino acid sequence of SEQ ID NO: 26, and an a-chain CDR3 comprising or consisting of the amino acid sequence of SEQ ID NO: 27, or

(f) an a-chain CDR1 comprising or consisting of the amino acid sequence of SEQ ID NO:

31 , an a-chain CDR2 comprising or consisting of the amino acid sequence of SEQ ID NO: 32, and an a-chain CDR3 comprising or consisting of the amino acid sequence of SEQ ID NO: 33, or

(g) an a-chain CDR1 comprising or consisting of the amino acid sequence of SEQ ID NO: 37, an a-chain CDR2 comprising or consisting of the amino acid sequence of SEQ ID NO: 38, and an a-chain CDR3 comprising or consisting of the amino acid sequence of SEQ ID NO: 39, or

(h) an a-chain CDR1 comprising or consisting of the amino acid sequence of SEQ ID NO: 43, an a-chain CDR2 comprising or consisting of the amino acid sequence of SEQ ID NO: 44, and an a-chain CDR3 comprising or consisting of the amino acid sequence of SEQ ID NO: 45, or

(i) an a-chain CDR1 comprising or consisting of the amino acid sequence of SEQ ID NO: 49, an a-chain CDR2 comprising or consisting of the amino acid sequence of SEQ ID NO: 50, and an a-chain CDR3 comprising or consisting of the amino acid sequence of SEQ ID NO: 51 , or

(j) an a-chain CDR1 comprising or consisting of the amino acid sequence of SEQ ID NO: 55, an a-chain CDR2 comprising or consisting of the amino acid sequence of SEQ ID NO: 56, and an a-chain CDR3 comprising or consisting of the amino acid sequence of SEQ ID NO: 57, or

(k) an a-chain CDR1 comprising or consisting of the amino acid sequence of SEQ ID NO: 61 , an a-chain CDR2 comprising or consisting of the amino acid sequence of SEQ ID NO: 62, and an a-chain CDR3 comprising or consisting of the amino acid sequence of SEQ ID NO: 63, or

(l) an a-chain CDR1 comprising or consisting of the amino acid sequence of SEQ ID NO: 67, an a-chain CDR2 comprising or consisting of the amino acid sequence of SEQ ID NO: 68, and an a-chain CDR3 comprising or consisting of the amino acid sequence of SEQ ID NO: 69, or

(m) an a-chain CDR1 comprising or consisting of the amino acid sequence of SEQ ID NO: 73, an a-chain CDR2 comprising or consisting of the amino acid sequence of SEQ ID NO: 74, and an a-chain CDR3 comprising or consisting of the amino acid sequence of SEQ ID NO: 75. [0012] According to a third aspect, the invention provides a T cell receptor (TCR) having antigenic specificity for Preferentially Expressed Antigen in Melanoma (PRAME), wherein the TCR comprises:

(a) a p-chain CDR1 comprising or consisting of the amino acid sequence of SEQ ID NO:

4, a p-chain CDR2 comprising or consisting of the amino acid sequence of SEQ ID NO: 5, and a p-chain CDR3 comprising or consisting of the amino acid sequence of SEQ ID NO: 6; or

(b) a p-chain CDR1 comprising or consisting of the amino acid sequence of SEQ ID NO: 10, a p-chain CDR2 comprising or consisting of the amino acid sequence of SEQ ID NO: 11 , and a p-chain CDR3 comprising or consisting of the amino acid sequence of SEQ ID NO: 12; or

(c) a p-chain CDR1 comprising or consisting of the amino acid sequence of SEQ ID NO: 16, a p-chain CDR2 comprising or consisting of the amino acid sequence of SEQ ID NO: 17, and a p-chain CDR3 comprising or consisting of the amino acid sequence of SEQ ID NO: 18; or

(d) a p-chain CDR1 comprising or consisting of the amino acid sequence of SEQ ID NO: 22, a p-chain CDR2 comprising or consisting of the amino acid sequence of SEQ ID NO: 23, and a p-chain CDR3 comprising or consisting of the amino acid sequence of SEQ ID NO: 24; or

(e) a p-chain CDR1 comprising or consisting of the amino acid sequence of SEQ ID NO: 28, a p-chain CDR2 comprising or consisting of the amino acid sequence of SEQ ID NO: 29, and a p-chain CDR3 comprising or consisting of the amino acid sequence of SEQ ID NO: 30, or

(f) a p-chain CDR1 comprising or consisting of the amino acid sequence of SEQ ID NO: 34, a p-chain CDR2 comprising or consisting of the amino acid sequence of SEQ ID NO: 35, and a p-chain CDR3 comprising or consisting of the amino acid sequence of SEQ ID NO: 36, or

(g) a p-chain CDR1 comprising or consisting of the amino acid sequence of SEQ ID NO: 40, a p-chain CDR2 comprising or consisting of the amino acid sequence of SEQ ID NO: 41 , and a p-chain CDR3 comprising or consisting of the amino acid sequence of SEQ ID NO: 42, or

(h) a p-chain CDR1 comprising or consisting of the amino acid sequence of SEQ ID NO: 46, a p-chain CDR2 comprising or consisting of the amino acid sequence of SEQ ID NO: 47, and a p-chain CDR3 comprising or consisting of the amino acid sequence of SEQ ID NO: 48, or

(i) a p-chain CDR1 comprising or consisting of the amino acid sequence of SEQ ID NO: 52, a p-chain CDR2 comprising or consisting of the amino acid sequence of SEQ ID NO: 53, and a p-chain CDR3 comprising or consisting of the amino acid sequence of SEQ ID NO: 54, or

(j) a p-chain CDR1 comprising or consisting of the amino acid sequence of SEQ ID NO: 58, a p-chain CDR2 comprising or consisting of the amino acid sequence of SEQ ID NO: 59, and a p-chain CDR3 comprising or consisting of the amino acid sequence of SEQ ID NO: 60, or

(k) a p-chain CDR1 comprising or consisting of the amino acid sequence of SEQ ID NO: 64, a p-chain CDR2 comprising or consisting of the amino acid sequence of SEQ ID NO: 65, and a p-chain CDR3 comprising or consisting of the amino acid sequence of SEQ ID NO: 66, or

(l) a p-chain CDR1 comprising or consisting of the amino acid sequence of SEQ ID NO: 70, a p-chain CDR2 comprising or consisting of the amino acid sequence of SEQ ID NO: 71 , and a p-chain CDR3 comprising or consisting of the amino acid sequence of SEQ ID NO: 72, or

(m) a p-chain CDR1 comprising or consisting of the amino acid sequence of SEQ ID NO: 76, a p-chain CDR2 comprising or consisting of the amino acid sequence of SEQ ID NO: 77, and a p-chain CDR3 comprising or consisting of the amino acid sequence of SEQ ID NO: 78. [0013] In a fourth aspect, the invention provides a T cell receptor (TCR) having antigenic specificity for Preferentially Expressed Antigen in Melanoma (PRAME), wherein the TCR comprises:

(a) an a-chain CDR1 comprising or consisting of the amino acid sequence of SEQ ID NO: 1 , an a-chain CDR2 comprising or consisting of the amino acid sequence of SEQ ID NO: 2, an a-chain CDR3 comprising or consisting of the amino acid sequence of SEQ ID NO: 3; a p-chain CDR1 comprising or consisting of the amino acid sequence of SEQ ID NO: 4, a p-chain CDR2 comprising or consisting of the amino acid sequence of SEQ ID NO: 5, and a p-chain CDR3 comprising or consisting of the amino acid sequence of SEQ ID NO: 6; or

(b) an a-chain CDR1 comprising or consisting of the amino acid sequence of SEQ ID NO: an a-chain CDR2 comprising or consisting of the amino acid sequence of SEQ ID NO: 8, an a-chain CDR3 comprising or consisting of the amino acid sequence of SEQ ID NO: 9; a p-chain CDR1 comprising or consisting of the amino acid sequence of SEQ ID NO: 10, a p-chain CDR2 comprising or consisting of the amino acid sequence of SEQ ID NO: 11 , and a p-chain CDR3 comprising or consisting of the amino acid sequence of SEQ ID NO: 12; or

(c) an a-chain CDR1 comprising or consisting of the amino acid sequence of SEQ ID NO: 13, an a-chain CDR2 comprising or consisting of the amino acid sequence of SEQ ID NO: 14, an a-chain CDR3 comprising or consisting of the amino acid sequence of SEQ ID NO: 15; a p-chain CDR1 comprising or consisting of the amino acid sequence of SEQ ID NO: 16, a p-chain CDR2 comprising or consisting of the amino acid sequence of SEQ ID NO: 17, and a p-chain CDR3 comprising or consisting of the amino acid sequence of SEQ ID NO: 18; or

(d) an a-chain CDR1 comprising or consisting of the amino acid sequence of SEQ ID NO: 19, an a-chain CDR2 comprising or consisting of the amino acid sequence of SEQ ID NO: 20, an a-chain CDR3 comprising or consisting of the amino acid sequence of SEQ ID NO: 21 ; a p-chain CDR1 comprising or consisting of the amino acid sequence of SEQ ID NO: 22, a p-chain CDR2 comprising or consisting of the amino acid sequence of SEQ ID NO: 23, and a p-chain CDR3 comprising or consisting of the amino acid sequence of SEQ ID NO: 24; or

(e) an a-chain CDR1 comprising or consisting of the amino acid sequence of SEQ ID NO: 25, an a-chain CDR2 comprising or consisting of the amino acid sequence of SEQ ID NO: 26, an a-chain CDR3 comprising or consisting of the amino acid sequence of SEQ ID NO: 27; a p-chain CDR1 comprising or consisting of the amino acid sequence of SEQ ID NO: 28, a p-chain CDR2 comprising or consisting of the amino acid sequence of SEQ ID NO: 39, and a p-chain CDR3 comprising or consisting of the amino acid sequence of SEQ ID NO: 30, or

(f) an a-chain CDR1 comprising or consisting of the amino acid sequence of SEQ ID NO: 31 , an a-chain CDR2 comprising or consisting of the amino acid sequence of SEQ ID NO: 32, an a-chain CDR3 comprising or consisting of the amino acid sequence of SEQ ID NO: 33; a p-chain CDR1 comprising or consisting of the amino acid sequence of SEQ ID NO: 34, a p-chain CDR2 comprising or consisting of the amino acid sequence of SEQ ID NO: 35, and a p-chain CDR3 comprising or consisting of the amino acid sequence of SEQ ID NO: 36, or

(g) an a-chain CDR1 comprising or consisting of the amino acid sequence of SEQ ID NO: 37, an a-chain CDR2 comprising or consisting of the amino acid sequence of SEQ ID NO: 38, an a-chain CDR3 comprising or consisting of the amino acid sequence of SEQ ID NO: 39; a p-chain CDR1 comprising or consisting of the amino acid sequence of SEQ ID NO: 40, a p-chain CDR2 comprising or consisting of the amino acid sequence of SEQ ID NO: 41 , and a p-chain CDR3 comprising or consisting of the amino acid sequence of SEQ ID NO: 42, or

(h) an a-chain CDR1 comprising or consisting of the amino acid sequence of SEQ ID NO: 43, an a-chain CDR2 comprising or consisting of the amino acid sequence of SEQ ID NO: 44, an a-chain CDR3 comprising or consisting of the amino acid sequence of SEQ ID NO: 45; a p-chain CDR1 comprising or consisting of the amino acid sequence of SEQ ID NO: 46, a p-chain CDR2 comprising or consisting of the amino acid sequence of SEQ ID NO: 47, and a p-chain CDR3 comprising or consisting of the amino acid sequence of SEQ ID NO: 48, or

(i) an a-chain CDR1 comprising or consisting of the amino acid sequence of SEQ ID NO: 49, an a-chain CDR2 comprising or consisting of the amino acid sequence of SEQ ID NO: 50, an a-chain CDR3 comprising or consisting of the amino acid sequence of SEQ ID NO: 51 ; a p-chain CDR1 comprising or consisting of the amino acid sequence of SEQ ID NO: 52, a p-chain CDR2 comprising or consisting of the amino acid sequence of SEQ ID NO: 53, and a p-chain CDR3 comprising or consisting of the amino acid sequence of SEQ ID NO: 54, or

(j) an a-chain CDR1 comprising or consisting of the amino acid sequence of SEQ ID NO: 55, an a-chain CDR2 comprising or consisting of the amino acid sequence of SEQ ID NO: 56, an a-chain CDR3 comprising or consisting of the amino acid sequence of SEQ ID NO: 57; a p-chain CDR1 comprising or consisting of the amino acid sequence of SEQ ID NO: 58, a p-chain CDR2 comprising or consisting of the amino acid sequence of SEQ ID NO: 59, and a p-chain CDR3 comprising or consisting of the amino acid sequence of SEQ ID NO: 60, or

(k) an a-chain CDR1 comprising or consisting of the amino acid sequence of SEQ ID NO: 61 , an a-chain CDR2 comprising or consisting of the amino acid sequence of SEQ ID NO: 62, an a-chain CDR3 comprising or consisting of the amino acid sequence of SEQ ID NO: 63; a p-chain CDR1 comprising or consisting of the amino acid sequence of SEQ ID NO: 64, a p-chain CDR2 comprising or consisting of the amino acid sequence of SEQ ID NO: 65, and a p-chain CDR3 comprising or consisting of the amino acid sequence of SEQ ID NO: 66, or

(l) an a-chain CDR1 comprising or consisting of the amino acid sequence of SEQ ID NO: 67, an a-chain CDR2 comprising or consisting of the amino acid sequence of SEQ ID NO: 68, an a-chain CDR3 comprising or consisting of the amino acid sequence of SEQ ID NO: 69; a p-chain CDR1 comprising or consisting of the amino acid sequence of SEQ ID NO: 70, a p-chain CDR2 comprising or consisting of the amino acid sequence of SEQ ID NO: 71 , and a p-chain CDR3 comprising or consisting of the amino acid sequence of SEQ ID NO: 72, or

(m) an a-chain CDR1 comprising or consisting of the amino acid sequence of SEQ ID NO: 73, an a-chain CDR2 comprising or consisting of the amino acid sequence of SEQ ID NO: 74, an a-chain CDR3 comprising or consisting of the amino acid sequence of SEQ ID NO: 75; a p-chain CDR1 comprising or consisting of the amino acid sequence of SEQ ID NO: 76, a p-chain CDR2 comprising or consisting of the amino acid sequence of SEQ ID NO: 77, and a p-chain CDR3 comprising or consisting of the amino acid sequence of SEQ ID NO: 78. [0014] In accordance with this aspect, there is provided a T cell receptor comprising: (a) an a-chain variable domain having at least 90% sequence identity to an amino acid sequence as set forth in SEQ ID No. 81 ; and a p-chain variable domain having at least 90% sequence identity to an amino acid sequence as set forth in SEQ ID No. 82; or (b) (a) an a-chain variable domain having at least 90% sequence identity to an amino acid sequence as set forth in SEQ ID No. 83; and a p-chain variable domain having at least 90% sequence identity to an amino acid sequence as set forth in SEQ ID No. 84; or

(c) an a-chain variable domain having at least 90% sequence identity to an amino acid sequence as set forth in SEQ ID No. 85; and a p-chain variable domain having at least 90% sequence identity to an amino acid sequence as set forth in SEQ ID No. 86; or

(d) an a-chain variable domain having at least 90% sequence identity to an amino acid sequence as set forth in SEQ ID No. 87; and a p-chain variable domain having at least 90% sequence identity to an amino acid sequence as set forth in SEQ ID No. 88; or

(e) an a-chain variable domain having at least 90% sequence identity to an amino acid sequence as set forth in SEQ ID No. 89; and a p-chain variable domain having at least 90% sequence identity to an amino acid sequence as set forth in SEQ ID No. 90; or

(f) an a-chain variable domain having at least 90% sequence identity to an amino acid sequence as set forth in SEQ ID No. 91 ; and a p-chain variable domain having at least 90% sequence identity to an amino acid sequence as set forth in SEQ ID No. 92; or

(g) an a-chain variable domain having at least 90% sequence identity to an amino acid sequence as set forth in SEQ ID No. 93; and a p-chain variable domain having at least 90% sequence identity to an amino acid sequence as set forth in SEQ ID No. 94; or

(h) an a-chain variable domain having at least 90% sequence identity to an amino acid sequence as set forth in SEQ ID No. 95; and a p-chain variable domain having at least 90% sequence identity to an amino acid sequence as set forth in SEQ ID No. 96; or (i) an a-chain variable domain having at least 90% sequence identity to an amino acid sequence as set forth in SEQ ID No. 97; and a p-chain variable domain having at least 90% sequence identity to an amino acid sequence as set forth in SEQ ID No. 98; or

(j) an a-chain variable domain having at least 90% sequence identity to an amino acid sequence as set forth in SEQ ID No. 99; and a p-chain variable domain having at least 90% sequence identity to an amino acid sequence as set forth in SEQ ID No. 100; or

(k) an a-chain variable domain having at least 90% sequence identity to an amino acid sequence as set forth in SEQ ID No. 101 ; and a p-chain variable domain having at least 90% sequence identity to an amino acid sequence as set forth in SEQ ID No. 102; or

(l) an a-chain variable domain having at least 90% sequence identity to an amino acid sequence as set forth in SEQ ID No. 103; and a p-chain variable domain having at least 90% sequence identity to an amino acid sequence as set forth in SEQ ID No. 104; or

(m) an a-chain variable domain having at least 90% sequence identity to an amino acid sequence as set forth in SEQ ID No. 105; and a p-chain variable domain having at least 90% sequence identity to an amino acid sequence as set forth in SEQ ID No. 106.

[0015] The antigen recognizing constructs and TCRs provided herein are capable of recognizing and specifically binding PRAME, for example PRAME in its major histocompatibility complex (MHC) bound form. In particular, the antigen recognizing constructs and TCRs as herewith provided showed high-affinity PRAME binding and improved reactivity in comparison with a clinical-stage PRAME TCR and other publicly disclosed PRAME TCRs.

[0016] In a fifth aspect, the invention provides a nucleic acid sequence encoding the antigen recognizing construct or the T cell receptor.

[0017] In a sixth aspect, the invention provides a vector comprising the nucleic acid sequence. [0018] In a seventh aspect, the invention provides a host cell comprising the antigen recognizing construct or the T cell receptor, the nucleic acid sequence, or the vector of the invention.

[0019] In an eighth aspect, the invention provides the antigen recognizing construct or the T cell receptor, the nucleic acid sequence, the vector, or the host cell, of the invention for use in medicine or for use in detection, diagnosis, prognosis, prevention and/or treatment of a disease.

[0020] In a ninth aspect, the invention provides a use of the antigen recognizing construct or the T cell receptor, the nucleic acid sequence, the vector, or the host cell, for the manufacture of a medicament for treating a disease.

[0021] In a tenth aspect, the invention provides a method of treating a disease, comprising the step of administering a therapeutically effective amount of the antigen recognizing construct or the T cell receptor, or the nucleic acid sequence, or the vector, or the host cell of the invention.

[0022] In an eleventh aspect, the present invention provides a pharmaceutical composition comprising the antigen recognizing construct or the T cell receptor, the nucleic acid sequence, the vector, or the host cell of the invention.

[0023] In a twelfth aspect, the present invention provides a kit for use in medicine comprising the antigen recognizing construct or the T cell receptor, or the nucleic acid sequence, or the vector, or the host cell of the invention.

BRIEF DESCRIPTION OF THE DRAWINGS

[0024] The invention will be better understood with reference to the detailed description when considered in conjunction with the non-limiting examples and the drawings, in which:

[0025] Fig. 1 shows a graphical representation of the results of an assay for determining functional avidity of human T cells transduced with TCR1 - TCR13 according to the present invention or human T cells transduced with a previously known, published TCR directed against PRAME. IFNy secretion in the culture supernatants upon T cell activation was measured by ELISA.

[0026] Fig. 2 shows a graphical representation of results of a cell line recognition assay of human T cells transduced with TCR1 - TCR13 according to the present invention, human T cells transduced with previously known, published TCRs directed against PRAME, or mock transduced cells, respectively. IFNy secretion in the culture supernatants upon T cell activation was measured by ELISA. The mean +/- SEM of three donors is shown.

[0027] Fig. 3 shows a graphical representation of results of an assay for determining functional expression of T cell receptors TCR6 and TCR12, respectively, according to the present invention.

[0028] Fig. 4 shows a graphical representation of results of a cell line recognition assay of human T cells transduced with TCR 6 or TCR 12, respectively, according to the present invention, human T cells transduced with previously known, published TCRs directed against PRAME, or mock transduced cells, respectively. IFNy secretion in the culture supernatants upon T cell activation was measured by ELISA. The mean +/- SEM of three donors is shown.

DETAILED DESCRIPTION OF THE INVENTION

[0029] As explained above, in a first aspect, the invention is directed to an antigen recognizing construct capable of binding to a Preferentially Expressed Antigen in Melanoma (PRAME) peptide, wherein the antigen recognizing construct comprises a complementary determining region 3 (CDR3) of the a-chain having at least 90% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 3, 9, 15, 21 , 27, 33, 39, 45, 51 , 57, 63, 69, and 75, and/or wherein the antigen recognizing construct comprises a complementary determining region 3 (CDR3) of the p-chain having at least 90% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 6, 12, 18, 24, 36, 42, 48, 54, 60, 66, 72 and 78.

[0030] There is further provided an antigen recognizing construct, wherein the antigen recognizing construct comprises a complementary determining region 3 (CDR3) of the a-chain having at least 90% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 3, 9, 15, 21 , 27, 33, 39, 45, 51 , 57, 63, 69, and 75, and/or wherein the antigen recognizing construct comprises a complementary determining region 3 (CDR3) of the p-chain having at least 90% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 6, 12, 18, 24, 30, 36, 42, 48, 54, 60, 66, 72 and 78.

[0031] It is herewith envisaged that the antigen recognizing construct may comprise a complementary determining region 3 (CDR3) of the a-chain which may have one or more conservative amino acid substitutions relative to the amino acid sequence as set forth in the group consisting of SEQ ID NOs: 3, 9, 15, 21 , 27, 33, 39, 45, 51 , 57, 63, 69, and 75.

[0032] It is further contemplated that the antigen recognizing construct may comprise a complementary determining region 3 (CDR3) of the p-chain which may have one or more conservative amino acid substitutions relative to the amino acid sequence as set forth in the group consisting of SEQ ID NOs: 6, 12, 18, 24, 30, 36, 42, 48, 54, 60, 66, 72 and 78. [0033] The antigen recognizing construct according to the first aspect of the invention may be an antibody, or fragment thereof, or a T cell receptor (TCR), or fragment thereof.

[0034] Thus, it is within the scope of the first aspect of the invention that the antigen recognizing construct is an antibody, or a fragment thereof. The term “antibody” in its various grammatical forms is used herein to refer to immunoglobulin molecules and immunologically active portions of immunoglobulin molecules, i.e. , molecules that contain an antibody combining site or a paratope. Such molecules are also referred to as "antigen binding fragments" of immunoglobulin molecules. The invention further provides an antibody, or antigen binding portion thereof, which specifically binds to the antigens described herein. The antibody can be any type of immunoglobulin that is known in the art. For instance, the antibody can be of any isotype, e.g., IgA, IgD, IgE, IgG, IgM, etc. The antibody can be monoclonal or polyclonal. The antibody can be a naturally-occurring antibody, e.g., an antibody isolated and/or purified from a mammal, e.g., mouse, rabbit, goat, horse, chicken, hamster, human, etc. Alternatively, the antibody can be a genetically- engineered antibody, e.g., a humanized antibody or a chimeric antibody. The antibody can be in monomeric or polymeric form.

[0035] A "T cell" or "T lymphocyte" is an immune system cell that matures in the thymus and produces T cell receptors (TOR). T cells can be naive ("TN"; not exposed to antigen; increased expression of CD62L, CCR7, CD28, CD3, CD127, and CD45RA, and decreased or no expression of CD45RO as compared to TCM), memory T cells (TM) (antigen experienced and long-lived), including stem cell memory T cells, and effector cells (antigen-experienced, cytotoxic). TM can be further divided into subsets of central memory T cells (TEM, expresses CD62L, CCR7, CD28, CD95, CD45RO, and CD127) and effector memory T cells (TEM express CD45RO, decreased expression of CD62L, CCR7, CD28, and CD45RA). Effector T cells (TE) refers to antigen-experienced CD8+ cytotoxic T lymphocytes that express CD45RA, have decreased expression of CD62L, CCR7, and CD28 as compared to TCM, and are positive for granzyme and perforin. CD4+ cells influence the activity of other immune cells by releasing cytokines. CD4+ T cells can activate and suppress an adaptive immune response, and which of those two functions is induced will depend on the presence of other cells and signals. T cells can be collected using known techniques, and the various subpopulations or combinations thereof can be enriched or depleted by known techniques, such as by affinity binding to antibodies, flow cytometry, or immunomagnetic selection. Other exemplary T cells include regulatory T cells, such as CD4+ CD25+ (Foxp3+) regulatory T cells and Treg17 cells, as well as Tr1 , Th3, and Qa-1 restricted T cells.

[0036] "T cell receptor" (TOR) refers to an immunoglobulin superfamily member having a variable binding domain, a constant domain, a transmembrane region, and a short cytoplasmic tail; see, e. g., Janeway el al., Immunobiology: The Immune System in Health and Disease, 3rd Ed., Current Biology Publications, p. 433, 1997) capable of specifically binding to an antigen peptide bound to an MHC receptor. A TCR can be found on the surface of a cell or in soluble form and generally is comprised of a heterodimer having a and p chains (also known as TCR a and TCR p, respectively), or y and 8 chains (also known as TCRy and TCR8, respectively). A polynucleotide encoding a binding protein of this disclosure, e.g., a TCR, can be codon optimized to enhance expression in a particular host cell, such, for example, as a cell of the immune system, a hematopoietic stem cell, a T cell, a primary T cell, a T cell line, a NK cell, or a natural killer T cell (Scholten el al., Clin. Immunol. 119:135, 2006).

[0037] The terms "complementarity determining region" and "CDR," are synonymous with "hypervariable region" or "HVR," and are known in the art to refer to sequences of amino acids within immunoglobulin or TCR variable regions, which confer antigen specificity and/or binding affinity and are separated from one another in primary amino acid sequence by framework regions. In general, there are three CDRs in each TCR a- chain variable region (aCDR1 , aCDR2, aCDR3) and three CDRs in each TCR p-chain variable region (PCDR1 , PCDR2, PCDR3). In TCRs, CDR3 is thought to be the main CDR responsible for recognizing processed antigen. In general, CDR1 and CDR2 interact mainly or exclusively with the MHC.

[0038] The term “capable of binding” means that said PRAME peptide is specifically bound by said TCR. The term “specific(ally) binding” generally indicates that a TCR binds via its antigen binding site more readily to its intended antigenic target than to a random, unrelated non-target antigen. Particularly the term “specifically binds” may indicate that the binding specificity of the TCR will be at least about 5-fold, or at least 10-fold, or at least 25-fold, or at least 50-fold, or at least 100-fold or more, greater for its antigenic target than its binding specificity for a non-target antigen.

[0039] It is understood that the expression “PRAME peptide” relates to a human PRAME peptide/protein having an amino acid sequence according to UniProtKB database entry P78395 ■ PRAME_HUMAN, as set forth e.g. in SEQ ID No.: 79. The antigen recognizing construct according to the first aspect of the invention, which binds the PRAME peptide, may be presented by a MHC I molecule. For example, the MHC I molecule may be an HLA-A molecule. The HLA-A molecule may be, for example, HLA- A2. In embodiments, the HLA-A molecule may be HLA-A*02:01 , HLA-A*02:02, HLA- A*02:03, HLA-A*02:04, HLA-A*02:05, HLA-A*02:06, HLA-A*02:07, HLA-A*68:02, or HLA- A*6901. According to a specific example, the HLA-A molecule may be HLA-A*02:01.

[0040] According to an embodiment the antigen recognizing construct may bind the amino acid sequence SLLQHLIGL (SEQ ID NO: 80) within the PRAME peptide. This epitope (“epitope SLL425-433 ) is known to be highly abundant on PRAM E-expressing tumor cells.

[0041] There are two classes of MHC molecules, MHC class I and MHC class II. Complexes of peptide and MHC class I are recognized by CD8-positive T cells bearing the appropriate T-cell receptor (TCR), whereas complexes of peptide and MHC class II molecules are recognized by CD4-positive T cells bearing the appropriate TCR. Since both types of responses, CD8 and CD4 T cell dependent responses, contribute jointly and synergistically to the anti-tumor effect, the identification and characterization of tumor- associated and tumor-specific antigens and corresponding T cell receptors is important in the development of cancer immunotherapies, such as vaccines and cell therapies.

[0042] A TCR as described herein is envisaged to bind to its antigenic target (for example a PRAME peptide presented on HLA-A*02:01 encoded molecules by antigen presenting cells) with a high functional avidity. The term “functional avidity” refers to the capability of TCR expressing cells (in particular T cells expressing native TCRs as described herein) to respond in vitro to a given concentration of a ligand, and is thought to correlate with the in vivo effector capacity of TCR expressing cells. By definition, TCR expressing cells with high functional avidity respond in in vitro tests to very low antigen doses, while such cells of lower functional avidity require higher amounts of antigen before they mount an immune response similar to that of high-avidity TCR expressing cells. The functional avidity can be therefore considered as a quantitative determinant of the activation threshold of a TCR expressing cell. It is determined by exposing such cells in vitro to different amounts of cognate antigen. TCR expressing cells with high functional avidity respond to low antigen doses. Hence, the TCR of the present invention is a high- avidity TCR causing a half-maximal relative IFN-gamma secretion (EC50 value) of less than 10'⁸⁵ M, as measured by an IFN-gamma immunoassay (Figure 1).

[0043] Thus, in accordance with an embodiment, the antigen recognizing construct may be a TCR, and T-cells expressing said T-cell receptor may have a functional avidity characterized by an EC50 of less than about 1 x 10'^{85 M} The functional avidity may be assessed by evaluating secretion of interferon-gamma by a population of T cells expressing the T cell receptor. In illustrated examples, the functional avidity may be characterized by an EC50 of less than about 1 x 10'⁸⁶ M or of less than about 1 x 10'^{8 7} M or of less than about 1 x 10'⁸⁸ M, or of less than about 1 x 10'⁸⁹ M, or of less than about 1 x 10'⁹ M. The EC50 value may be determined by the procedure as carried out in Example 1 as described herein.

[0044] According to an embodiment, the antigen recognizing construct, e.g. the T cell receptor, may comprise a complementary determining region 1 (CDR1) of the a-chain having at least 90% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 1 , 7, 13, 19, 25, 31 , 37, 43, 49, 55, 61 , 67, and 73, and/or the antigen recognizing construct, e.g. the T cell receptor may comprise a complementary determining region 1 (CDR1) of the p-chain having at least 90% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 4, 10, 16, 22, 28, 34, 40, 46, 52, 58, 64, 70 and 76.

[0045] It is herewith envisaged that the antigen recognizing construct may comprise a complementary determining region 1 (CDR1) of the a-chain which may have one or more conservative amino acid substitutions relative to the amino acid sequence as set forth in the group consisting of SEQ ID NOs: 1 , 7, 13, 19, 25, 31 , 37, 43, 49, 55, 61 , 67, and 73.

[0046] It is further contemplated that the antigen recognizing construct may comprise a complementary determining region 1 (CDR1) of the p-chain which may have one or more conservative amino acid substitutions relative to the amino acid sequence as set forth in the group consisting of SEQ ID NOs: 4, 10, 16, 22, 28, 34, 40, 46, 52, 58, 64, 70 and 76. [0047] According to an embodiment, the antigen recognizing construct, e.g. the T cell receptor, may comprise a complementary determining region 2 (CDR2) of the a-chain having at least 90% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 2, 8, 14, 20, 26, 32, 36, 44, 50, 56, 62, 68, and 74, and/or the T cell receptor may comprise a complementary determining region 2 (CDR2) of the p- chain having at least 90% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 5, 11 , 17, 23, 29, 35, 41 , 47, 53, 59, 65, 71 and 77.

[0048] It is herewith envisaged that the antigen recognizing construct may comprise a complementary determining region 2 (CDR2) of the a-chain which may have one or more conservative amino acid substitutions relative to the amino acid sequence as set forth in the group consisting of SEQ ID Nos 2, 8, 14, 20, 26, 32, 36, 44, 50, 56, 62, 68, and 74.

[0049] It is further contemplates that the antigen recognizing construct may comprise a complementary determining region 2 (CDR2) of the p-chain which may have one or more conservative amino acid substitutions relative to the amino acid sequence as set forth in the group consisting of SEQ ID NOs: 5, 11 , 17, 23, 29, 35, 41 , 47, 53, 59, 65, 71 and 77. [0050] It is also comprised in the first aspect of the invention that the antigen recognizing construct, e.g. the T cell receptor, may comprise a complementary determining region 3 (CDR3) of the a-chain having at least 95%, or at least 96%, or at least 97%, or at least 98%, or at least 99%, or 100% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 3, 9, 15, 21 , 27, 33, 39, 45, 51 , 57, 63, 69, and 75, and/or the antigen recognizing construct, e.g. the T cell receptor, may comprise a complementary determining region 3 (CDR3) of the p-chain having at least 95%, or at least 96%, or at least 97%, or at least 98%, or at least 99%, or 100% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 6, 12, 18, 24, 30, 36, 42, 48, 54, 60, 66, 72 and 78. The antigen recognizing construct, e.g. the T cell receptor, may comprise a complementary determining region 1 (CDR1) of the a-chain having at least 95%, or at least 96%, or at least 97%, or at least 98%, or at least 99%, or 100% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 1 , 7, 13, 19, 25, 31 , 37, 43, 49, 55, 61 , 67, and 73, and/or the antigen recognizing construct, e.g. the T cell receptor, may comprise a complementary determining region 1 (CDR1) of the p-chain having at least 95%, or at least 96%, or at least 97%, or at least 98%, or at least 99%, or 100% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 4, 10, 16, 22, 28, 34, 40, 46, 52, 58, 64, 70 and 76. The antigen recognizing construct, e.g. the T cell receptor, may comprise a complementary determining region 2 (CDR2) of the a-chain having at least 95%, or at least 96%, or at least 97%, or at least 98%, or at least 99%, or 100% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 2, 8, 14, 20, 26, 32, 36, 44, 50, 56, 62, 68, and 74, and/or the antigen recognizing construct, e.g. the T cell receptor, may comprise a complementary determining region 2 (CDR2) of the p-chain having at least 95%, or at least 96%, or at least 97%, or at least 98%, or at least 99%, or 100% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 5, 11 , 17, 23, 28, 35, 41, 47, 53, 59, 65, 71 and 77.

[0051] The antigen recognizing construct, e.g. the T cell receptor, according to the first aspect of the invention may comprise

(m) an a-chain CDR1 comprising or consisting of the amino acid sequence of SEQ ID NO: 73, an a-chain CDR2 comprising or consisting of the amino acid sequence of SEQ ID NO: 74, and an a-chain CDR3 comprising or consisting of the amino acid sequence of SEQ ID NO: 75. [0052] The antigen recognizing construct, e.g, the T cell receptor, according to the first aspect of the invention may comprise

(m) a p-chain CDR1 comprising or consisting of the amino acid sequence of SEQ ID NO: 76, a p-chain CDR2 comprising or consisting of the amino acid sequence of SEQ ID NO: 77, and a p-chain CDR3 comprising or consisting of the amino acid sequence of SEQ ID NO: 78.

[0053] The antigen recognizing construct, e.g. the T cell receptor, according to the first aspect of the invention may comprise

(b) an a-chain CDR1 comprising or consisting of the amino acid sequence of SEQ ID NO: 7, an a-chain CDR2 comprising or consisting of the amino acid sequence of SEQ ID NO: 8, an a-chain CDR3 comprising or consisting of the amino acid sequence of SEQ ID NO: 9; a p-chain CDR1 comprising or consisting of the amino acid sequence of SEQ ID NO: 10, a p-chain CDR2 comprising or consisting of the amino acid sequence of SEQ ID NO: 11 , and a p-chain CDR3 comprising or consisting of the amino acid sequence of SEQ ID NO: 12; or

(m) an a-chain CDR1 comprising or consisting of the amino acid sequence of SEQ ID NO: 73, an a-chain CDR2 comprising or consisting of the amino acid sequence of SEQ ID NO: 74, an a-chain CDR3 comprising or consisting of the amino acid sequence of SEQ ID NO: 75; a p-chain CDR1 comprising or consisting of the amino acid sequence of SEQ ID NO: 76, a p-chain CDR2 comprising or consisting of the amino acid sequence of SEQ ID NO: 77, and a p-chain CDR3 comprising or consisting of the amino acid sequence of SEQ ID NO: 78. [0054] In yet a further aspect, the invention provides a T cell receptor (TCR) having antigenic specificity for Preferentially Expressed Antigen in Melanoma (PRAME), wherein the TCR comprises:

(b) an a-chain CDR1 comprising or consisting of the amino acid sequence of SEQ ID NO:

7, an a-chain CDR2 comprising or consisting of the amino acid sequence of SEQ ID NO: 8, and an a-chain CDR3 comprising or consisting of the amino acid sequence of SEQ ID NO: 9; or

(g) an a-chain CDR1 comprising or consisting of the amino acid sequence of SEQ ID NO:

37, an a-chain CDR2 comprising or consisting of the amino acid sequence of SEQ ID NO: 38, and an a-chain CDR3 comprising or consisting of the amino acid sequence of SEQ ID NO: 39, or

(m) an a-chain CDR1 comprising or consisting of the amino acid sequence of SEQ ID NO: 73, an a-chain CDR2 comprising or consisting of the amino acid sequence of SEQ ID NO: 74, and an a-chain CDR3 comprising or consisting of the amino acid sequence of SEQ ID NO: 75. [0055] In yet a further aspect, the invention provides a T cell receptor (TCR) having antigenic specificity for Preferentially Expressed Antigen in Melanoma (PRAME), wherein the TCR comprises:

(a) a p-chain CDR1 comprising or consisting of the amino acid sequence of SEQ ID NO: 4, a p-chain CDR2 comprising or consisting of the amino acid sequence of SEQ ID NO: 5, and a p-chain CDR3 comprising or consisting of the amino acid sequence of SEQ ID NO: 6; or

(g) a p-chain CDR1 comprising or consisting of the amino acid sequence of SEQ ID NO:

40, a p-chain CDR2 comprising or consisting of the amino acid sequence of SEQ ID NO: 41 , and a p-chain CDR3 comprising or consisting of the amino acid sequence of SEQ ID NO: 42, or

(h) a p-chain CDR1 comprising or consisting of the amino acid sequence of SEQ ID NO:

46, a p-chain CDR2 comprising or consisting of the amino acid sequence of SEQ ID NO: 47, and a p-chain CDR3 comprising or consisting of the amino acid sequence of SEQ ID NO: 48, or

(i) a p-chain CDR1 comprising or consisting of the amino acid sequence of SEQ ID NO:

52, a p-chain CDR2 comprising or consisting of the amino acid sequence of SEQ ID NO: 53, and a p-chain CDR3 comprising or consisting of the amino acid sequence of SEQ ID NO: 54, or

(j) a p-chain CDR1 comprising or consisting of the amino acid sequence of SEQ ID NO:

58, a p-chain CDR2 comprising or consisting of the amino acid sequence of SEQ ID NO: 59, and a p-chain CDR3 comprising or consisting of the amino acid sequence of SEQ ID NO: 60, or

(k) a p-chain CDR1 comprising or consisting of the amino acid sequence of SEQ ID NO:

64, a p-chain CDR2 comprising or consisting of the amino acid sequence of SEQ ID NO: 65, and a p-chain CDR3 comprising or consisting of the amino acid sequence of SEQ ID NO: 66, or

(l) a p-chain CDR1 comprising or consisting of the amino acid sequence of SEQ ID NO:

70, a p-chain CDR2 comprising or consisting of the amino acid sequence of SEQ ID NO: 71 , and a p-chain CDR3 comprising or consisting of the amino acid sequence of SEQ ID NO: 72, (m) a p-chain CDR1 comprising or consisting of the amino acid sequence of SEQ ID NO: 76, a p-chain CDR2 comprising or consisting of the amino acid sequence of SEQ ID NO: 77, and a p-chain CDR3 comprising or consisting of the amino acid sequence of SEQ ID NO: 78. [0056] In a further aspect, the invention provides a T cell receptor (TCR) having antigenic specificity for Preferentially Expressed Antigen in Melanoma (PRAME), wherein the TCR may comprise:

(m) an a-chain CDR1 comprising or consisting of the amino acid sequence of SEQ ID NO: 73, an a-chain CDR2 comprising or consisting of the amino acid sequence of SEQ ID NO: 74, an a-chain CDR3 comprising or consisting of the amino acid sequence of SEQ ID NO: 75; a p-chain CDR1 comprising or consisting of the amino acid sequence of SEQ ID NO: 76, a p-chain CDR2 comprising or consisting of the amino acid sequence of SEQ ID NO: 77, and a p-chain CDR3 comprising or consisting of the amino acid sequence of SEQ ID NO: 78.

[0057] According to an embodiment, the T cell receptor may comprise: an a-chain variable domain having at least 70%, or at least 80%, or at least 90%, or at least 95%, or at least 96%, or at least 97%, or at least 98%, or at least 99%, or 100% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 81 , 83, 85, 87, 89, 91 , 93, 95, 97, 99, 101 , 103, and 105 and/or a p-chain variable domain having at least 70%, or at least 80%, or at least 90%, or at least 95%, or at least 96%, or at least 97%, or at least 98%, or at least 99%, or 100% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 82, 84, 86, 88, 90, 92, 94, 96, 98, 100, 102, 104 and 106.

[0058] According to an embodiment, the T cell receptor may comprise:

(a) an a-chain variable domain having at least 90% sequence identity to an amino acid sequence as set forth in SEQ ID No. 81 ; and a p-chain variable domain having at least 90% sequence identity to an amino acid sequence as set forth in SEQ ID No. 82; or

(b) (a) an a-chain variable domain having at least 90% sequence identity to an amino acid sequence as set forth in SEQ ID No. 83; and a p-chain variable domain having at least 90% sequence identity to an amino acid sequence as set forth in SEQ ID No. 84; or

(h) an a-chain variable domain having at least 90% sequence identity to an amino acid sequence as set forth in SEQ ID No. 95; and a p-chain variable domain having at least 90% sequence identity to an amino acid sequence as set forth in SEQ ID No. 96; or

(i) an a-chain variable domain having at least 90% sequence identity to an amino acid sequence as set forth in SEQ ID No. 97; and a p-chain variable domain having at least 90% sequence identity to an amino acid sequence as set forth in SEQ ID No. 98; or

(j) an a-chain variable domain having at least 90% sequence identity to an amino acid sequence as set forth in SEQ ID No. 99; and a p-chain variable domain having at least 90% sequence identity to an amino acid sequence as set forth in SEQ ID No. 100; or (k) an a-chain variable domain having at least 90% sequence identity to an amino acid sequence as set forth in SEQ ID No. 101 ; and a p-chain variable domain having at least 90% sequence identity to an amino acid sequence as set forth in SEQ ID No. 102; or

[0059] Included in the scope of the invention are functional variants of the inventive antigen recognizing constructs or TCRs described herein. The term “functional variant,” as used herein, refers to an antigen recognizing construct or TCR having substantial or significant sequence identity or similarity to a parent antigen recognizing construct or TCR, which functional variant retains the biological activity of the antigen recognizing construct or TCR of which it is a variant. Functional variants encompass, for example, those variants of the antigen recognizing construct or TCR described herein (the parent antigen recognizing construct or TCR) that retain the ability to specifically bind to a PRAME peptide for which the parent antigen recognizing construct or TCR has antigenic specificity to a similar extent, the same extent, or to a higher extent, as the parent antigen recognizing construct or TCR. In reference to the parent antigen recognizing construct or TCR, the functional variant can, for instance, be at least about 30%, 50%, 75%, 80%, 90%, 95%, 96%, 97%, 98%, 99% or more identical in amino acid sequence to the parent antigen recognizing construct or TCR.

[0060] The functional variant can, for example, comprise the amino acid sequence of the parent antigen recognizing construct or TCR with at least one conservative amino acid substitution. Conservative amino acid substitutions are known in the art, and include amino acid substitutions in which one amino acid having certain physical and/or chemical properties is exchanged for another amino acid that has the same chemical or physical properties. For instance, the conservative amino acid substitution can be an acidic amino acid substituted for another acidic amino acid (e.g., Asp or Glu), an amino acid with a nonpolar side chain substituted for another amino acid with a nonpolar side chain (e.g., Ala, Gly, Vai, He, Leu, Met, Phe, Pro, Trp, Vai, etc.), a basic amino acid substituted for another basic amino acid (Lys, Arg, etc.), an amino acid with a polar side chain substituted for another amino acid with a polar side chain (Asn, Cys, Gin, Ser, Thr, Tyr, etc.), etc.

[0061] The term "sequence identity" or "identity" as used in the present invention means the percentage of pair-wise identical residues, following homology alignment of a sequence of a polypeptide and or nucleic acid of the present invention with a sequence in question, with respect to the number of residues in the longer of these two sequences.

[0062] The percentage of sequence homology or sequence identity can, for example, be determined herein using the program BLASTP, version blastp 2.2.5 (November 16, 2002; cf. Altschul, S. F. et al. (1997) Nucl. Acids Res. 25, 3389-3402). The percentage of homology is based on the alignment of the entire polypeptide sequences (matrix: BLOSLIM 62; gap costs: 11.1 ; cutoff value set to 10-3) including the respective sequences. It is calculated as the percentage of numbers of "positives" (homologous amino acids) indicated as result in the BLASTP program output divided by the total number of amino acids selected by the program for the alignment.

[0063] In accordance with a further aspect, the invention is directed to a nucleic acid sequence encoding the antigen recognizing construct or the T cell receptor according to the the invention.

[0064] A used herein, “nucleic acid” refers to single or double-stranded molecules, which may be DNA or RNA, including genomic DNA, mRNA, cRNA, miRNA, tRNA, although without limitation thereto. A nucleic acid comprises a nucleotide sequence which typically includes nucleotides that comprise an A, G, C, T or U base. However, nucleotide sequences may include other bases such as inosine, methylcytosine, hydroxymethycytosine, methylinosine, methyladenosine and/or thioitridine, although without limitation thereto.

[0065] In a further aspect, the inventon is directed to a vector comprising the nucleic acid sequence according to the invention.

[0066] According to an embodiment, the vector may further comprise a nucleic acid sequence encoding a CD8 Co-receptor.

[0067] For example, the CD8 Co-receptor may be a wildtype CD8 Co-receptor, such as a wildtype CD8 a Co-receptor, or a chimeric CD8 Co-receptor.

[0068] The invention will be further explained in the following making reference to either, several or all of these aspects. If reference is only made to one of these aspects, it is understood by the person skilled in the art, that this reference nevertheless includes references to all other aspects of the invention, if applicable.

[0069] In a further aspect, the invention is directed to a host cell comprising the antigen recognizing construct or T cell receptor according to the respective aspect of the invention, or the nucleic acid sequence according to the respective aspect of the invention, or the vector according to the respective aspect of the invention.

[0070] In accordance with a further aspect, the invention is directed to the antigen recognizing construct or T cell receptor according to the respective aspects of the invention, or the nucleic acid sequence according to the respective aspect of the invention, or the vector according to the respective aspect of the invention, or the host cell according to the respective aspect of the invention, for use in medicine.

[0071] In a further aspect, the invention is directed to the antigen recognizing construct or T cell receptor according to the respective aspects of the invention, or the nucleic acid sequence according to the respective aspect of the invention, or the vector according to the respective aspect of the invention, or the host cell according to the respective aspect of the invention, for use in detection, diagnosis, prognosis, prevention and/or treatment of a disease.

[0072] The terms “treat” and “prevent” as well as words stemming therefrom, like “treatment” or “prevention”, as used herein, do not necessarily imply 100% or complete treatment or prevention. Rather, there are varying degrees of treatment or prevention of which one of ordinary skill in the art recognizes as having a potential benefit or therapeutic effect. In this respect, the inventive methods can provide any amount of any level of treatment or prevention of cancer in a mammal. Furthermore, the treatment or prevention provided by the inventive method can include treatment or prevention of one or more conditions or symptoms of the cancer being treated or prevented. For example, treatment or prevention can include promoting the regression of a tumor. Also, for purposes herein, “prevention” can encompass delaying the onset of the cancer, or a symptom or condition thereof.

[0073] According to an embodiment, the disease may be cancer.

[0074] For example, the disease may be a tumor that expresses PRAME.

Specifically, the disease may be an advanced-stage tumor that expresses PRAME.

[0075] According to an embodiment, the disease may be selected from the group consisting of melanoma, bladder carcinoma, colon carcinoma, and breast adenocarcinoma, sarcoma, prostate cancer, uterine cancer, uveal cancer, uveal melanoma, squamous head and neck cancer, synovial carcinoma, Ewing’s sarcoma, breast cancer such as triple negative breast cancer, thyroid cancer, testicular cancer, renal cancer, pancreatic cancer, ovarian cancer, esophageal cancer, non-small-cell lung cancer (NSCLC), non-Hodgkin’s lymphoma, multiple myeloma, melanoma, hepatocellular carcinoma, head and neck cancer, gastric cancer, endometrial cancer, colorectal cancer, cholangiocarcinoma, breast cancer, bladder cancer, myeloid leukemia, acute lymphoblastic leukemia, small-cell lung cancer (SCLC), sarcoma and osteosarcoma. [0076] According to an embodiment, the breast cancer may be selected from the group consisting of ductal breast cancer, tubular breast cancer, medullary breast cancer and combinations thereof.

[0077] According to an embodiment, the sarcoma cancer may be selected from the group consisting of chondrosarcoma cancer, osteosarcoma cancer and combinations thereof.

[0078] In an embodiment, the gastric cancer may be gastric adenocarcinoma cancer.

[0079] A further aspect of the invention is directed to a use of the antigen recognizing construct or the T cell receptor according to the respective aspects of the invention, or the nucleic acid sequence according to the respective aspect of the invention, or the vector according to the respective aspect of the invention, or the host cell according to the respective aspect of the invention, for the manufacture of a medicament for treating a disease. For example, the disease may be a cancer. The disease may be a malignant or benign tumor disease, but is not limited thereto. In yet other illustrative example, the disease may be a tumor that expresses PRAME. Without being limited to it, the disease may be an advanced-stage tumor that expresses PRAME.

[0080] In accordance with some embodiments, the disease may be selected from the group consisting of melanoma, bladder carcinoma, colon carcinoma, and breast adenocarcinoma, sarcoma, prostate cancer, uterine cancer, uveal cancer, uveal melanoma, squamous head and neck cancer, synovial carcinoma, Ewing’s sarcoma, breast cancer such as triple negative breast cancer, thyroid cancer, testicular cancer, renal cancer, pancreatic cancer, ovarian cancer, esophageal cancer, non-small-cell lung cancer (NSCLC), non-Hodgkin’s lymphoma, multiple myeloma, melanoma, hepatocellular carcinoma, head and neck cancer, gastric cancer, endometrial cancer, colorectal cancer, cholangiocarcinoma, breast cancer, bladder cancer, myeloid leukemia, acute lymphoblastic leukemia, small-cell lung cancer (SCLC), sarcoma and osteosarcoma.

[0081] According to an embodiment, the breast cancer may be selected from the group consisting of ductal breast cancer, tubular breast cancer, medullary breast cancer and combinations thereof.

[0082] According to an embodiment, the sarcoma cancer may be selected from the group consisting of chondrosarcoma cancer, osteosarcoma cancer and combinations thereof.

[0083] In an embodiment, the gastric cancer may be gastric adenocarcinoma cancer.

[0084] In a further aspect, the invention is directed to a method of treating a disease, comprising the step of administering a therapeutically effective amount of the antigen recognizing construct or the T cell receptor according to the respective aspects of the invention, or the nucleic acid sequence according to the respective aspect of the invention, or the vector according to the respective aspect of the invention, or the host cell according to the respective aspect of the invention.

[0085] For example, the disease may be cancer. According to an embodiment, the disease may be a malignant or benign tumor disease, but is not limited thereto. In yet other illustrative example, the disease may be a tumor that expresses PRAME. Without being limited to it, the disease is an advanced-stage tumor that expresses PRAME.

In accordance with an embodiment, the disease may be selected from the group consisting of melanoma, bladder carcinoma, colon carcinoma, and breast adenocarcinoma, sarcoma, prostate cancer, uterine cancer, uveal cancer, uveal melanoma, squamous head and neck cancer, synovial carcinoma, Ewing’s sarcoma, breast cancer such as triple negative breast cancer, thyroid cancer, testicular cancer, renal cancer, pancreatic cancer, ovarian cancer, esophageal cancer, non-small-cell lung cancer (NSCLC), non-Hodgkin’s lymphoma, multiple myeloma, melanoma, hepatocellular carcinoma, head and neck cancer, gastric cancer, endometrial cancer, colorectal cancer, cholangiocarcinoma, breast cancer, bladder cancer, myeloid leukemia, acute lymphoblastic leukemia, small-cell lung cancer (SCLC), sarcoma and osteosarcoma.

[0086] According to an embodiment, the breast cancer may be selected from the group consisting of ductal breast cancer, tubular breast cancer, medullary breast cancer and combinations thereof.

[0087] According to an embodiment, the sarcoma cancer may be selected from the group consisting of chondrosarcoma cancer, osteosarcoma cancer and combinations thereof.

[0088] In an embodiment, the gastric cancer may be gastric adenocarcinoma cancer.

[0089] According to an embodiment, the method may further comprise coadministering a CD8 Co-receptor. For example, the CD8 Co-receptor may be a wildtype CD8 Co-receptor such as a wildtype CD8 ap Co-receptor, or a chimeric CD8 Co-receptor. [0090] The present invention also provides a pharmaceutical composition comprising the antigen recognizing construct or the T cell receptor according to the respective aspects of the invention, or the nucleic acid sequence according to the respective aspect of the invention, or the vector according to the respective aspect of the invention, or the host cell according to the respective aspect of the invention.

[0091] Dealing now with the kit of the invention, such a kit is a kit for use in medicine comprising the antigen recognizing construct or the T cell receptor according to the respective aspects of the invention, or the nucleic acid sequence according to the respective aspect of the invention, or the vector according to the respective aspect of the invention, or the host cell according to the respective aspect of the invention. According to an embodiment, the kit may be a diagnostic kit for selecting a patient for treatment of a tumor, wherein cells of the tumor express PRAME.

[0092] The invention will be further illustrated by the following non-limiting Experimental Examples.

[0093] Sequences, as used herein, are depicted in below Table 1.

[0094] Table 1. Sequences as used herein.

Experimental Examples

[0095] Example 1 :

[0096] Peripheral blood leukocytes (PBLs) from a human donor (mix of both CD4 and CD8 T cells) were transduced with either TCR1-TCR13, respectively, according to the present invention, each in a respective construct together with wildtype CD8 ap Coreceptor, respectively, or with a reference TCR (“Ref. TCR A” relates to “TCR R11 P3D3_KE” being a publicly known PRAME-specific T-Cell Receptor from Immatics Biotechnologies GmbH as published in the international Patent application Publication No. WO 2018/172533 A2) in a construct together with wildtype CD8 ap Co-receptor. The transduced human T cells were co-cultured, respectively, with T2 cells loaded with different peptide concentrations (10^-7 M to 10'¹⁴ M) (PRAME peptide “SLLQHLIGL”). IFNy secretion in the culture supernatants upon T cell activation was measured by a sandwich ELISA. The results of this Example are shown in Fig. 1 included herein. Specifically, normalized IFNg secretion curves were used to calculate EC50 values. The mean +/- SEM of three donors is shown in Fig. 1.

[0097] The results are given herein in Fig. 1 . As visible, TCR1 - TCR13 according to the present invention as herein provided show increased functional avidity in comparison with the reference TCR (Ref. TCR A). TCR 1 - TCR 13 as herein provided thus represent valuable high-affinity PRAME specific TCRs for e.g. providing improved deep and durable responses in hard-to-treat indications.

[0098] Example 2:

[0099] Peripheral blood leukocytes (PBLs) from a human donor (mix of both CD4 and CD8 T cells) were transduced with either TCR1-TCR13 according to the present invention, each in a respective construct together with wildtype CD8 ap Co-receptor, respectively, or with reference TCRs (“Ref. TCR B” relates to “TCR 027-004” being a publicly known PRAME-specific T-Cell Receptor from Medigene Immunotherapies GmbH as published in the international Patent application Publication No. WO 2021/099360 A1 , and “Ref. TCR C” relates to “TCR PN42498”, being a publicly known PRAME-specific T- Cell Receptor from Regeneron Pharmaceuticals, Inc. as published in the international Patent application Publication No. WO 2021/15084 A1 ), respectively, in a construct together with wildtype CD8 Co-receptor. Mock-transduced cells served as control. The transduced human T cells were co-cultured, respectively, with PRAME and HLA- A*02:01 expressing cell lines NCI-H1703 (with natural HLA-A*02:01 expression), or NCI- H1435 (with transgenic HLA-A*02:01 expression), respectively. IFNy secretion in the culture supernatants upon T cell activation was measured by sandwich ELISA. The mean +/- SEM of three donors is shown in Fig. 2 as included herein.

[00100] The results are given herein in Fig. 2. As visible, human T-cells Cotransduced with CD8 Coreceptor and TCR1 - TCR13 according to the present invention as herein provided show increased IFNy secretion upon T cell activation in comparison with the reference TCRs (Ref. TCR B and Ref. TCR C, respectively). This effect is present in two different PRAME expressing cell lines.

[00101 ] Example 3:

[00102] _Human T cells (mix of both CD4 and CD8 T cells) from a human donor were transduced with either TCR 6 or TCR12 according to the present invention in a construct together with wildtype CD8 p Co-receptor, respectively, or with reference TCR A, (“Ref. TCR A” relates to “TCR R11 P3D3_KE”, being the publicly known PRAME-specific T-Cell Receptor from Immatics Biotechnologies GmbH as published in the international Patent application Publication No. WO 2018/172533 A2), or with reference TCR B (“Ref. TCR B” relates to “TCR 027-004”, being the publicly known PRAME-specific T-Cell Receptor from Medigene Immunotherapies GmbH as published in the international Patent application Publication No. WO 2021/099360 A1), or with reference TCR E (“Ref. TCR E” relates to “TCR 366”, being the publicly known PRAME-specific T-Cell Receptor from T-Scan Therapeutics, Inc., as published in the international Patent application Publication No. WO 2024/077134 A1), respectively, in respective constructs together with wildtype CD8 ap Coreceptor. MHC-Multimer binding was measured via flow cytometry. The mean +/- SEM of three donors is shown in Fig. 3 as included herein.

[00103] The results are depicted in Fig. 3, wherein the measured median fluorescence intensity (MFI) is shown. Both TCR 6 and TCR 12 showed increased MHC-Multimer binding in comparison with the reference TCRs.

[00104] Example 4:

[00105] Human T cells (mix of both CD4 and CD8 T cells) from a human donor were transduced with either TCR 6 or TCR12 according to the present invention in a construct together with wildtype CD8 a Co-receptor, respectively, or with reference TCR A, (“Ref. TCR A” relates to “TCR R11 P3D3_KE”, being the publicly known PRAME-specific T-Cell Receptor from Immatics Biotechnologies GmbH as published in the international Patent application Publication No. WO 2018/172533 A2), or with reference TCR B (“Ref. TCR B” relates to “TCR 027-004”, being the publicly known PRAME-specific T-Cell Receptor from Medigene Immunotherapies GmbH as published in the international Patent application Publication No. WO 2021/099360 A1), or with with reference TCR E (“Ref. TCR E” relates to “TCR 366”, being the publicly known PRAME-specific T-Cell Receptor from T-Scan Therapeutics, Inc., as published in the international Patent application Publication No. WO 2024/077134 A1), respectively, in respective constructs together with wildtype CD8 ap Coreceptor. Mock transduced cells served as control. The transduced T cells were cocultured with PRAME and HLA-A*02:01 expressing cell lines LI266, EJM, NCI-H1703, NCI- H1650, NCI-H1435 and Hs695T, respectively (EJM and NCI-H1435 have transgenic HLA- A*02:01 expression, all other cell lines have natural HLA-A*02:01 expression). IFNy secretion in the culture supernatants upon T cell activation was measured by ELISA. The mean +/- SEM of three donors is shown in Fig. 4.

[00106] The results are illustrated in Fig. 4. As visible, human T-cells co-transduced with wildtype CD8 a Co-receptor and TCR 6 or TCR 12, respectively, according to the present invention as herein provided show increased IFNy secretion upon T cell activation in comparison with the reference TCRs (Ref. TCR A, Ref. TCR B and Ref TCR E, respectively) co-transduced with wildtype CD8 ap Co-receptor. This effect is demonstrated in all six tested PRAME expressing cell lines.

[00107] A further publicly available reference TCR (“TCR clone 54 SLL”, the publicly known PRAME-specific T-Cell Receptor from Leiden University Medical Center as published in the international Patent application Publication No. 2016/142783 A2) could not be expressed as construct together with the wildtype CD8 p Co-receptor and was therefore not included in the Examples.

Materials and Methods

[00108] Animal immunizations

To generate CD8+ T cell responses to PRAME epitopes, humanized TCR/HLA class I transgenic mice according to the ABabDII mouse strain described in Li etal. (2010, Nature Medicine 16, 1029-1034), were used for immunizations with either full length human PRAME in adenoviral vector i.p., or with a PRAME peptide “SLLQHLIGL” plus adjuvant s.c. For peptide immunizations, the mice were subcutaneously primed and boosted monthly with the PRAME peptide “SLLQHLIGL” emulsified in adjuvant. For adenoviral immunizations, the mice received intraperitoneal injections of an adenoviral vector that expresses the full length human PRAME protein under control of the CMV promoter.

[00109] TCR identification and characterization

Expanded TCR clonotypes were screened in a reporter-based screening assay. Most functional TCRs were selected and cloned into a retroviral vector and transduced into human peripheral blood lymphocytes for detailed functional characterization. [00110] It will be readily apparent to a person skilled in the art that varying substitutions and modifications may be made to the invention disclosed herein without departing from the scope and spirit of the invention.

[00111] All patents and publications mentioned in the specification are indicative of the levels of those of ordinary skill in the art to which the invention pertains. All patents and publications are herein incorporated by reference to the same extent as if each individual publication was specifically and individually indicated to be incorporated by reference.

[00112] As used herein, the term "about" means ± 20% of the indicated range, value, or structure, unless otherwise indicated. It should be understood that the terms "a" and "an", as used herein, refer to "one or more" of the enumerated components. The use of the alternative (e.g., "or") should be understood to mean either one, both, or any combination thereof of the alternatives. As used herein, the terms "include", "have", and "comprise" are used synonymously, which terms and variants thereof are intended to be construed as non-limiting.

[00113] The inventions illustratively described herein may suitably be practiced in the absence of any element or elements, limitation or limitations, not specifically disclosed herein. Thus, for example, the terms "comprising", "including", "containing", etc. shall be read expansively and without limitation. Additionally, the terms and expressions employed herein have been used as terms of description and not of limitation, and there is no intention in the use of such terms and expressions of excluding any equivalents of the features shown and described or portions thereof, but it is recognized that various modifications are possible within the scope of the invention claimed. Thus, it should be understood that although the present invention has been specifically disclosed by preferred embodiments and optional features, modification and variation of the inventions embodied therein and herein disclosed may be resorted to by those skilled in the art, and that such modifications and variations are considered to be within the scope of this invention. The invention has been described broadly and generically herein. Each of the narrower species and subgeneric groupings falling within the generic disclosure also form part of the invention. This includes the generic description of the invention with a proviso or negative limitation removing any subject matter from the genus, regardless of whether or not the excised material is specifically recited herein. In addition, where features or aspects of the invention are described in terms of Markush groups, those skilled in the art will recognize that the invention is also thereby described in terms of any individual member or subgroup of members of the Markush group. Further embodiments of the invention will become apparent from the following claims.

Claims

Claims:

What is claimed is:

1. An antigen recognizing construct capable of binding to a Preferentially Expressed Antigen in Melanoma (PRAME) peptide, wherein the antigen recognizing construct comprises a complementary determining region 3 (CDR3) of the a-chain having at least 90% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 3, 9, 15, 21 , 27, 33, 39, 45, 51 , 57, 63, 69, and 75, and/or wherein the antigen recognizing construct comprises a complementary determining region 3 (CDR3) of the p-chain having at least 90% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 6, 12, 18, 24, 30, 36, 42, 48, 54, 60, 66, 72 and 78.

2. The antigen recognizing construct of claim 1 , wherein the antigen recognizing construct is an antibody, or fragment thereof, or a T cell receptor (TCR), or fragment thereof.

3. The antigen recognizing construct of any one of the preceding claims, wherein the antigen recognizing construct is a T cell receptor (TCR).

4. The antigen recognizing construct of claim 3, wherein the antigen recognizing construct is a T cell receptor, and wherein T-cells expressing said T-cell receptor are having a functional avidity characterized by an ECso of less than about 1 x 10'⁸⁵ M.

5. The antigen recognizing construct of claim 4, wherein the functional avidity is assessed by evaluating secretion of interferon-gamma by a population of T cells expressing the T cell receptor; such as determining the ECso value that is indicative of the functional avidity of the T-cell receptor by the procedure as carried out in Example 1 .

6. The antigen recognizing construct of any one of the preceding claims, wherein the antigen recognizing construct binds the peptide being presented by a MHC I molecule.

7. The antigen recognizing construct of claim 6, wherein the MHC I molecule is an HLA-A molecule.

8. The antigen recognizing construct of claim 7, wherein the HLA-A molecule is HLA- A*02:01.

9. The antigen recognizing construct of any one of the preceding claims, wherein the antigen recognizing construct binds the amino acid sequence SLLQHLIGL (SEQ ID NO: 80) within the PRAME peptide.

10. The antigen recognizing construct of any one of claims 3 to 5, wherein the T cell receptor comprises a complementary determining region 1 (CDR1) of the a-chain having at least 90% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 1 , 7, 13, 19, 25, 31 , 37, 43, 49, 55, 61 , 67, and 73, and/or wherein the T cell receptor comprises a complementary determining region 1 (CDR1) of the p-chain having at least 90% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 4, 10, 16, 22, 28, 34, 40, 46, 52, 58, 64, 70 and

76.

11 . The antigen recognizing construct of any one of claims 3 to 5 or 10, wherein the T cell receptor comprises a complementary determining region 2 (CDR2) of the a-chain having at least 90% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 2, 8, 14, 20, 26, 32, 36, 44, 50, 56, 62, 68, and 74, and/or wherein the T cell receptor comprises a complementary determining region 2 (CDR2) of the p-chain having at least 90% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 5, 11 , 17, 23, 29, 35, 41 , 47, 53, 59, 65, 71 and

77.

12. The antigen recognizing construct of any one of claims 3 to 5 or 10 to 11 , wherein the T cell receptor comprises a complementary determining region 3 (CDR3) of the a-chain having at least 95% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 3, 9, 15, 21 , 27, 33, 39, 45, 51 , 57, 63, 69, and 75, and/or wherein the T cell receptor comprises a complementary determining region 3 (CDR3) of the p-chain having at least 95% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 6, 12, 18, 24, 30, 36, 42, 48, 54, 60, 66, 72 and

78.

13. The antigen recognizing construct of any one of claims 3 to 5 or 10 to 12, wherein the T cell receptor comprises a complementary determining region 1 (CDR1) of the a-chain having at least 95% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 1 , 7, 13, 19, 25, 31 , 37, 43, 49, 55, 61 , 67, and 73, and/or wherein the T cell receptor comprises a complementary determining region 1 (CDR1) of the p-chain having at least 95% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 4, 10, 16, 22, 28, 34, 40, 46, 52, 58, 64, 70 and

76.

14. The antigen recognizing construct of any one of claims 3 to 5 or 10 to 13, wherein the T cell receptor comprises a complementary determining region 2 (CDR2) of the a-chain having at least 95% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 2, 8, 14, 20, 26, 32, 36, 44, 50, 56, 62, 68, and 74, and/or wherein the T cell receptor comprises a complementary determining region 2 (CDR2) of the p-chain having at least 95% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 5, 11 , 17, 23, 29, 35, 41 , 47, 53, 59, 65, 71 and

77.

15. The antigen recognizing construct of any one of claims 3 to 5 or 10 to 14, comprising:

(f) an a-chain CDR1 comprising or consisting of the amino acid sequence of SEQ ID NO: 31 , an a-chain CDR2 comprising or consisting of the amino acid sequence of SEQ ID NO: 32, and an a-chain CDR3 comprising or consisting of the amino acid sequence of SEQ ID NO: 33, or

(m) an a-chain CDR1 comprising or consisting of the amino acid sequence of SEQ ID NO: 73, an a-chain CDR2 comprising or consisting of the amino acid sequence of SEQ ID NO: 74, and an a-chain CDR3 comprising or consisting of the amino acid sequence of SEQ ID NO: 75.

16. The antigen recognizing construct of any one of claims 3 to 5 or 10 to 15, comprising:

(i) a p-chain CDR1 comprising or consisting of the amino acid sequence of SEQ ID NO: a p-chain CDR2 comprising or consisting of the amino acid sequence of SEQ ID NO: 53, and a p-chain CDR3 comprising or consisting of the amino acid sequence of SEQ ID NO: 54, or

17. The antigen recognizing construct of any one of claims 3 to 5 or 10 to 16, comprising: (a) an a-chain CDR1 comprising or consisting of the amino acid sequence of SEQ ID NO: 1 , an a-chain CDR2 comprising or consisting of the amino acid sequence of SEQ ID NO: 2, an a-chain CDR3 comprising or consisting of the amino acid sequence of SEQ ID NO: 3; a p-chain CDR1 comprising or consisting of the amino acid sequence of SEQ ID NO: 4, a p-chain CDR2 comprising or consisting of the amino acid sequence of SEQ ID NO: 5, and a p-chain CDR3 comprising or consisting of the amino acid sequence of SEQ ID NO: 6; or (b) an a-chain CDR1 comprising or consisting of the amino acid sequence of SEQ ID NO: 7, an a-chain CDR2 comprising or consisting of the amino acid sequence of SEQ ID NO: 8, an a-chain CDR3 comprising or consisting of the amino acid sequence of SEQ ID NO: 9; a p-chain CDR1 comprising or consisting of the amino acid sequence of SEQ ID NO: 10, a p-chain CDR2 comprising or consisting of the amino acid sequence of SEQ ID NO: 11 , and a p-chain CDR3 comprising or consisting of the amino acid sequence of SEQ ID NO: 12; or

(e) an a-chain CDR1 comprising or consisting of the amino acid sequence of SEQ ID NO: 25, an a-chain CDR2 comprising or consisting of the amino acid sequence of SEQ ID NO: 26, an a-chain CDR3 comprising or consisting of the amino acid sequence of SEQ ID NO: 27; a p-chain CDR1 comprising or consisting of the amino acid sequence of SEQ ID NO: 28, a p-chain CDR2 comprising or consisting of the amino acid sequence of SEQ ID NO: 39, and a p-chain CDR3 comprising or consisting of the amino acid sequence of SEQ ID NO: 30, or (f) an a-chain CDR1 comprising or consisting of the amino acid sequence of SEQ ID NO: 31 , an a-chain CDR2 comprising or consisting of the amino acid sequence of SEQ ID NO: 32, an a-chain CDR3 comprising or consisting of the amino acid sequence of SEQ ID NO: 33; a p-chain CDR1 comprising or consisting of the amino acid sequence of SEQ ID NO: 34, a p-chain CDR2 comprising or consisting of the amino acid sequence of SEQ ID NO: 35, and a p-chain CDR3 comprising or consisting of the amino acid sequence of SEQ ID NO: 36, or

(i) an a-chain CDR1 comprising or consisting of the amino acid sequence of SEQ ID NO: 49, an a-chain CDR2 comprising or consisting of the amino acid sequence of SEQ ID NO: 50, an a-chain CDR3 comprising or consisting of the amino acid sequence of SEQ ID NO: 51 ; a p-chain CDR1 comprising or consisting of the amino acid sequence of SEQ ID NO: 52, a p-chain CDR2 comprising or consisting of the amino acid sequence of SEQ ID NO: 53, and a p-chain CDR3 comprising or consisting of the amino acid sequence of SEQ ID NO: 54, or (j) an a-chain CDR1 comprising or consisting of the amino acid sequence of SEQ ID NO: 55, an a-chain CDR2 comprising or consisting of the amino acid sequence of SEQ ID NO: 56, an a-chain CDR3 comprising or consisting of the amino acid sequence of SEQ ID NO: 57; a p-chain CDR1 comprising or consisting of the amino acid sequence of SEQ ID NO: 58, a p-chain CDR2 comprising or consisting of the amino acid sequence of SEQ ID NO: 59, and a p-chain CDR3 comprising or consisting of the amino acid sequence of SEQ ID NO: 60, or

18. A T cell receptor (TCR) having antigenic specificity for Preferentially Expressed Antigen in Melanoma (PRAME), wherein the TCR comprises:

(d) an a-chain CDR1 comprising or consisting of the amino acid sequence of SEQ ID NO:

19, an a-chain CDR2 comprising or consisting of the amino acid sequence of SEQ ID NO: 20, and an a-chain CDR3 comprising or consisting of the amino acid sequence of SEQ ID NO: 21 ; or

(e) an a-chain CDR1 comprising or consisting of the amino acid sequence of SEQ ID NO:

25, an a-chain CDR2 comprising or consisting of the amino acid sequence of SEQ ID NO: 26, and an a-chain CDR3 comprising or consisting of the amino acid sequence of SEQ ID NO: 27, or

19. A T cell receptor (TCR) having antigenic specificity for Preferentially Expressed Antigen in Melanoma (PRAME), wherein the TCR comprises:

20. A T cell receptor (TCR) having antigenic specificity for Preferentially Expressed Antigen in Melanoma (PRAME), wherein the TCR comprises:

21 . A nucleic acid sequence encoding an antigen recognizing construct or a T cell receptor as defined in any one of claims 1 to 20.

22. A vector comprising a nucleic acid sequence as defined in claim 21.

23. The vector according to claim 22, further comprising a nucleic acid sequence encoding a CD8 Co-receptor.

24. The vector according to claim 23, wherein the CD8 Co-receptor is a wildtype CD8 Coreceptor, such as a wildtype CD8 a Co-receptor, or a chimeric CD8 Co-receptor.

25. A host cell comprising the antigen recognizing construct or T cell receptor according to any one of claims 1 to 20, or the nucleic acid sequence according to claim 21 , or the vector according to any one of claims 22-24.

26. The host cell according to claim 25, wherein the host cell comprises a T cell, a hematopoietic stem cell, a NK cell, a primary T cell, or a natural killer T cell.

27. The host cell according to claim 25 or 26, wherein the host cell is a T cell.

28. The antigen recognizing construct or the T cell receptor according to any one of claims 1 to 20, or the nucleic acid sequence according to claim 21 , or the vector according to any one of claims 22 - 24, or the host cell according to any one of claims 25-27, for use in medicine.

29. The antigen recognizing construct or the T cell receptor according to any one of claims 1 to 20, or the nucleic acid sequence according to claim 21 , or the vector according to any one of claims 22 - 24, or the host cell according to any one of claims 25-27, for use in detection, diagnosis, prognosis, prevention and/or treatment of a disease.

30. The antigen recognizing construct, or the T cell receptor, or the nucleic acid sequence, or the vector, or the host cell according to claim 29, wherein the disease is cancer.

31 . The antigen recognizing construct, or the T cell receptor, or the nucleic acid sequence, or the vector, or the host cell according to according to claim 29 or 30, wherein the disease is a tumor that expresses PRAME.

32. The antigen recognizing construct, or the T cell receptor, or the nucleic acid sequence, or the vector, or the host cell according to claim 29 or claim 30, wherein the disease is an advanced-stage tumor that expresses PRAME.

33. The antigen recognizing construct, or the T cell receptor, or the nucleic acid sequence, or the vector, or the host cell according to any one of claims 29 to 32, wherein the disease is selected from the group consisting of melanoma, bladder carcinoma, colon carcinoma, and breast adenocarcinoma, sarcoma, prostate cancer, uterine cancer, uveal cancer, uveal melanoma, squamous head and neck cancer, synovial carcinoma, Ewing’s sarcoma, breast cancer such as triple negative breast cancer, thyroid cancer, testicular cancer, renal cancer, pancreatic cancer, ovarian cancer, esophageal cancer, non-small-cell lung cancer (NSCLC), non-Hodgkin’s lymphoma, multiple myeloma, melanoma, hepatocellular carcinoma, head and neck cancer, gastric cancer, endometrial cancer, colorectal cancer, cholangiocarcinoma, breast cancer, bladder cancer, myeloid leukemia, acute lymphoblastic leukemia, small-cell lung cancer (SCLC), sarcoma and osteosarcoma.

34. The antigen recognizing construct, or the T cell receptor, or the nucleic acid sequence, or the vector, or the host cell according to claim 33, wherein the breast cancer is selected from the group consisting of ductal breast cancer, tubular breast cancer, medullary breast cancer and combinations thereof.

35. The antigen recognizing construct, or the T cell receptor, or the nucleic acid sequence, or the vector, or the host cell according to claim 33, wherein the gastric cancer is gastric adenocarcinoma cancer.

36. The antigen recognizing construct, or the T cell receptor, or the nucleic acid sequence, or the vector, or the host cell according to claim 33, wherein the sarcoma cancer is selected from the group consisting of chondrosarcoma cancer, osteosarcoma cancer and combinations thereof.

37. Use of the antigen recognizing construct or the T cell receptor according to any one of claims 1 to 20, or the nucleic acid sequence according to claim 21 , or the vector according to any one of claims 22 - 24, or the host cell according to any one of claims 25 - 27, for the manufacture of a medicament for treating a disease.

38. The use of the antigen recognizing construct or the T cell receptor, or the nucleic acid sequence, or the vector, or the host cell according to claim 37, wherein the disease is a cancer.

39. The use of the antigen recognizing construct or the T cell receptor, or the nucleic acid sequence, or the vector, or the host cell, according to claim 37 or claim 38, wherein the disease is a tumor that expresses PRAME.

40. The use of the antigen recognizing construct or the T cell receptor, or the nucleic acid sequence, or the vector, or the host cell according to claim 39, wherein the disease is an advanced-stage tumor that expresses PRAME.

41 . The use of the antigen recognizing construct or the T cell receptor, or the nucleic acid sequence, or the vector, or the host cell according to any one of claims 37 to 41 , wherein the disease is selected from the group consisting of melanoma, bladder carcinoma, colon carcinoma, and breast adenocarcinoma, sarcoma, prostate cancer, uterine cancer, uveal cancer, uveal melanoma, squamous head and neck cancer, synovial carcinoma, Ewing’s sarcoma, breast cancer such as triple negative breast cancer, thyroid cancer, testicular cancer, renal cancer, pancreatic cancer, ovarian cancer, esophageal cancer, non-small- cell lung cancer (NSCLC), non-Hodgkin’s lymphoma, multiple myeloma, melanoma, hepatocellular carcinoma, head and neck cancer, gastric cancer, endometrial cancer, colorectal cancer, cholangiocarcinoma, breast cancer, bladder cancer, myeloid leukemia, acute lymphoblastic leukemia, small-cell lung cancer (SCLC), sarcoma and osteosarcoma.

42. The use of the antigen recognizing construct or the T cell receptor, or the nucleic acid sequence, or the vector, or the host cell according to claim 41 , wherein the breast cancer is selected from the group consisting of ductal breast cancer, tubular breast cancer, medullary breast cancer and combinations thereof.

43. The use of the antigen recognizing construct or the T cell receptor, or the nucleic acid sequence, or the vector, or the host cell according to claim 41 , wherein the gastric cancer is gastric adenocarcinoma cancer.

44. The use of the antigen recognizing construct or the T cell receptor, or the nucleic acid sequence, or the vector, or the host cell according to claim 39, wherein the sarcoma cancer is selected from the group consisting of chondrosarcoma cancer, osteosarcoma cancer and combinations thereof.

45. A method of treating a disease, comprising the step of administering a therapeutically effective amount of the antigen recognizing construct or the T cell receptor according to any one of claims 1 to 20, or the nucleic acid sequence according to claim 21 , or the vector according to any one of claims 22 - 24, or the host cell according to any one of claims 25- 27.

46. The method of treating a disease according to claim 45, wherein the disease is a cancer.

47. The method of treating a disease according to claim 45 or claim 46, wherein the disease is a tumor that expresses PRAME.

48. The method of treating a disease according to claim 47, wherein the disease is an advanced-stage tumor that expresses PRAME.

49. The method of treating a disease according to any one of claims 45-48, wherein the disease is selected from the group consisting of melanoma, bladder carcinoma, colon carcinoma, and breast adenocarcinoma, sarcoma, prostate cancer, uterine cancer, uveal cancer, uveal melanoma, squamous head and neck cancer, synovial carcinoma, Ewing’s sarcoma, breast cancer such as triple negative breast cancer, thyroid cancer, testicular cancer, renal cancer, pancreatic cancer, ovarian cancer, esophageal cancer, non-small- cell lung cancer (NSCLC), non-Hodgkin’s lymphoma, multiple myeloma, melanoma, hepatocellular carcinoma, head and neck cancer, gastric cancer, endometrial cancer, colorectal cancer, cholangiocarcinoma, breast cancer, bladder cancer, myeloid leukemia, acute lymphoblastic leukemia, small-cell lung cancer (SCLC), sarcoma and osteosarcoma.

50. The method of treating a disease according to claim 49, wherein the breast cancer is selected from the group consisting of ductal breast cancer, tubular breast cancer, medullary breast cancer and combinations thereof.

51. The method of treating a disease according to claim 49, wherein the gastric cancer is gastric adenocarcinoma cancer.

52. The method of treating a disease according to claim 49, wherein the sarcoma cancer is selected from the group consisting of chondrosarcoma cancer, osteosarcoma cancer and combinations thereof.

53. The method of treating a disease according to any one of claims 45-52, further comprising co- administering a CD8 Co-receptor.

54. The method according to claim 53, wherein the CD8 Co-receptor is a wildtype CD8 Co-receptor such as a wildtype CD8 a Co-receptor or a chimeric CD8 Co-receptor.

55. A pharmaceutical composition comprising the antigen recognizing construct or the T cell receptor according to any one of claims 1 to 20, or the nucleic acid sequence according to claim 21 , or the vector according to any one of claim 22 - 24, or the host cell according to any one of claims 25-27.

56. A kit for use in medicine comprising the antigen recognizing construct or the T cell receptor according to any one of claims 1 to 20, or the nucleic acid sequence according to claim 21 , or the vector according to any one of claims 22-24, or the host cell according to any one of claims 25-27.

57. The kit according to claim 56, wherein the kit is a diagnostic kit for selecting a patient for treatment of a tumor, wherein cells of the tumor express PRAME.