EA041360B1 - BISPECIFIC MOLECULES BINDING TO ANTI-TNF RELATED APOPTOSIS-INDUCING RECEPTOR 2 LIGAND AND ANTI-CADHERIN 17 FOR CANCER TREATMENT - Google Patents

Description

The field to which the invention relates

The present invention relates to binding molecules that bind to TNF-associated apoptosis-inducing receptor 2 ligand (TRAILR2) and cadherin-17 (CDH17), and their use in medicine, to pharmaceutical compositions containing them, and methods of using them as agents. for the treatment and/or prevention of cancer.

Prerequisites for the invention

Cancer is a group of diseases usually based on abnormal cell proliferation and the ability of cancer cells to invade or spread throughout the body. It is a serious disease and is the leading cause of death worldwide.

Various treatments have been used in an attempt to manage or in some cases treat cancer, including surgery, chemotherapy, radiation therapy, and hormone therapy. Although advances have been made in recent years in the treatment of certain types of cancer, there is still a need to improve the treatment of this disease.

Antibody-based biological molecules offer the potential to be powerful therapeutic agents for cancer treatment. Antibodies are designed to recognize and bind to specific proteins on the surface of cells (their target antigens), and such proteins can only be present on the surface of specific cancer cells or on immune cells. This binding can elicit a range of different biological responses depending on the function of their target antigen protein as well as the structure of the antibody itself.

For example, some antibodies trigger the immune system to attack and destroy cancer cells, either by attracting immune cells to cancer cells or by directly affecting the activity of the immune system itself. Another type of antibody-based therapy binds to cancer cells to stop or reduce cell division, thereby slowing down or preventing abnormal cell proliferation. Other types of antibodies have drugs or radioactive particles attached to them and therefore deliver these therapeutics to the cancer cell itself.

Apoptosis is a controlled cellular mechanism when the body maintains cellular homeostasis in normal tissue compartments and removes damaged cells.

There are two main signaling pathways leading to apoptosis in mammalian cells: the intrinsic pathway and the extrinsic pathway. The intrinsic pathway is initiated at the mitochondrial level and plays an essential role in cell death caused by chemotherapy or radiation. In contrast, the extrinsic death pathway is initiated through death receptor-mediated signals on the cell surface.

Externally induced cell death has been extensively studied following signals mediated by various members of the tumor necrosis factor receptor (TNF) superfamily. The TNF superfamily is characterized by a sequence of two to five cysteine-rich extracellular repeats. Death receptors, belonging to the TNF superfamily, have a homologous intracellular death domain of about 80 amino acids, which is required for apoptotic signaling.

TNF-associated apoptosis-inducing ligand (TRAIL) is a naturally occurring protein ligand that interacts with two types of receptors: death receptors that trigger and decoy receptors that inhibit TRAIL-induced apoptosis. To date, four human receptors specific for TRAIL have been identified: death receptors, DR4 (DR4/TRAIL receptor 1/TRAILR1) and DR5 (TRAIL receptor 2/TRAIL-R2/KILLER), and decoy receptors, DcR1/TRAILR3/ TRID and DcR2/TRAILR4. TRAIL can also bind to osteoprotegerin (OPG), a soluble decoy receptor, with low affinity.

Targeting TRAIL receptors is considered a viable approach in developing cancer therapies because if an antibody-based molecule can bind to and activate the TRAIL receptor, i.e. a TRAIL receptor agonist molecule, it can also induce apoptosis in cancer cells. As shown in many preclinical studies, TRAIL signaling effectively induces apoptosis in numerous tumor cell lines, but not in most normal cells. However, it is reported that normal tissues, especially hepatocytes in the liver, are also susceptible to this mechanism of apoptosis induction. Therefore, if a molecule is used that activates the pathway too effectively, then serious side effects can be caused due to the induction of apoptosis in non-malignant cells as well. On the other hand, if only weakly activating molecules are used, they have been shown to have poor anticancer activity, although they are well tolerated.

Therefore, there is a need to develop therapeutically effective but safe antibody-based biological molecules that can function as TRAIL receptor molecule agonists. Thus, the object of the present invention was to provide TRAIL receptor agonist molecules with an improved therapeutic profile.

Brief summary of the invention

The present invention is based on the concept of combining an antigen-binding site that specifically binds to a TNF-bound, apoptosis-inducing receptor 2 ligand.

- 1 041360 (TRAILR2) with an antigen-binding site that specifically binds to cadherin-17 (CDH17) within a single binding molecule. As discussed in more detail below, one advantage of the molecule of the invention is that apoptosis is only promoted in cells that present TRAILR2 and CDH17 on their surface.

Therefore, the first aspect of the invention relates to a binding molecule having at least one antigen-binding site (first antigen-binding site) that specifically binds to a TNF-associated apoptosis-inducing receptor 2 ligand (TRAILR2) and at least one antigen-binding site (second antigen-binding site). region) that specifically binds to cadherin-17 (CDH17).

In a preferred embodiment of the inventive binding molecule, the molecule is bispecific and tetravalent.

In a preferred embodiment of the binding molecule of the invention, at least one antigen-binding site that specifically binds to cadherin-17 (CDH17) is an immunoglobulin (Ig) molecule (having the usual Y-shaped structure of a full-length antibody with two light and two heavy chains ) and at least one antigen-binding site that specifically binds to a TNF-associated apoptosis-inducing receptor 2 ligand (TRAILR2) contains one or more scFvs.

In a preferred embodiment of the binding molecule of the invention, one or more scFvs have a VL-VH orientation from N- to C-terminus.

In a preferred embodiment of the binding molecule of the invention, one or more scFv(s) are fused to the C-terminus of the heavy chain of the Ig molecule. For example, one scFv is fused to the C-terminus of one of the heavy chains of the Ig molecule, and one scFv is fused to the C-terminus of the other heavy chain of the Ig molecule. As such, the scFv specific for TRAILR2 is fused to each of the heavy chains of the Ig molecule, thereby forming a symmetrical, bispecific and tetravalent structure.

In a preferred embodiment of the binding molecule of the invention, the Ig molecule is IgG1KO. In another preferred embodiment of the binding molecule of the invention, the Ig molecule is IgG1FcRnmut.

In a preferred embodiment of the binding molecule of the invention, one or more scFvs are fused to an Ig molecule via a peptidic linker, preferably a peptidic linker of about 4 to 20 amino acids in length (e.g., any of 6, 9, 12, 15 amino acids) .

In a preferred embodiment of the binding molecule of the invention, the antigen-binding site that specifically binds to TRAILR2 (first antigen-binding site) is selected from the group consisting of antigen-binding sites (i)-(xiii):

i) an antigen-binding site comprising complementarity-determining regions (CDRs) of the heavy chain containing the amino acid sequences of SEQ ID NO: 1 (CDR1), SEQ ID NO: 2 (CDR2) and SEQ ID NO: 3 (CDR3), and light chain CDRs, containing the amino acid sequences of SEQ ID NO: 4 (CDR1), SEQ ID NO: 5 (CDR2) and SEQ ID NO: 6 (CDR3);

ii) an antigen-binding site comprising heavy chain CDRs containing the amino acid sequences of SEQ ID NO: 7 (CDR1), SEQ ID NO: 8 (CDR2) and SEQ ID NO: 9 (CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO : 10 (CDR1), SEQ ID NO: 11 (CDR2) and SEQ ID NO: 12 (CDR3);

iii) an antigen-binding site comprising heavy chain CDRs containing the amino acid sequences of SEQ ID NO: 13 (CDR1), SEQ ID NO: 14 (CDR2) and SEQ ID NO: 15 (CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO : 16 (CDR1), SEQ ID NO: 17 (CDR2) and SEQ ID NO: 18 (CDR3);

iv) an antigen-binding site comprising heavy chain CDRs containing the amino acid sequences of SEQ ID NO: 19 (CDR1), SEQ ID NO: 20 (CDR2) and SEQ ID NO: 21 (CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO : 22 (CDR1), SEQ ID NO: 23 (CDR2) and SEQ ID NO: 24 (CDR3);

v) an antigen-binding site comprising heavy chain CDRs containing the amino acid sequences of SEQ ID NO: 25 (CDR1), SEQ ID NO: 26 (CDR2) and SEQ ID NO: 27 (CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO : 28 (CDR1), SEQ ID NO: 29 (CDR2) and SEQ ID NO: 30 (CDR3);

vi) an antigen-binding site comprising heavy chain CDRs containing the amino acid sequences of SEQ ID NO: 31 (CDR1), SEQ ID NO: 32 (CDR2) and SEQ ID NO: 33 (CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO : 34 (CDR1), SEQ ID NO: 35 (CDR2) and SEQ ID NO: 36 (CDR3);

vii) an antigen-binding site comprising heavy chain CDRs containing the amino acid sequences of SEQ ID NO: 37 (CDR1), SEQ ID NO: 38 (CDR2) and SEQ ID NO: 39 (CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO : 40 (CDR1), SEQ ID NO: 41

- 2 041360 (CDR2) and SEQ ID NO: 42 (CDR3);

viii) an antigen-binding site comprising a heavy chain CDR containing the amino acid sequence of SEQ ID NO: 43 (CDR1), SEQ ID NO: 44 (CDR2) and SEQ ID NO: 48 (CDR3) and a light chain CDR containing the amino acid sequence of SEQ ID NO : 49 (CDR1), SEQ ID NO: 50 (CDR2) and SEQ ID NO: 54 (CDR3);

ix) an antigen-binding site comprising a heavy chain CDR containing the amino acid sequence of SEQ ID NO: 43 (CDR1), SEQ ID NO: 45 (CDR2) and SEQ ID NO: 48 (CDR3) and a light chain CDR containing the amino acid sequence of SEQ ID NO : 49 (CDR1), SEQ ID NO: 51 (CDR2) and SEQ ID NO: 55 (CDR3);

x) an antigen-binding site comprising a heavy chain CDR comprising the amino acid sequence of SEQ ID NO: 43 (CDR1), SEQ ID NO: 45 (CDR2), and SEQ ID NO: 48 (CDR3), and a light chain CDR comprising the amino acid sequences of SEQ ID NO: 49 (CDR1), SEQ ID NO: 52 (CDR2) and SEQ ID NO: 56 (CDR3);

xi) an antigen-binding site comprising a heavy chain CDR comprising the amino acid sequence of SEQ ID NO: 43 (CDR1), SEQ ID NO: 45 (CDR2), and SEQ ID NO: 48 (CDR3), and a light chain CDR comprising the amino acid sequences of SEQ ID NO: 49 (CDR1), SEQ ID NO: 50 (CDR2) and SEQ ID NO: 56 (CDR3);

xii) an antigen-binding site comprising a heavy chain CDR comprising the amino acid sequence of SEQ ID NO: 43 (CDR1), SEQ ID NO: 46 (CDR2), and SEQ ID NO: 48 (CDR3), and a light chain CDR comprising the amino acid sequences of SEQ ID NO: 49 (CDR1), SEQ ID NO: 51 (CDR2) and SEQ ID NO: 57 (CDR3); and xiii) an antigen-binding site comprising a heavy chain CDR comprising the amino acid sequence of SEQ ID NO: 43 (CDR1), SEQ ID NO: 47 (CDR2), and SEQ ID NO: 48 (CDR3), and a light chain CDR comprising the amino acid sequences of SEQ ID NO: 49 (CDR1), SEQ ID NO: 53 (CDR2) and SEQ ID NO: 57 (CDR3).

In a preferred embodiment of the binding molecule of the invention, the antigen binding site that specifically binds to TRAILR2 (first antigen binding site) is selected from the group consisting of antigen binding sites (i)-(xiv):

i) an antigen binding site comprising a heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 82 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 83;

ii) an antigen binding site comprising a heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 84 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 85;

iii) an antigen binding site comprising a heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 86 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 87;

iv) an antigen binding site comprising a heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 88 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 89;

v) an antigen binding site comprising a heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 90 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 91;

vi) an antigen binding site comprising a heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 92 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 93;

vii) an antigen binding site comprising a heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 94 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 95;

viii) an antigen binding site comprising a heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 96 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 97;

ix) an antigen binding site comprising a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 98 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 99;

x) an antigen binding site comprising a heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 100 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 101;

xi) an antigen binding site comprising a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 102 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 103;

xii) an antigen-binding site comprising a heavy chain variable domain containing

- 3 041360 amino acid sequence of SEQ ID NO: 104, and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 105;

xiii) an antigen binding site comprising a heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 106 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 107; and xiv) an antigen binding site comprising a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 108 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 109.

In a preferred embodiment of the binding molecule of the invention, the antigen-binding site that specifically binds to CDH17 (second antigen-binding site) is selected from the group consisting of antigen-binding sites (i)-(iv):

i) an antigen-binding site comprising heavy chain CDRs containing the amino acid sequences of SEQ ID NO: 58 (CDR1), SEQ ID NO: 59 (CDR2) and SEQ ID NO: 60 (CDR3), and light chain CDRs containing the amino acid sequences of SEQ ID NO: 61 (CDR1), SEQ ID NO: 62 (CDR2) and SEQ ID NO: 63 (CDR3);

ii) an antigen-binding site comprising heavy chain CDRs containing the amino acid sequences of SEQ ID NO: 64 (CDR1), SEQ ID NO: 65 (CDR2) and SEQ ID NO: 66 (CDR3), and light chain CDRs containing the amino acid sequences of SEQ ID NO: 67 (CDR1), SEQ ID NO: 68 (CDR2) and SEQ ID NO: 69 (CDR3);

iii) an antigen-binding site comprising heavy chain CDRs containing the amino acid sequences of SEQ ID NO: 70 (CDR1), SEQ ID NO: 71 (CDR2) and SEQ ID NO: 72 (CDR3), and light chain CDRs containing the amino acid sequences of SEQ ID NO: 73 (CDR1), SEQ ID NO: 74 (CDR2) and SEQ ID NO: 75 (CDR3); and iv) an antigen-binding site comprising heavy chain CDRs comprising the amino acid sequences of SEQ ID NO: 76 (CDR1), SEQ ID NO: 77 (CDR2), and SEQ ID NO: 78 (CDR3), and light chain CDRs comprising the amino acid sequences of SEQ ID NO: 79 (CDR1), SEQ ID NO: 80 (CDR2) and SEQ ID NO: 81 (CDR3).

In a preferred embodiment of the binding molecule of the invention, the antigen-binding site that specifically binds to CDH17 (second antigen-binding site) is selected from the group consisting of antigen-binding sites (i)-(v):

i) an antigen binding site comprising a heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 110 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 111;

ii) an antigen binding site comprising a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 112 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 113;

iii) an antigen binding site comprising a heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 114 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 115;

iv) an antigen binding site comprising a heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 116 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 117; And

v) an antigen binding site comprising a heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 118 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 119.

In some embodiments, the heavy chain CDR3 sequence of SEQ ID NO: 272 is used instead of the heavy chain CDR3 sequences of SEQ ID NO: 78 in the context of the binding molecules of the present invention.

In some embodiments, the binding molecules are presented in various aspects below, as defined by their heavy chain amino acid sequence (e.g., a modified scFv heavy chain fused to the C-terminus of an Ig heavy chain) as well as their light chain amino acid sequences including two heavy chains and two light chains, thereby forming a symmetrical tetravalent and bispecific structure.

Another aspect of the invention provides a binding molecule having (i) a heavy chain containing the amino acid sequence of SEQ ID NO: 159, or a heavy chain containing the amino acid sequence of SEQ ID NO: 160, or a heavy chain containing the amino acid sequence of SEQ ID NO: 161, or a heavy chain containing the amino acid sequence of SEQ ID NO: 162, or a heavy chain containing the amino acid sequence of SEQ ID NO: 163, or a heavy chain containing the amino acid sequence of SEQ ID NO: 164, or a heavy chain containing the amino acid sequence of SEQ ID NO: 165 , or a heavy chain containing the amino acid sequence of SEQ ID NO: 166, or a heavy chain containing the amino acid sequence of SEQ ID NO: 167, or a heavy chain containing

- 4 041360 amino acid sequence of SEQ ID NO: 168, or a heavy chain containing the amino acid sequence of SEQ ID NO: 169, or a heavy chain containing the amino acid sequence of SEQ ID NO: 170, or a heavy chain containing the amino acid sequence of SEQ ID

NO: 171, or a heavy chain containing the amino acid sequence of SEQ ID NO: 172, and (ii) a light chain containing the amino acid sequence of SEQ ID NO: 173.

Another aspect of the invention provides a binding molecule having (i) a heavy chain containing the amino acid sequence of SEQ ID NO: 174, or a heavy chain containing the amino acid sequence of SEQ ID NO: 175, or a heavy chain containing the amino acid sequence of SEQ ID NO: 176, or a heavy chain containing the amino acid sequence of SEQ ID NO: 177, or a heavy chain containing the amino acid sequence of SEQ ID NO: 178, or a heavy chain containing the amino acid sequence of SEQ ID NO: 179, or a heavy chain containing the amino acid sequence of SEQ ID NO: 180 , or a heavy chain containing the amino acid sequence of SEQ ID NO: 181, or a heavy chain containing the amino acid sequence of SEQ ID NO: 182, or a heavy chain containing the amino acid sequence of SEQ ID NO: 183, or a heavy chain containing the amino acid sequence of SEQ ID NO: : 184, or a heavy chain containing the amino acid sequence SEQ ID NO: 185, or a heavy chain containing the amino acid sequence of SEQ ID NO: 186, or a heavy chain containing the amino acid sequence of SEQ ID NO: 187, and (ii) a light chain containing the amino acid sequence of SEQ ID NO: 188.

In a further aspect of the present invention, a binding molecule is provided having (i) a heavy chain comprising the amino acid sequence of SEQ ID NO: 189, or a heavy chain comprising the amino acid sequence of SEQ ID NO: 190, or a heavy chain comprising the amino acid sequence of SEQ ID NO: 191, or a heavy chain containing the amino acid sequence of SEQ ID NO: 192, or a heavy chain containing the amino acid sequence of SEQ ID NO: 193, or a heavy chain containing the amino acid sequence of SEQ ID NO: 194, or a heavy chain containing the amino acid the sequence of SEQ ID NO: 195, or a heavy chain containing the amino acid sequence of SEQ ID NO: 196, or a heavy chain containing the amino acid sequence of SEQ ID NO. 197, or a heavy chain containing the amino acid sequence of SEQ ID NO: 198, or a heavy chain containing the amino acid sequence of SEQ ID NO: 199, or a heavy chain containing the amino acid sequence of SEQ ID NO: 200, or a heavy chain containing the amino acid sequence of SEQ ID NO: 201, or a heavy chain containing the amino acid sequence of SEQ ID NO: 202, and (ii) a light chain containing the amino acid sequence of SEQ ID NO: 203.

Another aspect of the invention provides a binding molecule having (i) a heavy chain containing the amino acid sequence of SEQ ID NO: 204, or a heavy chain containing the amino acid sequence of SEQ ID NO: 205, or a heavy chain containing the amino acid sequence of SEQ ID NO: 206, or a heavy chain containing the amino acid sequence of SEQ ID NO: 207, or a heavy chain containing the amino acid sequence of SEQ ID NO: 208, or a heavy chain containing the amino acid sequence of SEQ ID NO: 209, or a heavy chain containing the amino acid sequence of SEQ ID NO: 210 , or a heavy chain containing the amino acid sequence of SEQ ID NO: 211, or a heavy chain containing the amino acid sequence of SEQ ID NO: 212, or a heavy chain containing the amino acid sequence of SEQ ID NO: 213, or a heavy chain containing the amino acid sequence of SEQ ID NO: : 214, or a heavy chain containing the amino acid sequence SEQ ID NO: 215, or a heavy chain containing the amino acid sequence of SEQ ID NO: 216, or a heavy chain containing the amino acid sequence of SEQ ID NO: 217; or a heavy chain containing the amino acid sequence of SEQ ID NO: 271, and (ii) a light chain containing the amino acid sequence of SEQ ID NO: 218.

In a further aspect of the present invention, a binding molecule is provided having (i) a heavy chain comprising the amino acid sequence of SEQ ID NO: 145, or a heavy chain comprising the amino acid sequence of SEQ ID NO: 147, or a heavy chain comprising the amino acid sequence of SEQ ID NO: 149, or a heavy chain containing the amino acid sequence of SEQ ID NO: 151, or a heavy chain containing the amino acid sequence of SEQ ID NO: 153, or a heavy chain containing the amino acid sequence of SEQ ID NO: 155, or a heavy chain containing the amino acid the sequence of SEQ ID NO: 157, or a heavy chain containing the amino acid sequence of SEQ ID NO: 219, or a heavy chain containing the amino acid sequence of SEQ ID NO: 220, or a heavy chain containing the amino acid sequence of SEQ ID NO: 221, or a heavy chain, containing the amino acid sequence of SEQ ID NO: 222, or a heavy chain containing a neighing amino acid sequence of SEQ ID NO: 223, or a heavy chain containing the amino acid sequence of SEQ ID

- 5 041360

NO: 224, or a heavy chain containing the amino acid sequence of SEQ ID NO: 225; or a heavy chain containing the amino acid sequence of SEQ ID NO: 226, or a heavy chain containing the amino acid sequence of SEQ ID NO: 227, or a heavy chain containing the amino acid sequence of SEQ ID NO: 228, or a heavy chain containing the amino acid sequence of SEQ ID NO: 229, or a heavy chain containing the amino acid sequence of SEQ ID NO: 230, or a heavy chain containing the amino acid sequence of SEQ ID NO: 231, or a heavy chain containing the amino acid sequence of SEQ ID NO: 232, and (ii) a light chain containing the amino acid the sequence of SEQ ID NO: 233.

Another aspect of the invention relates to a nucleic acid molecule encoding a binding molecule in accordance with the invention or to an expression vector containing such a nucleic acid molecule.

Another aspect of the invention relates to a host cell containing a nucleic acid molecule according to the invention in functional association with an expression control sequence. In addition, the present invention provides a host cell containing an expression vector comprising a nucleic acid molecule encoding a binding molecule, as described in this application.

Another aspect of the invention relates to a method for obtaining a binding molecule in accordance with the invention, including (a) culturing a host cell in accordance with the invention under conditions that allow the expression of the molecule;

(b) isolation of the molecule.

Another aspect of the invention relates to a binding molecule in accordance with the invention, for use in medicine.

Another aspect of the invention relates to a binding molecule according to the invention for use in the treatment of cancer, preferably colorectal cancer (CRC), stomach cancer (GC), pancreatic cancer (PC), esophageal cancer or neuroendocrine tumors.

Another aspect of the invention relates to a pharmaceutical composition containing a binding molecule in accordance with the invention together with a pharmaceutically acceptable carrier and optionally one or more other active substances.

Yet another aspect of the invention relates to a method of treating cancer comprising administering an effective amount of a binding molecule according to the invention to a patient in need thereof.

The present invention further provides antibodies that specifically bind to CDH17, comprising (i) heavy chain CDRs comprising the amino acid sequences of SEQ ID NO: 58 (CDR1), SEQ ID NO: 59 (CDR2), and SEQ ID NO: 60 (CDR3), and light chain CDRs containing the amino acid sequences of SEQ ID NO: 61 (CDR1), SEQ ID NO: 62 (CDR2) and SEQ ID NO: 63 (CDR3), (ii) heavy chain CDRs containing the amino acid sequences of SEQ ID NO: 64 (CDR1), SEQ ID NO: 65 (CDR2) and SEQ ID NO: 66 (CDR3), and light chain CDRs containing the amino acid sequences of SEQ ID NO: 67 (CDR1), SEQ ID NO: 68 (CDR2) and SEQ ID NO: 69 (CDR3), (iii) heavy chain CDRs containing the amino acid sequences of SEQ ID NO: 70 (CDR1), SEQ ID NO: 71 (CDR2) and SEQ ID NO: 72 (CDR3), and light chain CDRs containing amino acid sequences of SEQ ID NO: 73 (CDR1), SEQ ID NO: 74 (CDR2), and SEQ ID NO: 75 (CDR3), or (iv) heavy chain CDRs comprising the amino acid sequences of SEQ ID NO: 76 (CDR1), SEQ ID N O: 77 (CDR2) and SEQ ID NO: 78 (CDR3), and light chain CDRs containing the amino acid sequences of SEQ ID NO: 79 (CDR1), SEQ ID NO: 80 (CDR2) and SEQ ID NO: 81 (CDR3) .

Brief description of the figures

Fig. 1a and b. Schematic representation of the binding molecule according to the invention.

Depicted is an example of a binding molecule according to the invention containing (i) an Ig molecule specifically binding to CDH17, which contains two heavy and two light chains, and (ii) two scFv molecules specifically binding to TRAILR2. The N-terminus of the scFv is fused to the C-terminus of each of the heavy chains of the Ig molecule, thereby forming a symmetrical, bispecific and tetravalent antibody molecule.

Fig. 2. Surface expression analysis of TRAILR2 (TR2) and CDH17 protein in Colo-205 cells. Results are presented as mean PE values relative to cell-only controls. Data are representative of four independent experiments.

Fig. 3. Effect of antibody incubation on cell viability. Colo-205 cells were treated for 24 h with different concentrations of (i) anti-TRAILR2 alone (anti-TR2v1), (ii) anti-CDH17v1 alone, (iii) TRAILR2/CDH17 bispecific molecule (CDH17v1/TR2v1), or

- 6 041360 (iv) an equivalent combination of individual antibodies TRAILR2 (aHTu-TR2v1) and CDH17v1.

Data are expressed as mean relative values compared to untreated controls plus standard deviation. Two independent analyzes were performed in duplicate for each condition.

Fig. 4. Apoptosis induction assay (caspase activation assay) after antibody incubation. Colo-205 cells were treated for (A) 2 h or (B) 6 h with various concentrations of:

(i) anti-TRAILR2 (anti-TR2v1) only, (ii) anti-CDH17v1 only, and (iii) CDH17v1/TR2v1 bispecific molecule.

Measured (A) activation of caspase-8 or (B) caspase-3. Data are expressed as mean relative fold changes from untreated control plus standard deviation. Three independent analyzes were performed in duplicate for each condition.

Fig. 5. Specificity analysis of the CDH17v1/TR2v1 bispecific molecule. The inventors have created an additional bispecific molecule that recognizes human TR2 and trinitrophenol (TNP) instead of CDH17. Colo-205 cells were treated for 24 h with different concentrations:

(i) anti-TNP antibodies only, (ii) anti-CDH17v1 only, (iii) anti-TRAILR2 (anti-TR2v1), (iv) TNP/TR2v1 or (v) CDH17v1/TR2v1 bispecific molecule only, and cell viability was measured .

Data are expressed as mean relative values compared to untreated controls plus standard deviation. Two independent analyzes were performed in duplicate for each condition.

Fig. 6. Effect of antibody incubation on negative CDH17 cells. (A) Surface expression analysis of TRAILR2 (TR2) protein and CDH17 in HepG2 cells. Results are presented as mean PE values relative to cell-only controls. Data are representative of 3 independent experiments. (B) HepG2 cells were treated for 72 h with different concentrations:

(i) anti-TRAILR2 alone (anti-TR2v1), (ii) a TRAILR2/CDH17 bispecific molecule (CDH17v1/TR2v1), or (iii) anti-TRAILR2 in the presence of a cross-linker, goat anti-human IgG (anti-TR2v1 (cross-linking fc)).

Data are expressed as mean relative values compared to untreated controls plus standard deviation. Five independent analyzes were performed in duplicate for each condition.

Fig. 7. Effect of bispecific molecules containing various IgG scaffolds on cell viability. Colo-205 cells were treated for 24 h with various concentrations of antiTRAILR2 alone (anti-TR2v1 or anti-TR2v2) and various bispecific, tetravalent molecules recognizing human TRAILR2 and human CDH17 containing different IgG1 (A and C) and IgG4Pro (B) backbones. . Data are expressed as mean relative values compared to untreated controls plus standard deviation. Two independent analyzes were performed in duplicate for each condition.

Fig. 8. Effect of scFv variants on cell viability. Colo-205 cells were treated for 24 h with different concentrations of different bispecific molecules containing different scFv variants. Data are expressed as mean relative values compared to untreated controls plus standard deviation. Two independent analyzes were performed in duplicate for each condition.

Fig. 9. Effect of bispecific, tetravalent molecules recognizing human TRAILR2 (TR2) and human CDH17 on cell viability. (A-C) Colo-205 cells were treated for 24 hours with different concentrations of different bispecific molecules containing different CDH17 binders and scFv variants. (D) HepG2 cells were treated for 72 h with different concentrations of different bispecific molecules containing different CDH17 binders and scFv variants. Only anti-TR2v2 (A-D) and anti-CDH17H2 (CD) were used as controls. Data are expressed as mean relative values compared to untreated controls plus standard deviation. At least two independent analyzes were performed in duplicate for each condition.

Fig. 10. Effect of a bispecific molecule recognizing human TRAILR2 (TR2) and human CD44v6 on cell viability. (A) Surface expression analysis of TRAILR2 (TR2) protein and CD44v6 in Colo-205 cells. Results are presented as mean PE values relative to cell-only controls. Both proteins showed significant expression compared to their respective IgG controls. Data are representative of 3 independent experiments. (B) Colo-205 cells were treated for 72 h with different concentrations of anti-TRAILR2 (anti-TR2v1) only, anti-CD44v6 antibody and bispecific molecule

CD44v6/TR2v1. Data are expressed as mean relative values compared to untreated controls plus standard deviation. Two independent analyzes were performed in duplicate for each condition.

Fig. 11. Effect of antibody incubation on the viability of CRC cells. (A and B) Surface expression analysis of TRAILR2 (TR2) and CDH17 protein in CL-34 (A) and SK-CO-1 (B) cells. Results are presented as mean PE values relative to cell-only controls plus standard deviation. Data are representative of 2 independent experiments. CL-34 (C) and SK-CO-1 (D) cells were treated for 120 h and 96 h respectively with different concentrations of (i) anti-TRAILR2 (anti-TR2v1) alone, (ii) anti-CDH17v1 alone, or (iii) ) CDH17v1/TR2v1.

Data are expressed as mean relative values compared to untreated controls plus standard deviation.

Two independent analyzes were performed in duplicate for each condition.

Fig. 12. Efficacy of a tetravalent molecule recognizing human TRAILR2 (TR2) and human CDH17: bispecific molecule TRAILR2/CDH17 (black circles) (CDH17H2/TR2v2-(Ig G1 KO) (A and B) or CDH17H2/TR2v2-(Ig G1 FcRnmut) (C and D)) or vehicle control (white circles) were administered to mice bearing Colo205 (A and C) or Gp2d (B and D) tumor cells. Tumor volume (mm ³ ) was measured after administration on the indicated days and the data was expressed as the median of tumor volumes. Each group included 8-10 animals.

Fig. 13. Surface expression analysis of CDH17 protein in Colo-205 cells using novel CDH17 antibody molecules. Results are presented as mean APC values relative to control samples containing only cells. Representative data from a duplicated experiment.

Fig. 14. Expression of CDH17 and TRAILR2 protein in primary CRC and liver metastases KPP. Representative images for TRAILR2 and CDH17 using the respective primary antibodies followed by reagent detection and incubation in DAB solution.

Detailed description of the invention

As described above, one approach to treating cancer has been to induce apoptosis in the cancer cell using molecules that specifically bind to and activate the TNF-associated apoptosis-inducing ligand (TRAIL) receptor-mediated apoptotic pathway. As shown in many preclinical studies, TRAIL signaling effectively induces apoptosis in numerous tumor cell lines, but not in most normal cells. However, it is reported that normal tissues, especially hepatocytes in the liver, are also susceptible to this mechanism of apoptosis induction.

TRAIL binds with high affinity to four different cell surface receptors. Two of them, TRAILR1 and TRAILR2, are able to initiate TRAIL-induced apoptosis through the interaction of its intracellular death domain with various adapter proteins and pro-caspase 8. Clustering of TRAILR1 or TRAILR2 molecules through the TRAIL ligand facilitates autocatalytic cleavage and activation of pro-caspase 8, which, in turn, leads to the induction of apoptosis. TRAILR3 and TRAILR4 are decoy receptors, and although their extracellular domains are capable of binding TRAIL, the intracellular portions of the receptors do not contain a domain capable of inducing apoptosis upon binding TRAIL.

Overexpression of TRAIL decoy receptors can render cancer cells insensitive to the presence of a TRAIL ligand. Specific targeting of non-baiting, death-inducing TRAIL receptors avoids this problem and represents a more effective tumor treatment. The present invention is directed to the development of TRAILR2 agonist molecules. TRAILR2 is widely expressed in a wide range of cancers.

Several TRAILR2-specific agonist antibodies have been developed for the treatment of cancer, including lexatumumab (HGS-ETR2). However, these agonist antibodies are not sufficiently effective in clinical practice. This is probably due to the lack of sufficient clustering of the TRAILR2 receptor and, therefore, the inability to effectively induce apoptosis in cancer cells.

In another attempt to stimulate TRAILR2-mediated apoptosis in cancer cells, a new tetrameric TRAILR2-binding nanobody TAS266 has been developed. In preclinical experiments, it has shown antitumor efficacy superior to conventional TRAILR2-targeted antibodies. However, it has been reported that hepatocytes in the liver may be susceptible to TRAILR2-mediated apoptosis and therefore the off-target increase in TRAILR2 clustering that is stimulated by TAS266 carries the risk of potential toxicity. Indeed, the phase I clinical trial of TAS266 has been terminated.

- 8 041360

Therefore, if a molecule is used that agonizes the pathway too effectively, then serious side effects can be caused as apoptosis is induced in non-malignant cells. On the other hand, if only weakly agonizing molecules are used, they have been shown to have poor anti-cancer activity, although they are well tolerated.

In the past, attempts have been made to create bispecific molecules consisting of a TRAIL receptor binding site and a site that binds specific cancer cell molecules on the cell surface, called an anchor target. Theoretically, such bispecific molecules bind exclusively to cancer cells expressing the anchor target and induce apoptosis through their TRAIL receptor binding site.

A number of different anchor targets have been proposed as suitable combination partners for the TRAIL receptor binding molecule.

For example, CEA (CEAM5HUMAN) and CRIPTO have been proposed as suitable anchor targets in WO 2011039126A1. CEA and CRIPTO are linked to the cell membrane via the GPI anchor. However, CEA and CRIPTO are both easily cleaved from the cell surface and enter the bloodstream from tumors. Indeed, serum CEA levels are used as a biomarker for cancer screening. As can be understood, CEA and CRIPTO in serum can bind to a bispecific molecule that has a dual binding to the TRAIL receptor and these anchor targets. Therefore, serum CEA and CRIPTO will not only compete with membrane-bound anchor proteins, but may also increase the likelihood of TRAIL receptor activation in cells not expressing CEA or CRIPTO, and thus may cause unwanted side effects.

FAP (fibroblast activation protein) has also been proposed as an anchor target. However, FAP is only expressed on activated fibroblast cells that are located in the tumor stroma. FAP is not expressed in epithelial cancer cells. Therefore, a bispecific FAP molecule will only function to promote apoptosis in those cancer cells that are in close physical contact with an activated fibroblast (Brhnker et al., Molecular Cancer Therapeutics (2016), 15(5):946-957). Tumor cells that are not in direct contact with the activated fibroblast will not be affected by this treatment and will continue to proliferate. Therefore, there are clear disadvantages to using FAP as an anchor target for TRAIL receptor mediated apoptosis in cancer cells. Furthermore, since activated fibroblast cells are also found at sites of tissue remodeling, including hepatic fibrosis, pulmonary fibrosis, arteriosclerosis, and arthritis, a bispecific molecule targeting FAP and TRAIL receptors could potentially anchor to the surface of activated fibroblasts and induce apoptosis on adjacent normal TRAIL- sensitive cells in the liver or other organs.

Similarly, MCSP (melanoma-associated chondroitin sulfate proteoglycan) and ROBO4 (ring homologue 4) have been proposed as anchor targets (He Yuan et al., The Journal of Investigative Dermatology (2016), 136(2):541-544; WO2011039126A1 ). In addition to being expressed on the cell surface of melanomas, MCSP is mainly expressed in newly formed vessels. ROBO4 is specifically expressed in endothelial cells. Both are described at sites of angiogenesis in various types of tumors. Therefore, as with FAP, and with the sole exception of MSCP-expressing melanomas, for efficiency, a bispecific molecule targeting MCSP or ROBO4 and the TRAIL receptor will only function to stimulate apoptosis in cancer cells if they are in close physical contact. with endothelial cells. This is not always possible, as the tumor rapidly outgrows its blood supply as it grows. For this reason, again, there are disadvantages when using these molecules as anchor targets.

In addition, a bispecific antibody targeting TRAILR2 and LTeR (lymphotoxin-beta receptor) has been reported to potently inhibit tumor growth in mouse tumor xenograft models at levels comparable to or greater than the combination of the respective parental antibodies (Michaelson et al., mAbs (2009), 1(2): 128-141). Mouse LTeR signaling has been shown to be critical for liver regeneration, where LTeR is expressed on mature hepatocytes (Anders R.A. et al., J Immunol (2005), 175(2): 1295-1300). Similar to FAP, LTeR signaling is widely upregulated during chronic liver inflammation in patients with viral and non-viral hepatitis, cholangitis, and HCC. In particular, its expression in hepatocytes has been described (Haybaeck J. et al., Cancer Cell (2009), 16(4): 295-308). Targeting the LTeR and TRAIL receptor could potentially anchor on the surface of LTeR-expressing hepatocytes that are sensitive to TRAILR2 activation and thus could potentially cause liver toxicity.

Finally, TENASCIN C has also been proposed as a suitable anchor target. TENASCIN C is a secreted protein and therefore does not attach to the cell membrane. This represents another example of an indirect mechanism for the induction of apoptosis in cancer cells. As with anchoring strategies with MCSP or ROBO4, the TENASCIN C fragment must be in the correct orientation with respect to TRAILR2 molecules on the cancer cell membrane for apoptosis to occur. In addition, expression of TENASCIN C is also upregulated in chronic liver disease and treatment with a bispecific molecule containing a TRAIL agonist is expected to worsen the condition.

Therefore, there are currently problems in therapeutic modulation of the apoptotic pathway mediated by the TRAIL receptor, especially in cancer cells.

Therefore, the inventors set out to identify anchor proteins that are not present in significant amounts in serum and have been localized in cancer cells that co-express TRAILR2. It is important to note that the inventors chose to select anchor proteins that were not expressed in the liver due to the potential liver toxicity described above.

The present inventors have identified cadherin-17 as a potentially suitable anchor target that can be used in combination with a TRAIL receptor binding molecule.

Cadherin-17 (CDH17) is a member of the superfamily of cadherins, genes encoding calcium-dependent, membrane-bound glycoproteins. The encoded protein is cadherin-like, consisting of an extracellular region containing 7 cadherin domains and a transmembrane region but lacking a cytoplasmic domain conserved among other members of the cadherin superfamily.

The inventors analyzed the expression of CDH17 in non-tumor tissues. IHC studies revealed homogeneous staining of the CDH17 epithelium in the mucosal epithelium of the small intestine and colon. Heterogeneous staining of the epithelium was also found in the gallbladder, stomach, and intrapancreatic duct epithelium.

To date, no representative studies have been published revealing the co-expression of TRAILR2 and CDH17 in tumors. High predominance was consistently shown for CDH17 expression (88-100%) as well as for TRAILR2 (87-100%) in CRC. In gastric cancer, CDH17 has also been demonstrated in 56-90% of all cases, with a significantly higher incidence of intestinal types compared to diffuse ones, while TRAILR2 has been shown to be constitutively expressed at high levels in primary and metastatic gastric carcinomas. In PCa, CDH17 expression can be shown in 50-82% of all cases, and TRAILR2 in 81% (Gallmeier et al., PLoS One. 2013;8(2):e56760; Ito et al., Virchows Arch. 2005 Oct; 447(4):717-22 Koornstra et al., Eur J Cancer 2005 May;41(8): 1195-202 Koyama et al., J Cancer Res Clin Oncol 2002 Feb;128(2):73 -9; Luque-Garcia et al., Proteomics. 2010 Mar; 10(5):940-52; Panarelli et al., Am J Clin Pathol. 2012 Aug; 138(2):211-22; Su et al. , Mod Pathol 2008 Nov 21(11): 1379-86 Takamura et al, Cancer Sci 2003 May 94(5):425-30 van Geelen et al, J Clin Oncol 2006 Nov 1; 24(31):4998-5004).

Therefore, the inventors conducted this study and showed that TRAILR2 and CDH17 are co-expressed in various tumors (e.g., colorectal cancer (CRC), gastric cancer (GC) and pancreatic cancer (PCa)), with little or no co-expression in non-malignant cells. Notably, CDH17 was not detected in normal liver tissue or hepatocytes with a reported sensitivity to TRAILR2 activation.

It is important to note that, prior to the present invention, it has not been disclosed or even remotely intended to provide binding molecules that target these two antigens.

Accordingly, the inventors have obtained binding molecules comprising at least one antigen-binding site that specifically binds to TRAILR2 and at least one antigen-binding site that specifically binds to CDH17.

From the accompanying experimental data, it appears that such molecules are capable of inducing apoptosis in vitro in cells where both CDH17 and TRAILR2 are expressed and in a therapeutically acceptable amount of the molecule according to the invention. It is important that, as shown in the examples, the same molecules practically do not induce apoptosis in cells expressing TRAILR2, but not CDH17. Therefore, the binding molecules of the present invention have the potential to be therapeutically effective in cancers in which cancer cells express both CDH17 and TRAILR2. In one aspect, the binding molecules of the present invention do not affect CDH17-negative liver cells, thereby reducing the risk of liver toxicity. In addition, binding molecules according to the invention (for example, binding molecules with (i) two heavy chains, each containing a heavy chain variable region specific for CDH17, IgG and scFv constant domains specific for TRAILR2, and (ii) two light chains each containing a light chain variable region specific for CDH17) have been shown to be stable with monomer content above 95%, further supporting their therapeutic utility.

Therefore, the CDH17/TRAILR2 binding molecules according to the invention have clear advantages over the molecules known in the art and can be used to treat cancer, including colorectal cancer (CRC), gastric cancer (GC) and pancreatic cancer (PCa). ).

In addition, the ability of CDH17 to enhance the agonist effect of the TRAIL receptor binding molecule to TRAILR2 is not common to all cell surface proteins. This is known because the inventors investigated whether CD44v6 might also be a suitable anchor target. CD44v6 is a spliced variant of the ubiquitously expressed CD44 surface glycoprotein, and is known to be a tumor-associated antigen with a preferred expression pattern in tumors over normal tissues. However, as shown in the accompanying Examples, CD44v6 does not function as an anchor protein because it does not enhance the agonist effect of the TRAIL receptor binding molecule. Therefore, the usefulness of CDH17 as an anchor target is not predictable simply based on the protein expression profile.

In a first aspect of the invention, there is provided a binding molecule having at least one antigen-binding site (first antigen-binding site) that specifically binds to a TNF-associated apoptosis-inducing receptor 2 ligand (TRAILR2) and at least one antigen-binding site (second antigen-binding site) that binds specifically to cadherin-17 (CDH17).

As stated above, prior to the present invention, it has not been disclosed or even partly expected to provide binding molecules that can specifically bind to TRAILR2 and CDH17. However, individually, each protein and its associated genes are known in the art and are well represented in biological databases.

For the avoidance of doubt, by TNF-associated apoptosis-inducing receptor 2 ligand (TRAILR2), the inventors mean the human protein shown in UniProt O14763 http://www.uniprot.org/uniprot/O14763 and the nucleic acid sequence encoding this protein.

For the avoidance of doubt, by cadherin-17 (CDH17) the inventors mean the human protein presented in UniProt Q12864 http://www.uniprot.org/uniprot/Q12864 and the nucleic acid sequence encoding this protein.

The present invention relates to binding molecules that have binding specificities for at least two different targets. In relation to the present invention, binding molecules are derived from antibodies. Techniques for making binding molecules include, but are not limited to, recombinant co-expression of two immunoglobulin heavy chain and light chain pairs having different specificities (see Milstein and Cuello, Nature 305:537 (1983)), WO 93/08829 and Traunecker et al. , EMBO J. 10: 3655 (1991)), and the ridge-to-bottom technique (see, for example, US Pat. No. 5,731,168). Binding molecules in accordance with the invention can also be obtained by constructing proteins using the effect of electrostatic interaction to obtain Fc-heterodimeric antibody molecules (WO 2009/089004A1); crosslinking two or more antibodies or fragments (see, for example, US patent No. 4676980 and Brennan et al., Science, 229: 81 (1985)); the use of leucine zippers to generate bispecific antibodies (see, for example, Kostelny et al., Immunol., 148(5):1547-1553 (1992)); using diabody technology to generate bispecific antibody fragments (see, for example, Hollinger et al., Proc. Natl. Acad. Sci. USA, 90:6444-6448 (1993)); and the use of single chain Fv dimers (sFv) (see, for example, Gruber et al.,/Immunol., 152:5368 (1994)); and by making trispecific antibodies as described, for example, in Tutt et al.,/Immunol. 147:60 (1991).

Terms not specifically defined in the present invention should have the meanings to be given to them by a person skilled in the art in light of the disclosure and context. However, as used in the specification, unless otherwise indicated, the following terms have the meanings indicated and the following conventions are followed.

As used herein, the term antigen binding site includes a heavy chain variable domain (VH) and a light chain variable domain (V _L ) derived from an antibody. In this case, each variable domain contains 3 CDRs. In one aspect, the antigen-binding site according to the present invention, or certain parts of the protein, as a rule, is obtained from an antibody. The generalized structure of antibodies or immunoglobulin molecules is well known to those skilled in the art.

Antibodies or immunoglobulin molecules (also known as immunoglobulins, Ig for short) are gamma globulin proteins that can be found in the blood or other bodily fluids of vertebrates and are used by the immune system to identify and neutralize foreign objects such as bacteria and viruses. They are usually composed of basic structural units - each with two large heavy chains and two small light chains - to form, for example, single unit monomers, two unit dimers, or five unit pentamers. Antibodies can bind through non-covalent interaction with other molecules or structures, known as antigens. This binding is specific in the sense that the antibody will only bind to a particular structure with high affinity. The unique part of an antigen recognized by an antibody is called an epitope or antigenic determinant. The portion of an antibody that binds to an epitope is sometimes referred to as the paratope and is located in the so-called variable domain or variable region (Fv) of the antibody. The variable domain includes three so-called complementarity-determining regions (CDRs) separated by framework regions.

- 11 041360 (FR).

In the context of this invention, reference to CDR is based on the CCG definition, also referred to as IMGT (Lefranc MP, Pommie C, Ruiz M, Giudicelli V, Foulquier E, Truong L, Thouvenin-Contet V, Lefranc G. IMGT unique numbering for immunoglobulin and T cell receptor variable domains and Ig superfamily V-like domains Dev Comp Immunol 2003 Jan 27(1):55-77 Giudicelli V, Brochet X, Lefranc MP IMGT/VQUEST: IMGT standardized analysis of the immunoglobulin (IG) and T cell receptor (TR) nucleotide sequences Cold Spring Harb Protoc 2011;2011(6):695-715 Alternative CDR definition based on Chothia (Chothia and Lesk, J. Mol. Biol. 1987, 196: 901-917) , together with Kabat (E.A. Kabat, T.T. Wu, H. Bilofsky, M. Reid-Miller and H. Perry, Sequence of Proteins of Immunological Interest, National Institutes of Health, Bethesda (1983)).

Used in the present invention, the terms variable domains or variable region or Fv mean each of the pair of light and heavy chains, which is directly involved in the binding of an antibody to an antigen. The light chain variable domain is abbreviated VL and the heavy chain variable domain is abbreviated VH. The light and heavy chain variable domains have the same general structure, and each domain contains four framework (FR) regions, the sequences of which are conserved, connected by three HVRs (or CDRs). The framework regions adopt a beta sheet conformation and the CDRs can form loops connecting the beta sheet structure. The CDRs in each strand are held in their three-dimensional structure by the framework regions and, together with the CDRs from the other strand, form an antigen-binding site. The CDR3 regions of the heavy and light chains of an antibody play a particularly important role in the binding specificity/affinity of the antibodies according to the invention and therefore constitute another object of the invention. The term constant domains or constant region as used in the present invention refers to the sum of antibody domains other than the variable region. Such constant domains and regions are well known in the art and are, for example, described in Kabat et al. (Sequence of proteins of immunological interest, US Public Health Services, NIH Bethesda, MD, Publication No. 91).

The Fc portion of an antibody is not directly involved in antibody-antigen binding, but exhibits various effector functions. The Fc portion of an antibody is a term well known to the person skilled in the art and is defined based on the cleavage of antibodies with papain. Depending on the amino acid sequence of the constant region of their heavy chains, antibodies or immunoglobulins are divided into classes: IgA, IgD, IgE, IgG and IgM. According to the heavy chain constant regions, the various classes of immunoglobulins are named α, δ, ε, γ, and μ, respectively. Some of these can be further divided into subclasses (isotypes), such as IgG1, IgG2, IgG3, and IgG4, IgA1 and IgA2. The Fc portion of an antibody is directly involved in ADCC (antibody dependent cell mediated cytotoxicity) and CDC (complement dependent cytotoxicity) based on complement activation, Clq binding and Fc receptor binding. Complement activation (CDC) is initiated by the binding of complement factor Clq to the Fc portion of most IgG antibody subclasses. Although the effect of the antibody on the complement system is conditional, binding to Clq is caused by certain binding sites in the Fc portion. Such binding sites are known in the art and are described, for example, in Boakle et al., Nature 282 (1975) 742-743, Lukas et al., J. Immunol. 127 (1981) 2555-2560, Brunhouse and Cebra, Mol. Immunol. 16 (1979) 907-917, Burton et al., Nature 288 (1980) 338-344, Thommesen et al., Mol. Immunol. 37 (2000) 995-1004, Idusogie et al., J. Immunol. 164 (2000) 4178-4184, Hezareh et al., J. Virology 75 (2001) 12161-12168, Morgan et al., Immunology 86 (1995) 319-324, EP 0307434. Such binding sites are, for example, L234, L235, D270, N297, E318, K320, K322, P331 and P329 (numbered according to the Kabat EC code, see below). The most important among these residues in mediating IgG1 Clq and Fcgamma receptor binding are L234 and L235 (Hezareh et al., J. Virology 75 (2001) 12161-12168). Antibodies of the IgG1 and IgG3 subclass usually show complement activation and binding of Clq and C3, while IgG2 and IgG4 do not activate the complement system and do not bind Clq and C3.

In addition, antibodies have been developed in the art and made into versatile tools in medicine and technology. Thus, in the context of the present invention, the terms antibody molecule or antibody (used herein as synonyms) include not only antibodies, as they may be found in nature, for example, including two light chains and two heavy chains, or simply two heavy chains, as in camelid species, but also encompass all molecules containing at least one paratope with antigen binding specificity and structural similarity to an immunoglobulin variable domain.

Thus, an antibody can encompass a monoclonal antibody, a human antibody, a humanized antibody, a chimeric antibody, an antibody fragment, in particular an Fv, Fab, Fab' or F(ab') ₂ fragment, a single chain antibody, in particular a single chain variable fragment (scFv), small-sized modular protein (SMIP) immunopharmaceutical, domain antibody, nanobody, diabody. An antibody may have an effector function, such as ADCC or CDC, which is usually mediated by the Fc portion (antibody constant region) of the antibody, or it may not have an effector function, for example, due to the absence of an Fc portion or the presence of a blocked, masked Fc portion, essentially, the Fc portion that is not recognized or insufficiently recognized by the immune

- 12 041360 cells or components of the immune system, such as the complement system. Monoclonal antibodies (mAbs) are monospecific antibodies that are identical in amino acid sequence. They can be obtained using hybridoma technology from a hybrid cell line (called a hybridoma) that is a clone of the fusion of a specific antibody-producing B cell with a myeloma (B cell cancer) cell (Kohler G, Milstein C. Continuous cultures of fused cells secreting antibody of predefined specificity Nature 1975;256:495-7.). Alternatively, monoclonal antibodies can be generated by recombinant expression in host cells (Norderhaug L, Olafsen T, Michaelsen TE, Sandlie I. (May 1997). Versatile vectors for transient and stable expression of recombinant antibody molecules in mammalian cells. J Immunol Methods 204 (1): 7787; see also below). A recombinant antibody or recombinant binding molecule is an antibody or binding molecule that has been produced using a recombinantly engineered host cell. It can be isolated or cleared.

For use in humans, it is often desirable to reduce the immunogenicity of antibodies originally derived from other species, such as the mouse. This can be done by constructing chimeric antibodies or by a process called humanization. In this context, a chimeric antibody is understood to mean an antibody containing a portion of a sequence (eg, variable domain) derived from one species (eg, mouse) fused to a portion of the sequence (eg, constant domains) derived from another species (eg, human). A humanized antibody is an antibody containing a variable domain originally derived from a non-human species in which certain amino acids have been mutated to make the overall sequence of that variable domain more similar to that of a human variable domain. Methods for chimerizing and humanizing antibodies are well known in the art (Billetta R, Lobuglio AF. Chimeric antibodies. Int Rev Immunol. 1993; 10(2-3): 165-76; Riechmann L, Clark M, Waldmann H, Winter G (1988 ) Reshaping human antibodies for therapy, Nature: 332:323).

A humanized antibody refers to an antibody containing amino acid residues from non-human hypervariable regions (HVRs) and amino acid residues from human FRs. In some embodiments, a humanized antibody will include substantially all of at least one, and typically two, variable domains in which all or substantially all of the HVRs (e.g., complementarity determining regions (CDRs)) correspond to domains of the antibody, non-human, and all or substantially all of the framework regions (FR) correspond to those of a human antibody. A humanized antibody may optionally comprise at least a portion of an antibody constant region derived from a human antibody. A humanized form of an antibody, such as a non-human antibody, refers to an antibody that has been humanized.

In addition, technologies have been developed to create antibodies based on sequences derived from the human genome, for example, by phage display or the use of transgenic animals (WO 90/05144; D. Marks, H.R. Hoogenboom, T.P. Bonnert, J. McCafferty, A.D. Griffiths and G. Winter (1991) By-passing immunization Human antibodies from V-gene libraries displayed on phage J. Mol. Biol., 222, 581-597; Knappik et al., J. Mol. Biol. 296: 57- 86, 2000; S. Carmen and L. Jermutus, Concepts in antibody phage display. Briefings in Functional Genomics and Proteomics 2002 1(2):189-203; Lonberg N, Huszar D. Human antibodies from transgenic mice. Int Rev Immunol. 1995; 13(1):65-93; Bruggemann M, Taussig MJ. Production of human antibody repertoires in transgenic mice. Curr Opin Biotechnol. 1997 Aug; 8(4):455-8.). Such antibodies are human antibodies in the context of the present invention.

The antibody may also include immunoglobulin fragments that retain antigen-binding properties, such as Fab, Fab' or F(ab') ₂ fragments. Such fragments can be obtained by fragmentation of immunoglobulins, for example, proteolytic cleavage or recombinant expression of such fragments. For example, immunoglobulin digestion can be performed using routine methods, such as using papain or pepsin (WO 94/29348). Digestion of antibodies with papain typically produces two identical antigen-binding fragments, so-called Fab fragments, each with one antigen-binding site and a residual Fc fragment. Treatment with pepsin gives F(ab') ₂ . In Fab molecules, each variable domain is fused to a constant domain of an immunoglobulin, preferably of human origin. Thus, the heavy chain variable domain can be attached to the CH1 domain (so-called Fd fragment), and the light chain variable domain can be fused to the CL domain. Fab molecules can be produced by recombinant expression of the appropriate nucleic acids in host cells, see below.

A number of technologies have been developed to place the variable domains of immunoglobulins, or molecules derived from such variable domains, in a different molecular context. They should also be considered as antibodies in accordance with the present invention. Typically, these antibody molecules are smaller than immunoglobulins and may contain a single amino acid chain or multiple amino acid chains. For example, a single chain variable fragment (scFv) is a fusion of the heavy and light chain variable regions of an immunoglobulin

- 13 041360 lines linked together with a short linker, usually serine (S) or glycine (G) (WO 88/01649; WO 91/17271; Huston et al.; International Reviews of Immunology, vol. 10, 1993, 195-217 ). Single domain antibodies or nanobodies contain an antigen binding site in a single Ig-like domain (WO 94/04678; WO 03/050531, Ward et al., Nature. 1989 Oct 12; 341(6242):544-6; Revets et al., Expert Opin Biol Ther 5(1):111-24, 2005). One or more single domain antibodies with binding specificity for the same or a different antigen may be linked together. Diabodies are bivalent antibody molecules consisting of two amino acid chains containing two variable domains (WO 94/13804, Holliger et al., Proc Natl Acad Sci USA. 1993 Jul 15;90(14):6444-8). Other examples of antibody-like molecules are antibodies of the immunoglobulin superfamily (IgSF; Srinivasan and Roeske, Current Protein Pept. Sci. 2005, 6(2):185-96). Another concept leads to a so-called small modular protein (SMIP) immunopharmaceutical that contains an Fv domain associated with single chain hinge and effector domains, lacking a constant CH1 domain (WO 02/056910).

With respect to the present invention, a first aspect of the invention relates to a binding molecule comprising at least one antigen-binding site that specifically binds to a TNF-associated apoptosis-inducing ligand receptor 2 (TRAILR2) and at least one antigen-binding site that specifically binds to cadherin- 17 (CDH17).

As used herein, the term binding or specific binding refers to the binding of an antibody or antigen-binding site (eg, in the binding molecule described herein) to an antigen epitope in an in vitro assay. Affinity is the interaction between one antigen-binding site on an antibody molecule and one epitope. It is expressed by the equilibrium association constant K _a =ko _n /k _of f or the equilibrium dissociation constant K _d =k _off /k _on .

An epitope is a region of an antigen that is bound by an antibody or antigen-binding fragment. The term epitope includes any polypeptide determinant capable of specifically binding to an antibody or antigen-binding fragment. In some embodiments, epitope determinants include reactive surface groupings of molecules such as amino acids, glycan side chains, phosphoryl, or sulfonyl, and in some embodiments, may have specific three-dimensional structural characteristics and/or specific charge characteristics. Conformational and non-conformational epitopes differ in that binding to the former, but not to the latter, is lost in the presence of denaturing solvents. As used herein, the terms binding and specific binding refer to the binding of an antibody or antigen-binding fragment to an antigen epitope in an in vitro assay, preferably a plasmon resonance assay (BIAcore®, GE-Healthcare Uppsala, Sweden) with purified wild-type antigen.

In one aspect, a binding molecule in accordance with the present invention binds to TRAILR2 or CDH17 target antigens with an affinity as determined, for example, using a surface plasmon resonance assay (Malmqvist M., Surface plasmon resonance for detection and measurement of antibody-antigen affinity and kinetics, Curr Opin Immunol. 1993 Apr;5(2):282-6.), with a KD value in the range of 1 pM to 100 μM, preferably 1 pM to 1 μM. Antibody affinity can also be measured using Kinetic Exclusion Assay (KinExA) technology (Darling, R.J., and Brault P-A., Kinetic exclusion assay technology: Characterization of Molecular Interactions. ASSAY and Drug Development Technologies. 2004, Dec 2(6): 647 -657).

The binding affinity of an antibody molecule can be increased by a process known as affinity maturation (Marks et al., 1992, Biotechnology 10:779-783; Barbas, et al., 1994, Proc. Nat. Acad. Sci, USA 91:3809 -3813; Shier et al., 1995, Gene 169:147-155). Therefore, affinity matured antibodies are also included in the present invention.

As used herein, the term binding or specific binding refers to the binding of an antibody to an antigen epitope in an in vitro assay, preferably a surface plasmon resonance assay (SPR, BIAcore, GE-Healthcare Uppsala, Sweden). Binding affinity is defined in terms of kon (rate constant for association of an antibody from an antibody/antigen complex), k _off (dissociation constant), and K _D (k _off /k _on ). Specific binding generally refers to the formation of a complex between a receptor molecule and its ligands. In the context of antibody-antigen binding, high affinity antibodies typically bind their target antigens with an affinity of 10 ⁻⁹ M or less.

In one embodiment, a binding molecule according to the present invention can induce TRAILR2 mediated apoptosis in one or more cancer cell types, such as the Colo205 colorectal cancer cell line, with greater than 50% inhibition of cell viability at a concentration of 1 nM or less, and even more preferably less than 0.01 nM.

In another embodiment, a binding molecule according to the present invention cannot induce TRAILR2 mediated apoptosis in CDH17-negative liver-derived cells, such as the HepG2 cell line, with inhibition of cell viability less than

- 14 041360 than 50% at concentrations up to 1 nM, or more preferably up to 10 nM and even more preferably up to 100 nM.

In a further preferred embodiment, at least one antigen-binding site that specifically binds to cadherin-17 (CDH17) is an immunoglobulin (Ig) molecule (having the usual Y-shaped structure of a full length antibody containing two heavy and two light chains) and at least one antigen-binding site that specifically binds to a TNF-associated apoptosis-inducing ligand receptor 2 (TRAILR2) comprises one or more scFv, scFab, Fab, or Fv-binding elements. Preferably, an antigen-binding site that specifically binds to a TNF-associated apoptosis-inducing receptor 2 ligand (TRAILR2) contains one or more scFv(ob).

A single chain Fv fragment (scFv) is a polypeptide comprising an antibody heavy chain variable domain (VH), a linker, and an antibody light chain variable domain (VL), wherein said antibody domains and said linker are in one of the following orders from N-terminal to C-terminal directions:

a) VH-linker-VL,

b) VL-linker-VH; and where the specified linker is a polypeptide from 15 to 25 amino acids in length, preferably 20 amino acids.

In addition, these single chain Fv fragments can be further stabilized by incorporating disulfide bonds between the VH and VL domains, within the VH domain, or within the VL domain by incorporating cysteine residues. The term N-terminus denotes the first amino acid of the polypeptide chain, while the term C-terminus denotes the last amino acid of the C-terminus of the polypeptide chain. Therefore, an embodiment of the invention is that one or more scFv(ob) contain additional cysteine residues for the formation of disulfide bonds.

In one embodiment of the invention, the stability of the scFv fragment can be increased by including two cysteine residues in close three-dimensional proximity to form a disulfide bond within the scFv (referred to in this application as scFvss). If the scFv is derived from TRv1 V region sequences (as described below), then exemplary potential sites where such stabilizing disulfide bonds could be constructed include:

(a) between VL position 99 and VH position 45, (b) between VL position 102 and VH position 44, (c) between VL positions 4 and 100, and (d) between VH positions 6 and 112.

To ensure stabilization by engineered disulfide bonds, the residues at these positions are preferably replaced by cysteine residues.

As shown in the accompanying examples, the inventors have shown that scFv TRAILR2, having an N- to C-terminal VL-VH orientation, can function in the binding molecules of the invention to induce apoptosis in target cells. While TRAILR2 scFv having an N- to C-terminal VH-VL orientation may also function, activity may be reduced in this orientation. Therefore, a preferred embodiment of the invention is one in which the order is VL-VH from the N- to the C-terminus.

Another preferred embodiment of the invention is one in which one or more scFv(ob) specific to TRAILR2 are fused to an Ig molecule (e.g. IgG1, IgG1(KO), human IgG1FcRnmut, IgG4Pro) specifically binding to CDH17 via a linker, preferably a peptide linker having a length of about 4 to 20 amino acids (eg, any of 6, 9, 12, or 15). Preferably the scFv is fused to the C-terminus of the heavy chain of the Ig molecule. Preferably the Ig molecule is IgG.

Methods for linking scFv molecules to the C-terminus of the heavy chain of an IgG molecule or linking variable domains in scFv molecules are well known in the art. Typically, a small linker sequence of the amino acids glycine and serine (called a GS mini-linker) is used. The number of amino acids in the linker can vary from 4 (GGGS) (SEQ ID NO: 144), 6 (GGSGGS) (SEQ ID NO: 141), 10 (GGGGSGGGGS) (SEQ ID NO: 142) or more. In practice, typically a linker is formed by combining a nucleic acid molecule encoding the IgG of interest (which in this case would include a nucleic acid encoding a heavy chain variable domain for the CDH17 binding site and IgG type constant domains) with a nucleic acid encoding the desired scFv (which in this case will include a nucleic acid encoding a heavy and light chain variable domain, either in the VL-VH or VH-VL orientation for the TRAILR2 binding site), interspersed with a nucleic acid molecule encoding a linker sequence (e.g., a GS mini-linker of any of 5, 10, 15 or 20 amino acids, preferably a linker of SEQ ID NO: 143). Then, as further explained below, this complete HC-scFv-encoding nucleic acid molecule is placed in an expression vector and introduced into the appropriate host cells such that a complete single IgG scFv-scFv heavy chain polypeptide is formed.

Preferably, the GS mini-linker between the scFV molecule and the heavy chain C-terminus of the IgG molecule is GGSGGS (SEQ ID NO: 141).

In one embodiment, the present invention relates to a binding molecule, which is a multispecific binding protein comprising (i) an Ig molecule that specifically binds to CDH17 with two heavy and two light chains, (ii) two scFv molecules (scFv(ob)), each of which binds specifically to TRAILR2.

Preferably, each heavy chain of the Ig molecule has one scFv fused to its C-terminus, thereby forming a bispecific tetravalent binding protein.

In one embodiment, the present invention relates to a binding molecule (also referred to in the present invention as a multispecific binding protein or a modified Ig molecule) having:

(i) two heavy chains, each containing N- to C-terminus:

heavy chain variable domain specific for CDH17 (eg, murine, humanized, or human VH domain) IgG constant domains (eg, human IgG1 or IgG4) peptidic linker (eg, GS mini-linker) and scFv specific for TRAILR2 (eg, scFv N- to C-terminal VH domain (eg, murine, humanized, or human VH domain), linker, and VL domain (eg, murine, humanized, or human VL domain), or vice versa, VL domain, linker, and VH domain); and (ii) two light chains, each containing an N to a C terminus:

a light chain variable domain specific for CDH17 (eg murine, humanized or human VL domain), a light chain constant domain (eg human kappa chain).

An antibody molecule or binding molecule as described herein may be fused (as a fusion protein) or otherwise linked (via covalent or non-covalent bonds) to other molecular entities that have the desired effect on the properties of the antibody molecule. For example, it may be desirable to improve the pharmacokinetic properties of the antibodies or binding molecules described in the present invention, stability, for example, in body fluids such as blood, in particular in the case of single chain antibodies or domain antibodies. In this regard, a number of technologies have been developed, in particular to extend the half-life of such antibody molecules in the circulation, such as pegylation (WO 98/25971; WO 98/48837; WO 2004081026), fusion or covalent attachment of an antibody molecule to another antibody molecule, having an affinity for a serum protein such as albumin (WO 2004041865; WO 2004003019), or expressing the antibody molecule as a fusion protein with all or part of a serum protein such as albumin or transferrin (WO 01/79258).

Because the Fc region of an antibody interacts with a number of Fc receptors, resulting in a number of important functionalities (called effector functions), in some embodiments, the antibody is a full-length antibody or an antibody that contains a portion of the Fc region, the latter, as the antibody exhibits specific binding both to the corresponding part of the antigen and to Fc receptors and complements. The choice of the type and length of the constant region depends on whether effector functions such as complement fixation or antibody-dependent cell-mediated cytotoxicity are desirable features and on the desired pharmacological properties of the antibody protein.

In one embodiment of the invention, binding to a Clq complement product or Fc-gamma receptor binding molecule in the present invention is abolished by using an IgG4 constant region or an IgG1 constant region with L-A site-directed mutagenesis at positions 234 and 235.

In one embodiment of the invention, the binding molecule according to the invention may have an Fc region, or a corresponding portion thereof, which has been designed to avoid inadvertent crosslinking by soluble Fc gamma receptors or the Clq complement. In one embodiment, such a binding molecule or antibody variant has a much lower affinity for Fc-gamma receptors and Clq complement than the parent antibody. (Hereinafter, unless otherwise indicated, the term parent in the context of an antibody molecule or in the context of an IgG or Fc region refers to the unengineered antibody molecule, Fc region or IgG, respectively, from which the mutated (engineered) molecule is derived). Therefore, one embodiment of the invention is that the Ig molecule contains an Fc variant having reduced affinity for Fc gamma receptors or complement receptors, or both, compared to the wild-type Fc region. Such an Ig molecule is referred to in the present invention as IgG1(KO).

Another embodiment of the invention is that the binding molecule according to the invention contains an Fc region, or a corresponding region thereof, which has been designed to modify serum levels (half-life) by optimizing its interaction with the neonatal Fc receptor (FcRn), e.g. , a point mutation in the CH2 domain at position H31OA). Such an Ig molecule is referred to in the present invention as IgG1FcRnmut.

- 16 041360

Another embodiment of the invention is that the binding molecule contains an Ig molecule that contains an IgG4 variant in the hinge region that eliminates the exchange of heavy chains with other IgG4 molecules. Such an Ig molecule is referred to here as IgG4Pro.

The present invention provides a binding molecule having at least one antigen-binding site that specifically binds to a TNF-associated apoptosis-inducing ligand receptor 2 (TRAILR2) and at least one antigen-binding site that specifically binds to cadherin-17 (CDH17).

Methods for obtaining binding sites that bind to specific target antigens are well known in the art. One of skill in the art can easily use these methods to design an antigen-binding site having the desired specificity for TRAILR2 or CDH17 target antigens.

Methods for generating antibodies and antibody fragments are well known in the art. For example, antibodies can be generated using any of several methods that utilize in vivo induction of antibody molecule production, screening of immunoglobulin libraries (Orlandi et al., 1989. Proc. Natl. Acad. Sci. U.S.A. 86:3833-3837; Winter et al., 1991, Nature 349:293-299) or generation of monoclonal antibody molecules by cell lines in culture. These include, but are not limited to, the hybridoma technique, the human B-cell hybridoma technique, and the Epstein-Barr virus (EBV) hybridoma technique (Kohler et al., 1975. Nature 256:4950497; Kozbor et al., 1985. J. Immunol Cote et al., 1983. Proc. Natl. Acad. Sci. USA 80:2026-2030; Cole et al., 1984. Mol. Cell. Biol. 62:109-120).

Using methods known in the art and described herein, it would be common for one skilled in the art to generate antibodies having a binding site with the required specificity for the TRAILR2 and/or CDH17 target antigens and binding molecules described herein. Isolation of binding domains from such antibodies is common practice, and indeed, additional information on the methods that can be used to obtain antibodies and binding molecules, as described herein, is presented in the attached examples.

The authors of the present invention have received binding TRAILR2/CDH17 molecules in accordance with the invention, which are discussed in the attached examples.

The inventors have made a number of antigen-binding sites specific for TRAILR2 and named them TR#1, TR#2, TR#3, TR#7, TR#8, TR#10, TR#12, TR2v1, TR2v2, TR2v3, TR2v4, TR2v5 , TR2v6 and TR2v7. The inventors also made a number of antigen-binding sites specific for CDH17 and named them CDH17A6, CDH17E2, CDH17H2, CDH17E9 and CDH17v1. In any of the embodiments below, the binding molecule comprises at least one antigen-binding site that specifically binds to CDH17 (any of CDH17A6, CDH17E2, CDH17H2, CDH17E9, and CDH17v1) and at least one antigen-binding site that specifically binds to TRAILR2 ( any of TR#1, TR#2, TR#3, TR#7, TR#8, TR#10, TR#12, TR2v1, TR2v2, TR2v3, TR2v4, TR2v5, TR2v6 and TR2v7). Preferably, at least one antigen-binding site specific for CDH17 is an Ig molecule (consisting of two heavy and two light chains) and at least one antigen-binding site specifically binding to TRAILR2 contains two scFv(s)), where one scFv bound to the C-terminus of one of the heavy chains; and one scFv is bound to the C-terminus of the other heavy chain of the Ig molecule, forming a bispecific and tetravalent binding protein.

In some embodiments, the antigen binding site (any of TR#1, TR#2, TR#3, TR#7, TR#8, TR#10, TR#12, TR2v1, TR2v2, TR2v3, TR2v4, TR2v5, TR2v6, TR2v7 , CDH17A6, CDH17E2, CDH17H2, CDH17E9, and CDH17v1) is a humanized antigen binding site (e.g., containing a humanized VH/VL domain) containing amino acid residues from non-human hypervariable regions (HVRs; e.g. human framework sequences. In some embodiments, an antigen binding site (any of TR#1, TR#2, TR#3, TR#7, TR#8, TR#10, TR#12, TR2v1, TR2v2, TR2v3, TR2v4, TR2v5, TR2v6, TR2v7, CDH17A6, CDH17E2, CDH17H2, CDH17E9, and CDH17v1) is a human antigen binding site (eg, containing a human VH/VL domain) comprising CDR and FR sequences, both derived from human genome sequences.

The amino acid sequences of specific antigen-binding sites are given in the description (Table 3) and the sequence listing.

Below are the details of preferred embodiments of the invention, which include specific antigen-binding sites for TRAILR2 and/or CDH17.

For the avoidance of doubt, each of the specific embodiments listed below for the first aspect of the invention may also be considered an independent aspect of the invention.

In a preferred embodiment of the binding molecule of the invention, the antigen-binding site specific for TRAILR2 comprises heavy chain CDRs comprising the amino acid sequences of SEQ ID NO: 1 (CDR1), SEQ ID NO: 2 (CDR2), and SEQ ID NO:

(CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO: 4 (CDR1),

SEQ ID NO: 5 (CDR2) and SEQ ID NO: 6 (CDR3). In this embodiment, the antigen-binding site specific for TRAILR2 is TR#1.

Preferably, the antigen-binding site specific for TRAILR2 comprises a heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 82 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 83. In this embodiment, the antigen-binding site specific for TRAILR2 is TR#1. For example, an antigen-binding site specific for TRAILR2 contains an scFv molecule containing the amino acid sequence of SEQ ID NO: 134 (VL-VH orientation).

In a preferred embodiment of the binding molecule of the invention, the antigen-binding site specific for TRAILR2 comprises heavy chain CDRs comprising the amino acid sequences of SEQ ID NO: 7 (CDR1), SEQ ID NO: 8 (CDR2), and SEQ ID NO: 9 ( CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO: 10 (CDR1), SEQ ID NO: 11 (CDR2) and SEQ ID NO: 12 (CDR3). In this embodiment, the antigen-binding site specific for TRAILR2 is TR#2.

Preferably, the antigen-binding site specific for TRAILR2 comprises a heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 84 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 85. In this embodiment, the antigen-binding site specific for TRAILR2 is TR#2. For example, an antigen-binding site specific for TRAILR2 contains an scFv molecule containing the amino acid sequence of SEQ ID NO: 135 (VL-VH orientation).

In a preferred embodiment of the binding molecule of the invention, the antigen-binding site specific for TRAILR2 comprises heavy chain CDRs comprising the amino acid sequences of SEQ ID NO: 13 (CDR1), SEQ ID NO: 14 (CDR2), and SEQ ID NO: 15 ( CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO: 16 (CDR1), SEQ ID NO: 17 (CDR2) and SEQ ID NO: 18 (CDR3). In this embodiment, the antigen-binding site specific for TRAILR2 is TR#3.

Preferably, the antigen-binding site specific for TRAILR2 comprises a heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 86 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 87. In this embodiment, the antigen-binding site specific for TRAILR2 is TR#3. For example, an antigen-binding site specific for TRAILR2 contains an scFv molecule containing the amino acid sequence of SEQ ID NO: 136 (VL-VH orientation).

In a preferred embodiment of the binding molecule of the invention, the antigen-binding site specific for TRAILR2 comprises heavy chain CDRs comprising the amino acid sequences of SEQ ID NO: 19 (CDR1), SEQ ID NO: 20 (CDR2), and SEQ ID NO: 21 ( CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO: 22 (CDR1), SEQ ID NO: 23 (CDR2) and SEQ ID NO: 24 (CDR3). In this embodiment, the antigen-binding site specific for TRAILR2 is TR#7.

Preferably, the antigen-binding site specific for TRAILR2 comprises a heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 88 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 89. In this embodiment, the antigen-binding site specific for TRAILR2 is TR#7. For example, an antigen-binding site specific for TRAILR2 contains an scFv molecule containing the amino acid sequence of SEQ ID NO: 137 (VL-VH orientation).

In a preferred embodiment of the binding molecule of the invention, the antigen-binding site specific for TRAILR2 comprises heavy chain CDRs comprising the amino acid sequences of SEQ ID NO: 25 (CDR1), SEQ ID NO: 26 (CDR2), and SEQ ID NO: 27 ( CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO: 28 (CDR1), SEQ ID NO: 29 (CDR2) and SEQ ID NO: 30 (CDR3). In this embodiment, the antigen-binding site specific for TRAILR2 is TR#8.

Preferably, the antigen-binding site specific for TRAILR2 comprises a heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 90 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 91. In this embodiment, the antigen-binding site specific for TRAILR2 is TR#8. For example, an antigen-binding site specific for TRAILR2 contains an scFv molecule containing the amino acid sequence of SEQ ID NO: 138 (VL-VH orientation).

In a preferred embodiment of the binding molecule of the invention, the antigen-binding site specific for TRAILR2 comprises heavy chain CDRs comprising the amino acid sequences of SEQ ID NO: 31 (CDR1), SEQ ID NO: 32 (CDR2), and SEQ ID NO: 33 ( CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO: 34

- 18 041360 (CDR1), SEQ ID NO: 35 (CDR2) and SEQ ID NO: 36 (CDR3). In this embodiment, the antigen-binding site specific for TRAILR2 is TR#10.

Preferably, the antigen-binding site specific for TRAILR2 comprises a heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 92 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 93. In this embodiment, the antigen-binding site specific for TRAILR2 is TR#10. For example, an antigen-binding site specific for TRAILR2 contains an scFv molecule containing the amino acid sequence of SEQ ID NO: 139 (VL-VH orientation).

In a preferred embodiment of the binding molecule of the invention, the antigen-binding site specific for TRAILR2 comprises heavy chain CDRs comprising the amino acid sequences of SEQ ID NO: 37 (CDR1), SEQ ID NO: 38 (CDR2), and SEQ ID NO: 39 ( CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO: 40 (CDR1), SEQ ID NO: 41 (CDR2) and SEQ ID NO: 42 (CDR3). In this embodiment, the antigen-binding site specific for TRAILR2 is TR#12.

Preferably, the antigen-binding site specific for TRAILR2 comprises a heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 94 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 95. In this embodiment, the antigen-binding site specific for TRAILR2 is TR#12. For example, an antigen-binding site specific for TRAILR2 contains an scFv molecule containing the amino acid sequence of SEQ ID NO: 140 (VL-VH orientation).

In a preferred embodiment of the binding molecule of the invention, the antigen-binding site specific for TRAILR2 comprises heavy chain CDRs comprising the amino acid sequences of SEQ ID NO: 43 (CDR1), SEQ ID NO: 44 (CDR2), and SEQ ID NO: 48 ( CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO: 49 (CDR1), SEQ ID NO: 50 (CDR2) and SEQ ID NO: 54 (CDR3). In this embodiment, the antigen-binding site specific for TRAILR2 is TR2v1 or TR2v2.

Preferably, the antigen binding site specific for TRAILR2 comprises a heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 96 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 97.

In this embodiment, the antigen-binding site specific for TRAILR2 is TR2v1. For example, an antigen-binding site specific for TRAILR2 contains an scFv molecule containing the amino acid sequence SEQ ID NO: 124 (VL-VH orientation), SEQ ID NO: 131 (VH-VL orientation), SEQ ID NO: 132 (VH-VL orientation). with disulfide bonds, scFvss) or SEQ ID NO: 133 (VL-VH orientation with disulfide bonds, scFV).

Preferably, the antigen-binding site specific for TRAILR2 comprises a heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 98 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 99. In this embodiment, the antigen-binding site specific for TRAILR2 is TR2v2. For example, an antigen-binding site specific for TRAILR2 contains an scFv molecule containing the amino acid sequence of SEQ ID NO: 125 (VL-VH orientation).

In a preferred embodiment of the binding molecule of the invention, the antigen-binding site specific for TRAILR2 comprises heavy chain CDRs comprising the amino acid sequences of SEQ ID NO: 43 (CDR1), SEQ ID NO: 45 (CDR2) and SEQ ID NO: 48 ( CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO: 49 (CDR1), SEQ ID NO: 51 (CDR2) and SEQ ID NO: 55 (CDR3). In this embodiment, the antigen-binding site specific for TRAILR2 is TR2v3.

Preferably, the antigen-binding site specific for TRAILR2 comprises a heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 100 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 101. In this embodiment, the antigen-binding site specific for TRAILR2 is TR2v3. For example, an antigen-binding site specific for TRAILR2 contains an scFv molecule containing the amino acid sequence of SEQ ID NO: 126 (VL-VH orientation).

In a preferred embodiment of the binding molecule of the invention, the antigen-binding site specific for TRAILR2 comprises heavy chain CDRs comprising the amino acid sequences of SEQ ID NO: 43 (CDR1), SEQ ID NO: 45 (CDR2) and SEQ ID NO: 48 ( CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO: 49 (CDR1), SEQ ID NO: 52 (CDR2) and SEQ ID NO: 56 (CDR3). In this embodiment, the antigen-binding site specific for TRAILR2 is TR2v4.

Preferably, the antigen-binding site specific for TRAILR2 comprises a heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 102 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 103. In this embodiment, the antigen-binding site specific for TRAILR2 is

- 19 041360 a TR2v4. For example, an antigen-binding site specific for TRAILR2 contains an scFv molecule containing the amino acid sequence of SEQ ID NO: 127 (VL-VH orientation).

In a preferred embodiment of the binding molecule of the invention, the antigen-binding site specific for TRAILR2 comprises heavy chain CDRs comprising the amino acid sequences of SEQ ID NO: 43 (CDR1), SEQ ID NO: 45 (CDR2), and SEQ ID NO: 48 ( CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO: 49 (CDR1), SEQ ID NO: 50 (CDR2) and SEQ ID NO: 56 (CDR3). In this embodiment, the antigen-binding site specific for TRAILR2 is TR2v5.

Preferably, the antigen-binding site specific for TRAILR2 comprises a heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 104 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 105. In this embodiment, the antigen-binding site specific for TRAILR2 is TR2v5. For example, an antigen-binding site specific for TRAILR2 contains an scFv molecule containing the amino acid sequence of SEQ ID NO: 128 (VL-VH orientation).

In a preferred embodiment of the binding molecule of the invention, the antigen-binding site specific for TRAILR2 comprises heavy chain CDRs comprising the amino acid sequences of SEQ ID NO: 43 (CDR1), SEQ ID NO: 46 (CDR2), and SEQ ID NO: 48 ( CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO: 49 (CDR1), SEQ ID NO: 51 (CDR2) and SEQ ID NO: 57 (CDR3). In this embodiment, the antigen-binding site specific for TRAILR2 is TR2v6.

Preferably, the antigen-binding site specific for TRAILR2 comprises a heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 106 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 107. In this embodiment, the antigen-binding site specific for TRAILR2 is TR2v6. For example, an antigen-binding site specific for TRAILR2 contains an scFv molecule containing the amino acid sequence of SEQ ID NO: 129 (VL-VH orientation).

In a preferred embodiment of the binding molecule of the invention, the antigen-binding site specific for TRAILR2 comprises heavy chain CDRs comprising the amino acid sequences of SEQ ID NO: 43 (CDR1), SEQ ID NO: 47 (CDR2), and SEQ ID NO: 48 ( CDR3) and has light chain CDRs containing the amino acid sequences of SEQ ID NO: 49 (CDR1), SEQ ID NO: 53 (CDR2) and SEQ ID NO: 57 (CDR3). In this embodiment, the antigen-binding site specific for TRAILR2 is TR2v7.

Preferably, the antigen-binding site specific for TRAILR2 comprises a heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 108 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 109. In this embodiment, the antigen-binding site specific for TRAILR2 is TR2v7. For example, an antigen-binding site specific for TRAILR2 contains an scFv molecule containing the amino acid sequence of SEQ ID NO: 130 (VL-VH orientation).

In a preferred embodiment of the binding molecule of the invention, the antigen-binding site specific for CDH17 comprises heavy chain CDRs comprising the amino acid sequences of SEQ ID NO: 58 (CDR1), SEQ ID NO: 59 (CDR2), and SEQ ID NO: 60 ( CDR3) and has light chain CDRs containing the amino acid sequences of SEQ ID NO: 61 (CDR1), SEQ ID NO: 62 (CDR2) and SEQ ID NO: 63 (CDR3). In this embodiment, the CDH17 specific antigen binding site is CDH17E9.

Preferably, the antigen-binding site specific for CDH17 comprises a heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 110 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 111. In this embodiment, the antigen-binding site specific for CDH17 is CDH17E9.

In a preferred embodiment of the binding molecule of the invention, the antigen-binding site specific for CDH17 comprises heavy chain CDRs comprising the amino acid sequences of SEQ ID NO: 64 (CDR1), SEQ ID NO: 65 (CDR2), and SEQ ID NO: 66 ( CDR3), and has light chain CDRs containing the amino acid sequences of SEQ ID NO: 67 (CDR1), SEQ ID NO: 68 (CDR2) and SEQ ID NO: 69 (CDR3). In this embodiment, the CDH17 specific antigen binding site is CDH17A6.

Preferably, the antigen binding site specific for CDH17 comprises a heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 112 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 113. In this embodiment, the antigen binding site specific for CDH17 is is CDH17A6.

In a preferred embodiment of the binding molecule of the invention, the antigen-binding site specific for CDH17 comprises heavy chain CDRs comprising the amino acid sequences of SEQ ID NO: 70 (CDR1), SEQ ID NO: 71 (CDR2), and SEQ ID NO:

(CDR3) and has light chain CDRs containing the amino acid sequences of SEQ ID NO: 73 (CDR1), SEQ ID NO: 74 (CDR2) and SEQ ID NO: 75 (CDR3). In this embodiment, the CDH17 specific antigen binding site is CDH17E2 or CDH17H2.

Preferably, the antigen-binding site specific for CDH17 comprises a heavy chain variable domain containing the amino acid sequence of SEQ ID NO:114 and a light chain variable domain containing the amino acid sequence of SEQ ID NO:115. In this embodiment, the antigen-binding site specific for CDH17 is CDH17E2.

Preferably, the antigen-binding site specific for CDH17 comprises a heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 116 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 117. In this embodiment, the antigen-binding site specific for CDH17 is CDH17H2.

In a preferred embodiment of the binding molecule of the invention, the antigen-binding site specific for CDH17 comprises heavy chain CDRs comprising the amino acid sequences of SEQ ID NO: 76 (CDR1), SEQ ID NO: 77 (CDR2), and SEQ ID NO: 78 ( CDR3), and has light chain CDRs containing the amino acid sequences of SEQ ID NO: 79 (CDR1), SEQ ID NO: 80 (CDR2) and SEQ ID NO: 81 (CDR3). In this embodiment, the antigen-binding site specific for CDH17 is CDH17v1.

Preferably, the antigen-binding site specific for CDH17 comprises a heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 118 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 119. In this embodiment, the antigen-binding site specific for CDH17 is is CDH17v1.

In a preferred embodiment of the binding molecule of the invention, the antigen-binding site specific for TRAILR2 comprises heavy chain CDRs comprising the amino acid sequences of SEQ ID NO: 1 (CDR1), SEQ ID NO: 2 (CDR2), and SEQ ID NO: 3 ( CDR3), and light chain CDRs containing the amino acid sequences of SEQ ID NO: 4 (CDR1), SEQ ID NO: 5 (CDR2), and SEQ ID NO: 6 (CDR3) and an antigen-binding site specific for CDH17 contains a heavy chain CDR, containing the amino acid sequences of SEQ ID NO: 58 (CDR1), SEQ ID NO: 59 (CDR2) and SEQ ID NO: 60 (CDR3), and light chain CDRs containing the amino acid sequences of SEQ ID NO: 61 (CDR1), SEQ ID NO : 62 (CDR2) and SEQ ID NO: 63 (CDR3). In this embodiment, the antigen-binding site specific for TRAILR2 is TR#1 and the antigen-binding site specific for CDH17 is CDH17E9.

Preferably, the antigen-binding site specific for TRAILR2 comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 82 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 83 and the antigen-binding site specific for CDH17 comprises a heavy chain variable domain, containing the amino acid sequence of SEQ ID NO: 110, and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 111. In this embodiment, the antigen-binding site specific for TRAILR2 is TR#1 (e.g., scFv SEQ ID NO: 134) and the antigen-binding site specific for CDH17 is CDH17E9.

In a preferred embodiment of the binding molecule of the invention, the antigen-binding site specific for TRAILR2 comprises heavy chain CDRs comprising the amino acid sequences of SEQ ID NO: 1 (CDR1), SEQ ID NO: 2 (CDR2), and SEQ ID NO: 3 ( CDR3), and light chain CDRs containing the amino acid sequences of SEQ ID NO: 4 (CDR1), SEQ ID NO: 5 (CDR2), and SEQ ID NO: 6 (CDR3) and an antigen-binding site specific for CDH17 contains a heavy chain CDR, containing the amino acid sequences of SEQ ID NO: 64 (CDR1), SEQ ID NO: 65 (CDR2) and SEQ ID NO: 66 (CDR3), and light chain CDRs containing the amino acid sequences of SEQ ID NO: 67 (CDR1), SEQ ID NO : 68 (CDR2) and SEQ ID NO: 69 (CDR3). In this embodiment, the antigen-binding site specific for TRAILR2 is TR#1 and the antigen-binding site specific for CDH17 is CDH17A6.

Preferably, the antigen-binding site specific for TRAILR2 comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 82 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 83, and the antigen-binding site specific for CDH17 comprises a heavy chain variable domain. chain containing the amino acid sequence of SEQ ID NO: 112; and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 113. In this embodiment, the antigen-binding site specific for TRAILR2 is TR#1 (e.g., scFv SEQ ID NO: 134) and the antigen-binding site specific for CDH17 is

- 21 041360

CDH17A6.

In a preferred embodiment of the binding molecule of the invention, the antigen-binding site specific for TRAILR2 comprises heavy chain CDRs comprising the amino acid sequences of SEQ ID NO: 1 (CDR1), SEQ ID NO: 2 (CDR2), and SEQ ID NO: 3 ( CDR3), and light chain CDRs containing the amino acid sequences of SEQ ID NO: 4 (CDR1), SEQ ID NO: 5 (CDR2), and SEQ ID NO: 6 (CDR3) and an antigen-binding site specific for CDH17 contains a heavy chain CDR, containing the amino acid sequences of SEQ ID NO: 70 (CDR1), SEQ ID NO: 71 (CDR2) and SEQ ID NO: 72 (CDR3), and light chain CDRs containing the amino acid sequences of SEQ ID NO: 73 (CDR1), SEQ ID NO : 74 (CDR2) and SEQ ID NO: 75 (CDR3). In this embodiment, the antigen-binding site specific for TRAILR2 is TR#1 and the antigen-binding site specific for CDH17 is CDH17E2 or CDH17H2.

Preferably, the antigen-binding site specific for TRAILR2 comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 82 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 83 and the antigen-binding site specific for CDH17 comprises a heavy chain variable domain , containing the amino acid sequence of SEQ ID NO: 114, and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 115. In this embodiment, the antigen-binding site specific for TRAILR2 is TR#1 (for example, scFv SEQ ID NO: 134 ) and the antigen-binding site specific for CDH17 is CDH17E2.

Preferably, the antigen binding site specific for TRAILR2 comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 82 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 83 and the antigen binding site specific for CDH17 comprises a heavy chain variable domain , containing the amino acid sequence of SEQ ID NO: 116, and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 117. In this embodiment, the antigen-binding site specific for TRAILR2 is TR#1 (for example, scFv SEQ ID NO: 134 ), and the antigen-binding site specific for CDH17 is CDH17H2.

In a preferred embodiment of the binding molecule of the invention, the antigen-binding site specific for TRAILR2 comprises heavy chain CDRs comprising the amino acid sequences of SEQ ID NO: 1 (CDR1), SEQ ID NO: 2 (CDR2), and SEQ ID NO: 3 ( CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO: 4 (CDR1), SEQ ID NO: 5 (CDR2), and SEQ ID NO: 6 (CDR3), and an antigen-binding site specific for CDH17 contains a heavy chain CDR, containing the amino acid sequences of SEQ ID NO: 76 (CDR1), SEQ ID NO: 77 (CDR2) and SEQ ID NO: 78 (CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO: 79 (CDR1), SEQ ID NO: 80 (CDR2) and SEQ ID NO: 81 (CDR3). In this embodiment, the antigen-binding site specific for TRAILR2 is TR#1 and the antigen-binding site specific for CDH17 is CDH17v1.

Preferably, the antigen-binding site specific for TRAILR2 comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 82 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 83, and the antigen-binding site specific for CDH17 comprises a heavy chain variable domain. chain containing the amino acid sequence of SEQ ID NO: 118; and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 119. In this embodiment, the antigen-binding site specific for TRAILR2 is TR#1 (e.g., scFv SEQ ID NO: 134) and the antigen-binding site specific for CDH17 is CDH17v1.

In a preferred embodiment of the binding molecule of the invention, the antigen-binding site specific for TRAILR2 comprises heavy chain CDRs comprising the amino acid sequences of SEQ ID NO: 7 (CDR1), SEQ ID NO: 8 (CDR2), and SEQ ID NO: 9 ( CDR3), and light chain CDRs containing the amino acid sequences of SEQ ID NO: 10 (CDR1), SEQ ID NO: 11 (CDR2), and SEQ ID NO: 12 (CDR3 and antigen-binding site specific for CDH17 contain heavy chain CDRs containing amino acid sequences of SEQ ID NO: 58 (CDR1), SEQ ID NO: 59 (CDR2) and SEQ ID NO: 60 (CDR3), and has light chain CDRs containing the amino acid sequences of SEQ ID NO: 61 (CDR1), SEQ ID NO : 62 (CDR2) and SEQ ID NO: 63 (CDR3) In this embodiment, the TRAILR2 specific antigen binding site is TR#2 and the CDH17 specific antigen binding site is CDH17E9.

Preferably, the antigen-binding site specific for TRAILR2 comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 84 and wa

- 22 041360 light chain ribable domain containing the amino acid sequence of SEQ ID NO: 85, and an antigen-binding site specific for CDH17, contains a heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 110, and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 111. In this embodiment, the TRAILR2 specific antigen binding site is TR#2 (eg, scFv SEQ ID NO: 135) and the CDH17 specific antigen binding site is CDH17E9.

In a preferred embodiment of the binding molecule of the invention, the antigen-binding site specific for TRAILR2 comprises heavy chain CDRs comprising the amino acid sequences of SEQ ID NO: 7 (CDR1), SEQ ID NO: 8 (CDR2), and SEQ ID NO: 9 ( CDR3), and light chain CDRs containing the amino acid sequences of SEQ ID NO: 10 (CDR1), SEQ ID NO: 11 (CDR2), and SEQ ID NO: 12 (CDR3 and antigen-binding site specific for CDH17 contain heavy chain CDRs containing amino acid sequences of SEQ ID NO: 64 (CDR1), SEQ ID NO: 65 (CDR2) and SEQ ID NO: 66 (CDR3), and light chain CDRs containing the amino acid sequences of SEQ ID NO: 67 (CDR1), SEQ ID NO: 68 (CDR2) and SEQ ID NO: 69 (CDR3) In this embodiment, the TRAILR2 specific antigen binding site is TR#2 and the CDH17 specific antigen binding site is CDH17A6.

Preferably, the antigen-binding site specific for TRAILR2 comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 84 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 85, and the antigen-binding site specific for CDH17 comprises a heavy chain variable domain. chain containing the amino acid sequence of SEQ ID NO: 112; and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 113. In this embodiment, the antigen-binding site specific for TRAILR2 is TR#2 (e.g., scFv SEQ ID NO: 135) and the antigen-binding site specific for CDH17 is CDH17A6.

In a preferred embodiment of the binding molecule of the invention, the antigen-binding site specific for TRAILR2 comprises heavy chain CDRs comprising the amino acid sequences of SEQ ID NO: 7 (CDR1), SEQ ID NO: 8 (CDR2), and SEQ ID NO: 9 ( CDR3), and light chain CDRs containing the amino acid sequences of SEQ ID NO: 10 (CDR1), SEQ ID NO: 11 (CDR2) and SEQ ID NO: 12 (CDR3) and an antigen-binding site specific for CDH17 contains a heavy chain CDR, containing the amino acid sequences of SEQ ID NO: 70 (CDR1), SEQ ID NO: 71 (CDR2) and SEQ ID NO: 72 (CDR3), and light chain CDRs containing the amino acid sequences of SEQ ID NO: 73 (CDR1), SEQ ID NO : 74 (CDR2) and SEQ ID NO: 75 (CDR3). In this embodiment, the antigen-binding site specific for TRAILR2 is TR#2 and the antigen-binding site specific for CDH17 is CDH17E2 or CDH17H2.

Preferably, the antigen-binding site specific for TRAILR2 comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 84 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 85, and the antigen-binding site specific for CDH17 comprises a heavy chain variable domain. chain containing the amino acid sequence of SEQ ID NO: 114; and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 115. In this embodiment, the antigen-binding site specific for TRAILR2 is TR#2 (e.g., scFv SEQ ID NO: 135), and the antigen-binding site specific for CDH17 is CDH17E2.

Preferably, the antigen-binding site specific for TRAILR2 comprises a heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 84 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 85, and the antigen-binding site specific for CDH17 comprises a heavy chain variable domain. chain containing the amino acid sequence of SEQ ID NO: 116; and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 117. In this embodiment, the antigen-binding site specific for TRAILR2 is TR#2 (e.g., scFv SEQ ID NO: 135), and the antigen-binding site specific for CDH17 is CDH17H2.

In a preferred embodiment of the binding molecule of the invention, the antigen-binding site specific for TRAILR2 comprises heavy chain CDRs comprising the amino acid sequences of SEQ ID NO: 7 (CDR1), SEQ ID NO: 8 (CDR2), and SEQ ID NO: 9 ( CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO: 10 (CDR1), SEQ ID NO: 11 (CDR2), and SEQ ID NO: 12 (CDR3 and an antigen-binding site specific for CDH17 contain heavy chain CDRs containing amino acids SEQ ID NO: 76 (CDR1), SEQ ID NO: 77 (CDR2) and SEQ ID NO: 78 (CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO: 79 (CDR1), SEQ ID NO: 80 (CDR2) and SEQ ID NO: 81 (CDR3) In this embodiment, the TRAILR2 specific antigen binding site is TR#2 and the CDH17 specific antigen binding site is CDH17v1.

Preferably, the antigen-binding site specific for TRAILR2 comprises a heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 84 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 85, and the antigen-binding site specific for CDH17 comprises a heavy chain variable domain. chain containing the amino acid sequence of SEQ ID NO: 118; and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 119. In this embodiment, the antigen-binding site specific for TRAILR2 is TR#2 (e.g., scFv SEQ ID NO: 135) and the antigen-binding site specific for CDH17 is CDH17v1.

In a preferred embodiment of the binding molecule of the invention, the antigen-binding site specific for TRAILR2 comprises heavy chain CDRs comprising the amino acid sequences of SEQ ID NO: 13 (CDR1), SEQ ID NO: 14 (CDR2), and SEQ ID NO: 15 ( CDR3), and light chain CDRs containing the amino acid sequences of SEQ ID NO: 16 (CDR1), SEQ ID NO: 17 (CDR2), and SEQ ID NO: 18 (CDR3), and an antigen-binding site specific for CDH17 contains a heavy chain CDR containing the amino acid sequences of SEQ ID NO: 58 (CDR1), SEQ ID NO: 59 (CDR2) and SEQ ID NO: 60 (CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO: 61 (CDR1), SEQ ID NO : 62 (CDR2) and SEQ ID NO: 63 (CDR3). In this embodiment, the antigen-binding site specific for TRAILR2 is TR#3 and the antigen-binding site specific for CDH17 is CDH17E9.

Preferably, the antigen-binding site specific for TRAILR2 comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 86 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 87, and the antigen-binding site specific for CDH17 comprises a heavy chain variable domain. chain containing the amino acid sequence of SEQ ID NO: 110 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 111. In this embodiment, the antigen-binding site specific for TRAILR2 is TR#3 (e.g., scFv SEQ ID NO: 136 ) and the antigen-binding site specific for CDH17 is CDH17E9.

In a preferred embodiment of the binding molecule of the invention, the antigen-binding site specific for TRAILR2 comprises heavy chain CDRs comprising the amino acid sequences of SEQ ID NO: 13 (CDR1), SEQ ID NO: 14 (CDR2), and SEQ ID NO: 15 ( CDR3), and light chain CDRs containing the amino acid sequences of SEQ ID NO: 16 (CDR1), SEQ ID NO: 17 (CDR2), and SEQ ID NO: 18 (CDR3), and an antigen-binding site specific for CDH17 contains a heavy chain CDR containing the amino acid sequences of SEQ ID NO: 64 (CDR1), SEQ ID NO: 65 (CDR2) and SEQ ID NO: 66 (CDR3), and light chain CDRs containing the amino acid sequences of SEQ ID NO: 67 (CDR1), SEQ ID NO: 68 (CDR2) and SEQ ID NO: 69 (CDR3). In this embodiment, the antigen-binding site specific for TRAILR2 is TR#3 and the antigen-binding site specific for CDH17 is CDH17A6.

Preferably, the antigen-binding site specific for TRAILR2 comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 86 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 87 and the antigen-binding site specific for CDH17 comprises a heavy chain variable domain, containing the amino acid sequence of SEQ ID NO: 112 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 113. In this embodiment, the antigen-binding site specific for TRAILR2 is TR#3 (e.g., scFv SEQ ID NO: 136) and the antigen-binding site specific for CDH17 is CDH17A6.

In a preferred embodiment of the binding molecule of the invention, the antigen-binding site specific for TRAILR2 comprises heavy chain CDRs comprising the amino acid sequences of SEQ ID NO: 13 (CDR1), SEQ ID NO: 14 (CDR2), and SEQ ID NO: 15 ( CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO: 16 (CDR1), SEQ ID NO: 17 (CDR2) and SEQ ID NO: 18 (CDR3) and an antigen-binding site specific for CDH17 contains heavy chain CDRs containing amino acid sequences of SEQ ID NO: 70 (CDR1), SEQ ID NO: 71 (CDR2) and SEQ ID NO: 72 (CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO: 73 (CDR1), SEQ ID NO: 74 (CDR2) and SEQ ID NO: 75 (CDR3). In this embodiment, the TRAILR2 specific antigen binding site is TR#3 and the CDH17 specific antigen binding site is CDH17E2 or CDH17H2.

Preferably, the antigen-binding site specific for TRAILR2 comprises a heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 86 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 87, and the antigen-binding site specific for CDH17 comprises a heavy chain variable domain. chain containing the amino acid sequence of SEQ ID NO: 114; and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 115. In this embodiment, the antigen-binding site specific for TRAILR2 is TR#3 (e.g., scFv SEQ ID NO: 136) and the antigen-binding site specific for CDH17 is CDH17E2.

Preferably, the antigen-binding site specific for TRAILR2 comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 86 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 87 and the antigen-binding site specific for CDH17 comprises a heavy chain variable domain, containing the amino acid sequence of SEQ ID NO: 116, and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 117. In this embodiment, the antigen-binding site specific for TRAILR2 is TR#3 (e.g., scFv SEQ ID NO: 136) , and the antigen-binding site specific for CDH17 is CDH17H2.

In a preferred embodiment of the binding molecule of the invention, the antigen-binding site specific for TRAILR2 comprises heavy chain CDRs comprising the amino acid sequences of SEQ ID NO: 13 (CDR1), SEQ ID NO: 14 (CDR2), and SEQ ID NO: 15 ( CDR3), and light chain CDRs containing the amino acid sequences of SEQ ID NO: 16 (CDR1), SEQ ID NO: 17 (CDR2), and SEQ ID NO: 18 (CDR3), and an antigen-binding site specific for CDH17 contains a heavy chain CDR containing the amino acid sequences of SEQ ID NO: 76 (CDR1), SEQ ID NO: 77 (CDR2) and SEQ ID NO: 78 (CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO: 79 (CDR1), SEQ ID NO : 80 (CDR2) and SEQ ID NO: 81 (CDR3). In this embodiment, the antigen-binding site specific for TRAILR2 is TR#3 and the antigen-binding site specific for CDH17 is CDH17v1.

Preferably, the antigen-binding site specific for TRAILR2 comprises a heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 86 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 87, and the antigen-binding site specific for CDH17 comprises a heavy chain variable domain. chain containing the amino acid sequence of SEQ ID NO: 118 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 119. In this embodiment, the antigen-binding site specific for TRAILR2 is TR#3 (e.g., scFv SEQ ID NO: 136 ) and the antigen-binding site specific for CDH17 is CDH17v1.

In a preferred embodiment of the binding molecule of the invention, the antigen-binding site specific for TRAILR2 comprises heavy chain CDRs comprising the amino acid sequences of SEQ ID NO: 19 (CDR1), SEQ ID NO: 20 (CDR2), and SEQ ID NO: 21 ( CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO: 22 (CDR1), SEQ ID NO: 23 (CDR2) and SEQ ID NO: 24 (CDR3) and an antigen-binding site specific for CDH17 contains heavy chain CDRs containing amino acid sequences of SEQ ID NO: 58 (CDR1), SEQ ID NO: 59 (CDR2) and SEQ ID NO: 60 (CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO: 61 (CDR1), SEQ ID NO: 62 (CDR2) and SEQ ID NO: 63 (CDR3). In this embodiment, the antigen-binding site specific for TRAILR2 is TR#7 and the antigen-binding site specific for CDH17 is CDH17E9.

Preferably, the antigen-binding site specific for TRAILR2 comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 88 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 89 and the antigen-binding site specific for CDH17 comprises a heavy chain variable domain, containing the amino acid sequence of SEQ ID NO: 110 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 111. In this embodiment, the antigen-binding site specific for TRAILR2 is TR#7 (e.g., scFv SEQ ID NO: 137) and the antigen-binding site specific for CDH17 is CDH17E9.

In a preferred embodiment of the binding molecule of the invention, the antigen-binding site specific for TRAILR2 comprises heavy chain CDRs comprising the amino acid sequences of SEQ ID NO: 19 (CDR1), SEQ ID NO: 20 (CDR2), and SEQ ID

- 25 041360

NO: 21 (CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO: 22 (CDR1), SEQ ID NO: 23 (CDR2) and SEQ ID NO: 24 (CDR3) and an antigen-binding site specific for CDH17 contains a CDR heavy chain containing the amino acid sequences of SEQ ID NO: 64 (CDR1), SEQ ID NO: 65 (CDR2) and SEQ ID NO: 66 (CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO: 67 (CDR1), SEQ ID NO: 68 (CDR2) and SEQ ID NO: 69 (CDR3). In this embodiment, the antigen-binding site specific for TRAILR2 is TR#7 and the antigen-binding site specific for CDH17 is CDH17A6.

Preferably, the antigen-binding site specific for TRAILR2 comprises a heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 88 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 89, and the antigen-binding site specific for CDH17 comprises a heavy chain variable domain. chain containing the amino acid sequence of SEQ ID NO: 112; and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 113. In this embodiment, the antigen-binding site specific for TRAILR2 is TR#7 (e.g., scFv SEQ ID NO: 137) and the antigen-binding site specific for CDH17 is CDH17A6.

In a preferred embodiment of the binding molecule of the invention, the antigen-binding site specific for TRAILR2 comprises heavy chain CDRs comprising the amino acid sequences of SEQ ID NO: 19 (CDR1), SEQ ID NO: 20 (CDR2), and SEQ ID NO: 21 ( CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO: 22 (CDR1), SEQ ID NO: 23 (CDR2) and SEQ ID NO: 24 (CDR3) and an antigen-binding site specific for CDH17 contains heavy chain CDRs containing amino acid sequences of SEQ ID NO: 70 (CDR1), SEQ ID NO: 71 (CDR2) and SEQ ID NO: 72 (CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO: 73 (CDR1), SEQ ID NO: 74 (CDR2) and SEQ ID NO: 75 (CDR3). In this embodiment, the antigen-binding site specific for TRAILR2 is TR#7 and the antigen-binding site specific for CDH17 is CDH17E2 or CDH17H2.

Preferably, the antigen-binding site specific for TRAILR2 comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 88 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 89 and the antigen-binding site specific for CDH17 comprises a heavy chain variable domain, containing the amino acid sequence of SEQ ID NO: 114 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 115. In this embodiment, the antigen-binding site specific for TRAILR2 is TR#7 (e.g., scFv SEQ ID NO: 137) and the antigen-binding site specific for CDH17 is CDH17E2.

Preferably, the antigen-binding site specific for TRAILR2 comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 88 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 89 and the antigen-binding site specific for CDH17 comprises a heavy chain variable domain, containing the amino acid sequence of SEQ ID NO: 116 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 117. In this embodiment, the antigen-binding site specific for TRAILR2 is TR#7 (e.g., scFv SEQ ID NO: 137) and the antigen-binding site specific for CDH17 is CDH17H2.

In a preferred embodiment of the binding molecule of the invention, the antigen-binding site specific for TRAILR2 comprises heavy chain CDRs comprising the amino acid sequences of SEQ ID NO: 19 (CDR1), SEQ ID NO: 20 (CDR2), and SEQ ID NO: 21 ( CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO: 22 (CDR1), SEQ ID NO: 23 (CDR2) and SEQ ID NO: 24 (CDR3) and an antigen-binding site specific for CDH17 contains heavy chain CDRs containing amino acid sequences of SEQ ID NO: 76 (CDR1), SEQ ID NO: 77 (CDR2) and SEQ ID NO: 78 (CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO: 79 (CDR1), SEQ ID NO: 80 (CDR2) and SEQ ID NO: 81 (CDR3). In this embodiment, the antigen-binding site specific for TRAILR2 is TR#7 and the antigen-binding site specific for CDH17 is CDH17v1.

Preferably, the antigen-binding site specific for TRAILR2 comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 88 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 89 and the antigen-binding site specific for CDH17 comprises a heavy chain variable domain, containing the amino acid sequence of SEQ ID NO: 118 and the variable domain of the light chain,

- 26 041360 containing the amino acid sequence of SEQ ID NO: 119. In this embodiment, the antigen-binding site specific for TRAILR2 is TR#7 (for example, scFv

SEQ ID NO: 137) and the antigen-binding site specific for CDH17 is

CDH17v1.

In a preferred embodiment of the binding molecule of the invention, the antigen-binding site specific for TRAILR2 comprises heavy chain CDRs comprising the amino acid sequences of SEQ ID NO: 25 (CDR1), SEQ ID NO: 26 (CDR2), and SEQ ID NO: 27 ( CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO: 28 (CDR1), SEQ ID NO: 29 (CDR2) and SEQ ID NO: 30 (CDR3) and an antigen-binding site specific for CDH17 contains heavy chain CDRs containing amino acid sequences of SEQ ID NO: 58 (CDR1), SEQ ID NO: 59 (CDR2) and SEQ ID NO: 60 (CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO: 61 (CDR1), SEQ ID NO: 62 (CDR2) and SEQ ID NO: 63 (CDR3). In this embodiment, the antigen-binding site specific for TRAILR2 is TR#8 and the antigen-binding site specific for CDH17 is CDH17E9.

Preferably, the antigen-binding site specific for TRAILR2 comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 90 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 91 and the antigen-binding site specific for CDH17 comprises a heavy chain variable domain, containing the amino acid sequence of SEQ ID NO: 110 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 111. In this embodiment, the antigen-binding site specific for TRAILR2 is TR#8 (e.g., scFv SEQ ID NO: 138) and the antigen-binding site specific for CDH17 is CDH17E9.

In a preferred embodiment of the binding molecule of the invention, the antigen-binding site specific for TRAILR2 comprises heavy chain CDRs comprising the amino acid sequences of SEQ ID NO: 25 (CDR1), SEQ ID NO: 26 (CDR2), and SEQ ID NO: 27 ( CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO: 28 (CDR1), SEQ ID NO: 29 (CDR2) and SEQ ID NO: 30 (CDR3) and an antigen-binding site specific for CDH17 contains heavy chain CDRs containing amino acid sequences of SEQ ID NO: 64 (CDR1), SEQ ID NO: 65 (CDR2) and SEQ ID NO: 66 (CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO: 67 (CDR1), SEQ ID NO: 68 (CDR2) and SEQ ID NO: 69 (CDR3). In this embodiment, the antigen-binding site specific for TRAILR2 is TR#8 and the antigen-binding site specific for CDH17 is CDH17A6.

Preferably, the antigen-binding site specific for TRAILR2 comprises a heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 90 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 91, and the antigen-binding site specific for CDH17 comprises a heavy chain variable domain. chain containing the amino acid sequence of SEQ ID NO: 112; and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 113. In this embodiment, the antigen-binding site specific for TRAILR2 is TR#8 (e.g., scFv SEQ ID NO: 138) and the antigen-binding site specific for CDH17 is CDH17A6.

In a preferred embodiment of the binding molecule of the invention, the antigen-binding site specific for TRAILR2 comprises heavy chain CDRs comprising the amino acid sequences of SEQ ID NO: 25 (CDR1), SEQ ID NO: 26 (CDR2), and SEQ ID NO: 27 ( CDR3), and light chain CDRs containing the amino acid sequences of SEQ ID NO: 28 (CDR1), SEQ ID NO: 29 (CDR2), and SEQ ID NO: 30 (CDR3), and an antigen-binding site specific for CDH17 contains a heavy chain CDR containing the amino acid sequences of SEQ ID NO: 70 (CDR1), SEQ ID NO: 71 (CDR2) and SEQ ID NO: 72 (CDR3), and light chain CDRs containing the amino acid sequences of SEQ ID NO: 73 (CDR1), SEQ ID NO: 74 (CDR2) and SEQ ID NO: 75 (CDR3). In this embodiment, the antigen-binding site specific for TRAILR2 is TR#8 and the antigen-binding site specific for CDH17 is CDH17E2 or CDH17H2.

Preferably, the antigen-binding site specific for TRAILR2 comprises a heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 90 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 91, and the antigen-binding site specific for CDH17 comprises a heavy chain variable domain. chain containing the amino acid sequence of SEQ ID NO: 114, and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 115. In this embodiment, the antigen-binding site specific for TRAILR2 is TR#8 (e.g., scFv

- 27 041360

SEQ ID NO: 138) and the antigen-binding site specific for CDH17 is

CDH17E2.

Preferably, the antigen-binding site specific for TRAILR2 comprises a heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 90 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 91, and the antigen-binding site specific for CDH17 comprises a heavy chain variable domain. chain containing the amino acid sequence of SEQ ID NO: 116; and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 117. In this embodiment, the antigen-binding site specific for TRAILR2 is TR#8 (e.g., scFv SEQ ID NO: 138) and the antigen-binding site specific for CDH17 is CDH17H2.

In a preferred embodiment of the binding molecule of the invention, the antigen-binding site specific for TRAILR2 comprises heavy chain CDRs comprising the amino acid sequences of SEQ ID NO: 25 (CDR1), SEQ ID NO: 26 (CDR2), and SEQ ID NO: 27 ( CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO: 28 (CDR1), SEQ ID NO: 29 (CDR2) and SEQ ID NO: 30 (CDR3) and an antigen-binding site specific for CDH17 contains heavy chain CDRs containing amino acid sequences of SEQ ID NO: 76 (CDR1), SEQ ID NO: 77 (CDR2) and SEQ ID NO: 78 (CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO: 79 (CDR1), SEQ ID NO: 80 (CDR2) and SEQ ID NO: 81 (CDR3). In this embodiment, the antigen-binding site specific for TRAILR2 is TR#8 and the antigen-binding site specific for CDH17 is CDH17v1.

Preferably, the antigen-binding site specific for TRAILR2 comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 90 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 91 and the antigen-binding site specific for CDH17 comprises a heavy chain variable domain, containing the amino acid sequence of SEQ ID NO: 118 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 118. In this embodiment, the antigen-binding site specific for TRAILR2 is TR#8 (e.g., scFv SEQ ID NO: 138) and the antigen-binding site specific for CDH17 is CDH17v1.

In a preferred embodiment of the binding molecule of the invention, the antigen-binding site specific for TRAILR2 comprises heavy chain CDRs comprising the amino acid sequences of SEQ ID NO: 31 (CDR1), SEQ ID NO: 32 (CDR2), and SEQ ID NO: 33 ( CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO: 34 (CDR1), SEQ ID NO: 35 (CDR2) and SEQ ID NO: 36 (CDR3) and an antigen-binding site specific for CDH17 contains heavy chain CDRs containing amino acid sequences of SEQ ID NO: 58 (CDR1), SEQ ID NO: 59 (CDR2) and SEQ ID NO: 60 (CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO: 61 (CDR1), SEQ ID NO: 62 (CDR2) and SEQ ID NO: 63 (CDR3). In this embodiment, the antigen-binding site specific for TRAILR2 is TR#10 and the antigen-binding site specific for CDH17 is CDH17E9.

Preferably, the antigen-binding site specific for TRAILR2 comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 92 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 93 and the antigen-binding site specific for CDH17 comprises a heavy chain variable domain, containing the amino acid sequence of SEQ ID NO: 110 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 111. In this embodiment, the antigen-binding site specific for TRAILR2 is TR#10 (for example, scFv SEQ ID NO: 139) and the antigen-binding site specific for CDH17 is CDH17E9.

In a preferred embodiment of the binding molecule of the invention, the antigen-binding site specific for TRAILR2 comprises heavy chain CDRs comprising the amino acid sequences of SEQ ID NO: 31 (CDR1), SEQ ID NO: 32 (CDR2), and SEQ ID NO: 33 ( CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO: 34 (CDR1), SEQ ID NO: 35 (CDR2) and SEQ ID NO: 36 (CDR3) and an antigen-binding site specific for CDH17 contains heavy chain CDRs containing amino acid sequences of SEQ ID NO: 64 (CDR1), SEQ ID NO: 65 (CDR2) and SEQ ID NO: 66 (CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO: 67 (CDR1), SEQ ID NO: 68 (CDR2) and SEQ ID NO: 69 (CDR3). In this embodiment, the TRAILR2 specific antigen binding site is TR#10 and the CDH17 specific antigen binding site is CDH17A6.

- 28 041360

Preferably, the antigen-binding site specific for TRAILR2 comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 92 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 93 and the antigen-binding site specific for CDH17 comprises a heavy chain variable domain, containing the amino acid sequence of SEQ ID NO: 112 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 113. In this embodiment, the antigen-binding site specific for TRAILR2 is TR#10 (e.g., scFv SEQ ID NO: 139) and the antigen-binding site specific for CDH17 is CDH17A6.

Preferably, the antigen-binding site specific for TRAILR2 comprises heavy chain CDRs containing the amino acid sequences of SEQ ID NO: 31 (CDR1), SEQ ID NO: 32 (CDR2) and SEQ ID NO: 33 (CDR3) and light chain CDRs containing the amino acid sequences SEQ ID NO: 34 (CDR1), SEQ ID NO: 35 (CDR2) and SEQ ID NO: 36 (CDR3) and the antigen-binding site specific for CDH17 contains heavy chain CDRs containing the amino acid sequences of SEQ ID NO: 70 (CDR1) , SEQ ID NO: 71 (CDR2) and SEQ ID NO: 72 (CDR3) and has light chain CDRs containing the amino acid sequences of SEQ ID NO: 73 (CDR1), SEQ ID NO: 74 (CDR2) and SEQ ID NO: 75 (CDR3). In this embodiment, the TRAILR2 specific antigen binding site is TR#10 and the CDH17 specific antigen binding site is CDH17E2 or CDH17H2.

Preferably, the antigen-binding site specific for TRAILR2 comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 92 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 93 and the antigen-binding site specific for CDH17 comprises a heavy chain variable domain, containing the amino acid sequence of SEQ ID NO: 114 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 115. In this embodiment, the antigen-binding site specific for TRAILR2 is TR#10 (e.g., scFv SEQ ID NO: 139) and the antigen-binding site specific for CDH17 is CDH17E2.

Preferably, the antigen-binding site specific for TRAILR2 comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 92 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 93 and the antigen-binding site specific for CDH17 comprises a heavy chain variable domain, containing the amino acid sequence of SEQ ID NO: 116 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 117. In this embodiment, the antigen-binding site specific for TRAILR2 is TR#10 (for example, scFv SEQ ID NO: 139) and the antigen-binding site specific for CDH17 is CDH17H2.

Preferably, the antigen-binding site specific for TRAILR2 comprises heavy chain CDRs containing the amino acid sequences of SEQ ID NO: 31 (CDR1), SEQ ID NO: 32 (CDR2) and SEQ ID NO: 33 (CDR3) and light chain CDRs containing the amino acid sequences SEQ ID NO: 34 (CDR1), SEQ ID NO: 35 (CDR2) and SEQ ID NO: 36 (CDR3) and the antigen-binding site specific for CDH17 contains heavy chain CDRs containing the amino acid sequences of SEQ ID NO: 76 (CDR1) , SEQ ID NO: 77 (CDR2) and SEQ ID NO: 78 (CDR3) and has light chain CDRs containing the amino acid sequences of SEQ ID NO: 79 (CDR1), SEQ ID NO: 80 (CDR2) and SEQ ID NO: 81 (CDR3). In this embodiment, the TRAILR2 specific antigen binding site is TR#10 and the CDH17 specific antigen binding site is CDH17v1.

Preferably, the antigen-binding site specific for TRAILR2 comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 92 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 93 and the antigen-binding site specific for CDH17 comprises a heavy chain variable domain, containing the amino acid sequence of SEQ ID NO: 118 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 119. In this embodiment, the antigen-binding site specific for TRAILR2 is TR#10 (e.g., scFv SEQ ID NO: 139) and the antigen-binding site specific for CDH17 is CDH17v1.

In a preferred embodiment of the binding molecule of the invention, the antigen-binding site specific for TRAILR2 comprises heavy chain CDRs comprising the amino acid sequences of SEQ ID NO: 37 (CDR1), SEQ ID NO: 38 (CDR2), and SEQ ID NO: 39 ( CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO: 40 (CDR1), SEQ ID NO: 41 (CDR2) and SEQ ID NO: 42 (CDR3) and an antigen-binding site specific for CDH17 contains heavy chain CDRs containing amino acid sequences

- 29 041360

ID NO: 58 (CDR1), SEQ ID NO: 59 (CDR2) and SEQ ID NO: 60 (CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO: 61 (CDR1), SEQ ID NO: 62 (CDR2) and SEQ ID NO: 63 (CDR3). In this embodiment, the antigen-binding site specific for TRAILR2 is TR#12 and the antigen-binding site specific for CDH17 is CDH17E9.

Preferably, the antigen-binding site specific for TRAILR2 comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 94 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 95 and the antigen-binding site specific for CDH17 comprises a heavy chain variable domain, containing the amino acid sequence of SEQ ID NO: 110 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 111. In this embodiment, the antigen-binding site specific for TRAILR2 is TR#12 (e.g., scFv SEQ ID NO: 140) and the antigen-binding site specific for CDH17 is CDH17E9.

In a preferred embodiment of the binding molecule of the invention, the antigen-binding site specific for TRAILR2 comprises heavy chain CDRs comprising the amino acid sequences of SEQ ID NO: 37 (CDR1), SEQ ID NO: 38 (CDR2), and SEQ ID NO: 39 ( CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO: 40 (CDR1), SEQ ID NO: 41 (CDR2) and SEQ ID NO: 42 (CDR3) and an antigen-binding site specific for CDH17 contains heavy chain CDRs containing amino acid sequences of SEQ ID NO: 64 (CDR1), SEQ ID NO: 65 (CDR2) and SEQ ID NO: 66 (CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO: 67 (CDR1), SEQ ID NO: 68 (CDR2) and SEQ ID NO: 69 (CDR3). In this embodiment, the antigen-binding site specific for TRAILR2 is TR#12 and the antigen-binding site specific for CDH17 is CDH17A6.

Preferably, the antigen-binding site specific for TRAILR2 comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 94 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 95 and the antigen-binding site specific for CDH17 comprises a heavy chain variable domain, containing the amino acid sequence of SEQ ID NO: 112 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 113. In this embodiment, the antigen-binding site specific for TRAILR2 is TR#12 (e.g., scFv SEQ ID NO: 140) and the antigen-binding site specific for CDH17 is CDH17A6.

In a preferred embodiment of the binding molecule of the invention, the antigen-binding site specific for TRAILR2 comprises heavy chain CDRs comprising the amino acid sequences of SEQ ID NO: 37 (CDR1), SEQ ID NO: 38 (CDR2), and SEQ ID NO: 39 ( CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO: 40 (CDR1), SEQ ID NO: 41 (CDR2) and SEQ ID NO: 42 (CDR3) and an antigen-binding site specific for CDH17 contains heavy chain CDRs containing amino acid sequences of SEQ ID NO: 70 (CDR1), SEQ ID NO: 71 (CDR2) and SEQ ID NO: 72 (CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO: 73 (CDR1), SEQ ID NO: 74 (CDR2) and SEQ ID NO: 75 (CDR3). In this embodiment, the antigen-binding site specific for TRAILR2 is TR#12 and the antigen-binding site specific for CDH17 is CDH171E2 or CDH171H2.

Preferably, the antigen-binding site specific for TRAILR2 comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 94 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 95 and the antigen-binding site specific for CDH17 comprises a heavy chain variable domain, containing the amino acid sequence of SEQ ID NO: 114 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 115. In this embodiment, the antigen-binding site specific for TRAILR2 is TR#12 (e.g., scFv SEQ ID NO: 140) and the antigen-binding site specific for CDH17 is CDH17E2.

Preferably, the antigen-binding site specific for TRAILR2 comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 94 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 95 and the antigen-binding site specific for CDH17 comprises a heavy chain variable domain, containing the amino acid sequence of SEQ ID NO: 116 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 117. In this embodiment, the antigen-binding site specific for TRAILR2 is TR#12 (for example, scFv SEQ ID NO: 140) and antigen-binding site specific for CDH17 is

- 30 041360

CDH17H2.

In a preferred embodiment of the binding molecule of the invention, the antigen-binding site specific for TRAILR2 comprises heavy chain CDRs comprising the amino acid sequences of SEQ ID NO: 37 (CDR1), SEQ ID NO: 38 (CDR2), and SEQ ID NO: 39 ( CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO: 40 (CDR1), SEQ ID NO: 41 (CDR2) and SEQ ID NO: 42 (CDR3) and an antigen-binding site specific for CDH17 contains heavy chain CDRs containing amino acid sequences of SEQ ID NO: 76 (CDR1), SEQ ID NO: 77 (CDR2) and SEQ ID NO: 78 (CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO: 79 (CDR1), SEQ ID NO: 80 (CDR2) and SEQ ID NO: 81 (CDR3). In this embodiment, the antigen-binding site specific for TRAILR2 is TR#12 and the antigen-binding site specific for CDH17 is CDH17v1.

Preferably, the antigen-binding site specific for TRAILR2 comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 94 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 95 and the antigen-binding site specific for CDH17 comprises a heavy chain variable domain, containing the amino acid sequence of SEQ ID NO: 118 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 119. In this embodiment, the antigen-binding site specific for TRAILR2 is TR#12 (e.g., scFv SEQ ID NO: 140) and the antigen-binding site specific for CDH17 is CDH17v1.

In a preferred embodiment of the binding molecule of the invention, the antigen-binding site specific for TRAILR2 comprises heavy chain CDRs comprising the amino acid sequences of SEQ ID NO: 43 (CDR1), SEQ ID NO: 44 (CDR2), and SEQ ID NO: 48 ( CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO: 49 (CDR1), SEQ ID NO: 50 (CDR2) and SEQ ID NO: 54 (CDR3) and an antigen-binding site specific for CDH17 contains heavy chain CDRs containing amino acid sequences of SEQ ID NO: 58 (CDR1), SEQ ID NO: 59 (CDR2) and SEQ ID NO: 60 (CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO: 61 (CDR1), SEQ ID NO: 62 (CDR2) and SEQ ID NO: 63 (CDR3). In this embodiment, the antigen-binding site specific for TRAILR2 is TRv1 or TRv2 and the antigen-binding site specific for CDH17 is CDH17E9.

Preferably, the antigen-binding site specific for TRAILR2 comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 96 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 97 and the antigen-binding site specific for CDH17 comprises a heavy chain variable domain, containing the amino acid sequence of SEQ ID NO: 110 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 111. In this embodiment, the antigen-binding site specific for TRAILR2 is TRv1 (for example, scFv SEQ ID NO: 124, SEQ ID NO : 131, SEQ ID NO: 132, SEQ ID NO: 133) and the antigen-binding site specific for CDH17 is CDH17E9.

Preferably, the antigen-binding site specific for TRAILR2 comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 98 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 99 and the antigen-binding site specific for CDH17 comprises a heavy chain variable domain, containing the amino acid sequence of SEQ ID NO: 110 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 111. In this embodiment, the antigen-binding site specific for TRAILR2 is TRv2 (for example, scFv SEQ ID NO: 125) and the antigen-binding site , specific for CDH17, is CDH17E9.

In a preferred embodiment of the binding molecule of the invention, the antigen-binding site specific for TRAILR2 comprises heavy chain CDRs comprising the amino acid sequences of SEQ ID NO: 43 (CDR1), SEQ ID NO: 44 (CDR2), and SEQ ID NO: 48 ( CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO: 49 (CDR1), SEQ ID NO: 50 (CDR2) and SEQ ID NO: 54 (CDR3) and an antigen-binding site specific for CDH17 contains heavy chain CDRs containing amino acid sequences of SEQ ID NO: 64 (CDR1), SEQ ID NO: 65 (CDR2) and SEQ ID NO: 66 (CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO: 67 (CDR1), SEQ ID NO: 68 (CDR2) and SEQ ID NO: 69 (CDR3). In this embodiment, the antigen-binding site specific for TRAILR2 is TRv1 or TRv2 and the antigen-binding site specific for CDH17 is CDH17A6.

Preferably, the antigen-binding site specific for TRAILR2 contains the variant

- 31 041360 a heavy chain white domain containing the amino acid sequence of SEQ ID NO: 96 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 97 and an antigen-binding site specific for CDH17, contains a heavy chain variable domain containing the amino acid sequence of SEQ ID NO : 112 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 113. In this embodiment, the antigen-binding site specific for TRAILR2 is TRv1 (e.g., scFv SEQ ID NO: 124, SEQ ID NO: 131, SEQ ID NO : 132, SEQ ID NO: 133) and the antigen-binding site specific for CDH17 is CDH17A6.

Preferably, the antigen-binding site specific for TRAILR2 comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 98 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 99 and the antigen-binding site specific for CDH17 comprises a heavy chain variable domain, containing the amino acid sequence of SEQ ID NO: 112 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 113. In this embodiment, the antigen-binding site specific for TRAILR2 is TRv2 (for example, scFv SEQ ID NO: 125) and the antigen-binding site , specific for CDH17, is CDH17A6.

In a preferred embodiment of the binding molecule of the invention, the antigen-binding site specific for TRAILR2 comprises heavy chain CDRs comprising the amino acid sequences of SEQ ID NO: 43 (CDR1), SEQ ID NO: 44 (CDR2), and SEQ ID NO: 48 ( CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO: 49 (CDR1), SEQ ID NO: 50 (CDR2) and SEQ ID NO: 54 (CDR3) and an antigen-binding site specific for CDH17 contains heavy chain CDRs containing amino acid sequences of SEQ ID NO: 70 (CDR1), SEQ ID NO: 71 (CDR2) and SEQ ID NO: 72 (CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO: 73 (CDR1), SEQ ID NO: 74 (CDR2) and SEQ ID NO: 75 (CDR3). In this embodiment, the TRAILR2 specific antigen binding site is TRv1 or TRv2 and the CDH17 specific antigen binding site is CDH17E2 or CDH17H2.

Preferably, the antigen-binding site specific for TRAILR2 comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 96 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 97 and the antigen-binding site specific for CDH17 comprises a heavy chain variable domain, containing the amino acid sequence of SEQ ID NO: 114 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 115. In this embodiment, the antigen-binding site specific for TRAILR2 is TRv1 (for example, scFv SEQ ID NO: 124, SEQ ID NO : 131, SEQ ID NO: 132, SEQ ID NO: 133) and the antigen-binding site specific for CDH17 is CDH17E2.

Preferably, the antigen-binding site specific for TRAILR2 comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 98 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 99 and the antigen-binding site specific for CDH17 comprises a heavy chain variable domain, containing the amino acid sequence of SEQ ID NO: 114 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 115. In this embodiment, the antigen-binding site specific for TRAILR2 is TRv2 (for example, scFv SEQ ID NO: 125) and the antigen-binding site , specific for CDH17, is CDH17E2.

Preferably, the antigen-binding site specific for TRAILR2 comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 96 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 97 and the antigen-binding site specific for CDH17 comprises a heavy chain variable domain, containing the amino acid sequence of SEQ ID NO: 116 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 117. In this embodiment, the antigen-binding site specific for TRAILR2 is TRv1 (for example, scFv SEQ ID NO: 124, SEQ ID NO : 131, SEQ ID NO: 132, SEQ ID NO: 133) and the antigen-binding site specific for CDH17 is CDH17H2.

Preferably, the antigen-binding site specific for TRAILR2 comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 98 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 99 and the antigen-binding site specific for CDH17 comprises a heavy chain variable domain, containing the amino acid sequence of SEQ ID NO: 116 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 117. In this embodiment, the antigen-binding site specific for TRAILR2 is TRv2 (e.g., scFv

- 32 041360

SEQ ID NO: 125) and the antigen-binding site specific for CDH17 is

CDH17H2.

In a preferred embodiment of the binding molecule of the invention, the antigen-binding site specific for TRAILR2 comprises heavy chain CDRs comprising the amino acid sequences of SEQ ID NO: 43 (CDR1), SEQ ID NO: 44 (CDR2), and SEQ ID NO: 48 ( CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO: 49 (CDR1), SEQ ID NO: 50 (CDR2) and SEQ ID NO: 54 (CDR3) and an antigen-binding site specific for CDH17 contains heavy chain CDRs containing amino acid sequences of SEQ ID NO: 76 (CDR1), SEQ ID NO: 77 (CDR2) and SEQ ID NO: 78 (CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO: 79 (CDR1), SEQ ID NO: 80 (CDR2) and SEQ ID NO: 81 (CDR3). In this embodiment, the antigen-binding site specific for TRAILR2 is TRv1 or TRv2 and the antigen-binding site specific for CDH17 is CDH17v1.

Preferably, the antigen-binding site specific for TRAILR2 comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 96 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 97 and the antigen-binding site specific for CDH17 comprises a heavy chain variable domain, containing the amino acid sequence of SEQ ID NO: 118 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 119. In this embodiment, the antigen-binding site specific for TRAILR2 is TRv1 (for example, scFv SEQ ID NO: 124, SEQ ID NO : 131, SEQ ID NO: 132, SEQ ID NO: 133) and the antigen-binding site specific for CDH17 is CDH17v1.

Preferably, the antigen-binding site specific for TRAILR2 comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 98 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 99 and the antigen-binding site specific for CDH17 comprises a heavy chain variable domain, containing the amino acid sequence of SEQ ID NO: 118 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 119. In this embodiment, the antigen-binding site specific for TRAILR2 is TRv2 (for example, scFv SEQ ID NO: 125) and the antigen-binding site , specific for CDH17, is CDH17v1.

In a preferred embodiment of the binding molecule of the invention, the antigen-binding site specific for TRAILR2 comprises heavy chain CDRs comprising the amino acid sequences of SEQ ID NO: 43 (CDR1), SEQ ID NO: 45 (CDR2), and SEQ ID NO: 48 ( CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO: 49 (CDR1), SEQ ID NO: 51 (CDR2) and SEQ ID NO: 55 (CDR3) and an antigen-binding site specific for CDH17 contains heavy chain CDRs containing amino acid sequences of SEQ ID NO: 58 (CDR1), SEQ ID NO: 59 (CDR2) and SEQ ID NO: 60 (CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO: 61 (CDR1), SEQ ID NO: 62 (CDR2) and SEQ ID NO: 63 (CDR3). In this embodiment, the antigen-binding site specific for TRAILR2 is TRv3 and the antigen-binding site specific for CDH17 is CDH17E9.

Preferably, the antigen-binding site specific for TRAILR2 comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 100 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 101 and the antigen-binding site specific for CDH17 comprises a heavy chain variable domain, containing the amino acid sequence of SEQ ID NO: 110 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 111. In this embodiment, the antigen-binding site specific for TRAILR2 is TRv3 (for example, scFv SEQ ID NO: 126) and the antigen-binding site , specific for CDH17, is CDH17E9.

In a preferred embodiment of the binding molecule of the invention, the antigen-binding site specific for TRAILR2 comprises heavy chain CDRs comprising the amino acid sequences of SEQ ID NO: 43 (CDR1), SEQ ID NO: 45 (CDR2), and SEQ ID NO: 48 ( CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO: 49 (CDR1), SEQ ID NO: 51 (CDR2) and SEQ ID NO: 55 (CDR3) and an antigen-binding site specific for CDH17 contains heavy chain CDRs containing amino acid sequences of SEQ ID NO: 64 (CDR1), SEQ ID NO: 65 (CDR2) and SEQ ID NO: 66 (CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO: 67 (CDR1), SEQ ID NO: 68 (CDR2) and SEQ ID NO: 69 (CDR3). In this embodiment, the antigen-binding site specific for TRAILR2 is TRv3 and the antigen-binding site specific for CDH17 is CDH17A6.

- 33 041360

Preferably, the antigen-binding site specific for TRAILR2 comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 100 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 101 and the antigen-binding site specific for CDH17 comprises a heavy chain variable domain, containing the amino acid sequence of SEQ ID NO: 112 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 113. In this embodiment, the antigen-binding site specific for TRAILR2 is TRv3 (for example, scFv SEQ ID NO: 126) and the antigen-binding site , specific for CDH17, is CDH17A6.

In a preferred embodiment of the binding molecule of the invention, the antigen-binding site specific for TRAILR2 comprises heavy chain CDRs comprising the amino acid sequences of SEQ ID NO: 43 (CDR1), SEQ ID NO: 45 (CDR2), and SEQ ID NO: 48 ( CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO: 49 (CDR1), SEQ ID NO: 51 (CDR2) and SEQ ID NO: 55 (CDR3) and an antigen-binding site specific for CDH17 contains heavy chain CDRs containing amino acid sequences of SEQ ID NO: 70 (CDR1), SEQ ID NO: 71 (CDR2) and SEQ ID NO: 72 (CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO: 73 (CDR1), SEQ ID NO: 74 (CDR2) and SEQ ID NO: 75 (CDR3). In this embodiment, the antigen-binding site specific for TRAILR2 is TRv3 and the antigen-binding site specific for CDH17 is CDH17E2 or CDH17H2.

Preferably, the antigen-binding site specific for TRAILR2 comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 100 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 101 and the antigen-binding site specific for CDH17 comprises a heavy chain variable domain, containing the amino acid sequence of SEQ ID NO: 114 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 115. In this embodiment, the antigen-binding site specific for TRAILR2 is TRv3 (for example, scFv SEQ ID NO: 126) and the antigen-binding site , specific for CDH17, is CDH17E2.

Preferably, the antigen-binding site specific for TRAILR2 comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 100 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 101 and the antigen-binding site specific for CDH17 comprises a heavy chain variable domain, containing the amino acid sequence of SEQ ID NO: 116 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 117. In this embodiment, the antigen-binding site specific for TRAILR2 is TRv3 (for example, scFv SEQ ID NO: 126) and the antigen-binding site , specific for CDH17, is CDH17H2.

In a preferred embodiment of the binding molecule of the invention, the antigen-binding site specific for TRAILR2 comprises heavy chain CDRs comprising the amino acid sequences of SEQ ID NO: 43 (CDR1), SEQ ID NO: 45 (CDR2), and SEQ ID NO: 48 ( CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO: 49 (CDR1), SEQ ID NO: 51 (CDR2) and SEQ ID NO: 55 (CDR3) and an antigen-binding site specific for CDH17 contains heavy chain CDRs containing amino acid sequences of SEQ ID NO: 76 (CDR1), SEQ ID NO: 77 (CDR2) and SEQ ID NO: 78 (CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO: 79 (CDR1), SEQ ID NO: 80 (CDR2) and SEQ ID NO: 81 (CDR3). In this embodiment, the antigen-binding site specific for TRAILR2 is TRv3 and the antigen-binding site specific for CDH17 is CDH17v1.

Preferably, the antigen-binding site specific for TRAILR2 comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 100 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 101 and the antigen-binding site specific for CDH17 comprises a heavy chain variable domain, containing the amino acid sequence of SEQ ID NO: 118 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 119. In this embodiment, the antigen-binding site specific for TRAILR2 is TRv3 (for example, scFv SEQ ID NO: 126) and the antigen-binding site , specific for CDH17, is CDH17v1.

In a preferred embodiment of the binding molecule of the invention, the antigen-binding site specific for TRAILR2 comprises heavy chain CDRs comprising the amino acid sequences of SEQ ID NO: 43 (CDR1), SEQ ID NO: 45 (CDR2), and SEQ ID NO: 48 ( CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO: 49

- 34 041360 (CDR1), SEQ ID NO: 52 (CDR2) and SEQ ID NO: 56 (CDR3) and the antigen-binding site specific for CDH17 contains heavy chain CDRs containing the amino acid sequences of SEQ ID NO: 58 (CDR1), SEQ ID NO: 59 (CDR2) and SEQ ID NO: 60 (CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO: 61 (CDR1), SEQ ID NO: 62 (CDR2) and SEQ ID NO: 63 (CDR3) . In this embodiment, the antigen-binding site specific for TRAILR2 is TRv4 and the antigen-binding site specific for CDH17 is CDH17E9.

Preferably, the antigen-binding site specific for TRAILR2 comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 102 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 103 and the antigen-binding site specific for CDH17 comprises a heavy chain variable domain, containing the amino acid sequence of SEQ ID NO: 110 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 111. In this embodiment, the antigen-binding site specific for TRAILR2 is TRv4 (for example, scFv SEQ ID NO: 127) and the antigen-binding site , specific for CDH17, is CDH17E9.

In a preferred embodiment of the binding molecule of the invention, the antigen-binding site specific for TRAILR2 comprises heavy chain CDRs comprising the amino acid sequences of SEQ ID NO: 43 (CDR1), SEQ ID NO: 45 (CDR2) and SEQ ID NO: 48 ( CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO: 49 (CDR1), SEQ ID NO: 52 (CDR2) and SEQ ID NO: 56 (CDR3) and an antigen-binding site specific for CDH17 contains heavy chain CDRs containing amino acid sequences of SEQ ID NO: 64 (CDR1), SEQ ID NO: 65 (CDR2) and SEQ ID NO: 66 (CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO: 67 (CDR1), SEQ ID NO: 68 (CDR2) and SEQ ID NO: 69 (CDR3). In this embodiment, the antigen-binding site specific for TRAILR2 is TRv4 and the antigen-binding site specific for CDH17 is CDH17A6.

Preferably, the antigen-binding site specific for TRAILR2 comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 102 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 103 and the antigen-binding site specific for CDH17 comprises a heavy chain variable domain, containing the amino acid sequence of SEQ ID NO: 112 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 113. In this embodiment, the antigen-binding site specific for TRAILR2 is TRv4 (for example, scFv SEQ ID NO: 127) and the antigen-binding site , specific for CDH17, is CDH17A6.

In a preferred embodiment of the binding molecule of the invention, the antigen-binding site specific for TRAILR2 comprises heavy chain CDRs comprising the amino acid sequences of SEQ ID NO: 43 (CDR1), SEQ ID NO: 45 (CDR2), and SEQ ID NO: 48 ( CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO: 49 (CDR1), SEQ ID NO: 52 (CDR2) and SEQ ID NO: 56 (CDR3) and an antigen-binding site specific for CDH17 contains heavy chain CDRs containing amino acid sequences of SEQ ID NO: 70 (CDR1), SEQ ID NO: 71 (CDR2) and SEQ ID NO: 72 (CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO: 73 (CDR1), SEQ ID NO: 74 (CDR2) and SEQ ID NO: 75 (CDR3). In this embodiment, the antigen-binding site specific for TRAILR2 is TRv4 and the antigen-binding site specific for CDH17 is CDH17E2 or CDH17H2.

Preferably, the antigen-binding site specific for TRAILR2 comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 102 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 103 and the antigen-binding site specific for CDH17 comprises a heavy chain variable domain, containing the amino acid sequence of SEQ ID NO: 114 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 115. In this embodiment, the antigen-binding site specific for TRAILR2 is TRv4 (for example, scFv SEQ ID NO: 127) and the antigen-binding site , specific for CDH17, is CDH17E2.

Preferably, the antigen-binding site specific for TRAILR2 comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 102 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 103 and the antigen-binding site specific for CDH17 comprises a heavy chain variable domain, containing the amino acid sequence of SEQ ID NO: 116 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 117. In this embodiment, the antigen-binding site specific for TRAILR2 is TRv4 (e.g., scFv

SEQ ID NO: 127) and the antigen-binding site specific for CDH17 is

CDH17H2.

In a preferred embodiment of the binding molecule of the invention, the antigen-binding site specific for TRAILR2 comprises heavy chain CDRs comprising the amino acid sequences of SEQ ID NO: 43 (CDR1), SEQ ID NO: 45 (CDR2), and SEQ ID NO: 48 ( CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO: 49 (CDR1), SEQ ID NO: 52 (CDR2) and SEQ ID NO: 56 (CDR3) and an antigen-binding site specific for CDH17 contains heavy chain CDRs containing amino acid sequences of SEQ ID NO: 76 (CDR1), SEQ ID NO: 77 (CDR2) and SEQ ID NO: 78 (CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO: 79 (CDR1), SEQ ID NO: 80 (CDR2) and SEQ ID NO: 81 (CDR3). In this embodiment, the antigen-binding site specific for TRAILR2 is TRv4 and the antigen-binding site specific for CDH17 is CDH17v1.

Preferably, the antigen-binding site specific for TRAILR2 comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 102 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 103 and the antigen-binding site specific for CDH17 comprises a heavy chain variable domain, containing the amino acid sequence of SEQ ID NO: 118 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 119. In this embodiment, the antigen-binding site specific for TRAILR2 is TRv4 (for example, scFv SEQ ID NO: 127) and the antigen-binding site , specific for CDH17, is CDH17v1.

In a preferred embodiment of the binding molecule of the invention, the antigen-binding site specific for TRAILR2 comprises heavy chain CDRs comprising the amino acid sequences of SEQ ID NO: 43 (CDR1), SEQ ID NO: 45 (CDR2), and SEQ ID NO: 48 ( CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO: 49 (CDR1), SEQ ID NO: 50 (CDR2) and SEQ ID NO: 56 (CDR3) and an antigen-binding site specific for CDH17 contains heavy chain CDRs containing amino acid sequences of SEQ ID NO: 58 (CDR1) SEQ ID NO: 59 (CDR2) and SEQ ID NO: 60 (CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO: 61 (CDR1), SEQ ID NO: 62 ( CDR2) and SEQ ID NO: 63 (CDR3). In this embodiment, the antigen-binding site specific for TRAILR2 is TRv5 and the antigen-binding site specific for CDH17 is CDH17E9.

Preferably, the antigen-binding site specific for TRAILR2 comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 104 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 105 and the antigen-binding site specific for CDH17 comprises a heavy chain variable domain, containing the amino acid sequence of SEQ ID NO: 110 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 111. In this embodiment, the antigen-binding site specific for TRAILR2 is TRv5 (for example, scFv SEQ ID NO: 128) and the antigen-binding site , specific for CDH17, is CDH17E9.

In a preferred embodiment of the binding molecule of the invention, the antigen-binding site specific for TRAILR2 comprises heavy chain CDRs comprising the amino acid sequences of SEQ ID NO: 43 (CDR1), SEQ ID NO: 45 (CDR2), and SEQ ID NO: 48 ( CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO: 49 (CDR1), SEQ ID NO: 50 (CDR2) and SEQ ID NO: 56 (CDR3) and an antigen-binding site specific for CDH17 contains heavy chain CDRs containing amino acid sequences of SEQ ID NO: 64 (CDR1) SEQ ID NO: 65 (CDR2) and SEQ ID NO: 66 (CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO: 67 (CDR1), SEQ ID NO: 68 ( CDR2) and SEQ ID NO: 69 (CDR3). In this embodiment, the antigen-binding site specific for TRAILR2 is TRv5 and the antigen-binding site specific for CDH17 is CDH17A6.

Preferably, the antigen-binding site specific for TRAILR2 comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 104 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 105 and the antigen-binding site specific for CDH17 comprises a heavy chain variable domain, containing the amino acid sequence of SEQ ID NO: 112 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 113. In this embodiment, the antigen-binding site specific for TRAILR2 is TRv5 (for example, scFv SEQ ID NO: 128) and the antigen-binding site , specific for CDH17, is

- 36 041360

CDH17A6.

In a preferred embodiment of the binding molecule of the invention, the antigen-binding site specific for TRAILR2 comprises heavy chain CDRs comprising the amino acid sequences of SEQ ID NO: 43 (CDR1), SEQ ID NO: 45 (CDR2) and SEQ ID NO: 48 ( CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO: 49 (CDR1), SEQ ID NO: 50 (CDR2) and SEQ ID NO: 56 (CDR3) and an antigen-binding site specific for CDH17 contains heavy chain CDRs containing amino acid sequences of SEQ ID NO: 70 (CDR1), SEQ ID NO: 71 (CDR2) and SEQ ID NO: 72 (CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO: 73 (CDR1), SEQ ID NO: 74 (CDR2) and SEQ ID NO: 75 (CDR3). In this embodiment, the antigen-binding site specific for TRAILR2 is TRv5 and the antigen-binding site specific for CDH17 is CDH17E2 or CDH17H2.

Preferably, the antigen-binding site specific for TRAILR2 comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 104 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 105 and the antigen-binding site specific for CDH17 comprises a heavy chain variable domain, containing the amino acid sequence of SEQ ID NO: 114 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 115. In this embodiment, the antigen-binding site specific for TRAILR2 is TRv5 (for example, scFv SEQ ID NO: 128) and the antigen-binding site , specific for CDH17, is CDH17E2.

Preferably, the antigen-binding site specific for TRAILR2 comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 104 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 105 and the antigen-binding site specific for CDH17 comprises a heavy chain variable domain, containing the amino acid sequence of SEQ ID NO: 116 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 117. In this embodiment, the antigen-binding site specific for TRAILR2 is TRv5 (for example, scFv SEQ ID NO: 128) and the antigen-binding site , specific for CDH17, is CDH17H2.

In a preferred embodiment of the binding molecule of the invention, the antigen-binding site specific for TRAILR2 comprises heavy chain CDRs comprising the amino acid sequences of SEQ ID NO: 43 (CDR1), SEQ ID NO: 45 (CDR2), and SEQ ID NO: 48 ( CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO: 49 (CDR1), SEQ ID NO: 50 (CDR2) and SEQ ID NO: 56 (CDR3) and an antigen-binding site specific for CDH17 contains heavy chain CDRs containing amino acid sequences of SEQ ID NO: 76 (CDR1), SEQ ID NO: 77 (CDR2) and SEQ ID NO: 78 (CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO: 79 (CDR1), SeQ ID NO: 80 (CDR2) and SEQ ID NO: 81 (CDR3). In this embodiment, the antigen-binding site specific for TRAILR2 is TRv5 and the antigen-binding site specific for CDH17 is CDH17v1.

Preferably, the antigen-binding site specific for TRAILR2 comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 104 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 105 and the antigen-binding site specific for CDH17 comprises a heavy chain variable domain, containing the amino acid sequence of SEQ ID NO: 118 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 119. In this embodiment, the antigen-binding site specific for TRAILR2 is TRv5 (for example, scFv SEQ ID NO: 128) and the antigen-binding site , specific for CDH17, is CDH17v1.

In a preferred embodiment of the binding molecule of the invention, the antigen-binding site specific for TRAILR2 comprises heavy chain CDRs comprising the amino acid sequences of SEQ ID NO: 43 (CDR1), SEQ ID NO: 46 (CDR2), and SEQ ID NO: 48 ( CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO: 49 (CDR1), SEQ ID NO: 51 (CDR2) and SEQ ID NO: 57 (CDR3) and an antigen-binding site specific for CDH17 contains heavy chain CDRs containing amino acid sequences of SEQ ID NO: 58 (CDR1) SEQ ID NO: 59 (CDR2) and SEQ ID NO: 60 (CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO: 61 (CDR1), SEQ ID NO: 62 ( CDR2) and SEQ ID NO: 63 (CDR3). In this embodiment, the antigen-binding site specific for TRAILR2 is TRv6 and the antigen-binding site specific for CDH17 is CDH17E9.

Preferably, the antigen-binding site specific for TRAILR2 contains the variant

- 37 041360 heavy chain white domain containing the amino acid sequence of SEQ ID NO: 106 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 107 and an antigen-binding site specific for CDH17, contains a heavy chain variable domain containing the amino acid sequence of SEQ ID NO : 110 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 111. In this embodiment, the antigen-binding site specific for TRAILR2 is TRv6 (e.g., scFv SEQ ID NO: 129) and the antigen-binding site specific for CDH17 is is CDH17E9.

In a preferred embodiment of the binding molecule of the invention, the antigen-binding site specific for TRAILR2 comprises heavy chain CDRs comprising the amino acid sequences of SEQ ID NO: 43 (CDR1), SEQ ID NO: 46 (CDR2), and SEQ ID NO: 48 ( CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO: 49 (CDR1), SEQ ID NO: 51 (CDR2) and SEQ ID NO: 57 (CDR3) and an antigen-binding site specific for CDH17 contains heavy chain CDRs containing amino acid sequences of SEQ ID NO: 64 (CDR1) SEQ ID NO: 65 (CDR2) and SEQ ID NO: 66 (CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO: 67 (CDR1), SEQ ID NO: 68 ( CDR2) and SEQ ID NO: 69 (CDR3). In this embodiment, the antigen-binding site specific for TRAILR2 is TRv6 and the antigen-binding site specific for CDH17 is CDH17A6.

Preferably, the antigen-binding site specific for TRAILR2 comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 106 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 107 and the antigen-binding site specific for CDH17 comprises a heavy chain variable domain, containing the amino acid sequence of SEQ ID NO: 112 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 113. In this embodiment, the antigen-binding site specific for TRAILR2 is TRv6 (for example, scFv SEQ ID NO: 129) and the antigen-binding site , specific for CDH17, is CDH17A6.

In a preferred embodiment of the binding molecule of the invention, the antigen-binding site specific for TRAILR2 comprises heavy chain CDRs comprising the amino acid sequences of SEQ ID NO: 43 (CDR1), SEQ ID NO: 46 (CDR2), and SEQ ID NO: 48 ( CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO: 49 (CDR1), SEQ ID NO: 51 (CDR2) and SEQ ID NO: 57 (CDR3) and an antigen-binding site specific for CDH17 contains heavy chain CDRs containing amino acid sequences of SEQ ID NO: 70 (CDR1), SEQ ID NO: 71 (CDR2) and SEQ ID NO: 72 (CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO: 73 (CDR1), SEQ ID NO: 74 (CDR2) and SEQ ID NO: 75 (CDR3). In this embodiment, the antigen-binding site specific for TRAILR2 is TRv6 and the antigen-binding site specific for CDH17 is CDH17E2 or CDH17H2.

Preferably, the antigen-binding site specific for TRAILR2 comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 106 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 107 and the antigen-binding site specific for CDH17 comprises a heavy chain variable domain, containing the amino acid sequence of SEQ ID NO: 114 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 115. In this embodiment, the antigen-binding site specific for TRAILR2 is TRv6 (for example, scFv SEQ ID NO: 129) and the antigen-binding site , specific for CDH17, is CDH17E2.

Preferably, the antigen-binding site specific for TRAILR2 comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 106 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 107 and the antigen-binding site specific for CDH17 comprises a heavy chain variable domain, containing the amino acid sequence of SEQ ID NO: 116 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 117. In this embodiment, the antigen-binding site specific for TRAILR2 is TRv6 (for example, scFv SEQ ID NO: 129) and the antigen-binding site , specific for CDH17, is CDH17H2.

In a preferred embodiment of the binding molecule of the invention, the antigen-binding site specific for TRAILR2 comprises heavy chain CDRs comprising the amino acid sequences of SEQ ID NO: 43 (CDR1), SEQ ID NO: 46 (CDR2), and SEQ ID NO: 48 ( CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO: 49 (CDR1), SEQ ID NO: 51 (CDR2) and SEQ ID NO: 57 (CDR3) and an antigen-binding site, specific

- 38 041360 for CDH17, contains heavy chain CDRs containing the amino acid sequences of SEQ ID NO: 76 (CDR1), SEQ ID NO: 77 (CDR2) and SEQ ID NO: 78 (CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO: 79 (CDR1), SeQ ID NO: 80 (CDR2) and SEQ ID NO: 81 (CDR3). In this embodiment, the antigen-binding site specific for TRAILR2 is TRv6 and the antigen-binding site specific for CDH17 is CDH17v1.

Preferably, the antigen-binding site specific for TRAILR2 comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 106 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 107 and the antigen-binding site specific for CDH17 comprises a heavy chain variable domain, containing the amino acid sequence of SEQ ID NO: 118 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 119. In this embodiment, the antigen-binding site specific for TRAILR2 is TRv6 (for example, scFv SEQ ID NO: 129) and the antigen-binding site , specific for CDH17, is CDH17v1.

In a preferred embodiment of the binding molecule of the invention, the antigen-binding site specific for TRAILR2 comprises heavy chain CDRs comprising the amino acid sequences of SEQ ID NO: 43 (CDR1), SEQ ID NO: 47 (CDR2), and SEQ ID NO: 48 ( CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO: 49 (CDR1), SEQ ID NO: 53 (CDR2) and SEQ ID NO: 57 (CDR3) and an antigen-binding site specific for CDH17 contains heavy chain CDRs containing amino acid sequences of SEQ ID NO: 58 (CDR1) SEQ ID NO: 59 (CDR2) and SEQ ID NO: 60 (CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO: 61 (CDR1), SEQ ID NO: 62 ( CDR2) and SEQ ID NO: 63 (CDR3). In this embodiment, the antigen-binding site specific for TRAILR2 is TRv7 and the antigen-binding site specific for CDH17 is CDH17E9.

Preferably, the antigen-binding site specific for TRAILR2 comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 108 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 109 and the antigen-binding site specific for CDH17 comprises a heavy chain variable domain, containing the amino acid sequence of SEQ ID NO: 110 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 111. In this embodiment, the antigen-binding site specific for TRAILR2 is TRv7 (for example, scFv SEQ ID NO: 130) and the antigen-binding site , specific for CDH17, is CDH17E9.

In a preferred embodiment of the binding molecule of the invention, the antigen-binding site specific for TRAILR2 comprises heavy chain CDRs comprising the amino acid sequences of SEQ ID NO: 43 (CDR1), SEQ ID NO: 47 (CDR2), and SEQ ID NO: 48 ( CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO: 49 (CDR1), SEQ ID NO: 53 (CDR2) and SEQ ID NO: 57 (CDR3) and an antigen-binding site specific for CDH17 contains heavy chain CDRs containing amino acid sequences of SEQ ID NO: 64 (CDR1) SEQ ID NO: 65 (CDR2) and SEQ ID NO: 66 (CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO: 67 (CDR1), SEQ ID NO: 68 ( CDR2) and SEQ ID NO: 69 (CDR3). In this embodiment, the antigen-binding site specific for TRAILR2 is TRv7 and the antigen-binding site specific for CDH17 is CDH17A6.

Preferably, the antigen-binding site specific for TRAILR2 comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 108 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 109 and the antigen-binding site specific for CDH17 comprises a heavy chain variable domain, containing the amino acid sequence of SEQ ID NO: 112 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 113. In this embodiment, the antigen-binding site specific for TRAILR2 is TRv7 (for example, scFv SEQ ID NO: 130) and the antigen-binding site , specific for CDH17, is CDH17A6.

In a preferred embodiment of the binding molecule of the invention, the antigen-binding site specific for TRAILR2 comprises heavy chain CDRs comprising the amino acid sequences of SEQ ID NO: 43 (CDR1), SEQ ID NO: 47 (CDR2), and SEQ ID NO: 48 ( CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO: 49 (CDR1), SEQ ID NO: 53 (CDR2) and SEQ ID NO: 57 (CDR3) and an antigen-binding site specific for CDH17 contains heavy chain CDRs containing amino acid sequences of SEQ ID NO: 70 (CDR1), SEQ ID NO: 71 (CDR2) and SEQ ID NO: 72 (CDR3) and light chain CDRs containing

- 39 041360 amino acid sequences of SEQ ID NO: 73 (CDR1), SEQ ID NO: 74 (CDR2) and SEQ ID NO: 75 (CDR3). In this embodiment, the antigen-binding site specific for TRAILR2 is TRv7 and the antigen-binding site specific for CDH17 is CDH17E2 or CDH17H2.

Preferably, the antigen-binding site specific for TRAILR2 comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 108 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 109 and the antigen-binding site specific for CDH17 comprises a heavy chain variable domain, containing the amino acid sequence of SEQ ID NO: 114 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 115. In this embodiment, the antigen-binding site specific for TRAILR2 is TRv7 (for example, scFv SEQ ID NO: 130) and the antigen-binding site , specific for CDH17, is CDH17E2.

Preferably, the antigen-binding site specific for TRAILR2 comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 108 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 109 and the antigen-binding site specific for CDH17 comprises a heavy chain variable domain, containing the amino acid sequence of SEQ ID NO: 116 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 117. In this embodiment, the antigen-binding site specific for TRAILR2 is TRv7 (for example, scFv SEQ ID NO: 130) and the antigen-binding site , specific for CDH17, is CDH17H2.

In a preferred embodiment of the binding molecule of the invention, the antigen-binding site specific for TRAILR2 comprises heavy chain CDRs comprising the amino acid sequences of SEQ ID NO: 43 (CDR1), SEQ ID NO: 47 (CDR2), and SEQ ID NO: 48 ( CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO: 49 (CDR1), SEQ ID NO: 53 (CDR2) and SEQ ID NO: 57 (CDR3) and an antigen-binding site specific for CDH17 contains heavy chain CDRs containing amino acid sequences of SEQ ID NO: 76 (CDR1), SEQ ID NO: 77 (CDR2) and SEQ ID NO: 78 (CDR3) and light chain CDRs containing the amino acid sequences of SEQ ID NO: 79 (CDR1), SEQ ID NO: 80 (CDR2) and SEQ ID NO: 81 (CDR3). In this embodiment, the antigen-binding site specific for TRAILR2 is TRv7 and the antigen-binding site specific for CDH17 is CDH17v1.

Preferably, the antigen-binding site specific for TRAILR2 comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 108 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 109 and the antigen-binding site specific for CDH17 comprises a heavy chain variable domain, containing the amino acid sequence of SEQ ID NO: 118 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 119. In this embodiment, the antigen-binding site specific for TRAILR2 is TRv7 (for example, scFv SEQ ID NO: 130) and the antigen-binding site , specific for CDH17, is CDH17v1.

The above are specific combinations of antigen-binding sites specific for TRAILR2 and CDH17 (e.g., the TRAILR2 binding site of any of TR#1, TR#2, TR#3, TR#7, TR#8, TR#10, TR#12, TR2v1, TR2v2, TR2v3, TR2v4, TR2v5, TR2v6 and TR2v7 in combination with the CDH17 binding site of any of CDH17E9, CDH17A6, CDH17E2, CDH17H2 and CDH17v1) that can be used in a binding molecule according to the invention (e.g. bispecific and tetravalent binding molecules).

In some embodiments of the combinations described above, the CDH17 antigen binding site comprises a heavy chain variable domain specific for CDH17 (e.g., the VH of the CDH17E9, CDH17A6, CDH17E2, CDH17H2, or CDH17v1 antigen binding site) and is fused to a human heavy chain constant region, e.g., an IgG constant region. , IgG1, IgG2, IgG3, IgG4, IgA, IgE or IgM. Preferably, the human IgG1 heavy chain constant region is used.

Another embodiment of the invention is that the antigen binding site for CDH17 further comprises a light chain variable domain specific for CDH17 (e.g., VL of the antigen binding site CDH17E9, CDH17A6, CDH17E2, CDH17H2, or CDH17v1) and is fused to a human kappa or lambda light chain constant region . Preferably, the human kappa light chain constant region is used.

In some embodiments, the binding molecule comprises an Ig molecule (comprising two VH domains each fused to heavy chain constant regions, e.g., IgG1 constant regions, and two VL domains, each fused to light chain constant regions) containing any of CDH17 specific antigen-binding sites (CDH17E9, CDH17A6,

- 40 041360

CDH17E2, CDH17H2, or CDH17v1) and two scFvs containing any of the TRAILR2 specific antigen-binding sites (TR#1, TR#2, TR#3, TR#7, TR#8, TR#10, TR#12, TR2v1, TR2v2, TR2v3, TR2v4,

TR2v5, TR2v6, or TR2v7), where each of the scFv(s) is fused to the C-terminus of an IgG molecule (e.g., one scFv with one heavy chain and another scFv with a different heavy chain, each via a peptide linker), thereby forming a bispecific tetravalent binding molecule.

An exemplary sequence for heavy chain constant regions of human IgG1 wild type is shown in SEQ ID NO: 120, IgG1 KO is shown in SEQ ID NO: 121, wild type IgG4 Pro is shown in SEQ ID NO: 122. IgG1 FcRnmut is shown in SEQ ID NO: 270 Preferably the heavy chain constant region is IgGIKO as shown in SEQ ID NO: 121 or IgG1FcRnmut as shown in SEQ ID NO: 270.

An exemplary sequence for the human kappa light chain constant region is shown in SEQ ID NO: 123.

Below are the binding molecules (also referred to in the present invention as multispecific binding proteins) in accordance with the invention. Each of the specific molecules/proteins of the invention comprises modified immunoglobulin molecules wherein (i) the immunoglobulin heavy chain comprises a heavy chain variable domain amino acid sequence that specifically binds to CDH17 (e.g., the VH of any of CDH17E9, CDH17A6, CDH17E2, CDH17H2, and CDH17v1), immunoglobulin heavy chain constant domains, and scFv that specifically binds to TRAILR2 containing the amino acid sequence of light chain and heavy chain variable domains (e.g., VL and VH of any of TR#1, TR#2, TR#3, TR #7, TR#8, TR#10, TR#12, TR2v1, TR2v2, TR2v3, TR2v4, TR2v5, TR2v6 and TR2v7), and wherein the scFv is linked to the C-terminus of the Ig constant domains, and (ii) the immunoglobulin light chain contains a light chain variable domain amino acid sequence that specifically binds to CDH17 (eg, a VL of any of CDH17E9, CDH17A6, CDH17E2, CDH17H2, and CDH17v1) and a light chain constant domain. Preferably, the modified immunoglobulin molecules comprise two immunoglobulin heavy chains (eg, modified heavy chains) and two immunoglobulin light chains.

In some embodiments, the binding molecules set forth in various aspects below, which are defined by their heavy chain amino acid sequence (e.g., a modified heavy chain with scFv fused to the C-terminus of an Ig heavy chain) as well as their light chain amino acid sequences, comprise two heavy chains. chains and two light chains, thus forming a symmetrical tetravalent and bispecific structure.

Another aspect of the invention relates to a binding molecule containing a heavy chain containing the amino acid sequence of SEQ ID NO: 159 and a light chain containing the amino acid sequence of SEQ ID NO: 173. In this aspect, the antigen-binding site specific for CDH17 is CDH17E9 and the antigen-binding site TRAILR2 specific is TR#1.

Another aspect of the invention relates to a binding molecule containing a heavy chain containing the amino acid sequence of SEQ ID NO: 160 and a light chain containing the amino acid sequence of SEQ ID NO: 173. In this aspect, the antigen-binding site specific for CDH17 is CDH17E9 and the antigen-binding site TRAILR2 specific is TR#2.

Another aspect of the invention relates to a binding molecule containing a heavy chain containing the amino acid sequence of SEQ ID NO: 161 and a light chain containing the amino acid sequence of SEQ ID NO: 173. In this aspect, the antigen-binding site specific for CDH17 is CDH17E9 and the antigen-binding site TRAILR2 specific is TR#3.

Another aspect of the invention relates to a binding molecule containing a heavy chain containing the amino acid sequence of SEQ ID NO: 162 and a light chain containing the amino acid sequence of SEQ ID NO: 173. In this aspect, the antigen-binding site specific for CDH17 is CDH17E9, and the antigen-binding site specific for TRAILR2 is TR#7.

Another aspect of the invention relates to a binding molecule containing a heavy chain containing the amino acid sequence of SEQ ID NO: 163 and a light chain containing the amino acid sequence of SEQ ID NO: 173. In this aspect, the antigen-binding site specific for CDH17 is CDH17E9 and antigen-binding the TRAILR2 specific region is TR#8.

Another aspect of the invention relates to a binding molecule containing a heavy chain containing the amino acid sequence of SEQ ID NO: 164 and a light chain containing the amino acid sequence of SEQ ID NO: 173. In this aspect, the antigen-binding site specific for CDH17 is CDH17E9 and antigen-binding the TRAILR2 specific region is TR#10.

- 41 041360

Another aspect of the invention relates to a binding molecule containing a heavy chain containing the amino acid sequence of SEQ ID NO: 165 and a light chain containing the amino acid sequence of SEQ ID NO: 173. In this aspect, the antigen-binding site specific for CDH17 is CDH17E9 and the antigen-binding area specific to

TRAILR2 is TR#12.

Another aspect of the invention relates to a binding molecule containing a heavy chain containing the amino acid sequence of SEQ ID NO: 166 and a light chain containing the amino acid sequence of SEQ ID NO: 173. In this aspect, the antigen-binding site specific for CDH17 is CDH17E9 and the antigen-binding site , specific for TRAILR2, is TRv1.

Another aspect of the invention relates to a binding molecule containing a heavy chain containing the amino acid sequence of SEQ ID NO: 167 and a light chain containing the amino acid sequence of SEQ ID NO: 173. In this aspect, the antigen-binding site specific for CDH17 is CDH17E9 and the antigen-binding site , specific for TRAILR2, is TRv2.

Another aspect of the invention relates to a binding molecule containing a heavy chain containing the amino acid sequence of SEQ ID NO: 168 and a light chain containing the amino acid sequence of SEQ ID NO: 173. In this aspect, the antigen-binding site specific for CDH17 is CDH17E9 and the antigen-binding site , specific to TRAILR2, is TRv3.

Another aspect of the invention relates to a binding molecule containing a heavy chain containing the amino acid sequence of SEQ ID NO: 169 and a light chain containing the amino acid sequence of SEQ ID NO: 173. In this aspect, the antigen-binding site specific for CDH17 is CDH17E9 and antigen-binding the TRAILR2 specific region is TRv4.

Another aspect of the invention relates to a binding molecule containing a heavy chain containing the amino acid sequence of SEQ ID NO: 170 and a light chain containing the amino acid sequence of SEQ ID NO: 173. In this aspect, the antigen-binding site specific for CDH17 is CDH17E9 and antigen-binding the TRAILR2 specific region is TRv5.

Another aspect of the invention relates to a binding molecule containing a heavy chain containing the amino acid sequence of SEQ ID NO: 171 and a light chain containing the amino acid sequence of SEQ ID NO: 173. In this aspect, the antigen-binding site specific for CDH17 is CDH17E9 and the antigen-binding the TRAILR2 specific region is TRv6.

Another aspect of the invention relates to a binding molecule containing a heavy chain containing the amino acid sequence of SEQ ID NO: 172 and a light chain containing the amino acid sequence of SEQ ID NO: 173. In this aspect, the antigen-binding site specific for CDH17 is CDH17E9 and antigen-binding the TRAILR2 specific region is TRv7.

Another aspect of the invention relates to a binding molecule containing a heavy chain containing the amino acid sequence of SEQ ID NO: 174 and a light chain containing the amino acid sequence of SEQ ID NO: 188. In this aspect, the antigen-binding site specific for CDH17 is CDH17A6 and the antigen-binding the TRAILR2 specific region is TR#1.

Another aspect of the invention relates to a binding molecule containing a heavy chain containing the amino acid sequence of SEQ ID NO: 175 and a light chain containing the amino acid sequence of SEQ ID NO: 188. In this aspect, the antigen-binding site specific for CDH17 is CDH17A6 and the antigen-binding the TRAILR2 specific region is TR#2.

Another aspect of the invention relates to a binding molecule containing a heavy chain containing the amino acid sequence of SEQ ID NO: 176 and a light chain containing the amino acid sequence of SEQ ID NO: 188. In this aspect, the antigen-binding site specific for CDH17 is CDH17A6 and the antigen-binding site TRAILR2 specific is TR#3.

Another aspect of the invention relates to a binding molecule containing a heavy chain containing the amino acid sequence of SEQ ID NO: 177 and a light chain containing the amino acid sequence of SEQ ID NO: 188. In this aspect, the antigen-binding site specific for CDH17 is CDH17A6 and the antigen-binding site , specific to TRAILR2, is TR#7.

Another aspect of the invention relates to a binding molecule comprising a heavy chain containing the amino acid sequence of SEQ ID NO: 178 and a light chain containing the amino acid sequence of SEQ ID NO: 188. In this aspect, an antigen-binding site specific for CDH17 is is CDH17A6 and an antigen-binding site specific for

TRAILR2 is TR#8.

Another aspect of the invention relates to a binding molecule containing a heavy chain containing the amino acid sequence of SEQ ID NO: 179 and a light chain containing the amino acid sequence of SEQ ID NO: 188. In this aspect, the antigen-binding site specific for CDH17 is CDH17A6 and the antigen-binding the TRAILR2 specific region is TR#10.

Another aspect of the invention relates to a binding molecule containing a heavy chain containing the amino acid sequence of SEQ ID NO: 180 and a light chain containing the amino acid sequence of SEQ ID NO: 188. In this aspect, the antigen-binding site specific for CDH17 is CDH17A6 and the antigen-binding the TRAILR2 specific region is TR#12.

Another aspect of the invention relates to a binding molecule containing a heavy chain containing the amino acid sequence of SEQ ID NO: 181 and a light chain containing the amino acid sequence of SEQ ID NO: 188. In this aspect, the antigen-binding site specific for CDH17 is CDH17A6 and the antigen-binding the TRAILR2 specific region is TRv1.

Another aspect of the invention relates to a binding molecule containing a heavy chain containing the amino acid sequence of SEQ ID NO: 182 and a light chain containing the amino acid sequence of SEQ ID NO: 188. In this aspect, the antigen-binding site specific for CDH17 is CDH17A6 and the antigen-binding site , specific for TRAILR2, is TRv2.

Another aspect of the invention relates to a binding molecule containing a heavy chain containing the amino acid sequence of SEQ ID NO: 183 and a light chain containing the amino acid sequence of SEQ ID NO: 188. In this aspect, the antigen-binding site specific for CDH17 is CDH17A6 and the antigen-binding the TRAILR2 specific region is TRv3.

Another aspect of the invention relates to a binding molecule containing a heavy chain containing the amino acid sequence of SEQ ID NO: 184 and a light chain containing the amino acid sequence of SEQ ID NO: 188. In this aspect, the antigen-binding site specific for CDH17 is CDH17A6 and the antigen-binding the TRAILR2 specific region is TRv4.

Another aspect of the invention relates to a binding molecule containing a heavy chain containing the amino acid sequence of SEQ ID NO: 185 and a light chain containing the amino acid sequence of SEQ ID NO: 188. In this aspect, the antigen-binding site specific for CDH17 is CDH17A6 and the antigen-binding the TRAILR2 specific region is TRv5.

Another aspect of the invention relates to a binding molecule containing a heavy chain containing the amino acid sequence of SEQ ID NO: 186 and a light chain containing the amino acid sequence of SEQ ID NO: 188. In this aspect, the antigen-binding site specific for CDH17 is CDH17A6 and the antigen-binding the TRAILR2 specific region is TRv6.

Another aspect of the invention relates to a binding molecule containing a heavy chain containing the amino acid sequence of SEQ ID NO: 187 and a light chain containing the amino acid sequence of SEQ ID NO: 188. In this aspect, the antigen-binding site specific for CDH17 is CDH17A6 and the antigen-binding the TRAILR2 specific region is TRv7.

Another aspect of the invention relates to a binding molecule containing a heavy chain containing the amino acid sequence of SEQ ID NO: 189 and a light chain containing the amino acid sequence of SEQ ID NO: 203. In this aspect, the antigen-binding site specific for CDH17 is CDH17E2 and the antigen-binding the TRAILR2 specific region is TR#1.

Another aspect of the invention relates to a binding molecule containing a heavy chain containing the amino acid sequence of SEQ ID NO: 190 and a light chain containing the amino acid sequence of SEQ ID NO: 203. In this aspect, the antigen-binding site specific for CDH17 is CDH17E2 and antigen-binding the TRAILR2 specific region is TR#2.

Another aspect of the invention relates to a binding molecule containing a heavy chain containing the amino acid sequence of SEQ ID NO: 191 and a light chain containing the amino acid sequence of SEQ ID NO: 203. In this aspect, the antigen-binding site specific for CDH17 is CDH17E2 and the antigen-binding area specific to

- 43 041360

TRAILR2 is TR#3.

Another aspect of the invention relates to a binding molecule containing a heavy chain containing the amino acid sequence of SEQ ID NO: 192 and a light chain containing the amino acid sequence of SEQ ID NO: 203. In this aspect, the antigen-binding site specific for CDH17 is CDH17E2 and the antigen-binding the TRAILR2 specific region is TR#7.

Another aspect of the invention relates to a binding molecule containing a heavy chain containing the amino acid sequence of SEQ ID NO: 193 and a light chain containing the amino acid sequence of SEQ ID NO: 203. In this aspect, the antigen-binding site specific for CDH17 is CDH17E2 and antigen-binding the TRAILR2 specific region is TR#8.

Another aspect of the invention relates to a binding molecule containing a heavy chain containing the amino acid sequence of SEQ ID NO: 194 and a light chain containing the amino acid sequence of SEQ ID NO: 203. In this aspect, the antigen-binding site specific for CDH17 is CDH17E2 and the antigen-binding the TRAILR2 specific region is TR#10.

Another aspect of the invention relates to a binding molecule containing a heavy chain containing the amino acid sequence of SEQ ID NO. 195 and a light chain comprising the amino acid sequence of SEQ ID NO: 203. In this aspect, the CDH17 specific antigen binding site is CDH17E2 and the TRAILR2 specific antigen binding site is TR#12.

Another aspect of the invention relates to a binding molecule containing a heavy chain containing the amino acid sequence of SEQ ID NO: 196 and a light chain containing the amino acid sequence of SEQ ID NO: 203. In this aspect, the antigen-binding site specific for CDH17 is CDH17E2 and the antigen-binding the TRAILR2 specific region is TRv1.

Another aspect of the invention relates to a binding molecule containing a heavy chain containing the amino acid sequence of SEQ ID NO: 197 and a light chain containing the amino acid sequence of SEQ ID NO: 203. In this aspect, the antigen-binding site specific for CDH17 is CDH17E2 and antigen-binding the TRAILR2 specific region is TRv2.

Another aspect of the invention relates to a binding molecule containing a heavy chain containing the amino acid sequence of SEQ ID NO: 198 and a light chain containing the amino acid sequence of SEQ ID NO: 203. In this aspect, the antigen-binding site specific for CDH17 is CDH17E2 and the antigen-binding the TRAILR2 specific region is TRv3.

Another aspect of the invention relates to a binding molecule containing a heavy chain containing the amino acid sequence of SEQ ID NO: 199 and a light chain containing the amino acid sequence of SEQ ID NO: 203. In this aspect, the antigen-binding site specific for CDH17 is CDH17E2 and antigen-binding the TRAILR2 specific region is TRv4.

Another aspect of the invention relates to a binding molecule containing a heavy chain containing the amino acid sequence of SEQ ID NO: 200 and a light chain containing the amino acid sequence of SEQ ID NO: 203. In this aspect, the antigen-binding site specific for CDH17 is CDH17E2 and the antigen-binding the TRAILR2 specific region is TRv5.

Another aspect of the invention relates to a binding molecule containing a heavy chain containing the amino acid sequence of SEQ ID NO: 201 and a light chain containing the amino acid sequence of SEQ ID NO: 203. In this aspect, the antigen-binding site specific for CDH17 is CDH17E2 and the antigen-binding site , specific for TRAILR2, is TRv6.

Another aspect of the invention relates to a binding molecule containing a heavy chain containing the amino acid sequence of SEQ ID NO: 202 and a light chain containing the amino acid sequence of SEQ ID NO: 203. In this aspect, the antigen-binding site specific for CDH17 is CDH17E2 and the antigen-binding the TRAILR2 specific region is TRv7.

Another aspect of the invention relates to a binding molecule containing a heavy chain containing the amino acid sequence of SEQ ID NO: 204 and a light chain containing the amino acid sequence of SEQ ID NO: 218. In this aspect, the antigen-binding site specific for CDH17 is CDH17H2 and the antigen-binding the TRAILR2 specific region is TR#1.

Another aspect of the invention relates to a binding molecule comprising a heavy chain comprising the amino acid sequence of SEQ ID NO: 205 and a light chain comprising the amino acid sequence of SEQ ID NO: 218. In this aspect, an antigen-binding site specific for CDH17 is is CDH17H2 and an antigen-binding site specific for

TRAILR2 is TR#2.

Another aspect of the invention relates to a binding molecule containing a heavy chain containing the amino acid sequence of SEQ ID NO: 206 and a light chain containing the amino acid sequence of SEQ ID NO: 218. In this aspect, the antigen-binding site specific for CDH17 is CDH17H2 and the antigen-binding the TRAILR2 specific region is TR#3.

Another aspect of the invention relates to a binding molecule containing a heavy chain containing the amino acid sequence of SEQ ID NO: 207 and a light chain containing the amino acid sequence of SEQ ID NO: 218. In this aspect, the antigen-binding site specific for CDH17 is CDH17H2 and the antigen-binding the TRAILR2 specific region is TR#7.

Another aspect of the invention relates to a binding molecule containing a heavy chain containing the amino acid sequence of SEQ ID NO: 208 and a light chain containing the amino acid sequence of SEQ ID NO: 218. In this aspect, the antigen-binding site specific for CDH17 is CDH17H2 and the antigen-binding the TRAILR2 specific region is TR#8.

Another aspect of the invention relates to a binding molecule containing a heavy chain containing the amino acid sequence of SEQ ID NO: 209 and a light chain containing the amino acid sequence of SEQ ID NO: 218. In this aspect, the antigen-binding site specific for CDH17 is CDH17H2 and the antigen-binding the TRAILR2 specific region is TR#10.

Another aspect of the invention relates to a binding molecule containing a heavy chain containing the amino acid sequence of SEQ ID NO: 210 and a light chain containing the amino acid sequence of SEQ ID NO: 218. In this aspect, the antigen-binding site specific for CDH17 is CDH17H2 and the antigen-binding the TRAILR2 specific region is TR#12.

Another aspect of the invention relates to a binding molecule containing a heavy chain containing the amino acid sequence of SEQ ID NO: 211 and a light chain containing the amino acid sequence of SEQ ID NO: 218. In this aspect, the antigen-binding site specific for CDH17 is CDH17H2 and the antigen-binding the TRAILR2 specific region is TRv1.

Another aspect of the invention relates to a binding molecule containing a heavy chain containing the amino acid sequence of SEQ ID NO: 212 and a light chain containing the amino acid sequence of SEQ ID NO: 218. In this aspect, the antigen-binding site specific for CDH17 is CDH17H2 and the antigen-binding the TRAILR2 specific region is TRv2.

Another aspect of the invention relates to a binding molecule containing a heavy chain containing the amino acid sequence of SEQ ID NO: 271 and a light chain containing the amino acid sequence of SEQ ID NO: 218. In this aspect, the antigen-binding site specific for CDH17 is CDH17H2 and the antigen-binding the TRAILR2 specific region is TRv2.

Another aspect of the invention relates to a binding molecule containing a heavy chain containing the amino acid sequence of SEQ ID NO: 213 and a light chain containing the amino acid sequence of SEQ ID NO: 218. In this aspect, the antigen-binding site specific for CDH17 is CDH17H2 and the antigen-binding the TRAILR2 specific region is TRv3.

Another aspect of the invention relates to a binding molecule containing a heavy chain containing the amino acid sequence of SEQ ID NO: 214 and a light chain containing the amino acid sequence of SEQ ID NO: 218. In this aspect, the antigen-binding site specific for CDH17 is CDH17H2 and the antigen-binding the TRAILR2 specific region is TRv4.

Another aspect of the invention relates to a binding molecule containing a heavy chain containing the amino acid sequence of SEQ ID NO: 215 and a light chain containing the amino acid sequence of SEQ ID NO: 218. In this aspect, the antigen-binding site specific for CDH17 is CDH17H2 and the antigen-binding the TRAILR2 specific region is TRv5.

Another aspect of the invention relates to a binding molecule comprising a heavy chain containing the amino acid sequence of SEQ ID NO: 216 and a light chain containing the amino acid sequence of SEQ ID NO: 218. In this aspect, an antigen-binding site specific for CDH17 is is CDH17H2 and an antigen-binding site specific for

TRAILR2 is TRv6.

Another aspect of the invention relates to a binding molecule containing a heavy chain containing the amino acid sequence of SEQ ID NO: 217 and a light chain containing the amino acid sequence of SEQ ID NO: 218. In this aspect, the antigen-binding site specific for CDH17 is CDH17H2 and the antigen-binding the TRAILR2 specific region is TRv7.

Another aspect of the invention relates to a binding molecule containing a heavy chain containing the amino acid sequence of SEQ ID NO: 219 and a light chain containing the amino acid sequence of SEQ ID NO: 233. In this aspect, the antigen-binding site specific for CDH17 is CDH17v1 and antigen-binding the TRAILR2 specific region is TR#1.

Another aspect of the invention relates to a binding molecule containing a heavy chain containing the amino acid sequence of SEQ ID NO. 220 and a light chain comprising the amino acid sequence of SEQ ID NO: 233. In this aspect, the CDH17-specific antigen-binding site is CDH17v1 and the TRAILR2-specific antigen-binding site is TR#2.

Another aspect of the invention relates to a binding molecule containing a heavy chain containing the amino acid sequence of SEQ ID NO: 221 and a light chain containing the amino acid sequence of SEQ ID NO: 233. In this aspect, the antigen-binding site specific for CDH17 is CDH17v1 and antigen-binding the TRAILR2 specific region is TR#3.

Another aspect of the invention relates to a binding molecule containing a heavy chain containing the amino acid sequence of SEQ ID NO: 222 and a light chain containing the amino acid sequence of SEQ ID NO: 233. In this aspect, the antigen-binding site specific for CDH17 is CDH17v1 and antigen-binding the TRAILR2 specific region is TR#7.

Another aspect of the invention relates to a binding molecule containing a heavy chain containing the amino acid sequence of SEQ ID NO: 223, and a light chain containing the amino acid sequence of SEQ ID NO: 233. In this aspect, the antigen-binding site specific for CDH17 is CDH17v1 and antigen-binding the TRAILR2 specific region is TR#8.

Another aspect of the invention relates to a binding molecule containing a heavy chain containing the amino acid sequence of SEQ ID NO: 224 and a light chain containing the amino acid sequence of SEQ ID NO: 233. In this aspect, the antigen-binding site specific for CDH17 is CDH17v1 and antigen-binding the TRAILR2 specific region is TR#10.

Another aspect of the invention relates to a binding molecule containing a heavy chain containing the amino acid sequence of SEQ ID NO: 225 and a light chain containing the amino acid sequence of SEQ ID NO: 233. In this aspect, the antigen-binding site specific for CDH17 is CDH17v1 and antigen-binding the TRAILR2 specific region is TR#12.

Another aspect of the invention relates to a binding molecule containing a heavy chain containing the amino acid sequence of SEQ ID NO: 226 and a light chain containing the amino acid sequence of SEQ ID NO: 233. In this aspect, the antigen-binding site specific for CDH17 is CDH17v1 and antigen-binding the TRAILR2 specific region is TRv1.

Another aspect of the invention relates to a binding molecule containing (i) a heavy chain containing the amino acid sequence of SEQ ID NO: 145, and a light chain containing the amino acid sequence of SEQ ID NO: 146;

(ii) a heavy chain containing the amino acid sequence of SEQ ID NO: 147 and a light chain containing the amino acid sequence of SEQ ID NO: 148;

(iii) a heavy chain containing the amino acid sequence of SEQ ID NO: 153 and a light chain containing the amino acid sequence of SEQ ID NO: 154;

(iv) a heavy chain containing the amino acid sequence of SEQ ID NO: 155 and a light chain containing the amino acid sequence of SEQ ID NO: 156; or (v) a heavy chain containing the amino acid sequence of SEQ ID NO: 157 and a light chain containing the amino acid sequence of SEQ ID NO: 158.

In this aspect, the CDH17-specific antigen-binding site is CDH17v1 and the TRAILR2-specific antigen-binding site is TRv1.

Another aspect of the invention relates to a binding molecule containing a heavy chain containing the amino acid sequence of SEQ ID NO: 227 and a light chain containing the amino acid sequence of SEQ ID NO: 233. In this aspect, the antigen-binding site specific for CDH17 is CDH17v1 and antigen-binding site specific for

TRAILR2 is TRv2.

Another aspect of the invention relates to a binding molecule containing (i) a heavy chain containing the amino acid sequence of SEQ ID NO: 149 and a light chain containing the amino acid sequence of SEQ ID NO: 150, or (ii) a heavy chain containing the amino acid sequence of SEQ ID NO: 151, and a light chain containing the amino acid sequence of SEQ ID NO: 152.

In this aspect, the CDH17-specific antigen-binding site is CDH17v1 and the TRAILR2-specific antigen-binding site is TRv2.

Another aspect of the invention relates to a binding molecule containing a heavy chain containing the amino acid sequence of SEQ ID NO: 228, and a light chain containing the amino acid sequence of SEQ ID NO: 233. In this aspect, the antigen-binding site specific for CDH17 is CDH17v1 and antigen-binding the TRAILR2 specific region is TRv3.

Another aspect of the invention relates to a binding molecule containing a heavy chain containing the amino acid sequence of SEQ ID NO: 229 and a light chain containing the amino acid sequence of SEQ ID NO: 233. In this aspect, the antigen-binding site specific for CDH17 is CDH17v1 and antigen-binding the TRAILR2 specific region is TRv4.

Another aspect of the invention relates to a binding molecule containing a heavy chain containing the amino acid sequence of SEQ ID NO: 230 and a light chain containing the amino acid sequence of SEQ ID NO: 233. In this aspect, the antigen-binding site specific for CDH17 is CDH17v1 and antigen-binding the TRAILR2 specific region is TRv5.

Another aspect of the invention relates to a binding molecule containing a heavy chain containing the amino acid sequence of SEQ ID NO: 231 and a light chain containing the amino acid sequence of SEQ ID NO: 233. In this aspect, the antigen-binding site specific for CDH17 is CDH17v1 and antigen-binding the TRAILR2 specific region is TRv6.

Another aspect of the invention relates to a binding molecule containing a heavy chain containing the amino acid sequence of SEQ ID NO: 232 and a light chain containing the amino acid sequence of SEQ ID NO: 233. In this aspect, the antigen-binding site specific for CDH17 is CDH17v1 and antigen-binding the TRAILR2 specific region is TRv7.

In addition, the present invention provides antibody molecules (for example, a full-length antibody/immunoglobulin molecule having a Y-shaped structure with two heavy and two light chains, or fragments thereof, such as a fragment of Fv, Fab, Fab', or F(ab' ) ₂ , single chain antibody, single chain variable fragment (scFv)) that specifically bind to CDH17. In some embodiments, the antibody molecules specific for CDH17 are recombinant monoclonal antibodies, chimeric, humanized, or human antibodies.

In some embodiments, an antibody molecule specific for CDH17 comprises a heavy chain CDR comprising the amino acid sequences of SEQ ID NO: 58 (CDR1), SEQ ID NO: 59 (CDR2), and SEQ ID NO: 60 (CDR3), and a light chain CDR containing the amino acid sequences of SEQ ID NO: 61 (CDR1), SEQ ID NO: 62 (CDR2) and SEQ ID NO: 63 (CDR3).

In some embodiments, an antibody molecule specific for CDH17 comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 110 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 111.

In some embodiments, an antibody molecule specific for CDH17 comprises a heavy chain CDR comprising the amino acid sequences of SEQ ID NO: 64 (CDR1), SEQ ID NO: 65 (CDR2), and SEQ ID NO: 66 (CDR3), and a light chain CDR containing the amino acid sequences of SEQ ID NO: 67 (CDR1), SEQ ID NO: 68 (CDR2) and SEQ ID NO: 69 (CDR3).

In some embodiments, an antibody molecule specific for CDH17 comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 112 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 113.

In some embodiments, an antibody molecule specific for CDH17 comprises a heavy chain CDR comprising the amino acid sequences of SEQ ID NO: 70 (CDR1), SEQ ID NO: 71 (CDR2), and SEQ ID NO: 72 (CDR3), and a light chain CDR containing the amino acid sequences of SEQ ID NO: 73 (CDR1), SEQ ID NO: 74 (CDR2) and SEQ ID NO: 75 (CDR3).

In some embodiments, an antibody molecule specific for CDH17 comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 114, and

- 47 041360 light chain variable domain containing the amino acid sequence of SEQ ID NO: 115.

In some embodiments, an antibody molecule specific for CDH17 comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 116 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 117.

In some embodiments, an antibody molecule specific for CDH17 comprises a heavy chain CDR comprising the amino acid sequences of SEQ ID NO: 76 (CDR1), SEQ ID NO: 77 (CDR2), and SEQ ID NO: 78 (CDR3), and a light chain CDR containing the amino acid sequences of SEQ ID NO: 79 (CDR1), SEQ ID NO: 80 (CDR2) and SEQ ID NO: 81 (CDR3).

In some embodiments, an antibody molecule specific for CDH17 comprises a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 118 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 119.

In some embodiments, CDH17-specific antibodies as defined above further comprise human heavy chain constant domains (eg, an IgG constant domain) and a human light chain constant domain (e.g., a kappa or lambda light chain constant domain). In some embodiments, the heavy chain constant domain is wild-type human IgG1 (for example, as shown in SEQ ID NO: 120), IgG1 KO (for example, as shown in SEQ ID NO: 121), wild-type IgG4 Pro (for example, as shown in SEQ ID NO: 122), or IgG1 FcRnmut (eg, as shown in SEQ ID NO: 270). In some embodiments, the human light chain constant domain is human kappa (eg, as shown in SEQ ID NO: 123).

In some embodiments, the CDH17-specific antibody has a heavy chain comprising the sequence of any of SEQ ID NOs: 110, 112, 114, 116, or 118 fused to the sequence of any of SEQ ID NOs: 120, 121, 122, or 270, and a light chain containing the sequence of any of SEQ ID NOs: 111, 113, 115, 117, or 119 fused to the sequence of SEQ ID NO: 123 (for example, a light chain containing the sequence of any of SEQ ID NOs: 173, 188, 203, 218, or 233 ).

The CDH17-specific antibodies provided in the present invention can be used to label, localize, identify, or target cells expressing CDH17 (for example, in ELISA assays, FACS assay, immunohistology, or the like) by attaching a dye, drug, or other molecule with binding specificity for another antigen. The CDH17-specific antibodies described herein alone do not affect the viability of cells expressing CDH17. In some embodiments, CDH17-specific antibodies specifically bind to the surface of a CDH17-expressing cell and are used to localize and/or identify such cells. In some embodiments, anti-CDH17 antibodies provided herein are used to identify cells expressing CDH17 (eg, tumor cells). In some embodiments, anti-CDH17 antibodies of the present invention are used to deliver a drug or cytotoxic agent to a target cell (e.g., a tumor cell expressing CDH17) by attaching such drug or cytotoxic agent to said anti-CDH17 antibody, thus, for example , killing the specified target cell.

Another aspect of the invention relates to isolated nucleic acid molecules which encode a binding molecule according to the invention or an antibody molecule according to the invention, or an expression vector, such nucleic acid molecule(s).

In some embodiments, the binding molecules of the invention or the antibody molecule of the invention comprise antibody heavy chain and/or light chain polypeptides. As one skilled in the art will appreciate, nucleic acid molecules that encode heavy chain polypeptides, light chain polypeptides, or heavy chain polypeptides and light chain polypeptides can be readily prepared.

Nucleic acid molecules encoding the light chain and the heavy chain can be synthesized chemically and enzymatically by polymerase chain reaction (PCR) using methods known in the art (eg Gait, 1984) which can be used to produce a synthetic gene. Methods for generating synthetic genes from oligonucleotides are known in the art (eg, Stemmer et al., 1995; Ye et al., 1992; Hayden et Mandecki, 1988; Frank et al., 1987).

Nucleic acid molecules of the invention include, but are not limited to, DNA molecules encoding the polypeptide sequences shown in the sequence listing. The present invention also relates to nucleic acid molecules that hybridize to DNA molecules encoding the polypeptide sequences shown in the sequence listing under high stringency binding and washing conditions as defined in WO 2007/042309. Preferred molecules (in terms of mRNA) are those that have at least 75% or 80% (preferably at least 85%, more preferably at least 90% and most preferably at least 95%) homology or identity

- 48 041360 sequence with one of the DNA molecules described in the present invention. By way of example, in terms of antibody expression in eukaryotic cells, the DNA sequences presented in the sequence listing have been designed to match codon usage in eukaryotic cells. If it is desired to express antibodies in E. coli, then these sequences can be changed according to the use of the E. coli codon. Variants of DNA molecules in accordance with the invention can be designed in several different ways, as described, for example, in WO 2007/042309. As used herein, the terms identical or percent identity in the context of two or more nucleic acid sequences or polypeptides refers to two or more sequences or subsequences that are the same or have a certain percentage of nucleotides or amino acid residues that are the same when compared and aligned for maximum match. To determine percent identity, sequences are aligned for optimal comparison purposes (eg, gaps can be introduced into the first amino acid sequence or nucleic acid sequence for optimal alignment with the second amino acid or nucleic acid sequence). The amino acid residues or nucleotides at the corresponding amino acid positions or nucleotide positions are then compared. When a position in the first sequence is occupied by the same amino acid residue or nucleotide as the corresponding position in the second sequence, then the molecules at that position are identical. The percent identity between two sequences is a function of the number of identical positions separated by the sequences (ie, % identity = # identical positions/total # positions (eg, overlapping positions) x 100). In some embodiments, the two sequences being compared are the same length after spaces are introduced in the sequence, as appropriate (eg, except for an additional sequence outside the sequences being compared). For example, when variable region sequences are compared, leader sequences and/or constant region sequences are not taken into account. For sequence comparisons between two sequences, the corresponding CDR refers to the CDR at the same location in both sequences (eg, CDR-H1 of each sequence).

Determination of percent identity or percent similarity between two sequences can be performed using a mathematical algorithm. A preferred non-limiting example of a mathematical algorithm used to compare two sequences is the algorithm of Karlin and Altschul, 1990, Proc. Natl. Acad. sci. USA 87:2264-2268, modified as in Karlin and Altschul, 1993, Proc. Natl. Acad. sci. USA 90:5873-5877. Such an algorithm is included in the NBLAST and XBLAST programs by Altschul et al., 1990, J. Mol. Biol. 215:403-410. BLAST nucleotide searches can be performed using the NBLAST program, score=100, wordlength=12, to obtain nucleotide sequences homologous to the nucleic acid encoding the protein of interest. A BLAST protein search can be performed using the XBLAST program, score=50, wordlength=3, to obtain amino acid sequences homologous to the protein of interest. Gapped BLAST can be used to obtain gap alignments for comparison, as described in Altschul et al., 1997, Nucleic Acids Res. 25:3389-3402. Alternatively, PSI-Blast can be used to perform an iterative search that detects distant relationships between molecules (Id.). When using the BLAST, Gapped BLAST and PSI-Blast programs, the default parameters of the respective programs (eg XBLAST and NBLAST) can be used. Another preferred non-limiting example of a mathematical algorithm used for sequence comparison is the Myers and Miller algorithm, CABIOS (1989). Such an algorithm is included in the ALIGN program (version 2.0), part of the GCG sequence alignment software package. When using the ALIGN program to compare amino acid sequences, a PAM120 residue weight table, a gap length penalty of 12, and a gap penalty of 4 can be used. Additional algorithms for sequence analysis are known in the art and include ADVANCE and ADAM as described in Torellis and Robotti, 1994 , Comput. Appl. biosci. 10:3-5; and FASTA, described in Pearson and Lipman, 1988, Proc. Natl. Acad. sci. USA 85:2444-8. Within FASTA, ktup is a control option that sets the sensitivity and search speed. If ktup=2, then find similar regions in the two compared sequences by looking at pairs of aligned residues; if ktup=1, then individually aligned amino acids are examined, ktup can be set to 2 or 1 for protein sequences, or 1 to 6 for DNA sequences. By default, if ktup is not specified, it is 2 for proteins and 6 for DNA. Alternatively, protein sequence alignment can be performed using the CLUSTAL W algorithm as described in Higgins et al., 1996, Methods Enzymol. 266:383-402.

Another aspect of the invention relates to a method for obtaining a binding molecule or antibody molecule described in the present invention, including (a) culturing a host cell in accordance with the invention under conditions that allow the expression of the molecule; And

- 49 041360 (b) isolation of the molecule; and optionally (c) further purification and/or modification and/or formulation of the molecule.

An embodiment of this aspect according to the invention is that the preparation method further comprises the step (c) of further purification and/or modification and/or formulation of the binding molecule according to the invention.

To obtain binding molecules or antibodies according to the invention, DNA molecules encoding full length light and/or heavy chains or fragments thereof are inserted into an expression vector such that the sequences are operably linked to transcriptional and translational control sequences.

For the manufacture of binding molecules or antibodies in accordance with the invention, a person skilled in the art can choose from a wide variety of expression systems well known in the art, for example, reviewed by Kipriyanov and Le Gall, Curr Opin Drug Discov Devel. 2004 Mar;7(2):233-42.

Expression vectors include plasmids, retroviruses, cosmids derived from EBV episomes, and the like. The expression vector and expression control sequences are chosen to be compatible with the host cell. An antibody light chain gene and an antibody heavy chain gene or binding molecule heavy chain gene described in the present invention (eg, a gene containing an immunoglobulin heavy chain sequence fused at its C-terminus to an scFv sequence) can be inserted into separate vectors. In some embodiments, both light and heavy chain DNA sequences are inserted into the same expression vector. Suitable vectors are vectors that encode an operably completed human CH or CL immunoglobulin sequence with appropriate restriction sites designed such that any VH or VL sequence can be easily inserted and expressed as described above. The constant chain is usually kappa or lambda for an antibody light chain, for an antibody heavy chain it can be, without limitation, any IgG isotype (IgG1, IgG2, IgG3, IgG4) or other immunoglobulins, including allelic variants.

The recombinant expression vector may also encode a signal peptide that facilitates the secretion of an antibody chain (eg, heavy and light chain binding molecules or antibodies described in the present invention) from the host cell. The DNA encoding the antibody chain can be cloned into a vector such that the signal peptide binds in-frame to the amino terminus of the mature antibody DNA chain. The signal peptide can be an immunoglobulin signal peptide or a heterologous peptide from a non-immunoglobulin protein. Alternatively, the DNA sequence encoding the antibody chain (heavy and light chains of the binding molecules or antibodies described in the present invention) may already contain a signal peptide sequence.

In addition to DNA sequences encoding antibody chains (for example, the heavy and light chains of the binding molecules or antibodies described in the present invention), recombinant expression vectors carry regulatory sequences, including promoters, enhancers, termination and polyadenylation signals, and other expression control elements that control the expression of antibody chains in the host cell. Examples of promoter sequences (for example, for expression in mammalian cells) are promoters and/or enhancers derived from (CMV) (such as CMV Simian Virus 40 (SV40) (such as the SV40 promoter/enhancer), adenovirus (for example, the main late promoter adenovirus (AdMLP)), polyoma, and strong mammalian promoters such as native immunoglobulin and actin promoters Examples of polyadenylation signals are BGH polyA, late SV40 or early polyA, alternatively 3'UTRs of immunoglobulin genes, etc. can be used.

Recombinant expression vectors can also carry sequences that regulate replication of the vector in host cells (eg, origin of replication) and selectable marker genes. Nucleic acid molecules encoding the heavy chain or antigen-binding portion and/or light chain or antigen-binding portion thereof of the binding molecule or antibody described herein and vectors containing these DNA molecules can be introduced into host cells, e.g., bacterial cells. or higher eukaryotic cells, e.g., mammalian cells, according to transfection methods well known in the art, including liposome-mediated transfection, polycation-mediated transfection, protoplast fusion, microinjection, calcium phosphate precipitation, electroporation, or viral vector transfer.

Preferably, nucleic acid molecules encoding the heavy chain and light chain of the binding molecules or antibodies described in the present invention are present on two vectors that are co-transfected into a host cell, preferably a mammalian cell.

Therefore, an additional aspect provides a host cell containing an expression vector containing a nucleic acid molecule encoding a heavy chain and an expression vector containing a nucleic acid molecule encoding a light chain of the binding molecules or antibodies described in the present invention.

- 50 041360

Mammalian cell lines available as hosts for expression are well known in the art and include, among others, Chinese Hamster Ovary (CHO, CHO-DG44) cells, NSO, SP2/0 cells, HeLa cells, Baby Hamster Kidney (BHK ), monkey kidney (COS) cells, human carcinoma cells (eg, Hep G2), A549 cells, 3T3 cells, or derivatives/progeny of any such cell line. Other mammalian cells may be used, including, but not limited to, human, mouse, rat, monkey, and rodent cell lines, or other eukaryotic cells, including, but not limited to, yeast, insect, and plant cells, or prokaryotic cells such as bacteria. Binding molecules according to the invention are obtained by culturing the host cells for a period of time sufficient to allow expression of the binding molecule in the host cells.

The binding molecules and antibody molecules described in the present invention are preferably isolated from the culture medium as a secreted polypeptide, or they can be isolated from host cell lysates if, for example, expressed without a secretory signal. It is necessary to purify the binding or antibody molecules described in the present invention using standard protein purification methods used for recombinant and host cell proteins such that substantially homogeneous preparations of the binding molecule or antibody are obtained as described in the present invention. For example, prior art purification methods useful for preparing binding molecules and antibodies according to the invention include, as a first step, removing cells and/or cell debris particles from the culture medium or lysate. The binding molecule or antibody is then purified from contaminating soluble proteins, polypeptides, and nucleic acids, for example, by fractionation on immunoaffinity or ion exchange columns, ethanol precipitation, reverse phase HPLC, Sephadex chromatography, silica chromatography, or cation exchange resin. As a final step in the process for preparing a TRAILR2-CDH17 binding molecule or CDH17 antibody as described herein, the purified binding molecule or antibody can be dried, eg, lyophilized, as described below for therapeutic use.

Another aspect of the invention relates to a binding molecule or antibody according to the invention for use in medicine.

Another aspect of the invention relates to a binding molecule according to the invention (binding molecules with a TRAILR2 binding site of any of TR#1, TR#2, TR#3, TR#7, TR#8, TR#10, TR#12, TR2v1 , TR2v2, TR2v3, TR2v4, TR2v5, TR2v6, and TR2v7 and the CDH17 binding site of any of CDH17E9, CDH17A6, CDH17E2, CDH17H2, and CDH17v1) or an antibody specific for CDH17 (e.g., fused to a cytotoxic drug) for use in cancer treatment. Preferably, the cancer is colorectal cancer (CRC), gastric cancer (GC), pancreatic cancer (PC), liver cancer (including bile duct cancer), or neuroendocrine tumors.

As indicated above, the inventors have determined that the binding molecules described herein are of significant utility in inducing apoptosis in cancer cells and therefore can be used in the treatment of cancers that express both TRAILR2 and CDH17. Methods for identifying whether a particular tumor expresses TRAILR2 and CDH17 are well known in the art. For example, immunohistochemistry can be used to determine if tumor tissue expresses TRAILR2 and CDH17 (eg, using anti-CDH17 antibodies as described herein) and is therefore suitable for treatment with a binding molecule according to the invention.

In particular, the binding molecule according to the invention is suitable for the treatment of colorectal cancer (CRC).

Colorectal cancer (CRC) is a single malignant disease listed in the ICD-10 and is one of the leading causes of cancer morbidity and mortality worldwide. Approximately 25% of CRC patients have overt metastases, and 40-50% of newly diagnosed patients develop metastatic disease. Although recent beneficial effects in chemotherapy have increased the duration of survival with metastatic CRC, the majority of patients will still fall prey to this disease. Therefore, there is a huge need for additional therapeutic agents for the treatment of this disease.

Approximately 30-50% of colorectal cancers are known to have a mutated (abnormal) KRAS gene. KRAS mutations commonly found in neoplasms include those in exon 2 (codons 12 and 13) and exon 3 (codon 61) and can be analyzed by tumor biopsy. These include activation of mutations that result in continuous signal transduction, stimulation of downstream signaling pathways involved in cell growth, proliferation, invasion, and metastasis. It has been suggested that oncogenic K-Ras may confer resistance to TRAIL-induced apoptosis (Hoogwater et al., Gastroenterology. 2010 Jun; 138(7):2357-67). In some embodiments, the binding molecules described herein (binding molecules with a TRAILR2 binding site of any of TR#1, TR#2, TR#3, TR#7, TR#8, TR#10, TR#12, TR2v1, TR2v2, TR2v3, TR2v4, TR2v5, TR2v6 and TR2v7, and a binding site

- 51 041360 CDH17 expression of any of CDH17E9, CDH17A6, CDH17E2, CDH17H2 and CDH17v1) is used in the treatment of wild-type KRAS colorectal cancer (ie, patients with wild-type KRAS tumors). In some embodiments, the binding molecules described herein (binding molecules with a TRAILR2 binding site of any of TR#1, TR#2, TR#3, TR#7, TR#8, TR#10, TR#12, TR2v1, TR2v2, TR2v3, TR2v4, TR2v5, TR2v6, and TR2v7, and the CDH17 binding site of any of CDH17E9, CDH17A6, CDH17E2, CDH17H2, and CDH17v1) are used in the treatment of KRAS mutant colorectal cancer (i.e., patients with KRAS mutant tumors).

In a further aspect, the present invention relates to methods for treating or preventing cancer, which method comprises administering an effective amount of a binding molecule of the invention to a human (eg, a human suffering from or at risk of developing cancer).

The preferred route of administration is parenteral, by infusion or injection (intravenous, intramuscular, subcutaneous, intraperitoneal, intradermal), but other routes of administration such as inhalation, transdermal, intranasal, buccal, oral may also be used.

In a further aspect, a binding molecule according to the invention is used in conjunction with a device suitable for administering the binding molecule, such as a syringe, pen, micropump, or other device. In another aspect, the binding molecule according to the invention is contained in a kit of parts, for example also including a package insert with instructions for using the binding molecule.

A therapeutically effective amount of a molecule administered is the minimum amount necessary to prevent, ameliorate or treat the clinical symptoms of cancer, in particular the minimum amount that is effective for these disorders.

The dosage range of the binding molecule according to the invention, which is applied per day, is usually from 1 μg/kg to 100 mg/kg, preferably from 0.1 to 20 mg/kg.

The dosage range of the binding molecule of the invention applicable per day is usually 1 μg/kg to 100 mg/kg, preferably 0.1 to 20 mg/kg.

As a rule, for the treatment and/or alleviation of the diseases, disorders and conditions mentioned in the present invention, and depending on the specific disease, disorder or condition to be treated, the effectiveness of the specific binding molecule in accordance with the invention to be used, the specific route of administration and the specific pharmaceutical formulation or composition used, the antibody molecules of the invention are typically administered in an amount of 0.005 to 20.0 mg per kilogram of body weight and at a dose, preferably 0.05 to 10.0 mg/kg/dose, or more preferably 0.5 to 10 mg/kg/dose, either continuously (eg by infusion) or more preferably in unit doses. The administration interval may be, for example, twice a week, weekly or monthly, but may vary considerably, in particular depending on the above parameters. Thus, in some cases it may be sufficient to use the lower dose indicated above, while in other cases the upper limit may be exceeded. When using large amounts, it may be advisable to divide them into several smaller doses distributed throughout the day.

Depending on the specific binding molecule according to the invention and its specific pharmacokinetic and other properties, it can be administered daily, every second, third, fourth, fifth or sixth day, weekly, monthly, and the like. Administration regimens may include long-term treatment over several weeks. By long term is meant at least two weeks, and preferably months or years of treatment.

The effectiveness of the binding molecules of the invention and compositions containing them can be tested using any suitable in vitro assay, cell-based assay, in vivo assay and/or directly known animal model, or any combination thereof, depending on the specific disease being identified. Suitable assays and animal models will be understood by one of skill in the art and, for example, include the assays and animal models used in the examples below.

The actual pharmaceutically effective amount or therapeutic dosage will, of course, depend on factors known to those skilled in the art such as the age and body weight of the patient, the route of administration, and the severity of the disease. In any case, the binding molecule according to the invention will be administered in dosages and in such a way that a pharmaceutically effective amount can be delivered depending on the individual condition of the patient.

The binding molecules according to the invention can be used alone or in combination with other pharmacologically active substances, such as compounds known from the prior art or which are standard in treatment, such as, for example, cytostatic or cytotoxic substances, inhibitors of cell proliferation, anti-angiogenic substances , steroids, immunomodulators/checkpoint inhibitors, and the like.

Therefore, another aspect of the invention relates to a pharmaceutical composition containing

- 52 041360 binding molecule in accordance with the invention, together with a pharmaceutically acceptable carrier and optionally one or more other active substances.

Another aspect of the invention relates to a binding molecule in accordance with the invention for use in the treatment of cancer (for example, an individual suffering from cancer or at risk of developing cancer), and said therapy includes one or more pharmacologically active substances.

Another aspect of the invention relates to the use of one or more active substances in the manufacture of a medicament for the treatment of cancer and/or neoplastic diseases (for example, an individual suffering from cancer or at risk of developing cancer), and said drug contains a binding molecule in accordance with the invention .

Cytostatic and/or cytotoxic active substances that can be administered in combination with binding molecules according to the invention include, but are not limited to, hormones, hormone analogs and antihormones, aromatase inhibitors, LHRH agonists and antagonists, inhibitors of growth factors (growth factors such as such as platelet-derived growth factor (PDGF), fibroblast growth factor (FGF), vascular endothelial growth factor (VEGF), epidermal growth factor (EGF), insulin-like growth factors (IGF), human epidermal growth factor (HER, e.g. HER2, HER3, HER4) and hepatocyte growth factor (HGF)), inhibitors are e.g. anti-growth factor antibodies, anti-growth factor receptor antibodies and tyrosine kinase inhibitors such as e.g. imatinib, lapatinib, bosutinib and trastuzumab; antimetabolites (eg, antifolates such as methotrexate, raltitrexed, pyrimidine analogs such as 5-fluorouracil (5-FU), gemcitabine, irinotecan, doxorubicin, TAS-102, capecitabine and gemcitabine, purine and adenosine analogs such as mercaptopurine, thioguanine , cladribine and pentostatin, cytarabine (ara C), fludarabine); anticancer antibiotics (eg anthracyclines); platinum derivatives (eg cisplatin, oxaliplatin, carboplatin); alkylating agents (eg estramustine, meclorethamine, melphalan, chlorambucil, busulfan, dacarbazine, cyclophosphamide, ifosfamide, temozolomide, nitrosoureas such as, for example, carmustine and lomustine, thiotepa); antimitotic agents (eg vinca alkaloids such as, for example, vinblastine, vindesine, vinorelbine and vincristine; and taxanes such as paclitaxel, docetaxel); angiogenesis inhibitors, including bevacizumab, ramucirumab and aflibercept, tubulin inhibitors; DNA synthesis inhibitors, PARP inhibitors, topoisomerase inhibitors (eg epipodophyllotoxins such as etoposide and etopophos, teniposide, amsacrine, topotecan, irinotecan, mitoxantrone), serine/threonine kinase inhibitors (eg PDK1 inhibitors, Raf inhibitors, A- Raf, B-Raf inhibitors, C-Raf inhibitors, mTOR inhibitors, mTORC1/2 inhibitors, PI3K inhibitors, PI3Ka inhibitors, dual mTOR/PI3K inhibitors, STK33 inhibitors, AKT inhibitors, PLK1 inhibitors (such as volasertib), CDK inhibitors, including CDK9 inhibitors, aurorakinase inhibitors), tyrosine kinase inhibitors (eg, PTK2/FAK inhibitors), protein-protein interaction inhibitors, MEK inhibitors, ERK inhibitors, FLT3 inhibitors, BRD4 inhibitors, IGF-1R inhibitors, Bcl-xL inhibitors, Bcl-2 inhibitors , Bcl-2/Bcl-xL inhibitors, ErbB receptor inhibitors, BCR-ABL inhibitors, ABL inhibitors, Src inhibitors, rapamycin analogs (eg, everolimus, temsirolimus, ridaphorolimus, sirolimus), synthesis inhibitors androgens, androgen receptor inhibitors, DNMT inhibitors, HDAC inhibitors, ANG1/2 inhibitors, CYP17 inhibitors, radiopharmaceuticals, immunotherapies such as immune checkpoint inhibitors (e.g., CTLA4, PD1, PD-L1, LAG3, and TIM3 such as ipilimumab, nivolumab, pembrolizumab) and various chemotherapeutic agents such as amifostine, anagrelide, clodronate, filgrastin, interferon, interferon alfa, leucovorin, rituximab, procarbazine, levamisole, mesna, mitotane, pamidronate and porfimer; proteasome inhibitors (such as bortezomib); mimics Smac and BH3; agents that restore p53 functionality, including the mdm2-p53 antagonist; inhibitors of the Wnt signaling pathway/beta-catenin; and/or inhibitors of cyclin-dependent kinase 9.

Particular preference is given to treatment with binding molecules according to the invention in combination with a drug selected from the following:

(i) anti-VEGF antibodies (bevacizumab and other anti-angiogenic agents) with or without chemotherapy combination (including doxorubicin/cyclophosphamide combination and/or capecitabine/docetaxel combination in neoadjuvant regimen; taxane/platinum regimen for first and subsequent treatment) in patients with breast cancer;

(ii) chemotherapeutic agents used to treat CRC (including 5-fluorouracil, irinotecan, doxorubicin and TAS-102);

(iii) anti-EGFR antibodies (cetuximab and panitumumab in wild-type KRAS tumors) with or without chemotherapy combination (including irinotecan), anti-VEGF antibody combination (bevacizumab and other anti-angiogenic agents), or regorafenib combination, for example, for the treatment of patients with CRC;

(iv) immunotherapeutic agents, including anti-PD-1 and anti-PD-L1 agents, and agents

- 53 041360 against LAG3 such as pembrolizumab and nivolumab and antibodies as described in WO 2017/198741, for example for the treatment of CRC patients.

For use in therapy, the binding molecule or antibody of the invention is formulated into pharmaceutical compositions suitable for facilitating administration to animals or humans. Exemplary binding or antibody formulations described herein can be prepared by mixing the binding or antibody molecule with physiologically acceptable carriers, excipients, or stabilizers in the form of lyophilized or otherwise dried formulations, or aqueous solutions, or aqueous or non-aqueous suspensions. Carriers, excipients, modifiers or stabilizers are non-toxic at the dosages and concentrations used. These include buffer systems such as phosphate, citrate, acetate and other inorganic or organic acids and their salts; antioxidants including ascorbic acid and methionine; preservatives such as octadecyldimethylbenzylammonium chloride; hexamethonium chloride; benzalkonium chloride, benzethonium chloride; phenol, butyl or benzyl alcohol; alkyl parabens such as methyl or propyl paraben; catechin; resorcinol; cyclohexanol; 3-pentanol; and m-cresol; proteins such as serum albumin, gelatin or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone or polyethylene glycol (PEG); amino acids such as glycine, glutamine, asparagine, histidine, arginine or lysine; monosaccharides, disaccharides, oligosaccharides or polysaccharides and other carbohydrates including glucose, mannose, sucrose, trehalose, dextrins or dextrans; chelating agents such as EDTA; sugar alcohols such as mannitol or sorbitol; salt-forming counterions such as sodium; metal complexes (eg Zn-protein complexes); and/or ionic or nonionic surfactants such as TWEEN™ (polysorbates), PLURONICS™ or fatty acid esters, fatty acid ethers or sugar esters. The antibody formulation may also contain organic solvents such as ethanol or isopropanol. Excipients may also have the function of modifying release or absorption.

Generally, preference is given to aqueous solutions or suspensions. Generally, suitable formulations for therapeutic proteins such as binding molecules according to the invention are buffered protein solutions, such as solutions containing the protein at a suitable concentration (such as 0.001 to 400 mg/mL, preferably 0.005 to 200 mg/mL). ml, more preferably 0.01 to 200 mg/ml, more preferably 1.0-100 mg/ml, such as 1.0 to 10.0 mg/ml (intravenous) or 100 mg/ml (subcutaneous) , and an aqueous buffer such as phosphate buffered saline, pH 7.4, other phosphate buffers, pH 6.2 to 8.2, acetate buffers, pH 3.2 to 7.5, preferably pH 4.8 up to 5.5, histidine buffers, pH 5.5 to 7.0, succinate buffers, pH 3.2 to 6.6, and citrate buffers, pH 2.1 to 6.2, and a choice of salt (e.g. , NaCl), and/or stabilizing agents (such as, for example, sucrose, trehalose, lysine), and/or other polyalcohols (such as, for example, mannitol and glycerin) to ensure that the solution is isotonic and optionally detergents, for example to prevent aggregation (eg 0.02% Tween-20 or Tween-80).

Preferred buffered protein solutions for i.v. administrations are solutions containing about 10 mg/ml of a binding molecule of the invention dissolved in 10 mM citrate buffer, pH 5.5, 207 mM sucrose, 25 mM lysine HCl, and 0.02% polysorbate 20. Subcutaneous formulations may include significantly higher concentrations of the antibody of the invention, such as up to 100 mg/ml or even above 100 mg/ml. However, a person skilled in the art will understand that the ingredients and amounts indicated above represent only one preferred option. Alternatives and variations thereof will be immediately apparent to a person skilled in the art or can be easily understood from the above description.

The invention is now described using the following non-limiting examples.

Example 1A. The prevalence of both CDH17 and TRAILR2 in primary and metastatic colorectal cancer (CRC), as well as in primary gastric cancer (GC) and pancreatic cancer (PCa).

A study was conducted to evaluate the expression of the TRAILR2 and CDH17 protein in different types of tumors.

IHC for TRAILR2 and CDH17 in primary CRC was performed on fresh frozen and OCT-dipped samples. Sections with a thickness of 5 μm were prepared on a cryotome and applied to glass slides, followed by fixation in 4% PFA at room temperature and blocking solution (PBS/2% BSA). Sections were incubated with primary antibodies (clone anti-TRA[LR2 TR2.21 from Adipogen # AG-20B0028, C=10 µg/mL; anti-CDH17 (mulgG1) Clone 141713 from R&D Systems #MAB1032, C=1 µg/mL) 54 041360 followed by reagent detection (TRAILR2 and CDH17: EnVision+, HRP, Mouse, DAKO #4000) and incubation in DAB solution. Contrasting staining was performed using hematoxylin. If necessary, the washing steps were carried out in the FSB.

IHC for TRAILR2 and CDH17 on CRC metastases was performed on formalin-fixed and paraffin-embedded specimens. Sections with a thickness of 2 μm were prepared on a microtome, applied to glass slides, and deparaffinized. Demasking solution (TRAILR2: pH9, Vector Laboratories #H3301; CDH17: pH6, Vector Laboratories #H3300) was applied at 121°C/1 bar followed by blocking steps using 3% H ₂ O ₂ and subsequently normal goat serum ( Vector Laboratories #S-1000) in PBS/2% WCA. Sections were incubated with primary antibodies (anti-TRAILR2 (D4E9) XP® Rabbit mAb from Cell Signaling #8074, 1:20 dilution; anti-CDH17 (mulgG1) Clone 141713 from R&D Systems #MAB1032, C=1 µg/mL) for 1 hour at room temperature, followed by reagent detection (TRAILR2: EnVision+, HRP, Rabbit, DAKO #K4003; CDH17: EnVision+, HRP, Mouse, DAKO #4000) and incubation in DAB solution. Contrasting staining was performed using hematoxylin. If necessary, the washing steps were carried out in the FSB.

In FIG. 14 representative IHC images for both TRAILR2 and CDH17 are shown in primary CRC and in liver metastases from CRC. In primary CRC (n=26), CDH17 expression was observed in 100% of all cases and co-expression with TRAILR2 in 88%; in metastatic CRC (n=39; liver and lungs), CDH17 expression was observed in 100% of all cases, and co-expression with TRAILR2 in 100%; in primary gastric cancer (n=49), CDH17 expression was observed in 95% of all cases, and co-expression with TRAILR2 in 60%; in primary PaS (n=32), CDH17 expression was observed in 80% of all cases, and co-expression with TRAILR2 in 20%.

This study confirms the coexpression of CDH17 and TRAILR2 in a large subset of CRC, GC, and PCa.

Example 1B. Construction of binding molecules that recognize the human TRAIL receptor 2 (TRAILR2) and human cadherin17 (CDH17).

The present inventors have developed binding molecules that bind CDH17 and TRAILR2 and that induce apoptosis in cancer cells expressing both CDH17 and TRAILR2. The molecular construct used has an IgG antibody (referred to as a core antibody) that has specificity for a single target antigen, with scFvs of varying specificity linked to the C-terminus of the heavy chain. The design diagram is shown in Fig. 1.

Preferably the binding molecule is bispecific and tetravalent.

The bispecific molecule contains flexible peptide sequences between the heavy chain variable domains (VH) and the light chain variable domains (VL) of scFv, and the scFv domains are linked to the main IgG antibody via an additional series of linkers. In one configuration, the scFv is oriented such that the VL domain forms the N-terminal end of the scFv and thus fuses with the C-terminus of the main antibody heavy chain, while the VH forms the C-terminus of the scFv and indeed the entire heavy chain polypeptide. However, it should be understood that this N-VL-VH-C structure can be reversed, ie N-VH-VL-C.

To test the feasibility of this concept, a number of different bispecific molecules were prepared based on the format depicted in FIG. 1.

The following examples explain the methods used to create a bispecific molecule that binds CDH17 and TRAILR2, as well as the differences in the format and biological activity of these molecules.

Example 2 Generation of binding domains that recognize CDH17 and TRAILR2 using high throughput V gene recovery from hybridomas and cultured single B cells.

It is clear that in order to obtain bispecific molecules that bind to human CDH17 and TRAILR2, it is necessary to obtain variable domains that bind to individual target antigens.

To achieve this, clonal hybridomas or single B cells derived from mice immunized with CDH17 or TRAILR2 were cultured in vitro. Supernatants were screened for reactivity against human CDH17 or TRAILR2. The immunoglobulin (Ig) VH and VL genes were then amplified from the identified positive clones.

For RNA isolation from hybridomas, about 2x106 cells from single clones were precipitated and used as starting material. For single B cells, from 100 to 500 cells were used as starting material, which developed from single isolated B cells. RNA was isolated using RNeasy Plus (Qiagen, Hilden, Germany). The cDNA was then synthesized using the Smarter cDNA Synthesis Kit (Clontech, Mount View, CA) according to the manufacturer's instructions.

The Ig VH and VL genes were amplified using the primers listed in Table 1. 1. Each 50 µl PCR reaction consisted of 20 µM forward and reverse primer mixture, 25 µl Pfu premix (Agilent Technologies), 2 µl crude cDNA, and 21 µl doubly distilled H2O. PCR reaction

- 55 041360 started with a step of 94°C for 3 min, followed by 15 cycles of PCR (92°C for 1 min,

55°C for 1 min, 68°C for 1 min), then 25 cycles of PCR (94°C for 1 min, 50°C for min, 72°C for 1 min) and ended with stage 72° C for 7 min.

A second round of PCR was then performed with the primer set, barcode for 454 next generation sequencing. The reaction was started at 94°C for 3 min, then 35 cycles were repeated at 92°C for 1 min, 53°C for 1 min and 68°C for 1 min. PCR products VL and VH were pooled, gel purified, and then subjected to DNA sequencing (Roche GS-FLX, SeqWright, USA).

Using this methodology, a large number of VH and VL Ig gene pairs encoding binding domains with specificity for CDH17 or TRAILR2 were prepared. Some of the VH and VL sequences were further humanized using methods known in the art.

Table 1. Primer sequences

Stage PCR Primer name Sequence 5'-3' VL 1 tbsp. PCR 5'UPM long CTAATACGACTCACTATAGGGCAAGCAGTGGTATCAACGCAGAGT (SEQ ID NO: 234) 5' UPM short CTAATACGACTCACTATAGGGC (SEQ ID NO: 235) 3' MKS R CTGCTCACTGGATGGTGGGAAGATGG (SEQ ID NO: 236) VH 1 tbsp. PCR 5' UPM long CTAATACGACTCACTATAGGGCAAGCAGTGGTATCAACGCAGAGT (SEQ ID NO: 234) 5' UPM short CTAATACGACTCACTATAGGGC (SEQ ID NO: 235) 3' MHGC R GGGGCCAGTGGATAGACAGATGGGGG (SEQ ID NO: 237) 454 2nd VL PCR MuFl CGTATCGCCTCCCTCGCGCCATCAGACGAGTGCGTCTAATACGACT CACTATAGGGC (SEQ ID NO: 238) 454 2nd VL PCR MuF2 CGTATCGCCTCCCTCGCGCCATCAGACGCTCGACACTAATACGACT CACTATAGGGC (SEQ ID NO: 239) 454 2nd VL PCR MuF3 CGTATCGCCTCCCTCGCGCCATCAGAGACGCACTCCTAATACGACT CACTATAGGGC (SEQ ID NO: 240) 454 2nd VL PCR MuF4 CGTATCGCCTCCCTCGCGCCATCAGAGCACTGTAGCTAATACGACT CACTATAGGGC (SEQ ID NO: 241) 454 2nd VL PCR MuF5 CGTATCGCCTCCCTCGCGCCATCAGATCAGACACGCTAATACGACT CACTATAGGGC (SEQ ID NO: 242) 454 2nd VL PCR MuF6 CGTATCGCCTCCCTCGCGCCATCAGATATCGCGAGCTAATACGACT CACTATAGGGC (SEQ ID NO: 243) 454 2nd VL PCR MuF7 CGTATCGCCTCCCTCGCGCCATCAGCCGTGTCTCTACTAATACGACT CACTATAGGGC (SEQ ID NO: 244) 454 2nd VL PCR MuF8 CGTATCGCCTCCCTCGCGCCATCAGCTCGCGTGTCCTAATACGACT CACTATAGGGC (SEQ ID NO: 245) 454 2nd VL PCR MuKRl CTATGCGCCTTGCCAGCCCGCTCAGACGAGTGCGTCTGCTCACTGG ATGGTGGGAAGATGG(SEQ ID NO: 246) 454 2nd VL PCR MuKR2 CTATGCGCCTTGCCAGCCCGCTCAGACGCTCGACACTGCTCACTGG ATGGTGGGAAGATGG (SEQ ID NO: 247) 454 2nd VL PCR MuKR3 CTATGCGCCTTGCCAGCCCGCTCAGAGACGCACTCCTGCTCACTGG ATGGTGGGAAGATGG (SEQ ID NO: 248) 454 2nd VL PCR MuKR4 CTATGCGCCTTGCCAGCCCGCTCAGAGCACTGTAGCTGCTCACTGG ATGGTGGGAAGATGG (SEQ ID NO: 249) 454 2nd VL PCR MuKR5 CTATGCGCCTTGCCAGCCCGCTCAGATCAGACACGCTGCTCACTGG ATGGTGGGAAGATGG (SEQ ID NO: 250) 454 2nd VL PCR MuKR6 CTATGCGCCTTGCCAGCCCGCTCAGATATCGCGAGCTGCTCACTGG ATGGTGGGAAGATGG (SEQ ID NO: 251) 454 2nd VL PCR MuKR7 CTATGCGCCTTGCCAGCCCGCTCAGCGTGTCTCTACTGCTCACTGG ATGGTGGGAAGATGG (SEQ ID NO: 252) 454 2nd VL PCR MuKR8 CTATGCGCCTTGCCAGCCCGCTCAGCTCGCGTGTCCTGCTCACTGG ATGGTGGGAAGATGG (SEQ ID NO: 253) 454 second VHPCR MuFl CGTATCGCCTCCCTCGCGCCATCAGACGAGTGCGTCTAATACGACT CACTATAGGGC (SEQ ID NO: 254) 454 second VHPCR MuF2 CGTATCGCCTCCCTCGCGCCATCAGACGCTCGACACTAATACGACT CACTATAGGGC (SEQ ID NO: 255)

-56041360

454 second VHPCR MuF3 CGTATCGCCTCCCTCGCGCCATCAGAGACGCACTCCTAATACGACT CACTATAGGGC (SEQ ID NO: 256) 454 second VHPCR MuF4 CGTATCGCCTCCCTCGCGCCATCAGAGCACTGTAGCTAATACGACT CACTATAGGGC (SEQ ID NO: 257) 454 second VHPCR MuF5 CGTATCGCCTCCCTCGCGCCATCAGATCAGACACGCTAATACGACT CACTATAGGGC (SEQ ID NO: 258) 454 second VHPCR MuF6 CGTATCGCCTCCCTCGCGCCATCAGATATCGCGAGCTAATACGACT CACTATAGGGC (SEQ ID NO: 259) 454 second VH PCR MuF7 CGTATCGCCTCCCTCGCGCCATCAGCCGTGTCTCTACTAATACGACT CACTATAGGGC (SEQ ID NO: 260) 454 second VHPCR MuF8 CGTATCGCCTCCCTCGCGCCATCAGCTCGCCGTGTCCTAATACGACT CACTATAGGGC (SEQ ID NO: 261) 454 second VHPCR MuGIRL CTATGCGCCTTGCCAGCCCGCTCAGACGAGTGCGTGGGGCCAGTG GATAGACAGATGGGGG (SEQ ID NO: 262) 454 second VHPCR MuGlR2 CTATGCGCCTTGCCAGCCCGCTCAGACGCTCGACAGGGGCCAGTG GATAGACAGATGGGGG (SEQ ID NO: 263) 454 second VHPCR MuGlR3 CTATGCGCCTTGCCAGCCCGCTCAGAGACGCACTCGGGGCCAGTG GATAGACAGATGGGGG (SEQ ID NO: 264) 454 second VHPCR MuGlR4 CTATGCGCCTTGCCAGCCCGCTCAGAGCACTGTAGGGGGCCAGTG GATAGACAGATGGGGG (SEQ ID NO: 265) 454 second VHPCR MuGlR5 CTATGCGCCTTGCCAGCCCGCTCAGATCAGACACGGGGGCCAGTG GATAGACAGATGGGGG (SEQ ID NO: 266) 454 second VHPCR MuGlR6 CTATGCGCCTTGCCAGCCCGCTCAGATATCGCGAGGGGGCCAGTG GATAGACAGATGGGGG (SEQ ID NO: 267) 454 second VHPCR MuGlR7 CTATGCGCCTTGCCAGCCCGCTCAGCGTGTCTCTAGGGGCCAGTGG ATAGACAGATGGGGG (SEQ ID NO: 268) 454 second VHPCR MuGlR8 CTATGCGCCTTGCCAGCCCGCTCAGCTCGCGTGTCGGGGCCAGTGG ATAGACAGATGGGGG (SEQ ID NO: 269)

Example 3 High throughput design of bispecific molecules binding CDH17 and TRAILR2.

To construct a gene segment encoding a TRAILR2 scFv, the VL and VH gene pairs encoding TRAILR2-binding variable domains obtained in Example 2 or known in the art were combined with a gene segment encoding a flexible linker of the peptide postnoRaTenbHocTnGGGGSGGGGSGGGGSGGGGS (SEQ ID NO: 143) . χ /

The resulting segments of the scFv-encoding gene were in turn cloned in-frame up to the 3' end of the gene encoding the heavy chain of a human IgG antibody. These coding segments were synthesized by overlapping PCR methods and cloned into the pTT5 expression vector.

The VL and VH gene pairs encoding the CDH17 binding variable domains obtained in Example 2 were then formatted into the bispecific format described in Example 1. The VH genes were cloned into the pTT5 expression vector as an in-frame fusion at the 5' end of the gene encoding human Igy . The gene encoding TRAILR2-binding scFv was cloned in-frame at the 3' end of the same segment encoding Igy. Similarly, the VL genes were cloned into the pTT5 expression vector as an in-frame fusion with the gene encoding the human IgG kappa light chain.

The VL and VH gene pairs encoding the TRAILR2 binding variable domains obtained in Example 2 or known in the art were then used to generate antibody molecules (full-length antibody molecules containing two light and two heavy chains) that specifically bind to TRAILR2 (e.g. containing a TR2v1 or TRv2 antigen-binding site) using methods known in the art and used to detect/label cells or as control antibodies in the examples described below. Such antibodies include either human IgG1 WT or IgG1 constant domains (KO).

Likewise, the VL and VH gene pairs encoding the CDH17 binding variable domains obtained in Example 2 or known in the art were further used to generate antibody molecules (full-length antibody molecules containing two light and two heavy chains) that specifically bind to CDH17 (eg, containing the CDH17v1 or CDH17H2 antigen binding site) using methods known in the art and used to detect/label cells or as control antibodies in the examples described below. Such antibodies include either human IgG1 WT or IgG1 constant domains (KO).

The antibodies specific for either TRAILR2 or CDH17 described above and used as control antibodies in the following examples are conventional full-length antibodies with two heavy and light chains. The binding sites of such reference antibodies are defined by the same variable domains as the bispecific binding proteins to which they are compared. For example, in FIG. 3 a binding molecule with a CDH17-specific CDH17v1 binding site and a TRAILR2-specific TR2v1 binding site (referred to as CDH17v1/TR2v1 binding molecule) contains a VH sequence specific for CDH17 SEQ ID NO: 118 and

- 57 041360 VL sequence specific for CDH17 SEQ ID NO: 119 and VH sequence specific for TRAILR2 SEQ ID NO: 96 and VL sequence specific for TRAILR2 SEQ ID NO: 97 (see Table 2). Accordingly, the anti-CDH17v1 control antibody used in this example contains the VH sequence specific for CDH17 SEQ ID NO: 118 and the VL sequence specific for CDH17 SEQ ID NO: 119 and the control anti-TRv1 antibody contains the VH sequence specific for TRAILR2 SEQ ID NO: 96 and the VL sequence specific for TRAILR2 SEQ ID NO: 97 (see FIG. 3).

The In-Fusion® HD Cloning Kit (Clonetech, USA) was used in the above procedure for targeted cloning of the VH and VL genes. PCR primers for VL/VH were synthesized with 15 bp extensions complementary to the ends of the linearized vector. PCR was performed using the manufacturer's standard protocol and the amplicons were purified or processed with Cloning Enhancer and then cloned into the appropriate vector. After that, E. coli was transformed according to the manufacturer's instructions (Clonetech, USA). DNA minipreps were sequenced.

Each expression vector contains eukaryotic promoter elements for a gene encoding a chain, a gene encoding a signal sequence and a heavy or light chain, an expression cassette for a gene for a prokaryotic selection marker such as ampicillin, and a source of replication. These DNA plasmids were propagated in ampicillin resistant E. coli colonies and purified.

Example 4 Expression and purification of bispecific, tetravalent molecules recognizing human TRAILR2 and human CDH17.

The expression vectors obtained in Example 3 were transfected into CHO-E cells.

Transfected CHO-E cells growing in suspension in serum-free media were cultured in shake flasks, agitated at 140 rpm, 37°C and 5% CO2, and stored under exponential growth conditions. On the day of transfection, cells were chemically transfected with 1 mg light chain plasmid and 0.5 mg heavy chain plasmid. They were then seeded at 1 to 2x10 ⁶ cells/ml in 1 liter of Gibco® FreeStyle™ CHO expression medium (Life Technologies, NY, US).

Thereafter, the cells were incubated with orbital shaking for 10 to 12 days with a single feeding of 150 ml of commercial nutrient solution to ensure protein expression. Antibody/binding molecule titers in cell culture supernatants were determined using the Octet® instrument (Pall ForteBio, CA, US) and protA biosensor tips according to the manufacturer's instructions.

Recombinant binding molecules or antibodies were purified from the culture supernatant by protein A affinity chromatography using MabSelect™ (Amersham Biosciences) and stored in 60 mM NaOAc buffer (pH 5.0). The purity and degree of heterogeneity of the samples were assessed by mass spectrometry and analytical ultracentrifugation. All samples were confirmed to have >90% monomer content and <10% impurities prior to functional testing.

Example 5 Details of the resulting bispecific molecules recognizing human TRAILR2 molecules and human CDH17 molecules.

In the following examples, a number of different anti-TRAILR2/CDH17 bispecific antibody molecules were prepared in accordance with the invention. To avoid confusion, the characteristics and sequences of these molecules are given below.

Table 2 Details of the TRAILR2/CDH17 bispecific molecules according to the invention

Name TRAIL/R2 binding domain (VH and VL sequences) CDH17 binding domain (VH and VL sequences) Type and subsequence IgG Orientation and sequence of scFV CDH17vl/TR2vl TR2vl SEQ ID NOs 96 and 97. CDH17vl SEQ ID NOs 118 and 119. wild-type IgGl sequence SEQ ID NO 120 VL-VH SEQ ID NO: 124. CDH17vl/TR2vl(IgGl KO) TR2vl SEQ ID NOs 96 and 97. CDH17vl SEQ ID NOs 118 and 119. IgGlKO SEQ ID NO: 121 VL-VH SEQ ID NO: 124. CDH17vl/TR2v2(IgGl KO) TR2v2 SEQ ID NOs 98 and 99. CDH17vl SEQ ID NOs 118 and 119 IgGlKO SEQ ID NO: 121 VL-VH SEQ ID NO: 125. CDH17vl/TR2v2- (IgG4 Pro) TR2v2 SEQ ID NOs 98 and 99 CDH17vl SEQ ID NOs 118 and 119 IgG4 Pro sequence SEQ ID NO: 122 VL-VH SEQ ID NO: 125. CDH17vl/TR2vl(scFv VH-VL) TR2vl SEQ ID NOs 96 and 97. CDH17vl SEQ ID NOs 118 and 119 IgGlKO SEQ ID NO: 121 VH-VL SEQID NO. 131 CDH17vl/TR2vl(scFv VH-VL ss) TR2vl SEQ ID NOs 96 and 97. CDH17vl SEQ ID NOs 118 and 119 IgGlKO SEQ ID NO: 121 VH-VL SEQ ID NO. 132

- 58 041360

CDH17vl/TR2vl(scFv VL-VH ss) TR2vl SEQ ID NOs 96 and 97. CDH17vl SEQ ID NOs 118 and 119 IgGlKO SEQ ID NO: 121 VL-VH SEQ ID NO. 133 CDH17A6/TR2v2(IgGlKO) TR2v2 SEQ ID NOs 98 and 99 CDH17A6 SEQ ID NOs 112 and 113 IgGlKO SEQ ID NO: 121 VL-VH SEQ ID NO 125. CDH17E2/TR2v2(IgGlKO) TR2v2 SEQ ID NOs 98 and 99 CDH17E2 SEQ ID NOs 114 and 115 IgGlKO SEQ ID NO: 121 VL-VH SEQ ID NO. 125 CDH17E9/TR2v2- (IgGlKO) TR2v2 SEQ ID NOs 98 and 99 CDH17E9 SEQ ID NOs NO and 111 IgGlKO SEQ ID NO : 121 VL-VH SEQ ID NO. 125 CDH17H2/TR2v2(IgGlKO) TR2v2 SEQ ID NOs 98 and 99 CDH17H2 SEQ ID NOs 116 and 117 IgGlKO SEQ ID NO: 121 VL-VH SEQID NO. 125 CDH17H2/TR2v3- (IgGlKO TR2v3 SEQ ID NOs 100 and 101 CDH17H2 SEQ ID NOs 116 and 117 IgGlKO SEQ ID NO: 121 VL-VH SEQID NO. 126 CDH17H2/TR2v4(IgGlKO) TR2v4 SEQ ID NOs 102 and 103 CDH17H2 SEQ ID NOs 116 and 117 IgGlKO SEQ ID NO: 121 VL-VH SEQID NO. 127 CDH17H2/TR2v5(IgGlKO) TR2v5 SEQ ID NOs 104 and 105 CDH17H2 SEQ ID NOs 116 and 117 IgGlKO SEQ ID NO: 121 VL-VH SEQID NO. 128 CDH17H2/TR2v6(IgGlKO) TR2v6 SEQ ID NOs 106 and 107 CDH17H2 SEQ ID NOs 116 and 117 IgGlKO SEQ ID NO: 121 VL-VH SEQID NO. 129 CDH17H2/TR2v7(IgGlKO) TR2v2 SEQ ID NOs 108 and 109 CDH17H2 SEQ ID NOs 116 and 117 IgGlKO SEQ ID NO: 121 VL-VH SEQID NO. 130 CDH17vl/TR2#l(IgGl KO) TR2#1 SEQ ID NOs 82 and 83 CDH17vl SEQ ID NOs 118 and 119 IgGlKO SEQ ID NO: 121. VL-VH SEQID NO. 134 CDH17vl/TR2#2(IgGl KO) TR2#2 SEQ ID NOs 84 and 85 CDH17vl SEQ ID NOs 118 and 119 IgGlKO SEQ ID NO: 121. VL-VH SEQID NO. 135 CDH17vl/TR2#3(IgGl KO) TR2#3 SEQ ID NOs 86 and 87 CDH17vl SEQ ID NOs 118 and 119 IgGlKO SEQ ID NO: 121. VL-VH SEQ ID NO. 136 CDH17vl/TR2#7(IgGlKO) TR2#7 SEQ ID NOs 88 and 89 CDH17vl SEQ ID NOs 118 and 119 IgGlKO SEQ ID NO: 121. VL-VH SEQ ID NO. 137 CDH17vl/TR2#8(IgGlKO) TR2#8 SEQ ID NOs 90 and 91 CDH17vl SEQ ID NOs 118 and 119 IgGlKO SEQ ID NO: 121. VL-VH SEQ ID NO. 138 CDH17vl/TR2#10(IgGlKO) TR2#10 SEQ ID NOs 92 and 93 CDH17vl SEQ ID NOs 118 and 119 IgGlKO SEQ ID NO: 121. VL-VH SEQ ID NO. 139 CDH17H2/TR2v2(IgGl FcRnmut) TR2v2 SEQ ID NOs 98 and 99 CDH17H2 SEQ ID NOs 116 and 117 IgGl FcRnmut SEQ ID NO: 270 VL-VH SEQID NO. 125

-59041360

Table 3. Amino acid sequences and SEQ ID NOs CDRs, VH, VL, scFv, light and heavy chain binding molecules and antibodies according to the invention

SEQ ID number Short description of the sequence Subsequence SEQ ID NO: 1 TR2#1HCCDR1 DYTFTSYDIN SEQ ID NO:2 TR2#1HCCDR2 WIDPGSGNTKYNEKFKG SEQ ID NO:3 TR2#1HCCDR3 KNYGGSYAFTY SEQ ID NO:4 TR2#1LCCDR1 KASQDVITAVA SEQ ID NO:5 TR2#1LCCDR2 WASTRHT SEQ ID NO:6 TR2#1LCCDR3 QQHYSTPWT SEQ ID NO:7 TR2#2HCCDR1 GYTFTEYIIH SEQ ID NO:8 TR2#2HCCDR2 WFYPGSGSIKYNEKFKD SEQ ID NO:9 TR2#2HCCDR3 HEEGGYSAWFPY SEQ ID NO: 10 TR2#2LCCDR1 KASQDVSTAVA SEQ ID NO: 11 TR2#2LCCDR2 WASTRHT SEQ ID NO: 12 TR2#2LCCDR3 QQHYSTPYT SEQ ID NO: 13 TR2#3HCCDR1 GYTFTDYYYLN SEQ ID NO: 14 TR2#3HCCDR2 YIYPNNGDTSYNQKFKG SEQ ID NO: 15 TR2#3HCCDR3 GSNWIWYFDV SEQ ID NO: 16 TR2#3LCCDR1 RSSQSIVHSNGDTYLD SEQ ID NO: 17 TR2#3LCCDR2 KVSNRFS SEQ ID NO: 18 TR2#3LCCDR3 FRGSHIPPT SEQ ID NO: 19 TR2#7HCCDR1 GYSFTDYYIN SEQ ID NO:20 TR2#7HCCDR2 KIGPGSGNTYYNEKFEG SEQIDNO:21 TR2#7HCCDR3 TGPFAY SEQ ID NO:22 TR2#7LCCDR1 KASQNVGNNVA SEQ ID NO:23 TR2#7LCCDR2 YASNRYT SEQ ID NO:24 TR2#7LCCDR3 QQHYSSPLT SEQ ID NO:25 TR2#8HCCDR1 GFSLSTSGMGVS SEQ ID NO:26 TR2#8HCCDR2 HIYWDDDKRYNPSLKS SEQ ID NO:27 TR2#8HCCDR3 RRLGPFAY SEQ ID NO:28 TR2#8LCCDR1 KASQDVGTAVA SEQ ID NO:29 TR2#8LCCDR2 WASTRHT SEQID NQ:30 TR2#8LCCDR3 QQYSSYPYT SEQIDNO:31 TR2#1OHCCDR1 GYTFTDYYYMN SEQ ID NO:32 TR2#10HCCDR2 YIYPNNGGTRYNQKFKG SEQ ID NO:33 TR2#10HCCDR3 GGNWNWYFDV

- 60 041360

SEQ ID NO:34 TR2#1OLCCDR1 RSSQNIVHSNGNTYLD SEQ ID NO:35 TR2#10LCCDR2 KVSNRFS SEQ ID NO:36 TR2#10LCCDR3 FQGSHVPPT SEQ ID NO:37 TR2#12HCCDR1 GYTFTEYIIH SEQ ID NO:38 TR2#12HCCDR2 WFYPGSGNIKYNEKFKD SEQ ID NO:39 TR2#12HCCDR3 HEGGSNFFPY SEQ ID NO:40 TR2#12LCCDR1 KASQDVSNAVA SEQ ID NO:41 TR2#12LCCDR2 WASTRHN SEQ ID NO:42 TR2#12LCCDR3 QQHYRTPYT SEQ ID NO:43 TRvl, TRv2, TRv3, TRv4, TRv5, TRv6 and TRv7 HCCDR1 GFTFDDYGMS SEQ ID NO:44 TRvl and TRv2 HCCDR2 GINWNGGSTGYADSVKG SEQ ID NO:45 TRv3, TRv4, TRv5 HCCDR2 GINWSGGSTGYADSVKG SEQ ID NO:46 TRv6 HCCDR2 GINWAGGSTGYADSVKG SEQ ID NO:47 TRv7 HCCDR2 GINWTGGSTGYADSVKG SEQ ID NO:48 TRvl, TRv2, TRv3, TRv4, TRv5, TRv6 and TRv7 HCCDR3 ILGAGRGWYFDL SEQ ID NO:49 TRvl, TRv2, TRv3, TRv4, TRv5, TRv6 and TRv7 LCCDR1 QGDSLRSYYAS SEQ ID NO:50 TRvl, TRv2 and TRv5 LCCDR2 GKNNRPS SEQIDNO:51 TRv3 and TRv6 LCCDR2 GKTNRPS SEQ ID NO:52 TRv4 LCCDR2 GKANRPS SEQ ID NO:53 TRv7 LCCDR2 GKDNRPS SEQ ID NO:54 TRvl and TRv2 LCCDR3 NSRDSSGNHW SEQ ID NO:55 TRv3 LCCDR3 NSRDSSGTHVV SEQ ID NO:56 TRv4 and TRv5 NSRDSSGHVV

- 61 041360

LCCDR3 SEQ ID NO:57 TRv6 and TRv7 LCCDR3 NSRDSSGDHW SEQ ID NO:58 CDH17E9HCCDR1 GYTLTDHTHIH SEQ ID NO:59 CDH17E9HCCDR2 YIYPRDDITQYNENFKA SEQ ID NO:60 CDH17E9HCCDR3 WSWHFGSNYYGLDY SEQIDNO:61 CDH17E9LCCDR1 KASQNVGTALA SEQ ID NO:62 CDH17E9LCCDR2 SASNRFT SEQ ID NO:63 CDH17E9LCCDR3 QQYSTYPLT SEQ ID NO:64 CDH17A6HCCDR1 GYTFTDHTTIH SEQ ID NO:65 CDH17A6HCCDR2 YIYPRDAITKYNEKFKG SEQ ID NO:66 CDH17A6HCCDR3 WGYYFGSTSYYFDF SEQ ID NO:67 CDH17A6LCCDR1 KASQNVGTGVV SEQ ID NO:68 CDH17A6LCCDR2 SPSNRYT SEQ ID NO:69 CDH17A6LCCDR3 QQYSTYPWT SEQ ID NO:70 CDH17E2HCCDR1 CDH17H2HCCDR1 GYTFTDHTTIH SEQIDNO:71 CDH17E2HCCDR2 CDH17H2HCCDR2 YIYPRDVITQYNEKFKG SEQ ID NO:72 CDH17E2HCCDR3 CDH17H2HCCDR3 WGYFYGSRSYYFDY SEQ ID NO:73 CDH17E2LCCDR1 CDH17H2LCCDR1 KASQNVGTAVA SEQ ID NO:74 CDH17E2LCCDR2 CDH17H2LCCDR2 SASNRYT SEQ ID NO:75 CDH17E2LCCDR3 CDH17H2LCCDR3 QQYSSYPWT SEQ ID NO:76 CDH17vlHCCDRl GYTLTDHTHIH SEQ ID NO:77 CDH17vlHCCDR2 YIYPRDGITGYNEKFKG SEQ ID NO:78 CDH17vlHCCDR3 WGYSYRNYAYYYDY SEQ ID NO:79 CDH17vlLCCDRl KSSQSLLHS SNQKNYLA SEQ ID NO:80 CDH17vlLCCDR2 WASTRES SEQIDNO:81 CDH17vlLCCDR3 QQYYSYPWT SEQ ID NO:82 TR2#1VH QVQLQQSGELVKPGASVKLSCKAFDYTFTSYD INWVKQRPGQGLEWIGWIDPGSGNTKYNEKFK GKATLTADKSSSTAYMHLSSLTSEDSAVYFCAR KNYGGSYAFTYWGQGTLVTVSA

- 62 041360

SEQ ID NO:83 TR2#1VL DIVMTQSHKFMSTSVGDRVSITCKASQDVITAV AWYQQKPGQSPKLLIYWASTRHTGVPDRFTGT GSGTDYTLTIS SVQ AEDL ALYYCQQHYSTP WTF GGGTKLEIK SEQ ID NO:84 TR2#2VH KVQLQQSGELVKTGTSVKLSCKASGYTFTEYII HWVKQRSGQGLEWIGWFYPGSGSIKYNEKFKD KATLTADKSSSTVYMELSRLTSEDSAVYFCARH EEGGYSAWFPYWGQGTLVTVSA SEQ ID NO:85 TR2#2VL DIVMTQSHKFMSTSVGDRVSITCKASQDVSTAV AWYQQKPGQSPKVLIYWASTRHTGVPDRFTGS GSGTDYILTISSVQAEDLALYYCQQHYSTPYTFG GGTKLEIK SEQ ID NO:86 TR2#3VH EVQLQQSGPELVKPGASVKISCKASGYTFTDYY YLNWVKQSHGKSLEWIGYIYPNNGDTSYNQKF KGKTTLTVDKSSSTAYMEFRSLTSEDSAVYYCT RGSNWIWYFDVWGTGTTVTVSS SEQ ID NO:87 TR2#3VL DVLMTQTPLSLPVSLGDQASISCRSSQSIVHSNG DTYLDWYLQKPGQSPKLLIYKVSNRFSGVPDRF SGSGSGTDFTLKISRVVAEDLGVYYCFRGSHIPP TFGAGTKLELK SEQ ID NO:88 TR2#7VH QVQLKQSGAELVKPGASVKISCKASGYSFTDYY INWVKQRPGQGLEWIGKIGPGSGNTYYNEKFEG KATLTADKSSSTAYMQLSSLTSEDSAVYFCAST GPFAYWGQGTLVTWSA SEQ ID NO:89 TR2#7VL SIVMTQTPKFLPVSAGDRVTMTCKASQNVGNN VAWYQQKPGQSPKLLISYASNRYTGVPDRFTGS GSGTDFTFTISSVQVEDLAVYFCQQHYSSPLTFG AGTKLELK SEQ ID NO:90 TR2#8VH QVTLKESGPGILQPSQTLSLTCSFSGFSLSTSGMG VSWIRQPSGKGLEWLAHIYWDDDKRYNPSLKS RLTISKDTSRNQVFLKITSVDTADTATYYCARRR LGPFAYWGQGTLVTVSA SEQIDNO:91 TR2#8VL DIVMTQSHKFMSTSVGDRVSITCKASQDVGTAV AWYQQKPGQSPKLLIYWASTRHTGVPDRFTGSG SGTDFTLTISNVQSEDLADYFCQQYSSYPYTFGG GTKLEIK SEQ ID NO:92 TR2#10VH EVQLQQSGPELVKPGASVKISCKASGYTFTDYY YMNWVKQSHGKSLEWIGYIYPNNGGTRYNQKF KGKATLTVDKSSSTAYMELRSLTSEDSAVYYCA RGGNWNWYFDVWGTGTTVTVSS SEQ ID NO:93 TR2#10VL DVLMTQTPLSLPVSLGDQASISCRSSQNIVHSNG NTYLDWYLQKPGQSPKLLIYKVSNRFSGVPDRF SGSGSGTDFTLKISRVAEDLGVYYCFQGSHVPP TFGAGTKLELK SEQ ID NO:94 TR2#12VH KVQLQQSGELVKPGASVKLSCKASGYTFTEYII HWVKQKSGQGLEWIGWFYPGSGNIKYNEKFKD KATLTADKSSSTVYMELSRLTSEDSAVYFCTRH EGGSNFFPYWGQGTLVTVSA SEQ ID NO:95 TR2#12VL DIVMTQSHKFMSTSVGDRVSITCKASQDVSNAV AWYQQKPGQSPKLLIYWASTRHNGVPDRFTGS

- 63 041360

GSGTDYILTISSVQAEDLALYYCQQHYRTPYTFG GGTKLEIK SEQ ID NO:96 TRvlVH EVQLVQSGGGVERPGGSLRLSCAASGFTFDDYG MS WVRQ APGKGLEWVSGINWNGGSTGYAD SV KGRVTISRDNAKNSLYLQMNSLRAEDTAVYYC AKILGAGRGWYFDLWGKGTTVTVSS SEQ ID NO:97 TRvlVL DIELTQDPAVSVALGQTVRITCQGDSLRSYYAS WYQQKPGQAPVLVIYGKNNRPSGIPDRFSGSSS GNTASLTITGAQAEDEADYYCNSRDSSGNHVVF GGGTKLTVL SEQ ID NO:98 TRv2VH EVQLVQSGGGVERPGGSLRLSCAASGFTFDDYG MS WVRQ APGKGLEWVSGINWNGGSTGYAD SV KGRVTISRDNAKNSLYLQMNSLRAEDTAVYYC AKILGAGRGWYFDLWGKGTTVTVSS SEQ ID NO:99 TRv2VL SSELTQDPAVSVALGQTVRITCQGDSLRSYYAS WYQQKPGQAPVLVIYGKNNRPSGIPDRFSGSSS GNTASLTITGAQAEDEADYYCNSRDSSGNHVVF GGGTKLTVL SEQ ID NO: 100 TRv3VH EVQLVQSGGGVERPGGSLRLSCAASGFTFDDYG MS WVRQ APGKGLEWVSGINWSGGSTGYAD S V KGRVTISRDNAKNSLYLOMNSLRAEDTAVYYC AKILGAGRGWYFDLWGKGTTVTVSS SEQ ID NO: 101 TRvIVL SSELTQDPAVSVALGQTVRITCQGDSLRSYYAS WYQQKPGQAPVLVIYGKTNRPSGIPDRFSGSSSG NTASLTITGAQAEDEADYYCNSRDSSGTHVVFG GGTKLTVL SEQ ID NO: 102 TRv4VH EVQLVQSGGGVERPGGSLRLSCAASGFTFDDYG MS WVRQ APGKGLEWVSGINWSGGSTGYAD SV KGRVTISRDNAKNSLYLOMNSLRAEDTAVYYC AKILGAGRGWYFDLWGKGTTVTVSS SEQ ID NO: 103 TRv4VL SSELTQDPAVSVALGQTVRITCQGDSLRSYYAS WYQQKPGQAPVLVIYGKANRPSGIPDRFSGSSS GNTASLTITGAQAEDEADYYCNSRDSSGAHWF GGGTKLTVL SEQ ID NO: 104 TRv5VH EVQLVQSGGGVERPGGSLRLSCAASGFTFDDYG MS WVRQ APGKGLEWVSGINWSGGSTGYAD SV KGRVTISRDNAKNSLYLOMNSLRAEDTAVYYC AKILGAGRGWYFDLWGKGTTVTVSS SEQ ID NO: 105 TRv5VL SSELTQDPAVSVALGQTVRITCQGDSLRSYYAS WYQQKPGQAPVLVIYGKNNRPSGIPDRFSGSSS GNTASLTITGAQAEDEADYYCNSRDSSGNHVVF GGGTKLTVL SEQ ID NO: 106 TRv6VH EVQLVQSGGGVERPGGSLRLSCAASGFTFDDYG MS WVRQ APGKGLEWVSGINWAGGSTGYAD SV KGRVTISRDNAKNSLYLOMNSLRAEDTAVYYC AKILGAGRGWYFDLWGKGTTVTVSS SEQ ID NO: 107 TRv6VL SSELTQDPAVSVALGQTVRITCQGDSLRSYYAS WYQQKPGQAPVLVIYGKTNRPSGIPDRFSGSSSG NTASLTITGAQAEDEADYYCNSRDSSGDHVVFG GGTKLTVL

-64041360

SEQ ID NO: 108 TRv7VH EVOLVOSGGGVERPGGSLRLSCAASGFTFDDYG MSWVRQAPGKGLEWVSGINWTGGSTGYADSV KGRVTISRDNAKNSLYLQMNSLRAEDTAVYYC AKILGAGRGWYFDLWGKGTTVTVSS SEQ ID NO: 109 TRv7VL SSELTQDPAVSVALGQTVRITCQGDSLRSYYAS WYQQKPGQAPVLVIYGKDNRPSGIPDRFSGSSS GNTASLTITGAQAEDEADYYCNSRDSSGDHVVF GGGTKLTVL SEQ ID NO: 110 CDH17E9VH QVQLQQSDAELVKPGASVKISCKVSGYTLTDHT IHWMKQRPDQGLDWIGYIYPRDDITQYNENFKA KSTLTADKSSSTAYMQLNSLTSEDSAVYFCARW SWHFGSNYYGLDYWGQGTSVTVSS SEQIDNO:111 CDH17E9VL DIVMTQSQTFMSTTVGDRVSITCKASQNVGTAL AWYQQKPRQSPKLLIYSASNRFTGVPDRFTGSG SGTDFTLTISNMRSEDLADYVCQQYSTYPLTFGS GTKLEIR SEQ ID NO: 112 CDH17A6VH QVQLQQSDAELVKPGASVKISCKVSGYTFTDHTI HWMKQRPEQGLEWIGYIYPRDAITKYNEKFKGK ATLTADKSSSTAYMOLNSLTSEDSAVYFCARWG YYFGSTSYYFDFWGQGTTLTVSS SEQ ID NO: 113 CDH17A6VL DIVMTQSQKFMSTTVGDRVSITCKASQNVGTGV VWYQQRPGQSPKLLIYSPSNRYTGVPDRFTGSG SGTDFTLTITNIQSEDLADYFCQQYSTYPWTFGG GTKLEIK SEQ ID NO: 114 CDH17E2VH QVQLQQSDAELVKPGASVKISCKVSGYTFTDHTI HWMKQRPEQGLEWIGYIYPRDVITQYNEKFKGK ATLTADKSSSTAYMOLNSLTSEDSAVYLCARW GYFYGSRSYYFDYWGQGTTLTVSS SEQ ID NO: 115 CDH17E2VL DIVMTQSOKFMSTTVGDRVNITCKASQNVGTAV AWYQQKPGQSPKLLIYSASNRYTGVPDRFTGSG SGTDFTLTITNMQSKDLADYFCQQYSSYPWTFG GGTKLEIK SEQ ID NO: 116 CDH17H2VH QVQLVQSGAEVKKPGSSVKVSCKVSGYTFTDHT IHWMKQRPGQGLEWIGYIYPRDVITQYNEKFKG KVTLTADKSTSTAYMELSSLRSEDTAVYLCARW GYFYGSRSYYFDYWGQGTTVTVSS SEQ ID NO: 117 CDH17H2VL DIQMTQSQSSLSASVGDRVGITCKASQNVGTAV AWYQQKPGKAPKLLIYSASNRYTGVPSRFSGSG SGTDFTLTISSLQPKDFATYFCQQYSSYPWTFGQ GTKLEIK SEQ ID NO: 118 CDH17vlVH EVQLQQSVAELVKPGASVKMSCKVSGYTLTDH TIHWMKQRPEQGLEWIGYIYPRDGITGYNEKFK GKATLTADTSSSTAYMOLNSLTSEDSAVYFCAR WGYSYRNYAYYYDYWGQGTTLTVSS SEQ ID NO: 119 CDH17vlVL DIVMSQSPSSLAVSVGEKVTMSCKSSQSLLHSSN QKNYLAWYQQKPGQSPKVLIYWASTRESGVPD RFTGSGSGTDFTLTITSVKSEDLAVYYCOQYYSY PWTFGGGTRLEIK SEQ ID NO: 120 IgGIWTHC ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFP EPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSV

- 65 041360

VTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKS CDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMIS RTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHN AKTKPREEQYNSTYRVVSVLTVLHQDWLNGKE YKCKVSNKALPAPIEKTISKAKGOPREPQVYTLP PSRDELTKNQVSLTCLVKGFYPSDIAVEWESNG QPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQ QGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO: 121 IgGlKOHC ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFP EPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSV VTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKS CDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMI SRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVH NAKTKPREEQYNSTYRVVSVLTVLHQDWLNGK EYKCKVSNKALPAPIEKTISKAKGQPREPQVYTL PPSREEMTKNQVSLTCLVKGFYPSDIAVEWESN GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRW QQGNVFSCSVMHEALHNHYTQKSLSLSPG SEQ ID NO: 122 IgG4ProHC ASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFP EPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSV VTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESK YGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRT PEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAK TKPREEQFNSTYRVVSVLTVLHQDWLNGKEYK CKVSNKGLPSSIEKTISKAKGQPREPQWTLPPSQ EEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPE NNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGN VFSCSVMHEALHNHYTQKSLSLSLGK SEQ ID NO: 123 Kappa light chain RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYP REAKVQWKVDNALQSGNSQESVTEQDSKDSTY SLSSTLTLSKADYEKHKVYACEVTHQGLSSPVT KSFNRGEC SEQ ID NO: 124 TR2vlscFvVLVH DIELTQDPAVSVALGQTVRITCQGDSLRSYYAS WYQQKPGQAPVLVIYGKNNRPSGIPDRFSGSSS GNTASLTITGAQAEDEADYYCNSRDSSGNHVVF GGGTKLTVLGQGGGGSGGGGSGGGGSGGGGSE VQLVQSGGGVERPGGSLRLSCAASGFTFDDYG MS WVRQ APGKGLEWVSGINWNGGSTGYAD SV KGRVTISRDNAKNSLYLQMNSLRAEDTAVYYC AKILGAGRGWYFDLWGKGTTVTVSS SEQ ID NO: 125 TR2v2scFvVLVH SSELTQDPAVSVALGQTVRITCQGDSLRSYYAS WYQQKPGQAPVLVIYGKNNRPSGIPDRFSGSSS GNTASLTITGAQAEDEADYYCNSRDSSGNHVVF GGGTKLTVLGGGGSGGGGSGGGGSGGGGSEVQ LVQSGGGVERPGGSLRLSCAASGFTFDDYGMS WVRQ APGKGLEWVSGINWNGGSTGYAD S VKG RVTISRDNAKNSLYLOMNSLRAEDTAVYYCAKI LGAGRGWYFDLWGKGTTVTVSS SEQ ID NO: 126 TR2v3scFvVLVH SSELTQDPAVSVALGQTVRITCQGDSLRSYYAS WYQQKPGQAPVLVIYGKTNRPSGIPDRFSGSSSG NTASLTITGAQAEDEADYYCNSRDSSGTHVVFG GGTKLTVLGGGGSGGGGSGGGGSGGGGSEVQL VQSGGGVERPGGSLRLSCAASGFTFDDYGMSW VRQAPGKGLEWVSGINVSGGSTGVADS

- 66 041360

TISRDNAKNSLYLQMNSLRAEDTAVYYCAKILG AGRGWYFDLWGKGTTVTVSS SEQ ID NO: 127 TR2v4scFvVLVH SSELTQDPAVSVALGQTVRITCQGDSLRSYYAS WYQQKPGQAPVLVIYGKANRPSGIPDRFSGSSS GNTASLTITGAQAEDEADYYCNSRDSSGAHWF GGGTKLTVLGGGGSGGGGSGGGGSGGGGSEVQ LVQSGGGVERPGGSLRLSCAASGFTFDDYGMS WVRQAPGKGLEWVSGINWSGGSTGYADSVKG RVTISRDNAKNSLYLOMNSLRAEDTAVYYCAKI LGAGRGWYFDLWGKGTTVTVSS SEQ ID NO: 128 TR2v5scFvVLVH SSELTQDPAVSVALGQTVRITCQGDSLRSYYAS WYQQKPGQAPVLVIYGKNNRPSGIPDRFSGSSS GNTASLTITGAQAEDEADYYCNSRDSSGNHVVF GGGTKLTVLGGGGSGGGGSGGGGSGGGGSEVQ LVQSGGGVERPGGSLRLSCAASGFTFDDYGMS WVRQ APGKGLEWVSGINWSGGSTGYAD S VKG RVTISRDNAKNSLYLOMNSLRAEDTAVYYCAKI LGAGRGWYFDLWGKGTTVTVSS SEQ ID NO: 129 TR2v6scFvVLVH SSELTQDPAVSVALGQTVRITCQGDSLRSYYAS WYQQKPGQAPVLVIYGKTNRPSGIPDRFSGSSSG NTASLTITGAQAEDEADYYCNSRDSSGDHVVFG GGTKLTVLGGGGSGGGGSGGGGSGGGGSEVQL VQSGGGVERPGGSLRLSCAASGFTFDDYGMSW VRQAPGKGLEWVSGINWAGGSTGYADSVKGRV TISRDNAKNSLYLQMNSLRAEDTAVYYCAKILG AGRGWYFDLWGKGTTVTVSS SEQ ID NO: 130 TR2v7scFvVLVH SSELTQDPAVSVALGQTVRITCQGDSLRSYYAS WYQQKPGQAPVLVIYGKDNRPSGIPDRFSGSSS GNTASLTITGAQAEDEADYYCNSRDSSGDHVVF GGGTKLTVLGGGGSGGGGSGGGGSGGGGSEVQ LVQSGGGVERPGGSLRLSCAASGFTFDDYGMS WVRQAPGKGLEWVSGINWTGGSTGYADSVKG RVTISRDNAKNSLYLQMNSLRAEDTAVYYCAKI LGAGRGWYFDLWGKGTTVTVSS SEQ ID NO: 131 TR2vlscFvVHVL EVQLVQSGGGVERPGGSLRLSCAASGFTFDDYG MS WVRQ APGKGLEWVSGINWNGGSTGYAD SV KGRVTISRDNAKNSLYLQMNSLRAEDTAVYYC AKILGAGRGWYFDLWGKGTTVTVSSGGGGSGG GGSGGGGSGGGGSDIELTQDPAVSVALGQTVRI TCQGDSLRSYYASWYQQKPGQAPVLVIYGKNN RPSGIPDRFSGSSSGNTASLTITGAQAEDEADYY CNSRDSSGNHVVFGGGTKLTVLGQ SEQ ID NO: 132 TR2v 1 scFv VH VLss EVQLVQSGGGVERPGGSLRLSCAASGFTFDDYG MS WVRQ APGKGLEWVSGINWNGGSTGYAD SV KGRVTISRDNAKNSLYLQMNSLRAEDTAVYYC AKILGAGRGWYFDLWGKGTTVTVSSGGGGSGG GGSGGGGSGGGGSDIELTQDPAVSVALGQTVRI TCQGDSLRSYYASWYQQKPGQAPVLVIYGKNN RPSGIPDRFSGSSSGNTASLTITGAQAEDEADYY CNSRDSSGNHVVFGGGTKLTVLGQ SEQ ID NO: 133 TR2v 1 scFv VL VHss DIELTQDPAVSVALGQTVRITCQGDSLRSYYAS WYQQKPGQAPVLVIYGKNNRPSGIPDRFSGSSS GNTASLTITGAQAEDEADYYCNSRDSSGNHVVF GCGTKLTVLGQGGGGSGGGGSGGGGSGGGGSE

- 67 041360

VQLVQSGGGVERPGGSLRLSCAASGFTFDDYG MS WVRQ APGKCLEWVSGINWNGGSTGYAD SV KGRVTISRDNAKNSLYLQMNSLRAEDTAVYYC AKILGAGRGWYFDLWGKGTTVTVSS SEQ ID NO: 134 TR2#lscFvVLVH DIVMTQSHKFMSTSVGDRVSITCKASQDVITAV AWYQQKPGQSPKLLIYWASTRHTGVPDRFTGT GSGTDYTLTIS SVQ AEDL ALYYCQQHYSTP WTF GGGTKLEIKGGGGSGGGGSGGGGSGGGGSQVQ LQQSGAELVKPGASVKLSCKAFDYTFTSYDINW VKQRPGQGLEWIGWIDPGSGNTKYNEKFKGKA TLTADKSSSTAYMHLSSLTSEDSAVYFCARKNY GGSYAFTYWGQGTLVTVA SEQ ID NO: 135 TR2#2scFvVLVH DIVMTQSHKFMSTSVGDRVSITCKASQDVSTAV AWYQQKPGQSPKVLIYWASTRHTGVPDRFTGS GSGTDYILTISSVQAEDLALYYCQQHYSTPYTFG GGTKLEIKGGGGSGGGGSGGGGSGGGGSKVQL QQSGAELVKTGTSVKLSCKASGYTFTEYIIHWV KQRSGQGLEWIGWFYPGSGSIKYNEKFKDKATL TADKSSSTVYMELSRLTSEDSAVYFCARHEEGG YSAWFPYWGQGTLVTVSA SEQ ID NO: 136 TR2#3scFvVLVH DVLMTQTPLSLPVSLGDQASISCRSSQSIVHSNG DTYLDWYLQKPGQSPKLLIYKVSNRFSGVPDRF SGSGSGTDFTLKISRVVAEDLGVYYCFRGSHIPP TFGAGTKLEKGGGGSGGGGSGGGGSGGGGSEV QLQQSGPELVKPGASVKISCKASGYTFTDYYYL NWVKQSHGKSLEWIGYIYPNNGDTSYNQKFKG KTTLTVDKSSSTAYMEFRSLTSEDSAVYYCTRG SNWIWYFDVWGTGTTVTVSS SEQ ID NO: 137 TR2#7scFvVLVH SIVMTQTPKFLPVSAGDRVTMTCKASQNVGNN VAWYQQKPGQSPKLLISYASNRYTGVPDRFTGS GSGTDFFTISSVQVEDLAVYFCQQHYSSPLTFGA GTKLELKGGGGSGGGGSGGGGSGGGGSQVQLK QSGAELVKPGASVKISCKASGYSFTDYYINWVK QRPGQGLEWIGKIGPGSGNTYYNEKFEGKATLT ADKSSSTAYMQLSSLTSEDSAVYFCASTGPFAY WGQGTLVTWSA SEQ ID NO: 138 TR2#8scFvVLVH DIVMTQSHKFMSTSVGDRVSITCKASQDVGTAV AWYQQKPGQSPKLLIYWASTRHTGVPDRFTGSG SGTDFTLTISNVQSEDLADYFCQQYSSYPYTFGG GTKLEIKGGGGSGGGGSGGGGSGGGGSQVTLK ESGPGILQPSQTLSLTCSFSGFSLSTSGMGVSWIR QPSGKGLEWLAHIYWDDDKRYNPSLKSRLTISK DTSRNQVFLKITSVDTADTATYYCARRRLGPFA YWGQGTLVTVSA SEQ ID NO: 139 TR2#10scFvVLVH DVLMTQTPLSLPVSLGDQASISCRSSQNIVHSNG NTYLDWYLQKPGQSPKLLIYKVSNRFSGVPDRF SGSGSGTDFTLKISRVEAEDLGVYYCFQGSHVPP TFGAGTKLELKGGGGSGGGGSGGGGSGGGGSE VQLQQSGPELVKPGASVKISCKASGYTFTDYYY MNWVKQSHGKSLEWIGYIYPNNGGTRYNQKFK GKATLTVDKSSSTAYMELRSLTSEDSAVYYCAR GGNWNWYFDVWGTGTTVTVSS SEQ ID NO: 140 TR2#12scFvVLVH DIVMTQSHKFMSTSVGDRVSITCKASQDVSNAV AWYQQKPGQSPKLLIYWASTRHNGVPDRFTGS

-68 041360

GSGTDYILTISSVQAEDLALYYCQQHYRTPYTFG GGTKLEIKGGGGSGGGGSGGGGSGGGGSKVQL QQSGAELVKPGASVKLSCKASGYTFTEYIIHWV KQKSGQGLEWIGWFYPGSGNIKYNEKFKDKATL TADKSSSTVYMELSRLTSEDSAVYFCTRHEGGS NFFPYWGQGTLVTVSA SEQ ID NO: 141 Linker between mAb and scFv GGSGGS SEQ ID NO: 142 Extended linker between mAb and scFv GGGGSGGGGS SEQ ID NO: 143 Linker VL and VH GGGGSGGGGSGGGGSGGGGS SEQ ID NO: 144 Linker from scFv to mAb GGGS SEQ ID NO: 145 CDH17vl/TR2vl (VL-VH, IgGIWT) NS EVQLQQSVAELVKPGASVKMSCKVSGYTLTDH TIHWMKQRPEQGLEWIGYIYPRDGITGYNEKFK GKATLTADTSSSTAYMQLNSLTSEDSAVYFCAR WGYSYRNYAYYYDYWGQGTTLTVSSASTKGPS VFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVS WNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSS SLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTH TCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEV TCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYKCKV SNKALPAPIEKTISKAKGQPREPQVYTLPPSRDEL TKNQVSLTCLVKGFYPSDIAVEWESNGQPENNY KTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFS CSVMHEALHNHYTQKSLSLSPGKGGSGGSDIEL TQDPAVSVALGQTVRITCQGDSLRSYYASWYQ QKPGQAPVLVIYGKNNRPSGIPDRFSGSSSGNTA SLTITGAQAEDEADYYCNSRDSSGNHVVFGGGT KLTVLGQGGGGSGGGGSGGGGSGGGGSEVQLV QSGGGVERPGGSLRLSCAASGFTFDDYGMSWV RQAPGKGLEWVSGINWNGGSTGYAD S VKGRVT ISRDNAKNSLYLQMNSLRAEDTAVYYCAKILGA GRGWYFDLWGKGTTVTVSS SEQ ID NO: 146 CDH17vl/TR2vl LC DIVMSQSPSSLAVSVGEKVTMSCKSSQSLLHSSN QKNYLAWYQQKPGQSPKVLIYWASTRESGVPD RFTGSGSGTDFTLTITSVKSEDLAVYYCQQYYSY PWTFGGGTRLEIKRTVAAPSVFIFPPSDEQLKSGT ASVVCLLNNFYPREAKVQWKVDNALQSGNSQE SVTEQECDSKDSTYSLSSSFTHNSKADYEK SEQ ID NO: 147 CDH17v l/TR2v 1 -(VL-VH, IgGl KO) HC EVQLQQSVAELVKPGASVKMSCKVSGYTLTDH TIHWMKQRPEQGLEWIGYIYPRDGITGYNEKFK GKATLTADTSSSTAYMQLNSLTSEDSAVYFCAR WGYSYRNYAYYYDYWGQGTTLTVSSASTKGPS VFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVS WNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSS SLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTH TCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEV TCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYKCKV SNKALPAPIEKTISKAKGQPREPQVYTLPPSREE MTKNQVSLTCLVKGFYPSDIAVEWESNGQPENN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVF

- 69 041360

SCSVMHEALHNHYTQKSLSLSPGGGSGGSDIELT QDPAVSVALGQTVRITCQGDSLRSYYASWYQQ KPGQAPVLVIYGKNNRPSGIPDRFSGSSSGNTAS LTITGAQAEDEADYYCNSRDSSGNHVVFGGGTK LTVLGQGGGGSGGGGSGGGGSGGGGSEVQLVQ SGGGVERPGGSLRLSCAASGFTFDDYGMSWVR QAPGKGLEWVSGINWNGGSTGYADSVKGRVTI SRDNAKNSLYLOMNSLRAEDTAVYYCAKILGA GRGWYFDLWGKGTTVTVSS SEQ ID NO: 148 CDH17vl/TR2vl-(IgGl KO) LC DIVMSQSPSSLAVSVGEKVTMSCKSSQSLLHSSN QKNYLAWYQQKPGQSPKVLIYWASTRESGVPD RFTGSGSGTDFTLTITSVKSEDLAVYYCOQYYSY PWTFGGGTRLEIKRTVAAPSVFIFPPSDEQLKSGT ASVVCLLNNFYPREAKVQWKVDNALQSGNSQE SVTEODSKDSDSPVSSTLVRGANACTY EKHKSFTLSKADYEKHQ SEQ ID NO: 149 CDH 17v l/TR2v2-(VL-VH, IgGl KO) HC EVQLQQSVAELVKPGASVKMSCKVSGYTLTDH TIHWMKQRPEQGLEWIGYIYPRDGITGYNEKFK GKATLTADTSSSTAYMOLNSLTSEDSAVYFCAR WGYSYRNYAYYYDYWGQGTTLTVSSASTKGPS VFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVS WNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSS SLGTOTYICNVNHKPSNTKVDKRVEPKSCDKTH TCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEV TCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYR VVSVLTVLHQDWLNGKEYKCKV SNKALPAPIEKTISKAKGOPREPQVYTLPPSREE MTKNQVSLTCLVKGFYPSDIAVEWESNGQPENN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVF SCSVMHEALHNHYTQKSLSLSPGGGSGGSSSEL TQDPAVSVALGQTVRITCQGDSLRSYYASWYQ QKPGQAPVLVIYGKNNRPSGIPDRFSGSSSGNTA SLTITGAQAEDEADYYCNSRDSSGNHVVFGGGT KLTVLGGGGSGGGGSGGGGSGGGGSEVQLVQS GGGVERPGGSLRLSCAASGFTFDDYGMSWVRQ APGKGLEWVSGINWNGGSTGYADSVKGRVTIS RDNAKNSLYLOMNSLRAEDTAVYYCAKILGAG RGWYFDL WGKGTT VT VS S SEQ ID NO: 150 CDH17vl/TR2v2-(IgGlKO) LC DIVMSQSPSSLAVSVGEKVTMSCKSSQSLLHSSN QKNYLAWYQQKPGQSPKVLIYWASTRESGVPD RFTGSGSGTDFTLTITSVKSEDLAVYYCOQYYSY PWTFGGGTRLEIKRTVAAPSVFIFPPSDEQLKSGT ASVVCLLNNFYPREAKVQWKVDNALQSGNSQE SVTEODSKDSDSPVSSTLVRGANACTY EKHKSFTLSKADYEKHQ SEQ ID NO: 151 CDH 17v l/TR2v2-(VL-VH, IgG4 Pro) HC EVQLQQSVAELVKPGASVKMSCKVSGYTLTDH TIHWMKQRPEQGLEWIGYIYPRDGITGYNEKFK GKATLTADTSSSTAYMOLNSLTSEDSAVYFCAR WGYSYRNYAYYYDYWGQGTTLTVSSASTKGPS VFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSW NSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSS LGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPP CPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVV VDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQ FNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKG

- 70 041360

LPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKN QVSLTCLVKGFYPSDIAVEWESNGQPENNYKTT PPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSV MHEALHNHYTQKSLSLSLGKGGSGGSSSELTQD PAVSVALGQTVRITCQGDSLRSYYASWYQQKPG QAPVLVIYGKNNRPSGIPDRFSGSSSGNTASLTIT GAQAEDEADYYCNSRDSSGNHVVFGGGTKLTV LGGGGSGGGGSGGGGSGGGGSEVQLVQSGGGV ERPGGSLRLSCAASGFTFDDYGMSWVRQAPGK GLEWVSGINWNGGSTGYAD S VKGRVTISRDNA KNSLYLQMNSLRAEDTAVYYCAKILGAGRGWY FDL WGKGTT VT VS S SEQ ID NO: 152 CDH17vl/TR2v2-(IgG4 Pro) LC DIVMSQSPSSLAVSVGEKVTMSCKSSQSLLHSSN QKNYLAWYQQKPGQSPKVLIYWASTRESGVPD RFTGSGSGTDFTLTITSVKSEDLAVYYCQQYYSY PWTFGGGTRLEIKRTVAAPSVFIFPPSDEQLKSGT ASVVCLLNNFYPREAKVQWKVDNALQSGNSQE SVTEQECDSKDSTYSLSSSFTHNSKADYEK SEQ ID NO: 153 CDH17vl/TR2vl-(scFv VHVL, IgGl KO) HC EVQLQQSVAELVKPGASVKMSCKVSGYTLTDH TIHWMKQRPEQGLEWIGYIYPRDGITGYNEKFK GKATLTADTSSSTAYMQLNSLTSEDSAVYFCAR WGYSYRNYAYYYDYWGQGTTLTVSSASTKGPS VFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVS WNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSS SLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTH TCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEV TCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYKCKV SNKALPAPIEKTISKAKGQPREPQVYTLPPSREE MTKNQVSLTCLVKGFYPSDIAVEWESNGQPENN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVF SCSVMHEALHNHYTQKSLSLSPGGGSGGSEVQL VQSGGGVERPGGSLRLSCAASGFTFDDYGMSW VRQAPGKGLEWVSGINWNGGSTGYADSVKGRV TISRDNAKNSLYLQMNSLRAEDTAVYYCAKILG AGRGWYFDLWGKGTTVTVSSGGGGSGGGGSG GGGSGGGGSDIELTQDPAVSVALGQTVRITCQG DSLRS YYASWYQQKPGQAPVLVIYGKNNRPSGI PDRFSGSSSGNTASLTITGAQAEDEADYYCNSRD SSGNHVVFGGGTKLTVLGQ SEQ ID NO: 154 CDH17vl/TR2vl-(scFv VH- VL-LC DIVMSQSPSSLAVSVGEKVTMSCKSSQSLLHSSN QKNYLAWYQQKPGQSPKVLIYWASTRESGVPD RFTGSGSGTDFTLTITSVKSEDLAVYYCQQYYSY PWTFGGGTRLEIKRTVAAPSVFIFPPSDEQLKSGT ASVVCLLNNFYPREAKVQWKVDNALQSGNSQE SVTEQECDSKDSTYSLSSSFTHNSKADYEK SEQ ID NO: 155 CDH17vl/TR2vl-(scFv VHVL ss) HC EVQLQQSVAELVKPGASVKMSCKVSGYTLTDH TIHWMKQRPEQGLEWIGYIYPRDGITGYNEKFK GKATLTADTSSSTAYMQLNSLTSEDSAVYFCAR WGYSYRNYAYYYDYWGQGTTLTVSSASTKGPS VFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVS WNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSS SLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTH

-71 041360

TCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEV TCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYKCKV SNKALPAPIEKTISKAKGQPREPO VYTLPPSREE MTKNQVSLTCLVKGFYPSDIAVEWESNGQPENN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVF SCSVMHEALHNHYTQKSLSLSPGGGSGGSEVQL VQSGGGVERPGGSLRLSCAASGFTFDDYGMSW VRQAPGKCLEWVSGINWNGGSTGYADSVKGRV TISRDNAKNSLYLOMNSLRAEDTAVYYCAKILG AGRGWYFDLWGKGTTVTVSSGGGGSGGGGSG GGGSGGGGSDIELTQDPAVSVALGQTVRITCQG DSLRS YYASWYQQKPGQAPVLVIYGKNNRPSGI PDRFSGSSSGNTASLTITGAQAEDEADYYCNSRD SSGNHVVFGCGTKLTVLGQ SEQ ID NO: 156 CDH17vl/TR2vl-(scFv VHVL ss) LC DIVMSQSPSSLAVSVGEKVTMSCKSSQSLLHSSN QKNYLAWYQQKPGQSPKVLIYWASTRESGVPD RFTGSGSGTDFTLTITSVKSEDLAVYYCOQYYSY PWTFGGGTRLEIKRTVAAPSVFIFPPSDEQLKSGT ASVVCLLNNFYPREAKVQWKVDNALQSGNSQE SVTEODSKDSDSPVSSTLVRGANACTY EKHKSFTLSKADYEKHQ SEQ ID NO: 157 CDH17vl/TR2vl-(scFv VL- VH ss, IgGl KO) HC EVQLQQSVAELVKPGASVKMSCKVSGYTLTDH TIHWMKQRPEQGLEWIGYIYPRDGITGYNEKFK GKATLTADTSSSTAYMOLNSLTSEDSAVYFCAR WGYSYRNYAYYYDYWGQGTTLTVSSASTKGPS VFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVS WNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSS SLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTH TCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEV TCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYKCKV SNKALPAPIEKTISKAKGQPREPO VYTLPPSREE MTKNQVSLTCLVKGFYPSDIAVEWESNGQPENN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVF SCSVMHEALHNHYTQKSLSLSPGGGSGGSDIELT QDPAVSVALGQTVRITCQGDSLRSYYASWYQQ KPGQAPVLVIYGKNNRPSGIPDRFSGSSSGNTAS LTITGAQAEDEADYYCNSRDSSGNHVVFGCGTK LTVLGQGGGGSGGGGSGGGGSGGGGSEVQLVQ SGGGVERPGGSLRLSCAASGFTFDDYGMSWVR QAPGKCLEWVSGINWNGGSTGYADSVKGRVTI SRDNAKNSLYLOMNSLRAEDTAVYYCAKILGA GRGWYFDLWGKGTTVTVSS SEQ ID NO: 158 CDH17vl/TR2vl-(scFv VLVH ss) LC DIVMSQSPSSLAVSVGEKVTMSCKSSQSLLHSSN QKNYLAWYQQKPGQSPKVLIYWASTRESGVPD RFTGSGSGTDFTLTITSVKSEDLAVYYCOQYYSY PWTFGGGTRLEIKRTVAAPSVFIFPPSDEQLKSGT ASVVCLLNNFYPREAKVQWKVDNALQSGNSQE SVTEQDSKDSPVVKSFTLVNSKADYEKVHKVDNALQSGNSQE SEQ ID NO: 159 CDH17E9/TR2#1 (IgGlKO)HC QVQLQQSDAELVKPGASVKISCKVSGYTLTDHT IHWMKQRPDQGLDWIGYIYPRDDITQYNENFKA KSTLTADKSSSTAYMQLNSLTSEDSAVYFCARW SWHFGSNYYGLDYWGQGTSVTVSSASTKGPSV

- 72 041360

FPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSW NSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSS LGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHT CPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVT CVVVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYKCKV SNKALPAPIEKTISKAKGQPREPQVYTLPPSREE MTKNQVSLTCLVKGFYPSDIAVEWESNGQPENN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVF SCSVMHEALHNHYTQKSLSLSPGGGSGGSDIVM TQSHKFMSTSVGDRVSITCKASQDVITAVAWYQ QKPGQSPKLLIYWASTRHTGVPDRFTGTGSGTD YTLTISSVQAEDLALYYCQQHYSTPWTFGGGTK LEIKGGGGSGGGGSGGGGSGGGGSQVQLQQSG AELVKPGASVKLSCKAFDYTFTSYDINWVKQRP GQGLEWIGWIDPGSGNTKYNEKFKGKATLTAD KSSSTAYMHLSSLTSEDSAVYFCARKNYGGSYA FTYWGQGTLVTVA SEQ TD NO: 160 CDH17E9/TR2#2 (IgGlKO)HC QVQLQQSDAELVKPGASVKTSCKVSGYTLTDHT IHWMKQRPDQGLDWIGYIYPRDDITQYNENFKA KSTLTADKSSSTAYMQLNSLTSEDSAVYFCARW SWHFGSNYYGLDYWGQGTSVTVSSASTKGPSV FPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSW NSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSS LGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHT CPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVT CVVVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYKCKV SNKALPAPIEKTISKAKGQPREPQVYTLPPSREE MTKNQVSLTCLVKGFYPSDIAVEWESNGQPENN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVF SCSVMHEALHNHYTQKSLSLSPGGGSGGSDIVM TQSHKFMSTSVGDRVSITCKASQDVSTAVAWY QQKPGQSPKVLIYWASTRHTGVPDRFTGSGSGT DYILTISSVQAEDLALYYCQQHYSTPYTFGGGTK LEIKGGGGSGGGGSGGGGSGGGGSKVQLQQSG AELVKTGTSVKLSCKASGYTFTEYIIHWVKQRS GQGLEWIGWFYPGSGSIKYNEKFKDKATLTADK SSSTVYMELSRLTSEDSAVYFCARHEEGGYSAW FPYWGQGTLVTVSA SEQ ID NO: 161 CDH17E9/TR2#3 (IgGlKO)HC QVQLQQSDAELVKPGASVKISCKVSGYTLTDHT IHWMKQRPDQGLDWIGYIYPRDDITQYNENFKA KSTLTADKSSSTAYMQLNSLTSEDSAVYFCARW SWHFGSNYYGLDYWGQGTSVTVSSASTKGPSV FPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSW NSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSS LGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHT CPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVT CVVVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYKCKV SNKALPAPIEKTISKAKGQPREPQVYTLPPSREE MTKNQVSLTCLVKGFYPSDIAVEWESNGQPENN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVF SCSVMHEALHNHYTQKSLSLSPGGGSGGSDVL MTQTPLSLPVSLGDQASISCRSSQSIVHSNGDTY LDWYLQKPGQSPKLLIYKVSNRFSGVPDRFSGS GSGTDFTLKISRVVAEDLGVYYCFRGSHIPPTFG

- 73 041360

AGTKLEKGGGGSGGGGSGGGGSGGGGSEVQLQ QSGPELVKPGASVKISCKASGYTFTDYYYLNWV KQSHGKSLEWIGYIYPNNGDTSYNQKFKGKTTL TVDKSSSTAYMEFRSLTSEDSAVYYCTRGSNWI WYFDVWGTGTTVTVSS SEQ ID NO: 162 CDH17E9/TR2#7 (IgGlKO)HC QVQLQQSDAELVKPGASVKISCKVSGYTLTDHT IHWMKQRPDQGLDWIGYIYPRDDITQYNENFKA KSTLTADKSSSTAYMQLNSLTSEDSAVYFCARW SWHFGSNYYGLDYWGQGTSVTVSSASTKGPSV FPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSW NSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSS LGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHT CPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVT CVVVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYKCKV SNKALPAPIEKTISKAKGOPREPQVYTLPPSREE MTKNQVSLTCLVKGFYPSDIAVEWESNGQPENN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVF SCSVMHEALHNHYTQKSLSLSPGGGSGGSSIVM TQTPKFLPVSAGDRVTMTCKASQNVGNNVAWY QQKPGQSPKLLISYASNRYTGVPDRFTGSGSGTD FFTISSVOVEDLAVYFCOOHYSSPLTFGAGTKLE LKGGGGSGGGGSGGGGSGGGGSQVQLKQSGAE LVKPGASVKISCKASGYSFTDYYINWVKQRPGQ GLEWIGKIGPGSGNTYYNEKFEGKATLTADKSS STAYMOLSSLTSEDSAVYFCASTGPFAYWGOGT LVTWSA SEQ ID NO: 163 CDH17E9/TR2#8 (IgGlKO)HC QVQLQQSDAELVKPGASVKISCKVSGYTLTDHT IHWMKQRPDQGLDWIGYIYPRDDITQYNENFKA KSTLTADKSSSTAYMQLNSLTSEDSAVYFCARW SWHFGSNYYGLDYWGQGTSVTVSSASTKGPSV FPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSW NSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSS LGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHT CPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVT CVVVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYKCKV SNKALPAPIEKTISKAKGOPREPQVYTLPPSREE MTKNQVSLTCLVKGFYPSDIAVEWESNGQPENN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVF SCSVMHEALHNHYTQKSLSLSPGGGSGGSDIVM TQSHKFMSTSVGDRVSITCKASQDVGTAVAWY QQKPGQSPKLLIYWASTRHTGVPDRFTGSGSGT DFTLTISNVQSEDLADYFCQQYSSYPYTFGGGTK LEIKGGGGSGGGGSGGGGSGGGGSQVTLKESGP GILQPSQTLSLTCSFSGFSLSTSGMGVSWIRQPSG KGLEWLAHIYWDDDKRYNPSLKSRLTISKDTSR NQVFLKITSVDTADTATYYCARRRLGPFAYWG QGTLVTVSA SEQ ID NO: 164 CDH17E9/TR2#10 (IgGlKO)HC QVQLQQSDAELVKPGASVKISCKVSGYTLTDHT IHWMKQRPDQGLDWIGYIYPRDDITQYNENFKA KSTLTADKSSSTAYMQLNSLTSEDSAVYFCARW SWHFGSNYYGLDYWGQGTSVTVSSASTKGPSV FPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSW NSGALTSGVHTFPAVLQSSGVLYSTICVPSNKSSK

- 74 041360

CPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVT CVVVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYR VVSVLTVLHQDWLNGKEYKCKV SNKALPAPIEKTISKAKGQPREPQVYTLPPSREE MTKNQVSLTCLVKGFYPSDIAVE WESNGQPENN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVF SCSVMHEALHNHYTQKSLSLSPGGGSGGSDVL MTQTPLSLPVSLGDQASISCRSSQNIVHSNGNTY LDWYLQKPGQSPKLLIYKVSNRFSGVPDRFSGS GSGTDFTLKISRVEAEDLGVYYCFQGSHVPPTFG AGTKLELKGGGGSGGGGSGGGGSGGGGSEVQL QQSGPELVKPGASVKISCKASGYTFTDYYYMN WVKQSHGKSLEWIGYIYPNNGGTRYNQKFKGK ATLTVDKSSSTAYMELRSLTSEDSAVYYCARGG NWNWYFDVWGTGTTVTVSS SEQ ID NO: 165 CDH17E9/TR2#12 (IgGlKO)HC QVQLQQSDAELVKPGASVKISCKVSGYTLTDHT IHWMKQRPDQGLDWIGYIYPRDDITQYNENFKA KSTLTADKSSSTAYMQLNSLTSEDSAVYFCARW SWHFGSNYYGLDYWGQGTSVTVSSASTKGPSV FPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSW NSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSS LGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHT CPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVT CVVVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYKCKV SNKALPAPIEKTISKAKGQPREPQVYTLPPSREE MTKNQVSLTCLVKGFYPSDIAVE WESNGQPENN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVF SCSVMHEALHNHYTQKSLSLSPGGGSGGSDIVM TQSHKFMSTSVGDRVSITCKASQDVSNAVAWY QQKPGQSPKLLIYWASTRHNGVPDRFTGSGSGT DYILTISSVQAEDLALYYCQQHYRTPYTFGGGT KLEIKGGGGSGGGGSGGGGSGGGGSKVQLQQS GAELVKPGASVKLSCKASGYTFTEYIIHWVKQK SGQGLEWIGWFYPGSGNIKYNEKFKDKATLTAD KSSSTVYMELSRLTSEDSAVYFCTRHEGGSNFFP YWGQGTLVTVSA SEQ ID NO: 166 CDH17E9/TR2vl (IgGlKO)HC QVQLQQSDAELVKPGASVKISCKVSGYTLTDHT IHWMKQRPDQGLDWIGYIYPRDDITQYNENFKA KSTLTADKSSSTAYMQLNSLTSEDSAVYFCARW SWHFGSNYYGLDYWGQGTSVTVSSASTKGPSV FPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSW NSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSS LGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHT CPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVT CVVVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYR VVSVLTVLHQDWLNGKEYKCKV SNKALPAPIEKTISKAKGQPREPQVYTLPPSREE MTKNQVSLTCLVKGFYPSDIAVE WESNGQPENN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVF SCSVMHEALHNHYTQKSLSLSPGGGSGGSDIELT QDPAVSVALGQTVRITCQGDSLRSYYASWYQQ KPGQAPVLVIYGKNNRPSGIPDRFSGSSSGNTAS LTITGAQAEDEADYYCNSRDSSGNHVVFGCGTK LTVLGGGGSGGGGSGGGGSGGGGSEVQLVQSG GGVERPGGSLRLSCAASGFTFDDYGMSWVRQA PGKCLEWVSGINWNGGSTGYADSVKGRVTISRD

-75 041360

NAKNSLYLOMNSLRAEDTAVYYCAKILGAGRG WYFDLWGKGTTVTVSS SEQ ID NO: 167 CDH17E9/TR2v2 (IgGlKO)HC QVQLQQSDAELVKPGASVKISCKVSGYTLTDHT IHWMKQRPDQGLDWIGYIYPRDDITQYNENFKA KSTLTADKSSSTAYMOLNSLTSEDSAVYFCARW SWHFGSNYYGLDYWGQGTSVTVSSASTKGPSV FPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSW NSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSS LGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHT CPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVT CVVVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYKCKV SNKALPAPIEKTISKAKGQPREPQ VYTLPPSREE MTKNQVSLTCLVKGFYPSDIAVE WESNGQPENN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVF SCSVMHEALHNHYTQKSLSLSPGGGSGGSSSEL TQDPAVSVALGQTVRITCQGDSLRSYYASWYQ QKPGQAPVLVIYGKNNRPSGIPDRFSGSSSGNTA SLTITGAQAEDEADYYCNSRDSSGNHVVFGGGT KLTVLGGGGSGGGGSGGGGSGGGGSEVQLVQS GGGVERPGGSLRLSCAASGFTFDDYGMSWVRQ APGKGLEWVSGINWNGGSTGYADSVKGRVTIS RDNAKNSLYLQMNSLRAEDTAVYYCAKILGAG RGWYFDL WGKGTT VTVS S SEQ ID NO: 168 CDH17E9/TR2v3 (IgGlKO)HC QVQLQQSDAELVKPGASVKISCKVSGYTLTDHT IHWMKQRPDQGLDWIGYIYPRDDITQYNENFKA KSTLTADKSSSTAYMQLNSLTSEDSAVYFCARW SWHFGSNYYGLDYWGQGTSVTVSSASTKGPSV FPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSW NSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSS LGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHT CPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVT CVVVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYKCKV SNKALPAPIEKTISKAKGQPREPQ VYTLPPSREE MTKNQVSLTCLVKGFYPSDIAVE WESNGQPENN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVF SCSVMHEALHNHYTQKSLSLSPGGGSGGSSSEL TQDPAVSVALGQTVRITCQGDSLRSYYASWYQ QKPGQAPVLVIYGKTNRPSGIPDRFSGSSSGNTA SLTITGAQAEDEADYYCNSRDSSGTHVVFGGGT KLTVLGGGGSGGGGSGGGGSGGGGSEVQLVQS GGGVERPGGSLRLSCAASGFTFDDYGMSWVRQ APGKGLEWVSGINWSGGSTGYADSVKGRVTISR DNAKNSLYLOMNSLRAEDTAVYYCAKILGAGR GWYFDLWGKGTTVTVSS SEQ ID NO: 169 CDH17E9/TR2v4 (IgGlKO)HC QVQLQQSDAELVKPGASVKISCKVSGYTLTDHT IHWMKQRPDQGLDWIGYIYPRDDITQYNENFKA KSTLTADKSSSTAYMQLNSLTSEDSAVYFCARW SWHFGSNYYGLDYWGQGTSVTVSSASTKGPSV FPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSW NSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSS LGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHT CPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVT CVVVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYKCKV

- 76 041360

SNKALPAPIEKTISKAKGQPREPQVYTLPPSREE MTKNOVSLTCLVKGFYPSDIAVEWESNGQPENN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVF SCSVMHEALHNHYTQKSLSLSPGGGSGGSSSEL TQDPAVSVALGQTVRITCQGDSLRSYYASWYQ QKPGQAPVLVIYGKANRPSGIPDRFSGSSSGNTA SLTITGAQAEDEADYYCNSRDSSGAHVVFGGGT KLTVLGGGGSGGGGSGGGGSGGGGSEVQLVQS GGGVERPGGSLRLSCAASGFTFDDYGMSWVRQ APGKGLEWVSGINWSGGSTGYADSVKGRVTISR DNAKNSLYLOMNSLRAEDTAVYYCAKILGAGR GWYFDLWGKGTTVTVSS SEQ ID NO: 170 CDH17E9/TR2v5 (IgGlKO)HC QVQLQQSDAELVKPGASVKISCKVSGYTLTDHT IHWMKQRPDQGLDWIGYIYPRDDITQYNENFKA KSTLTADKSSSTAYMQLNSLTSEDSAVYFCARW SWHFGSNYYGLDYWGQGTSVTVSSASTKGPSV FPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSW NSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSS LGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHT CPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVT CVVVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYKCKV SNKALPAPIEKTISKAKGQPREPQVYTLPPSREE MTKNQVSLTCLVKGFYPSDIAVEWESNGQPENN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVF SCSVMHEALHNHYTQKSLSLSPGGGSGGSSSEL TQDPAVSVALGQTVRITCQGDSLRSYYASWYQ QKPGQAPVLVIYGKNNRPSGIPDRFSGSSSGNTA SLTITGAQAEDEADYYCNSRDSSGNHVVFGGGT KLTVLGGGGSGGGGSGGGGSGGGGSEVQLVQS GGGVERPGGSLRLSCAASGFTFDDYGMSWVRQ APGKGLEWVSGINWSGGSTGYADSVKGRVTISR DNAKNSLYLOMNSLRAEDTAVYYCAKILGAGR GWYFDLWGKGTTVTVSS SEQ ID NO: 171 CDH17E9/TR2v6 (IgGlKO)HC QVQLQQSDAELVKPGASVKISCKVSGYTLTDHT IHWMKQRPDQGLDWIGYIYPRDDITQYNENFKA KSTLTADKSSSTAYMQLNSLTSEDSAVYFCARW SWHFGSNYYGLDYWGQGTSVTVSSASTKGPSV FPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSW NSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSS LGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHT CPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVT CVVVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYKCKV SNKALPAPIEKTISKAKGQPREPQVYTLPPSREE MTKNQVSLTCLVKGFYPSDIAVEWESNGQPENN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVF SCSVMHEALHNHYTQKSLSLSPGGGSGGSSSEL TQDPAVSVALGQTVRITCQGDSLRSYYASWYQ QKPGQAPVLVIYGKTNRPSGIPDRFSGSSSGNTA SLTITGAQAEDEADYYCNSRDSSGDHVVFGGGT KLTVLGGGGSGGGGSGGGGSGGGGSEVQLVQS GGGVERPGGSLRLSCAASGFTFDDYGMSWVRQ APGKGLEWVSGINWAGGSTGYADSVKGRVTIS RDNAKNSLYLOMNSLRAEDTAVYYCAKILGAG RGWYFDL WGKGTT VT VS S

- 77 041360

SEQ ID NO: 172 CDH17E9/TR2v7 (IgGlKO)HC QVQLQQSDAELVKPGASVKISCKVSGYTLTDHT IHWMKQRPDQGLDWIGYIYPRDDITQYNENFKA KSTLTADKSSSTAYMQLNSLTSEDSAVYFCARW SWHFGSNYYGLDYWGQGTSVTVSSASTKGPSV FPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSW NSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSS LGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHT CPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVT CVVVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYKCKV SNKALPAPIEKTISKAKGQPREPQ VYTLPPSREE MTKNQVSLTCLVKGFYPSDIAVEWESNGQPENN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVF SCSVMHEALHNHYTQKSLSLSPGGGSGGSSSEL TQDPAVSVALGQTVRITCQGDSLRSYYASWYQ QKPGQAPVLVIYGKDNRPSGIPDRFSGSSSGNTA SLTITGAQAEDEADYYCNSRDSSGDHVVFGGGT KLTVLGGGGSGGGGSGGGGSGGGGSEVQLVQS GGGVERPGGSLRLSCAASGFTFDDYGMSWVRQ APGKGLEWVSGINWTGGSTGYADSVKGRVTISR DNAKNSLYLQMNSLRAEDTAVYYCAKILGAGR GWYFDLWGKGTTVTVSS SEQ ID NO: 173 CDH17E9 LC DIVMTQSQTFMSTTVGDRVSITCKASQNVGTAL AWYQQKPRQSPKLLIYSASNRFTGVPDRFTGSG SGTDFTLTISNMRSEDLADYVCQQYSTYPLTFGS GTKLEIRRTVAAPSVFIFPPSDEQLKSGTASVVCL LNNFYPREAKVQWKVDNALQSGNSQESVTEQD SKECDSTYSLSSTLTLSKADYEKHKVYACEQ SEQ ID NO: 174 CDH17A6/TR2#1 (IgGlKO)HC QVQLQQSDAELVKPGASVKISCKVSGYTFTDHTI HWMKQRPEQGLEWIGYIYPRDAITKYNEKFKGK ATLTADKSSSTAYMQLNSLTSEDSAVYFCARWG YYFGSTSYYFDFWGQGTTLTVSSASTKGPSVFPL APSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGT QTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPP CPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCV VVDVSHEDPEVKFNWYVDGVEVHNAKTKPREE QYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK ALPAPIEKTISKAKGQPREPQVYTLPPSREEMTK NQVSLTCLVKGFYPSDIAVEWESNGQPENNYKT TPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCS VMHEALHNHYTQKSLSLSPGGGSGGSDIVMTQS HKFMSTSVGDRVSITCKASQDVITAVAWYQQKP GQSPKLLIYWASTRHTGVPDRFTGTGSGTDYTL TISSVQAEDLALYYCQQHYSTPWTFGGGTKLEIK GGGGSGGGGSGGGGSGGGGSQVQLQQSGAELV KPGASVKLSCKAFDYTFTSYDINWVKQRPGQGL EWIGWIDPGSGNTKYNEKFKGKATLTADKSSST AYMHLSSLTSEDSAVYFCARKNYGGSYAFTYW GQGTLVTVA SEQ ID NO: 175 CDH17A6/TR2#2 (IgGlKO)HC QVQLQQSDAELVKPGASVKISCKVSGYTFTDHTI HWMKQRPEQGLEWIGYIYPRDAITKYNEKFKGK ATLTADKSSSTAYMQLNSLTSEDSAVYFCARWG YYFGSTSYYFDFWGQGTTLTVSSASTKGPSVFPL APSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG

- 78 041360

ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGT QTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPP CPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCV VVDVSHEDPEVKFNWYVDGVEVHNAKTKPREE QYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK ALPAPIEKTISKAKGQPREPQVYTLPPSREEMTK NQVSLTCLVKGFYPSDIAVEWESNGQPENNYKT TPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCS VMHEALHNHYTQKSLSLSPGGGSGGSDIVMTQS HKFMSTSVGDRVSITCKASQDVSTAVAWYQQK PGQSPKVLIYWASTRHTGVPDRFTGSGSGTDYIL TISSVQAEDLALYYCQQHYSTPYTFGGGTKLEIK GGGGSGGGGSGGGGSGGGGSKVQLQQSGAELV KTGTSVKLSCKASGYTFIEYIIHWVKQRSGQGL EWIGWFYPGSGSIKYNEKFKDKATLTADKSSST VYMELSRLTSEDSAVYFCARHEEGGYSAWFPY WGQGTLVTVSA SEQ ID NO: 176 CDH17A6/TR2#3 (IgGlKO)HC QVQLQQSDAELVKPGASVKISCKVSGYTFTDHTI HWMKQRPEQGLEWIGYIYPRDAITKYNEKFKGK ATLTADKSSSTAYMQLNSLTSEDSAVYFCARWG YYFGSTSYYFDFWGQGTTLTVSSASTKGPSVFPL APSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGT QTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPP CPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCV VVDVSHEDPEVKFNWYVDGVEVHNAKTKPREE QYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK ALPAPIEKTISKAKGQPREPQVYTLPPSREEMTK NQVSLTCLVKGFYPSDIAVEWESNGQPENNYKT TPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCS VMHEALHNHYTQKSLSLSPGGGSGGSDVLMTQ TPLSLPVSLGDQASISCRSSQSIVHSNGDTYLDW YLQKPGQSPKLLIYKVSNRFSGVPDRFSGSGSGT DFTLKISRVVAEDLGVYYCFRGSHIPPTFGAGTK LEKGGGGSGGGGSGGGGSGGGGSEVQLQQSGP ELVKPGASVKISCKASGYTFTDYYYLNWVKQSH GKSLEWIGYIYPNNGDTSYNQKFKGKTTLTVDK SSSTAYMEFRSLTSEDSAVYYCTRGSNWIWYFD VWGTGTTVTVSS SEQ ID NO: 177 CDH17A6/TR2#7 (IgGlKO)HC QVQLQQSDAELVKPGASVKISCKVSGYTFTDHTI HWMKQRPEQGLEWIGYIYPRDAITKYNEKFKGK ATLTADKSSSTAYMQLNSLTSEDSAWFCARWG YYFGSTSYYFDFWGQGTTLTVSSASTKGPSVFPL APSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGT QTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPP CPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCV VVDVSHEDPEVKFNWYVDGVEVHNAKTKPREE QYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK ALPAPIEKTISKAKGQPREPQWTLPPSREEMTK NQVSLTCLVKGFYPSDIAVEWESNGQPENNYKT TPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCS VMHEALHNHYTQKSLSLSPGGGSGGSSIVMTQT PKFLPVSAGDRVTMTCKASQNVGNNVAWYQQ KPGQSPKLLISYASNRYTGVPDRFTGSGSGTDFF TISSVQVEDLAVYFCQQHYSSPLTFGAGTKLELK GGGGSGGGGSGGGGSGGGGSQVQLKQSGAELV

- 79 041360

KPGASVKISCKASGYSFTDYYINWVKQRPGQGL EWIGKIGPGSGNTYYNEKFEGKATLTADKSSST AYMQLSSLTSEDSAVYFCASTGPFAYWGQGTLV TWSA SEQ ID NO: 178 CDH17A6/TR2#8 (IgGlKO)HC QVQLQQSDAELVKPGASVKISCKVSGYTFTDHTI HWMKQRPEQGLEWIGYIYPRDAITKYNEKFKGK ATLTADKSSSTAYMOLNSLTSEDSAVYFCARWG YYFGSTSYYFDFWGQGTTLTVSSASTKGPSVFPL APSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGT QTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPP CPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCV VVDVSHEDPEVKFNWYVDGVEVHNAKTKPREE QYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK ALPAPIEKTISKAKGOPREPQVYTLPPSREEMTK NQVSLTCLVKGFYPSDIAVEWESNGQPENNYKT TPPVLDSDGSFFLYSKLTVDKSRWOQGNVFSCS VMHEALHNHYTQKSLSLSPGGGSGGSDIVMTQS HKFMSTSVGDRVSITCKASQDVGTAVAWYQQK PGQSPKLLIYWASTRHTGVPDRFTGSGSGTDFTL TISNVQSEDLADYFCQQYSSYPYTFGGGTKLEIK GGGGSGGGGSGGGGSGGGGSQVTLKESGPGILQ PSQTLSLTCSFSGFSLSTSGMGVSWIRQPSGKGL EWLAHIYWDDDKRYNPSLKSRLTISKDTSRNQV FLKITSVDTADTATYYCARRRLGPFAYWGQGTL VTVSA SEQ ID NO: 179 CDH17A6/TR2#10 (IgGlKO)HC QVQLQQSDAELVKPGASVKISCKVSGYTFTDHTI HWMKQRPEQGLEWIGYIYPRDAITKYNEKFKGK ATLTADKSSSTAYMOLNSLTSEDSAVYFCARWG YYFGSTSYYFDFWGQGTTLTVSSASTKGPSVFPL APSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGT QTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPP CPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCV VVDVSHEDPEVKFNWYVDGVEVHNAKTKPREE QYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK ALPAPIEKTISKAKGQPREPQVYTLPPSREEMTK NQVSLTCLVKGFYPSDIAVEWESNGQPENNYKT TPPVLDSDGSFFLYSKLTVDKSRWOQGNVFSCS VMHEALHNHYTQKSLSLSPGGGSGGSDVLMTQ TPLSLPVSLGDQASISCRSSQNIVHSNGNTYLDW YLQKPGQSPKLLIYKVSNRFSGVPDRFSGSGSGT DFTLKISRVEAEDLGVYYCFQGSHVPPTFGAGT KLELKGGGGSGGGGSGGGGSGGGGSEVQLQQS GPELVKPGASVKISCKASGYTFTDYYYMNWVK QSHGKSLEWIGYIYPNNGGTRYNQKFKGKATLT VDKSSSTAYMELRSLTSEDSAVYYCARGGNWN WYFDVWGTGTTVTVSS SEQ ID NO: 180 CDH17A6/TR2#12 (IgGlKO)HC QVQLQQSDAELVKPGASVKISCKVSGYTFTDHTI HWMKQRPEQGLEWIGYIYPRDAITKYNEKFKGK ATLTADKSSSTAYMOLNSLTSEDSAVYFCARWG YYFGSTSYYFDFWGQGTTLTVSSASTKGPSVFPL APSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGT QTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPP CPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCV

- 80 041360

VVDVSHEDPEVKFNWYVDGVEVHNAKTKPREE QYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK ALPAPIEKTISKAKGQPREPQVYTLPPSREEMTK NQVSLTCLVKGFYPSDIAVEWESNGQPENNYKT TPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCS VMHEALHNHYTQKSLSLSPGGGSGGSDIVMTQS HKFMSTSVGDRVSITCKASQDVSNAVAWYQQK PGQSPKLLIYWASTRHNGVPDRFTGSGSGTDYIL TISSVQAEDLALYYCQQHYRTPYTFGGGTKLEIK GGGGSGGGGSGGGGSGGGGSKVQLQQSGAELV KPGASVKLSCKASGYTFTEYIIHWVKQKSGQGL EWIGWFYPGSGNIKYNEKFKDKATLTADKSSST VYMELSRLTSEDSAVYFCTRHEGGSNFFPYWGQ GTLVTVSA SEQIDNO:181 CDH17A6/TR2vl (IgGlKO)HC QVQLQQSDAELVKPGASVKISCKVSGYTFTDHTI HWMKQRPEQGLEWIGYIYPRDAITKYNEKFKGK ATLTADKSSSTAYMQLNSLTSEDSAVYFCARWG YYFGSTSYYFDFWGQGTTLTVSSASTKGPSVFPL APSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGT QTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPP CPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCV VVDVSHEDPEVKFNWYVDGVEVHNAKTKPREE QYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK ALPAPIEKTISKAKGOPREPQVYTLPPSREEMTK NQVSLTCLVKGFYPSDIAVEWESNGQPENNYKT TPPVLDSDGSFFLYSKLTVDKSRWOOGNVFSCS VMHEALHNHYTQKSLSLSPGGGSGGSDIELTQD PAVSVALGQTVRITCQGDSLRSYYASWYQQKPG QAPVLVIYGKNNRPSGIPDRFSGSSSGNTASLTIT GAQAEDEADYYCNSRDSSGNHVVFGCGTKLTV LGGGGSGGGGSGGGGSGGGGSEVQLVQSGGGV ERPGGSLRLSCAASGFTFDDYGMSWVRQAPGK CLEWVSGINWNGGSTGY AD S VKGRVTISRDNA KNSLYLOMNSLRAEDTAVYYCAKILGAGRGWY FDL WGKGTT VT VS S SEQ ID NO: 182 CDH17A6/TR2v2 (IgGlKO)HC QVQLQQSDAELVKPGASVKISCKVSGYTFTDHTI HWMKQRPEQGLEWIGYIYPRDAITKYNEKFKGK ATLTADKSSSTAYMOLNSLTSEDSAVYFCARWG YYFGSTSYYFDFWGQGTTLTVSSASTKGPSVFPL APSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGT QTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPP CPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCV VVDVSHEDPEVKFNWYVDGVEVHNAKTKPREE QYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK ALPAPIEKTISKAKGOPREPQVYTLPPSREEMTK NQVSLTCLVKGFYPSDIAVEWESNGQPENNYKT TPPVLDSDGSFFLYSKLTVDKSRWOOGNVFSCS VMHEALHNHYTQKSLSLSPGGGSGGSSSELTQD PAVSVALGQTVRITCQGDSLRSYYASWYQQKPG QAPVLVIYGKNNRPSGIPDRFSGSSSGNTASLTIT GAQAEDEADYYCNSRDSSGNHVVFGGGTKLTV LGGGGSGGGGSGGGGSGGGGSEVQLVQSGGGV ERPGGSLRLSCAASGFTFDDYGMSWVRQAPGK GLEWVSGINWNGGSTGYAD S VKGRVTISRDNA KNSLYLOMNSLRAEDTAVYYCAKILGAGRGWY

- 81 041360

FDL WGKGTT VT VS S SEQ ID NO: 183 CDH17A6/TR2v3 (IgGlKO)HC QVQLQQSDAELVKPGASVKISCKVSGYTFTDHTI HWMKQRPEQGLEWIGYIYPRDAITKYNEKFKGK ATLTADKSSSTAYMQLNSLTSEDSAVYFCARWG YYFGSTSYYFDFWGQGTTLTVSSASTKGPSVFPL APSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGT QTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPP CPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCV VVDVSHEDPEVKFNWYVDGVEVHNAKTKPREE QYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK ALPAPIEKTISKAKGQPREPQVYTLPPSREEMTK NQVSLTCLVKGFYPSDIAVEWESNGQPENNYKT TPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCS VMHEALHNHYTQKSLSLSPGGGSGGSSSELTQD PAVSVALGQTVRITCQGDSLRSYYASWYQQKPG QAPVLVIYGKTNRPSGIPDRFSGSSSGNTASLTIT GAQAEDEADYYCNSRDSSGTHVVFGGGTKLTV LGGGGSGGGGSGGGGSGGGGSEVQLVQSGGGV ERPGGSLRLSCAASGFTFDDYGMSWVRQAPGK GLEWVSGINWSGGSTGYADSVKGRVTISRDNA KNSLYLOMNSLRAEDTAVYYCAKILGAGRGWY FDL WGKGTT VT VS S SEQ ID NO: 184 CDH17A6/TR2v4 (IgGlKO)HC QVQLQQSDAELVKPGASVKISCKVSGYTFTDHTI HWMKQRPEQGLEWIGYIYPRDAITKYNEKFKGK ATLTADKSSSTAYMOLNSLTSEDSAVYFCARWG YYFGSTSYYFDFWGQGTTLTVSSASTKGPSVFPL APSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGT QTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPP CPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCV VVDVSHEDPEVKFNWYVDGVEVHNAKTKPREE QYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK ALPAPIEKTISKAKGQPREPQVYTLPPSREEMTK NQVSLTCLVKGFYPSDIAVEWESNGQPENNYKT TPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCS VMHEALHNHYTQKSLSLSPGGGSGGSSSELTQD PAVSVALGQTVRITCQGDSLRSYYASWYQQKPG QAPVLVIYGKANRPSGIPDRFSGSSSGNTASLTIT GAQAEDEADYYCNSRDSSGAHVVFGGGTKLTV LGGGGSGGGGSGGGGSGGGGSEVQLVQSGGGV ERPGGSLRLSCAASGFTFDDYGMSWVRQAPGK GLEWVSGINWSGGSTGYADSVKGRVTISRDNA KNSLYLOMNSLRAEDTAVYYCAKILGAGRGWY FDL WGKGTT VT VS S SEQ ID NO: 185 CDH17A6/TR2v5 (IgGlKO)HC QVQLQQSDAELVKPGASVKISCKVSGYTFTDHTI HWMKQRPEQGLEWIGYIYPRDAITKYNEKFKGK ATLTADKSSSTAYMOLNSLTSEDSAVYFCARWG YYFGSTSYYFDFWGQGTTLTVSSASTKGPSVFPL APSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGT QTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPP CPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCV VVDVSHEDPEVKFNWYVDGVEVHNAKTKPREE QYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK ALPAPIEKTISKAKGOPREPQVYTLPPSREEMTK

-82 041360

NQVSLTCLVKGFYPSDIAVEWESNGQPENNYKT TPPVLDSDGSFFLYSKLTVDKSRWOOGNVFSCS VMHEALHNHYTQKSLSLSPGGGSGGSSSELTQD PAVSVALGQTVRITCQGDSLRSYYASWYQQKPG QAPVLVIYGKNNRPSGIPDRFSGSSSGNTASLTIT GAQAEDEADYYCNSRDSSGNHVVFGGGTKLTV LGGGGSGGGGSGGGGSGGGGSEVQLVQSGGGV ERPGGSLRLSCAASGFTFDDYGMSWVRQAPGK GLEWVSGINWSGGSTGYADSVKGRVTISRDNA KNSLYLQMNSLRAEDTAVYYCAKILGAGRGWY FDL WGKGTT VT VS S SEQ ID NO: 186 CDH17A6/TR2v6 (IgGlKO)HC QVQLQQSDAELVKPGASVKISCKVSGYTFTDHTI HWMKQRPEQGLEWIGYIYPRDAITKYNEKFKGK ATLTADKSSSTAYMOLNSLTSEDSAVYFCARWG YYFGSTSYYFDFWGQGTTLTVSSASTKGPSVFPL APSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGT OTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPP CPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCV VVDVSHEDPEVKFNWYVDGVEVHNAKTKPREE QYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK ALPAPIEKTISKAKGQPREPQVYTLPPSREEMTK NQVSLTCLVKGFYPSDIAVEWESNGQPENNYKT TPPVLDSDGSFFLYSKLTVDKSRWOOGNVFSCS VMHEALHNHYTQKSLSLSPGGGSGGSSSELTQD PAVSVALGQTVRITCQGDSLRSYYASWYQQKPG QAPVLVIYGKTNRPSGIPDRFSGSSSGNTASLTIT GAQAEDEADYYCNSRDSSGDHVVFGGGTKLTV LGGGGSGGGGSGGGGSGGGGSEVQLVQSGGGV ERPGGSLRLSCAASGFTFDDYGMSWVRQAPGK GLEWVSGINWAGGSTGYAD S VKGRVTISRDNA KNSLYLOMNSLRAEDTAVYYCAKILGAGRGWY FDL WGKGTT VT VS S SEQ ID NO: 187 CDH17A6/TR2v7 (IgGlKO)HC QVQLQQSDAELVKPGASVKISCKVSGYTFTDHTI HWMKQRPEQGLEWIGYIYPRDAITKYNEKFKGK ATLTADKSSSTAYMOLNSLTSEDSAVYFCARWG YYFGSTSYYFDFWGQGTTLTVSSASTKGPSVFPL APSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGT OTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPP CPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCV VVDVSHEDPEVKFNWYVDGVEVHNAKTKPREE QYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK ALPAPIEKTISKAKGQPREPQWTLPPSREEMTK NQVSLTCLVKGFYPSDIAVEWESNGQPENNYKT TPPVLDSDGSFFLYSKLTVDKSRWOOGNVFSCS VMHEALHNHYTQKSLSLSPGGGSGGSSSELTQD PAVSVALGQTVRITCQGDSLRSYYASWYQQKPG QAPVLVIYGKDNRPSGIPDRFSGSSSGNTASLTIT GAQAEDEADYYCNSRDSSGDHVVFGGGTKLTV LGGGGSGGGGSGGGGSGGGGSEVQLVQSGGGV ERPGGSLRLSCAASGFTFDDYGMSWVRQAPGK GLEWVSGINWTGGSTGYADSVKGRVTISRDNA KNSLYLQMNSLRAEDTAVYYCAKILGAGRGWY FDL WGKGTT VT VS S SEQ ID NO: 188 CDH17A6 DIVMTQSQKFMSTTVGDRVSITCKASQNVGTGV

- 83 041360

LC VWYQQRPGQSPKLLIYSPSNRYTGVPDRFTGSG SGTDFTLTITNIQSEDLADYFCQQYSTYPWTFGG GTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCL LNNFYPREAKVQWKVDNALQSGNSQESVTEQD SKDSTYSLSSTLTLSKADYEKHKVYACEVTHQG LSSPVTKSFNRGEC SEQ ID NO: 189 CDH17E2/TR2#1 (IgGlKO)HC QVQLQQSDAELVKPGASVKISCKVSGYTFTDHTI HWMKQRPEQGLEWIGYIYPRDVITQYNEKFKGK ATLTADKSSSTAYMQLNSLTSEDSAVYLCARW GYFYGSRSYYFDYWGQGTTLTVSSASTKGPSVF PLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN SGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSL GTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTC PPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTC VVVDVSHEDPEVKFNWYVDGVEVHNAKTKPRE EQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSN KALPAPIEKTISKAKGQPREPQVYTLPPSREEMT KNQVSLTCLVKGFYPSDIAVEWESNGQPENNYK TTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSC SVMHEALHNHYTQKSLSLSPGGGSGGSDIVMTQ SHKFMSTSVGDRVSITCKASQDVITAVAWYQQK PGQSPKLLIYWASTRHTGVPDRFTGTGSGTDYT LTISSVQAEDLALYYCQQHYSTPWTFGGGTKLEI KGGGGSGGGGSGGGGSGGGGSQVQLQQSGAEL VKPGASVKLSCKAFDYTFTSYDINWVKQRPGQG LEWIGWIDPGSGNTKYNEKFKGKATLTADKSSS TAYMHLSSLTSEDSAVYFCARKNYGGSYAFTY WGQGTLVTVA SEQ ID NO: 190 CDH17E2/TR2#2 (IgGlKO)HC QVQLQQSDAELVKPGASVKISCKVSGYTFTDHTI HWMKQRPEQGLEWIGYIYPRDVITQYNEKFKGK ATLTADKSSSTAYMQLNSLTSEDSAVYLCARW GYFYGSRSYYFDYWGQGTTLTVSSASTKGPSVF PLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN SGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSL GTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTC PPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTC VVVDVSHEDPEVKFNWYVDGVEVHNAKTKPRE EQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSN KALPAPIEKTISKAKGQPREPQVYTLPPSREEMT KNQVSLTCLVKGFYPSDIAVEWESNGQPENNYK TTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSC SVMHEALHNHYTQKSLSLSPGGGSGGSDIVMTQ SHKFMSTSVGDRVSITCKASQDVSTAVAWYQQ KPGQSPKVLIYWASTRHTGVPDRFTGSGSGTDYI LTISSVQAEDLALYYCQQHYSTPYTFGGGTKLEI KGGGGSGGGGSGGGGSGGGGSKVQLQQSGAEL VKTGTSVKLSCKASGYTFTEYIIHWVKQRSGQG LEWIGWFYPGSGSIKYNEKFKDKATLTADKSSS TVYMELSRLTSEDSAVYFCARHEEGGYSAWFPY WGQGTLVTVSA SEQIDNO:191 CDH17E2/TR2#3 (IgGlKO)HC QVQLQQSDAELVKPGASVKISCKVSGYTFTDHTI HWMKQRPEQGLEWIGYIYPRDVITQYNEKFKGK ATLTADKSSSTAYMQLNSLTSEDSAVYLCARW GYFYGSRSYYFDYWGQGTTLTVSSASTKGPSVF PLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN SGALTSGVHTVTSSVLQSSGSLLYSL

- 84 041360

GTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTC PPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTC VVVDVSHEDPEVKFNWYVDGVEVHNAKTKPRE EQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSN KALPAPIEKTISKAKGQPREPQVYTLPPSREEMT KNQVSLTCLVKGFYPSDIAVEWESNGQPENNYK TTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSC SVMHEALHNHYTOKSLSLSPGGGSGGSDVLMT QTPLSLPVSLGDQASISCRSSQSIVHSNGDTYLD WYLQKPGQSPKLLIYKVSNRFSGVPDRFSGSGS GTDFTLKISRWAEDLGVYYCFRGSHIPPTFGAG TKLEKGGGGSGGGGSGGGGSGGGGSEVQLQQS GPELVKPGASVKISCKASGYTFTDYYYLNWVKQ SHGKSLEWIGYIYPNNGDTSYNQKFKGKTTLTV DKSSSTAYMEFRSLTSEDSAVYYCTRGSNWIWY FD VWGTGTT VT VS S SEQ ID NO: 192 CDH17E2/TR2#7 (IgGlKO)HC QVQLQQSDAELVKPGASVKISCKVSGYTFTDHTI HWMKQRPEQGLEWIGYIYPRDVITQYNEKFKGK ATLTADKSSSTAYMQLNSLTSEDSAVYLCARW GYFYGSRSYYFDYWGQGTTLTVSSASTKGPSVF PLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN SGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSL GTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTC PPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTC VVVDVSHEDPEVKFNWYVDGVEVHNAKTKPRE EQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSN KALPAPIEKTISKAKGQPREPQVYTLPPSREEMT KNQVSLTCLVKGFYPSDIAVEWESNGQPENNYK TTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSC SVMHEALHNHYTQKSLSLSPGGGSGGSSIVMTQ TPKFLPVSAGDRVTMTCKASQNVGNNVAWYQ QKPGQSPKLLISYASNRYTGVPDRFTGSGSGTDF FTISSVQVEDLAVYFCQQHYSSPLTFGAGTKLEL KGGGGSGGGGSGGGGSGGGGSQVQLKQSGAEL VKPGASVKISCKASGYSFTDYYINWVKQRPGQG LEWIGKIGPGSGNTYYNEKFEGKATLTADKSSST AYMQLSSLTSEDSAVYFCASTGPFAYWGOGTLV TWSA SEQ ID NO: 193 CDH17E2/TR2#8 (IgGlKO)HC QVQLQQSDAELVKPGASVKISCKVSGYTFTDHTI HWMKQRPEQGLEWIGYIYPRDVITQYNEKFKGK ATLTADKSSSTAYMQLNSLTSEDSAVYLCARW GYFYGSRSYYFDYWGQGTTLTVSSASTKGPSVF PLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN SGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSL GTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTC PPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTC VVVDVSHEDPEVKFNWYVDGVEVHNAKTKPRE EQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSN KALPAPIEKTISKAKGQPREPQVYTLPPSREEMT KNQVSLTCLVKGFYPSDIAVEWESNGQPENNYK TTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSC SVMHEALHNHYTQKSLSLSPGGGSGGSDIVMTQ SHKFMSTSVGDRVSITCKASQDVGTAVAWYQQ KPGQSPKLLIYWASTRHTGVPDRFTGSGSGTDFT LTISNVQSEDLADYFCQQYSSYPYTFGGGTKLEI KGGGGSGGGGSGGGGSGGGGSQVTLKESGPGIL QPSQTLSLTCSFSGFSLSTSGMGVSWIRQPSGKG

- 85 041360

LEWLAHIYWDDDKRYNPSLKSRLTISKDTSRNQ VFLKITSVDTADTATYYCARRRLGPFAYWGQGT LVTVSA SEQ ID NO: 194 CDH17E2/TR2#10 (IgGlKO)HC QVQLQQSDAELVKPGASVKISCKVSGYTFTDHTI HWMKQRPEQGLEWIGYIYPRDVITQYNEKFKGK ATLTADKSSSTAYMOLNSLTSEDSAVYLCARW GYFYGSRSYYFDYWGQGTTLTVSSASTKGPSVF PLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN SGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSL GTOTYICNVNHKPSNTKVDKRVEPKSCDKTHTC PPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTC VVVDVSHEDPEVKFNWYVDGVEVHNAKTKPRE EQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSN KALPAPIEKTISKAKGOPREPQVYTLPPSREEMT KNQVSLTCLVKGFYPSDIAVEWESNGQPENNYK TTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSC SVMHEALHNHYTQKSLSLSPGGGSGGSDVLMT QTPLSLPVSLGDQASISCRSSQNIVHSNGNTYLD WYLQKPGQSPKLLIYKVSNRFSGVPDRFSGSGS GTDFTLKISRVEAEDLGVYYCFQGSHVPPTFGA GTKLELKGGGGSGGGGSGGGGSGGGGSEVQLQ QSGPELVKPGASVKISCKASGYTFTDYYYMNW VKQSHGKSLEWIGYIYPNNGGTRYNQKFKGKA TLTVDKSSSTAYMELRSLTSEDSAVYYCARGGN WNWYFDVWGTGTTVTVSS SEQ ID NO: 195 CDH17E2/TR2#12 (IgGlKO)HC QVQLQQSDAELVKPGASVKISCKVSGYTFTDHTI HWMKQRPEQGLEWIGYIYPRDVITQYNEKFKGK ATLTADKSSSTAYMQLNSLTSEDSAVYLCARW GYFYGSRSYYFDYWGQGTTLTVSSASTKGPSVF PLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN SGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSL GTOTYICNVNHKPSNTKVDKRVEPKSCDKTHTC PPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTC VVVDVSHEDPEVKFNWYVDGVEVHNAKTKPRE EQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSN KALPAPIEKTISKAKGOPREPQVYTLPPSREEMT KNQVSLTCLVKGFYPSDIAVEWESNGQPENNYK TTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSC SVMHEALHNHYTQKSLSLSPGGGSGGSDIVMTQ SHKFMSTSVGDRVSITCKASQDVSNAVAWYQQ KPGQSPKLLIYWASTRHNGVPDRFTGSGSGTDYI LTISSVQAEDLALYYCQQHYRTPYTFGGGTKLEI KGGGGSGGGGSGGGGSGGGGSKVQLQQSGAEL VKPGASVKLSCKASGYTFTEYIIHWVKQKSGQG LEWIGWFYPGSGNIKYNEKFKDKATLTADKSSS TVYMELSRLTSEDSAVYFCTRHEGGSNFFPYWG QGTLVTVSA SEQ ID NO: 196 CDH17E2/TR2vl (IgGlKO)HC QVQLQQSDAELVKPGASVKISCKVSGYTFTDHTI HWMKQRPEQGLEWIGYIYPRDVITQYNEKFKGK ATLTADKSSSTAYMOLNSLTSEDSAVYLCARW GYFYGSRSYYFDYWGQGTTLTVSSASTKGPSVF PLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN SGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSL GTOTYICNVNHKPSNTKVDKRVEPKSCDKTHTC PPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTC VVVDVSHEDPEVKFNWYVDGVEVHNAKTKPRE

-86041360

EQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSN KALPAPIEKTISKAKGQPREPQVYTLPPSREEMT KNQVSLTCLVKGFYPSDIAVEWESNGQPENNYK TTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSC SVMHEALHNHYTQKSLSLSPGGGSGGSDIELTQ DPAVSVALGQTVRITCQGDSLRSYYASWYQQKP GQAPVLVIYGKNNRPSGIPDRFSGSSSGNTASLTI TGAQAEDEADYYCNSRDSSGNHVVFGCGTKLT VLGGGGSGGGGSGGGGSGGGGSEVQLVQSGGG VERPGGSLRLSCAASGFTFDDYGMSWVRQAPG KCLEWVSGINWNGGSTGYADSVKGRVTISRDN AKNSLYLOMNSLRAEDTAVYYCAKILGAGRGW YFDL WGKGTT VTVS S SEQ ID NO: 197 CDH17E2/TR2v2 (IgGlKO)HC QVQLQQSDAELVKPGASVKISCKVSGYTFTDHTI HWMKQRPEQGLEWIGYIYPRDVITQYNEKFKGK ATLTADKSSSTAYMOLNSLTSEDSAVYLCARW GYFYGSRSYYFDYWGQGTTLTVSSASTKGPSVF PLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN SGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSL GTOTYICNVNHKPSNTKVDKRVEPKSCDKTHTC PPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTC VVVDVSHEDPEVKFNWYVDGVEVHNAKTKPRE EQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSN KALPAPIEKTISKAKGQPREPQVYTLPPSREEMT KNQVSLTCLVKGFYPSDIAVEWESNGQPENNYK TTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSC SVMHEALHNHYTQKSLSLSPGGGSGGSSSELTQ DPAVSVALGQTVRITCQGDSLRSYYASWYQQKP GQAPVLVIYGKNNRPSGIPDRFSGSSSGNTASLTI TGAQAEDEADYYCNSRDSSGNHVVFGGGTKLT VLGGGGSGGGGSGGGGSGGGGSEVQLVQSGGG VERPGGSLRLSCAASGFTFDDYGMSWVRQAPG KGLEWVSGINWNGGSTGYADSVKGRVTISRDN AKNSLYLOMNSLRAEDTAVYYCAKILGAGRGW YFDL WGKGTT VTVS S SEQ ID NO: 198 CDH17E2/TR2v3 HC (IgGlKO) QVQLQQSDAELVKPGASVKISCKVSGYTFTDHTI HWMKQRPEQGLEWIGYIYPRDVITQYNEKFKGK ATLTADKSSSTAYMQLNSLTSEDSAVYLCARW GYFYGSRSYYFDYWGQGTTLTVSSASTKGPSVF PLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN SGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSL GTOTYICNVNHKPSNTKVDKRVEPKSCDKTHTC PPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTC VVVDVSHEDPEVKFNWYVDGVEVHNAKTKPRE EQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSN KALPAPIEKTISKAKGOPREPQVYTLPPSREEMT KNQVSLTCLVKGFYPSDIAVEWESNGQPENNYK TTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSC SVMHEALHNHYTQKSLSLSPGGGSGGSSSELTQ DPAVSVALGQTVRITCQGDSLRSYYASWYQQKP GQAPVLVIYGKTNRPSGIPDRFSGSSSGNTASLTI TGAQAEDEADYYCNSRDSSGTHVVFGGGTKLT VLGGGGSGGGGSGGGGSGGGGSEVQLVQSGGG VERPGGSLRLSCAASGFTFDDYGMSWVRQAPG KGLEWVSGINWSGGSTGYADSVKGRVTISRDN AKNSLYLOMNSLRAEDTAVYYCAKILGAGRGW YFDL WGKGTT VTVS S

- 87 041360

SEQ ID NO: 199 CDH17E2/TR2v4 HC QVQLQQSDAELVKPGASVKISCKVSGYTFTDHTI HWMKQRPEQGLEWIGYIYPRDVITQYNEKFKGK ATLTADKSSSTAYMQLNSLTSEDSAVYLCARW GYFYGSRSYYFDYWGQGTTLTVSSASTKGPSVF PLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN SGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSL GTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTC PPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTC VVVDVSHEDPEVKFNWYVDGVEVHNAKTKPRE EQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSN KALPAPIEKTISKAKGQPREPQVYTLPPSREEMT KNQVSLTCLVKGFYPSDIAVEWESNGQPENNYK TTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSC SVMHEALHNHYTQKSLSLSPGGGSGGSSSELTQ DPAVSVALGQTVRITCQGDSLRSYYASWYQQKP GQAPVLVIYGKANRPSGIPDRFSGSSSGNTASLTI TGAQAEDEADYYCNSRDSSGAHVVFGGGTKLT VLGGGGSGGGGSGGGGSGGGGSEVQLVQSGGG VERPGGSLRLSCAASGFTFDDYGMSWVRQAPG KGLEWVSGINWSGGSTGYADSVKGRVTISRDN AKNSLYLQMNSLRAEDTAVYYCAKILGAGRGW YFDL WGKGTT VTVS S SEQ ID N0:200 CDH17E2/TR2v5 (IgGlKO)HC QVQLQQSDAELVKPGASVKISCKVSGYTFTDHTI HWMKQRPEQGLEWIGYIYPRDVITQYNEKFKGK ATLTADKSSSTAYMQLNSLTSEDSAVYLCARW GYFYGSRSYYFDYWGQGTTLTVSSASTKGPSVF PLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN SGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSL GTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTC PPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTC VVVDVSHEDPEVKFNWYVDGVEVHNAKTKPRE EQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSN KALPAPIEKTISKAKGQPREPQVYTLPPSREEMT KNQVSLTCLVKGFYPSDIAVEWESNGQPENNYK TTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSC SVMHEALHNHYTQKSLSLSPGGGSGGSSSELTQ DPAVSVALGQTVRITCQGDSLRSYYASWYQQKP GQAPVLVIYGKNNRPSGIPDRFSGSSSGNTASLTI TGAQAEDEADYYCNSRDSSGNHVVFGGGTKLT VLGGGGSGGGGSGGGGSGGGGSEVQLVQSGGG VERPGGSLRLSCAASGFTFDDYGMSWVRQAPG KGLEWVSGINWSGGSTGYADSVKGRVTISRDN AKNSLYLQMNSLRAEDTAVYYCAKILGAGRGW YFDLWGKGTTVTVSS SEQIDNO:201 CDH17E2/TR2v6 (IgGlKO)HC QVQLQQSDAELVKPGASVKISCKVSGYTFTDHTI HWMKQRPEQGLEWIGYIYPRDVITQYNEKFKGK ATLTADKSSSTAYMQLNSLTSEDSAVYLCARW GYFYGSRSYYFDYWGQGTTLTVSSASTKGPSVF PLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN SGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSL GTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTC PPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTC VVVDVSHEDPEVKFNWYVDGVEVHNAKTKPRE EQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSN KALPAPIEKTISKAKGQPREPQVYTLPPSREEMT KNQVSLTCLVKGFYPSDIAVEWESNGQPENNYK

- 88 041360

TTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSC SVMHEALHNHYTQKSLSLSPGGGSGGSSSELTQ DPAVSVALGQTVRITCQGDSLRSYYASWYQQKP GQAPVLVIYGKTNRPSGIPDRFSGSSSGNTASLTI TGAQAEDEADYYCNSRDSSGDHVVFGGGTKLT VLGGGGSGGGGSGGGGSGGGGSEVQLVQSGGG VERPGGSLRLSCAASGFTFDDYGMSWVRQAPG KGLEWVSGINWAGGSTGYADSVKGRVTISRDN AKNSLYLQMNSLRAEDTAVYYCAKILGAGRGW YFDL WGKGTT VTVS S SEQ ID NO:202 CDH17E2/TR2v7 (IgGlKO)HC QVQLQQSDAELVKPGASVKISCKVSGYTFTDHTI HWMKQRPEQGLEWIGYIYPRDVITQYNEKFKGK ATLTADKSSSTAYMQLNSLTSEDSAVYLCARW GYFYGSRSYYFDYWGQGTTLTVSSASTKGPSVF PLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN SGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSL GTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTC PPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTC VVVDVSHEDPEVKFNWYVDGVEVHNAKTKPRE EQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSN KALPAPIEKTISKAKGQPREPQVYTLPPSREEMT KNQVSLTCLVKGFYPSDIAVEWESNGQPENNYK TTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSC SVMHEALHNHYTQKSLSLSPGGGSGGSSSELTQ DPAVSVALGQTVRITCQGDSLRSYYASWYQQKP GQAPVLVIYGKDNRPSGIPDRFSGSSSGNTASLTI TGAQAEDEADYYCNSRDSSGDHVVFGGGTKLT VLGGGGSGGGGSGGGGSGGGGSEVQLVQSGGG VERPGGSLRLSCAASGFTFDDYGMSWVRQAPG KGLEWVSGINWTGGSTGYADSVKGRVTISRDN AKNSLYLOMNSLRAEDTAVYYCAKILGAGRGW YFDL WGKGTT VTVS S SEQ ID NO:203 CDH17E2 LC DIVMTQSOKFMSTTVGDRVNITCKASONVGTAV AWYQQKPGQSPKLLIYSASNRYTGVPDRFTGSG SGTDFTLTITNMQSKDLADYFCQQYSSYPWTFG GGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVV CLLNNFYPREAKVQWKVDNALQSGNSQESVTE ODSKDSSSTYSLSSTLTLECSSVGLSFETKHKVY SEQ ID NO:204 CDH17H2/TR2#1 (IgGlKO)HC QVQLVQSGAEVKKPGSSVKVSCKVSGYTFTDHT IHWMKQRPGQGLEWIGYIYPRDVITQYNEKFKG KVTLTADKSTSTAYMELSSLRSEDTAVYLCARW GYFYGSRSYYFDYWGQGTTVTVSSASTKGPSVF PLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN SGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSL GTOTYICNVNHKPSNTKVDKRVEPKSCDKTHTC PPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTC VVVDVSHEDPEVKFNWYVDGVEVHNAKTKPRE EQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSN KALPAPIEKTISKAKGQPREPQWTLPPSREEMT KNQVSLTCLVKGFYPSDIAVEWESNGQPENNYK TTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSC SVMHEALHNHYTQKSLSLSPGGGSGGSDIVMTQ SHKFMSTSVGDRVSITCKASQDVITAVAWYQQK PGQSPKLLIYWASTRHTGVPDRFTGTGSGTDYT LTISSVQAEDLALYYCQQHYSTPWTFGGGTKLEI

-89 041360

KGGGGSGGGGSGGGGSGGGGSQVQLQQSGAEL VKPGASVKLSCKAFDYTFTSYDINWVKQRPGQG LEWIGWIDPGSGNTKYNEKFKGKATLTADKSSS TAYMHLSSLTSEDSAVYFCARKNYGGSYAFTY WGQGTLVTVA SEQ ID NO:205 CDH17H2/TR2#2 (IgGlKO)HC QVQLVQSGAEVKKPGSSVKVSCKVSGYTFTDHT IHWMKQRPGQGLEWIGYIYPRDVITQYNEKFKG KVTLTADKSTSTAYMELSSLRSEDTAVYLCARW GYFYGSRSYYFDYWGQGTTVTVSSASTKGPSVF PLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN SGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSL GTOTYICNVNHKPSNTKVDKRVEPKSCDKTHTC PPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTC VVVDVSHEDPEVKFNWYVDGVEVHNAKTKPRE EQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSN KALPAPIEKTISKAKGOPREPQVYTLPPSREEMT KNQVSLTCL VKGFYPSDIA VEWESNGQPENNYK TTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSC SVMHEALHNHYTQKSLSLSPGGGSGGSDIVMTQ SHKFMSTSVGDRVSITCKASQDVSTAVAWYQQ KPGQSPKVLIYWASTRHTGVPDRFTGSGSGTDYI LTISSVQAEDLALYYCQQHYSTPYTFGGGTKLEI KGGGGSGGGGSGGGGSGGGGSKVQLQQSGAEL VKTGTSVKLSCKASGYTFTEYIIHWVKQRSGQG LEWIGWFYPGSGSIKYNEKFKDKATLTADKSSS TVYMELSRLTSEDSAVYFCARHEEGGYSAWFPY WGQGTLVTVSA SEQ ID NO:206 CDH17H2/TR2#3 (IgGlKO)HC QVQLVQSGAEVKKPGSSVKVSCKVSGYTFTDHT IHWMKQRPGQGLEWIGYIYPRDVITQYNEKFKG KVTLTADKSTSTAYMELSSLRSEDTAVYLCARW GYFYGSRSYYFDYWGQGTTVTVSSASTKGPSVF PLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN SGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSL GTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTC PPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTC VVVDVSHEDPEVKFNWYVDGVEVHNAKTKPRE EQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSN KALPAPIEKTISKAKGOPREPQVYTLPPSREEMT KNQVSLTCL VKGFYPSDIAVEWESNGQPENNYK TTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSC SVMHEALHNHYTQKSLSLSPGGGSGGSDVLMT QTPLSLPVSLGDQASISCRSSQSIVHSNGDTYLD WYLQKPGQSPKLLIYKVSNRFSGVPDRFSGSGS GTDFTLKISRWAEDLGVYYCFRGSHIPPTFGAG TKLEKGGGGSGGGGSGGGGSGGGGSEVQLQQS GPELVKPGASVKISCKASGYTFTDYYYLNWVKQ SHGKSLEWIGYIYPNNGDTSYNQKFKGKTTLTV DKSSSTAYMEFRSLTSEDSAVYYCTRGSNWIWY FDVWGTGTTVTVSS SEQ ID NO:207 CDH17H2/TR2#7 (IgGlKO)HC QVQLVQSGAEVKKPGSSVKVSCKVSGYTFTDHT IHWMKQRPGQGLEWIGYIYPRDVITQYNEKFKG KVTLTADKSTSTAYMELSSLRSEDTAVYLCARW GYFYGSRSYYFDYWGQGTTVTVSSASTKGPSVF PLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN SGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSL GTOTYICNVNHKPSNTKVDKRVEPKSCDKTHTC

- 90 041360

PPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTC VVVDVSHEDPEVKFNWYVDGVEVHNAKTKPRE EQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSN KALPAPIEKTISKAKGQPREPQVYTLPPSREEMT KNQVSLTCLVKGFYPSDIAVEWESNGQPENNYK TTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSC SVMHEALHNHYTQKSLSLSPGGGSGGSSIVMTQ TPKFLPVSAGDRVTMTCKASQNVGNNVAWYQ QKPGQSPKLLISYASNRYTGVPDRFTGSGSGTDF FTISSVQVEDLAVYFCQQHYSSPLTFGAGTKLEL KGGGGSGGGGSGGGGSGGGGSQVQLKQSGAEL VKPGASVKISCKASGYSFTDYYINWVKQRPGQG LEWIGKIGPGSGNTYYNEKFEGKATLTADKSSST AYMOLSSLTSEDSAVYFCASTGPFAYWGOGTLV TWSA SEQ ID NO:208 CDH17H2/TR2#8 (IgGlKO)HC QVQLVQSGAEVKKPGSSVKVSCKVSGYTFTDHT IHWMKQRPGQGLEWIGYIYPRDVITQYNEKFKG KVTLTADKSTSTAYMELSSLRSEDTAVYLCARW GYFYGSRSYYFDYWGQGTTVTVSSASTKGPSVF PLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN SGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSL GTOTYICNVNHKPSNTKVDKRVEPKSCDKTHTC PPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTC VVVDVSHEDPEVKFNWYVDGVEVHNAKTKPRE EQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSN KALPAPIEKTISKAKGOPREPQVYTLPPSREEMT KNQVSLTCLVKGFYPSDIAVEWESNGQPENNYK TTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSC SVMHEALHNHYTQKSLSLSPGGGSGGSDIVMTQ SHKFMSTSVGDRVSITCKASQDVGTAVAWYQQ KPGQSPKLLIYWASTRHTGVPDRFTGSGSGTDFT LTISNVQSEDLADYFCQQYSSYPYTFGGGTKLEI KGGGGSGGGGSGGGGSGGGGSQVTLKESGPGIL QPSQTLSLTCSFSGFSLSTSGMGVSWIRQPSGKG LEWLAHIYWDDDKRYNPSLKSRLTISKDTSRNQ VFLKITSVDTADTATYYCARRRLGPFAYWGQGT LVTVSA SEQ ID NO:209 CDH17H2/TR2#10 (IgGlKO)HC QVQLVQSGAEVKKPGSSVKVSCKVSGYTFTDHT IHWMKQRPGQGLEWIGYIYPRDVITQYNEKFKG KVTLTADKSTSTAYMELSSLRSEDTAVYLCARW GYFYGSRSYYFDYWGQGTTVTVSSASTKGPSVF PLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN SGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSL GTOTYICNVNHKPSNTKVDKRVEPKSCDKTHTC PPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTC VVVDVSHEDPEVKFNWYVDGVEVHNAKTKPRE EQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSN KALPAPIEKTISKAKGQPREPQVYTLPPSREEMT KNQVSLTCLVKGFYPSDIAVEWESNGQPENNYK TTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSC SVMHEALHNHYTQKSLSLSPGGGSGGSDVLMT QTPLSLPVSLGDQASISCRSSQNIVHSNGNTYLD WYLQKPGQSPKLLIYKVSNRFSGVPDRFSGSGS GTDFTLKISRVEAEDLGVYYCFQGSHVPPTFGA GTKLELKGGGGSGGGGSGGGGSGGGGSEVQLQ QSGPELVKPGASVKISCKASGYTFTDYYYMNW VKQSHGKSLEWIGYIYPNNGGTRYNQKFKGKA

- 91 041360

TLTVDKSSSTAYMELRSLTSEDSAVYYCARGGN WNWYFDVWGTGTTVTVSS SEQIDNO:210 CDH17H2/TR2#12 (IgGlKO)HC QVQLVQSGAEVKKPGSSVKVSCKVSGYTFTDHT IHWMKQRPGQGLEWIGYIYPRDVITQYNEKFKG KVTLTADKSTSTAYMELSSLRSEDTAVYLCARW GYFYGSRSYYFDYWGQGTTVTVSSASTKGPSVF PLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN SGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSL GTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTC PPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTC VVVDVSHEDPEVKFNWYVDGVEVHNAKTKPRE EQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSN KALPAPIEKTISKAKGQPREPQVYTLPPSREEMT KNQVSLTCLVKGFYPSDIAVEWESNGQPENNYK TTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSC SVMHEALHNHYTQKSLSLSPGGGSGGSDIVMTQ SHKFMSTSVGDRVSITCKASQDVSNAVAWYQQ KPGQSPKLLIYWASTRHNGVPDRFTGSGSGTDYI LTISSVQAEDLALYYCQQHYRTPYTFGGGTKLEI KGGGGSGGGGSGGGGSGGGGSKVQLQQSGAEL VKPGASVKLSCKASGYTFTEYIIHWVKQKSGQG LEWIGWFYPGSGNIKYNEKFKDKATLTADKSSS TVYMELSRLTSEDSAVYFCTRHEGGSNFFPYWG QGTLVTVSA SEQIDNO:211 CDH17H2/TR2vl HC QVQLVQSGAEVKKPGSSVKVSCKVSGYTFTDHT IHWMKQRPGQGLEWIGYIYPRDVITQYNEKFKG KVTLTADKSTSTAYMELSSLRSEDTAVYLCARW GYFYGSRSYYFDYWGQGTTVTVSSASTKGPSVF PLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN SGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSL GTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTC PPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTC VVVDVSHEDPEVKFNWYVDGVEVHNAKTKPRE EQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSN KALPAPIEKTISKAKGQPREPQVYTLPPSREEMT KNQVSLTCL VKGFYPSDIA VEWESNGQPENNYK TTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSC SVMHEALHNHYTQKSLSLSPGGGSGGSDIELTQ DPAVSVALGQTVRITCQGDSLRSYYASWYQQKP GQAPVLVIYGKNNRPSGIPDRFSGSSSGNTASLTI TGAQAEDEADYYCNSRDSSGNHVVFGCGTKLT VLGGGGSGGGGSGGGGSGGGGSEVQLVQSGGG VERPGGSLRLSCAASGFTFDDYGMSWVRQAPG KCLEWVSGINWNGGSTGYADSVKGRVTISRDN AKNSLYLQMNSLRAEDTAVYYCAKILGAGRGW YFDL WGKGTT VTVS S SEQIDNO:212 CDH17H2/TR2v2 (IgGlKO)HC QVQLVQSGAEVKKPGSSVKVSCKVSGYTFTDHT IHWMKQRPGQGLEWIGYIYPRDVITQYNEKFKG KVTLTADKSTSTAYMELSSLRSEDTAVYLCARW GYFYGSRSYYFDYWGQGTTVTVSSASTKGPSVF PLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN SGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSL GTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTC PPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTC VVVDVSHEDPEVKFNWYVDGVEVHNAKTKPRE EQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSN

-92 041360

KALPAPIEKTISKAKGQPREPQVYTLPPSREEMT KNQVSLTCLVKGFYPSDIAVEWESNGQPENNYK TTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSC SVMHEALHNHYTQKSLSLSPGGGSGGSSSELTQ DPAVSVALGQTVRITCQGDSLRSYYASWYQQKP GQAPVLVIYGKNNRPSGIPDRFSGSSSGNTASLTI TGAQAEDEADYYCNSRDSSGNHVVFGGGTKLT VLGGGGSGGGGSGGGGSGGGGSEVQLVQSGGG VERPGGSLRLSCAASGFTFDDYGMSWVROAPG KGLEWVSGINWNGGSTGYADSVKGRVTISRDN AKNSLYLQMNSLRAEDTAVYYCAKILGAGRGW YFDL WGKGTT VTVS S SEQIDNO:213 CDH17H2/TR2v3 (IgGlKO)HC QVQLVQSGAEVKKPGSSVKVSCKVSGYTFTDHT IHWMKQRPGQGLEWIGYIYPRDVITQYNEKFKG KVTLTADKSTSTAYMELSSLRSEDTAVYLCARW GYFYGSRSYYFDYWGQGTTVTVSSASTKGPSVF PLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN SGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSL GTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTC PPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTC VVVDVSHEDPEVKFNWYVDGVEVHNAKTKPRE EQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSN KALPAPIEKTISKAKGQPREPQVYTLPPSREEMT KNQVSLTCLVKGFYPSDIAVEWESNGQPENNYK TTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSC SVMHEALHNHYTQKSLSLSPGGGSGGSSSELTQ DPAVSVALGQTVRITCQGDSLRSYYASWYQQKP GQAPVLVIYGKTNRPSGIPDRFSGSSSGNTASLTI TGAQAEDEADYYCNSRDSSGTHVVFGGGTKLT VLGGGGSGGGGSGGGGSGGGGSEVQLVQSGGG VERPGGSLRLSCAASGFTFDDYGMSWVRQAPG KGLEWVSGINWSGGSTGYADSVKGRVTISRDN AKNSLYLQMNSLRAEDTAVYYCAKILGAGRGW YFDLWGKGTTVTVSS SEQIDNO:214 CDH17H2/TR2v4 (IgGlKO)HC QVQLVQSGAEVKKPGSSVKVSCKVSGYTFTDHT IHWMKQRPGQGLEWIGYIYPRDVITQYNEKFKG KVTLTADKSTSTAYMELSSLRSEDTAVYLCARW GYFYGSRSYYFDYWGQGTTVTVSSASTKGPSVF PLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN SGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSL GTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTC PPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTC VVVDVSHEDPEVKFNWYVDGVEVHNAKTKPRE EQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSN KALPAPIEKTISKAKGQPREPQVYTLPPSREEMT KNQVSLTCLVKGFYPSDIAVEWESNGQPENNYK TTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSC SVMHEALHNHYTQKSLSLSPGGGSGGSSSELTQ DPAVSVALGQTVRITCQGDSLRSYYASWYQQKP GQAPVLVIYGKANRPSGIPDRFSGSSSGNTASLTI TGAQAEDEADYYCNSRDSSGAHVVFGGGTKLT VLGGGGSGGGGSGGGGSGGGGSEVQLVQSGGG VERPGGSLRLSCAASGFTFDDYGMSWVRQAPG KGLEWVSGINWSGGSTGYADSVKGRVTISRDN AKNSLYLQMNSLRAEDTAVYYCAKILGAGRGW YFDL WGKGTT VTVS S

- 93 041360

SEQIDNO:215 CDH17H2/TR2v5 (IgGlKO)HC QVQLVQSGAEVKKPGSSVKVSCKVSGYTFTDHT IHWMKQRPGQGLEWIGYIYPRDVITQYNEKFKG KVTLTADKSTSTAYMELSSLRSEDTAVYLCARW GYFYGSRSYYFDYWGQGTTVTVSSASTKGPSVF PLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN SGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSL GTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTC PPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTC VVVDVSHEDPEVKFNWYVDGVEVHNAKTKPRE EQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSN KALPAPIEKTISKAKGQPREPQVYTLPPSREEMT KNQVSLTCLVKGFYPSDIAVEWESNGQPENNYK TTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSC SVMHEALHNHYTQKSLSLSPGGGSGGSSSELTQ DPAVSVALGQTVRITCQGDSLRSYYASWYQQKP GQAPVLVIYGKNNRPSGIPDRFSGSSSGNTASLTI TGAQAEDEADYYCNSRDSSGNHVVFGGGTKLT VLGGGGSGGGGSGGGGSGGGGSEVQLVQSGGG VERPGGSLRLSCAASGFTFDDYGMSWVRQAPG KGLEWVSGINWSGGSTGYADSVKGRVTISRDN AKNSLYLQMNSLRAEDTAVYYCAKILGAGRGW YFDL WGKGTT VTVS S SEQIDNO:216 CDH17H2/TR2v6 (IgGlKO)HC QVQLVQSGAEVKKPGSSVKVSCKVSGYTFTDHT IHWMKQRPGQGLEWIGYIYPRDVITQYNEKFKG KVTLTADKSTSTAYMELSSLRSEDTAVYLCARW GYFYGSRSYYFDYWGQGTTVTVSSASTKGPSVF PLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN SGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSL GTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTC PPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTC VVVDVSHEDPEVKFNWYVDGVEVHNAKTKPRE EQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSN KALPAPIEKTISKAKGQPREPQVYTLPPSREEMT KNQVSLTCLVKGFYPSDIAVEWESNGQPENNYK TTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSC SVMHEALHNHYTQKSLSLSPGGGSGGSSSELTQ DPAVSVALGQTVRITCQGDSLRSYYASWYQQKP GQAPVLVIYGKTNRPSGIPDRFSGSSSGNTASLTI TGAQAEDEADYYCNSRDSSGDHVVFGGGTKLT VLGGGGSGGGGSGGGGSGGGGSEVQLVQSGGG VERPGGSLRLSCAASGFTFDDYGMSWVRQAPG KGLEWVSGINWAGGSTGYADSVKGRVTISRDN AKNSLYLQMNSLRAEDTAVYYCAKILGAGRGW YFDL WGKGTT VTVS S SEQIDNO:217 CDH17H2/TR2v7 (IgGlKO)HC QVQLVQSGAEVKKPGSSVKVSCKVSGYTFTDHT IHWMKQRPGQGLEWIGYIYPRDVITQYNEKFKG KVTLTADKSTSTAYMELSSLRSEDTAVYLCARW GYFYGSRSYYFDYWGQGTTVTVSSASTKGPSVF PLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN SGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSL GTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTC PPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTC VVVDVSHEDPEVKFNWYVDGVEVHNAKTKPRE EQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSN KALPAPIEKTISKAKGQPREPQVYTLPPSREEMT KNQVSLTCLVKGFYPSDIAVEWESNGQPENNYK

- 94 041360

TTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSC SVMHEALHNHYTQKSLSLSPGGGSGGSSSELTQ DPAVSVALGQTVRITCQGDSLRSYYASWYQQKP GQAPVLVIYGKDNRPSGIPDRFSGSSSGNTASLTI TGAQAEDEADYYCNSRDSSGDHVVFGGGTKLT VLGGGGSGGGGSGGGGSGGGGSEVQLVQSGGG VERPGGSLRLSCAASGFTFDDYGMSWVRQAPG KGLEWVSGINWTGGSTGYADSVKGRVTISRDN AKNSLYLQMNSLRAEDTAVYYCAKILGAGRGW YFDL WGKGTT VTVS S SEQIDNO:218 CDH17H2 LC DIQMTQSQSSLSASVGDRVGITCKASQNVGTAV AWYQQKPGKAPKLLIYSASNRYTGVPSRFSGSG SGTDFTLTISSLQPKDFATYFCQQYSSYPWTFGQ GTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCL LNNFYPREAKVQWKVDNALQSGNSQESVTERGQD SKDSVTYSLSSTKSFTHNSKADY SEQIDNO:219 CDH17vl/TR2#l (IgGl KO) HC EVQLQQSVAELVKPGASVKMSCKVSGYTLTDH TIHWMKQRPEQGLEWIGYIYPRDGITGYNEKFK GKATLTADTSSSTAYMOLNSLTSEDSAVYFCAR WGYSYRNYAYYYDYWGQGTTLTVSSASTKGPS VFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVS WNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSS SLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTH TCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEV TCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYKCKV SNKALPAPIEKTISKAKGOPREPQVYTLPPSREE MTKNQVSLTCLVKGFYPSDIAVEWESNGQPENN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVF SCSVMHEALHNHYTQKSLSLSPGGGSGGSDIVM TQSHKFMSTSVGDRVSITCKASQDVITAVAWYQ QKPGQSPKLLIYWASTRHTGVPDRFTGTGSGTD YTLTISSVQAEDLALYYCQQHYSTPWTFGGGTK LEIKGGGGSGGGGSGGGGSGGGGSQVQLQQSG AELVKPGASVKLSCKAFDYTFTSYDINWVKQRP GQGLEWIGWIDPGSGNTKYNEKFKGKATLTAD KSSSTAYMHLSSLTSEDSAVYFCARKNYGGSYA FTYWGQGTLVTVA SEQ ID NO:220 CDH17vl/TR2#2 (IgGlKO)HC EVQLQQSVAELVKPGASVKMSCKVSGYTLTDH TIHWMKQRPEQGLEWIGYIYPRDGITGYNEKFK GKATLTADTSSSTAYMQLNSLTSEDSAVYFCAR WGYSYRNYAYYYDYWGQGTTLTVSSASTKGPS VFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVS WNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSS SLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTH TCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEV TCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYKCKV SNKALPAPIEKTISKAKGOPREPQVYTLPPSREE MTKNQVSLTCLVKGFYPSDIAVEWESNGQPENN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVF SCSVMHEALHNHYTQKSLSLSPGGGSGGSDIVM TQSHKFMSTSVGDRVSITCKASQDVSTAVAWY QQKPGQSPKVLIYWASTRHTGVPDRFTGSGSGT DYILTISSVQAEDLALYYCQQHYSTPYTFGGGTK

- 95 041360

LEIKGGGGSGGGGSGGGGSGGGGSKVQLQQSG AELVKTGTSVKLSCKASGYTFTEYIIHWVKQRS GQGLEWIGWFYPGSGSIKYNEKFKDKATLTADK SSSTVYMELSRLTSEDSAVYFCARHEEGGYSAW FPYWGQGTLVTVSA SEQIDNO:221 CDH17vl/TR2#3 (IgGlKO)HC EVQLQQSVAELVKPGASVKMSCKVSGYTLTDH TIHWMKQRPEQGLEWIGYIYPRDGITGYNEKFK GKATLTADTSSSTAYMOLNSLTSEDSAVYFCAR WGYSYRNYAYYYDYWGQGTTLTVSSASTKGPS VFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVS WNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSS SLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTH TCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEV TCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYKCKV SNKALPAPIEKTISKAKGOPREPQVYTLPPSREE MTKNQVSLTCLVKGFYPSDIAVEWESNGQPENN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVF SCSVMHEALHNHYTQKSLSLSPGGGSGGSDVL MTQTPLSLPVSLGDQASISCRSSQSIVHSNGDTY LDWYLQKPGQSPKLLIYKVSNRFSGVPDRFSGS GSGTDFTLKISRVVAEDLGVYYCFRGSHIPPTFG AGTKLEKGGGGSGGGGSGGGGSGGGGSEVQLQ QSGPELVKPGASVKISCKASGYTFTDYYYLNWV KQSHGKSLEWIGYIYPNNGDTSYNQKFKGKTTL TVDKSSSTAYMEFRSLTSEDSAVYYCTRGSNWI WYFDVWGTGTTVTVSS SEQ ID NO:222 CDH17vl/TR2#7 (IgGlKO)HC EVQLQQSVAELVKPGASVKMSCKVSGYTLTDH TIHWMKQRPEQGLEWIGYIYPRDGITGYNEKFK GKATLTADTSSSTAYMOLNSLTSEDSAVYFCAR WGYSYRNYAYYYDYWGQGTTLTVSSASTKGPS VFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVS WNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSS SLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTH TCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEV TCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYKCKV SNKALPAPIEKTISKAKGOPREPQVYTLPPSREE MTKNQVSLTCLVKGFYPSDIAVEWESNGQPENN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVF SCSVMHEALHNHYTQKSLSLSPGGGSGGSSIVM TQTPKFLPVSAGDRVTMTCKASQNVGNNVAWY QQKPGQSPKLLISYASNRYTGVPDRFTGSGSGTD FFTISSVOVEDLAVYFCOOHYSSPLTFGAGTKLE LKGGGGSGGGGSGGGGSGGGGSQVQLKQSGAE LVKPGASVKISCKASGYSFTDYYINWVKQRPGQ GLEWIGKIGPGSGNTYYNEKFEGKATLTADKSS STAYMOLSSLTSEDSAVYFCASTGPFAYWGQGT LVTWSA SEQ ID NO:223 CDH17vl/TR2#8 (IgGlKO)HC EVQLQQSVAELVKPGASVKMSCKVSGYTLTDH TIHWMKQRPEQGLEWIGYIYPRDGITGYNEKFK GKATLTADTSSSTAYMOLNSLTSEDSAVYFCAR WGYSYRNYAYYYDYWGQGTTLTVSSASTKGPS VFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVS WNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSS SLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTH

- 96 041360

TCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEV TCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYR VVSVLTVLHQDWLNGKEYKCKV SNKALPAPIEKTISKAKGQPREPQVYTLPPSREE MTKNQVSLTCLVKGFYPSDIAVEWESNGQPENN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVF SCSVMHEALHNHYTQKSLSLSPGGGSGGSDIVM TQSHKFMSTSVGDRVSITCKASQDVGTAVAWY QQKPGQSPKLLIYWASTRHTGVPDRFTGSGSGT DFTLTISNVQSEDLADYFCQQYSSYPYTFGGGTK LEIKGGGGSGGGGSGGGGSGGGGSQVTLKESGP GILQPSQTLSLTCSFSGFSLSTSGMGVSWIRQPSG KGLEWLAHIYWDDDKRYNPSLKSRLTISKDTSR NQVFLKITSVDTADTATYYCARRRLGPFAYWG QGTLVTVSA SEQ ID NO:224 CDH17vl/TR2#10 (IgGlKO)HC EVQLQQSVAELVKPGASVKMSCKVSGYTLTDH TIHWMKQRPEQGLEWIGYIYPRDGITGYNEKFK GKATLTADTSSSTAYMOLNSLTSEDSAVYFCAR WGYSYRNYAYYYDYWGQGTTLTVSSASTKGPS VFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVS WNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSS SLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTH TCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEV TCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYR VVSVLTVLHQDWLNGKEYKCKV SNKALPAPIEKTISKAKGQPREPQVYTLPPSREE MTKNQVSLTCLVKGFYPSDIAVEWESNGQPENN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVF SCSVMHEALHNHYTQKSLSLSPGGGSGGSDVL MTQTPLSLPVSLGDQASISCRSSQNIVHSNGNTY LDWYLQKPGQSPKLLIYKVSNRFSGVPDRFSGS GSGTDFTLKISRVEAEDLGVYYCFQGSHVPPTFG AGTKLELKGGGGSGGGGSGGGGSGGGGSEVQL QQSGPELVKPGASVKISCKASGYTFTDYYYMN WVKQSHGKSLEWIGYIYPNNGGTRYNQKFKGK ATLTVDKSSSTAYMELRSLTSEDSAWYCARGG NWNWYFDVWGTGTTVTVSS SEQ ID NO:225 CDH17vl/TR2#12 (IgGlKO)HC EVQLQQSVAELVKPGASVKMSCKVSGYTLTDH TIHWMKQRPEQGLEWIGYIYPRDGITGYNEKFK GKATLTADTSSSTAYMOLNSLTSEDSAVYFCAR WGYSYRNYAYYYDYWGQGTTLTVSSASTKGPS VFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVS WNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSS SLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTH TCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEV TCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYR VVSVLTVLHQDWLNGKEYKCKV SNKALPAPIEKTISKAKGQPREPQVYTLPPSREE MTKNQVSLTCLVKGFYPSDIAVEWESNGQPENN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVF SCSVMHEALHNHYTQKSLSLSPGGGSGGSDIVM TQSHKFMSTSVGDRVSITCKASQDVSNAVAWY QQKPGQSPKLLIYWASTRHNGVPDRFTGSGSGT DYILTISSVQAEDLALYYCQQHYRTPYTFGGGT KLEIKGGGGSGGGGSGGGGSGGGGSKVQLQQS GAELVKPGASVKLSCKASGYTFTEYIIHWVKQK SGQGLEWIGWFYPGSGNIKYNEKFKDKATLTAD

- 97 041360

KSSSTVYMELSRLTSEDSAVYFCTRHEGGSNFFP YWGQGTLVTVSA SEQ ID NO:226 CDH17vl/TR2vl (IgGlKO)HC EVQLQQSVAELVKPGASVKMSCKVSGYTLTDH TIHWMKQRPEQGLEWIGYIYPRDGITGYNEKFK GKATLTADTSSSTAYMOLNSLTSEDSAVYFCAR WGYSYRNYAYYYDYWGQGTTLTVSSASTKGPS VFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVS WNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSS SLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTH TCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEV TCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEOYNSTYRVVSVLTVLHODWLNGKEYKCKV SNKALPAPIEKTISKAKGOPREPQVYTLPPSREE MTKNQVSLTCLVKGFYPSDIAVEWESNGQPENN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVF SCSVMHEALHNHYTQKSLSLSPGGGSGGSDIELT QDPAVSVALGQTVRITCQGDSLRSYYASWYQQ KPGQAPVLVIYGKNNRPSGIPDRFSGSSSGNTAS LTITGAQAEDEADYYCNSRDSSGNHVVFGCGTK LTVLGGGGSGGGGSGGGGSGGGGSEVQLVQSG GGVERPGGSLRLSCAASGFTFDDYGMSWVRQA PGKCLEWVSGINWNGGSTGYADSVKGRVTISRD NAKNSLYLQMNSLRAEDTAVYYCAKILGAGRG WYFDLWGKGTTVTVSS SEQ ID NO:227 CDH17vl/TR2v2 (IgGlKO)HC EVQLQQSVAELVKPGASVKMSCKVSGYTLTDH TIHWMKQRPEQGLEWIGYIYPRDGITGYNEKFK GKATLTADTSSSTAYMOLNSLTSEDSAVYFCAR WGYSYRNYAYYYDYWGQGTTLTVSSASTKGPS VFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVS WNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSS SLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTH TCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEV TCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYKCKV SNKALPAPIEKTISKAKGOPREPQVYTLPPSREE MTKNQVSLTCLVKGFYPSDIAVEWESNGQPENN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVF SCSVMHEALHNHYTQKSLSLSPGGGSGGSSSEL TQDPAVSVALGQTVRITCQGDSLRSYYASWYQ QKPGQAPVLVIYGKNNRPSGIPDRFSGSSSGNTA SLTITGAQAEDEADYYCNSRDSSGNHVVFGGGT KLTVLGGGGSGGGGSGGGGSGGGGSEVQLVQS GGGVERPGGSLRLSCAASGFTFDDYGMSWVRQ APGKGLEWVSGINWNGGSTGYADSVKGRVTIS RDNAKNSLYLOMNSLRAEDTAVYYCAKILGAG RG WYFDLWGKGTTVTVSS SEQ ID NO:228 CDH17vl/TR2v3 (IgGlKO)HC EVQLQQSVAELVKPGASVKMSCKVSGYTLTDH TIHWMKQRPEQGLEWIGYIYPRDGITGYNEKFK GKATLTADTSSSTAYMOLNSLTSEDSAVYFCAR WGYSYRNYAYYYDYWGQGTTLTVSSASTKGPS VFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVS WNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSS SLGTOTYICNVNHKPSNTKVDKRVEPKSCDKTH TCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEV TCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYKCKV

- 98 041360

SNKALPAPIEKTISKAKGQPREPQVYTLPPSREE MTKNQVSLTCLVKGFYPSDIAVE WESNGQPENN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVF SCSVMHEALHNHYTQKSLSLSPGGGSGGSSSEL TQDPAVSVALGQTVRITCQGDSLRSYYASWYQ QKPGQAPVLVIYGKTNRPSGIPDRFSGSSSGNTA SLTITGAQAEDEADYYCNSRDSSGTHVVFGGGT KLTVLGGGGSGGGGSGGGGSGGGGSEVQLVQS GGGVERPGGSLRLSCAASGFTFDDYGMSWVRQ APGKGLEWVSGINWSGGSTGYADSVKGRVTISR DNAKNSLYLQMNSLRAEDTAVYYCAKILGAGR GWYFDLWGKGTTVTVSS SEQ ID NO:229 CDH17vl/TR2v4 (IgGlKO)HC EVQLQQSVAELVKPGASVKMSCKVSGYTLTDH TIHWMKQRPEQGLEWIGYIYPRDGITGYNEKFK GKATLTADTSSSTAYMQLNSLTSEDSAVYFCAR WGYSYRNYAYYYDYWGQGTTLTVSSASTKGPS VFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVS WNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSS SLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTH TCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEV TCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYKCKV SNKALPAPIEKTISKAKGQPREPQVYTLPPSREE MTKNQVSLTCLVKGFYPSDIAVE WESNGQPENN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVF SCSVMHEALHNHYTQKSLSLSPGGGSGGSSSEL TQDPAVSVALGQTVRITCQGDSLRSYYASWYQ QKPGQAPVLVIYGKANRPSGIPDRFSGSSSGNTA SLTITGAQAEDEADYYCNSRDSSGAHVVFGGGT KLTVLGGGGSGGGGSGGGGSGGGGSEVQLVQS GGGVERPGGSLRLSCAASGFTFDDYGMSWVRQ APGKGLEWVSGINWSGGSTGYADSVKGRVTISR DNAKNSLYLQMNSLRAEDTAVYYCAKILGAGR GWYFDLWGKGTTVTVSS SEQ ID NO:230 CDH17vl/TR2v5 (IgGl KO) HC EVQLQQSVAELVKPGASVKMSCKVSGYTLTDH TIHWMKQRPEQGLEWIGYIYPRDGITGYNEKFK GKATLTADTSSSTAYMQLNSLTSEDSAVYFCAR WGYSYRNYAYYYDYWGQGTTLTVSSASTKGPS VFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVS WNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSS SLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTH TCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEV TCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYKCKV SNKALPAPIEKTISKAKGQPREPQVYTLPPSREE MTKNQVSLTCLVKGFYPSDIAVE WESNGQPENN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVF SCSVMHEALHNHYTQKSLSLSPGGGSGGSSSEL TQDPAVSVALGQTVRITCQGDSLRSYYASWYQ QKPGQAPVLVIYGKNNRPSGIPDRFSGSSSGNTA SLTITGAQAEDEADYYCNSRDSSGNHVVFGGGT KLTVLGGGGSGGGGSGGGGSGGGGSEVQLVQS GGGVERPGGSLRLSCAASGFTFDDYGMSWVRQ APGKGLEWVSGINWSGGSTGYADSVKGRVTISR DNAKNSLYLQMNSLRAEDTAVYYCAKILGAGR GWYFDLWGKGTTVTVSS

- 99 041360

SEQIDNO:231 CDH17vl/TR2v6 (IgGl KO) HC EVQLQQSVAELVKPGASVKMSCKVSGYTLTDH TIHWMKQRPEQGLEWIGYIYPRDGITGYNEKFK GKATLTADTSSSTAYMQLNSLTSEDSAVYFCAR WGYSYRNYAYYYDYWGQGTTLTVSSASTKGPS VFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVS WNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSS SLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTH TCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEV TCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYKCKV SNKALPAPIEKTISKAKGQPREPQVYTLPPSREE MTKNQVSLTCLVKGFYPSDIAVEWESNGQPENN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVF SCSVMHEALHNHYTQKSLSLSPGGGSGGSSSEL TQDPAVSVALGQTVRITCQGDSLRSYYASWYQ QKPGQAPVLVIYGKTNRPSGIPDRFSGSSSGNTA SLTITGAQAEDEADYYCNSRDSSGDHVVFGGGT KLTVLGGGGSGGGGSGGGGSGGGGSEVQLVQS GGGVERPGGSLRLSCAASGFTFDDYGMSWVRQ APGKGLEWVSGINWAGGSTGYADSVKGRVTIS RDNAKNSLYLQMNSLRAEDTAVYYCAKILGAG RGWYFDL WGKGTT VT VS S SEQ ID NO:232 CDH17vl/TR2v7 (IgGlKO)HC EVQLQQSVAELVKPGASVKMSCKVSGYTLTDH TIHWMKQRPEQGLEWIGYIYPRDGITGYNEKFK GKATLTADTSSSTAYMQLNSLTSEDSAVYFCAR WGYSYRNYAYYYDYWGQGTTLTVSSASTKGPS VFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVS WNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSS SLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTH TCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEV TCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYKCKV SNKALPAPIEKTISKAKGQPREPQVYTLPPSREE MTKNQVSLTCLVKGFYPSDIAVEWESNGQPENN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVF SCSVMHEALHNHYTQKSLSLSPGGGSGGSSSEL TQDPAVSVALGQTVRITCQGDSLRSYYASWYQ QKPGQAPVLVIYGKDNRPSGIPDRFSGSSSGNTA SLTITGAQAEDEADYYCNSRDSSGDHVVFGGGT KLTVLGGGGSGGGGSGGGGSGGGGSEVQLVQS GGGVERPGGSLRLSCAASGFTFDDYGMSWVRQ APGKGLEWVSGINWTGGSTGYADSVKGRVTISR DNAKNSLYLQMNSLRAEDTAVYYCAKILGAGR GWYFDLWGKGTTVTVSS SEQ ID NO:233 CDH17vl LC DIVMSQSPSSLAVSVGEKVTMSCKSSQSLLHSSN QKNYLAWYQQKPGQSPKVLIYWASTRESGVPD RFTGSGSGTDFTLTITSVKSEDLAVYYCQQYYSY PWTFGGGTRLEIKRTVAAPSVFIFPPSDEQLKSGT ASVVCLLNNFYPREAKVQWKVDNALQSGNSQE SVTEQECDSKDSTYSLSSSFTHNSKADYEK SEQ ID NO:270 IgGlFcRnmut ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFP EPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSV VTVPSSSLGTQTYICNVNHKPSNIKVDKRVEPKS CDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMI SRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVH

- 100 041360

NAKTKPREEQYNSTYRVVSVLTVLAQDWLNGK EYKCKVSNKALPAPIEKTISKAKGQPREPQVYTL PPSREEMTKNQVSLTCLVKGFYPSDIAVEWESN GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRW QQGNVFSCSVMHEALHNHYTQKSLSLSPG SEQIDNO:271 CDH17H2/TRv2 (IgGlFcRnmut) HC QVQLVQSGAEVKKPGSSVKVSCKVSGYTFTDHT IHWMKQRPGQGLEWIGYIYPRDVITQYNEKFKG KVTLTADKSTSTAYMELSSLRSEDTAVYLCARW GYFYGSRSYYFDYWGQGTTVTVSSASTKGPSVF PLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN SGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSL GTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTC PPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTC VVVDVSHEDPEVKFNWYVDGVEVHNAKTKPRE EQYNSTYRVVSVLTVLAQDWLNGKEYKCKVSN KALPAPIEKTISKAKGQPREPQVYTLPPSREEMT KNQVSLTCLVKGFYPSDIAVEWESNGQPENNYK TTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSC SVMHEALHNHYTQKSLSLSPGGGSGGSSSELTQ DPAVSVALGQTVRITCQGDSLRSYYASWYQQKP GQAPVLVIYGKNNRPSGIPDRFSGSSSGNTASLTI TGAQAEDEADYYCNSRDSSGNHVVFGGGTKLT VLGGGGSGGGGSGGGGSGGGGSEVQLVQSGGG VERPGGSLRLSCAASGFTFDDYGMSWVRQAPG KGLEWVSGINWNGGSTGYADSVKGRVTISRDN AKNSLYLQMNSLRAEDTAVYYCAKILGAGRGW YFDL WGKGTT VTVS S SEQ ID NO:272 CDH17vlHCCDR3 GYSYRNYAYYYDY

Example 6 Increased apoptotic effect of bispecific molecules recognizing human TRAILR2 and human CDH17.

Introduction.

The above examples show the preparation of bispecific molecules recognizing human TRAILR2 and CDH17. In order to determine the effect of these molecules on the induction of apoptosis, the inventors decided to investigate whether the molecules could cause a decrease in cell viability, whether any such decrease was caused by apoptosis, and whether any apoptotic effect was mediated by bispecific TRAILR2/CDH17 binding molecules.

Development of cell analysis.

First, it was necessary to identify a cancer cell line that has TRAILR2 and CDH17 on the cell surface. The cell line Colo-205 is derived from human colon adenocarcinoma from the Caucasus. It is a well-known cell culture laboratory instrument.

The inventors selected it as a candidate cell line to evaluate the function of bispecific molecules that bind TRAILR2/CDH17.

Surface expression of TRAILR2 and CDH17 protein on Colo-205 cells was analyzed as follows. Cells were detached using accutase (Sigma A6964) and washed twice with FACS buffer (PBS, Gibco 14190; 3% FCS, Gibco 26140; and 0.09% NaN3 Sigma Aldrich S2002). Cells were counted using ViCell (Beckman Coulter Life Sciences) and the cell count was adjusted to 2-5x106 cells/ml. A cell suspension of 100 μl/well was seeded into 96-well round bottom plates.

After seeding the cells, the plate was centrifuged at 1200 rpm for 5 min and the supernatant was discarded. The cells were then incubated for 60 min at 4°C with 100 μl/well of the appropriate dilution of the primary antibody. Cells were washed twice with FACS buffer and 100 μl/well of the appropriate dilution of secondary/conjugated antibodies was added. Cells with secondary antibodies were incubated for 45 min at 4°C in the dark. After two washes with FACS buffer, cells were resuspended in 100 μl of FACS buffer per well and analyzed in FACS Canto (BD Biosciences).

CD262 (DR5) conjugated anti-human PE (eBioscience, 12-9908-42) was used to detect TRAILR2. For CDH17, an anti-CDH17 antibody (R&D Systems, MAB1032) was used, followed by a rabbit anti-mouse IgG PE secondary antibody (Dako, R0439). Mouse IgG1 isotype control PE (eBioscience, 12-4714-42) and mouse IgG1 isotype control (AbDserotec, MCA928EL), respectively, were used as controls.

In FIG. 2 shows surface expression of TRAILR2 and CDH17 protein in Colo-205 cells, both proteins showing significant expression.

Effect of TRAILR2/CDH17 bispecific molecules on Colo-205 cells.

Having identified Colo-205 cells as a suitable cancer cell line for evaluating the function of bispecific molecules that bind TRAILR2/CDH17, the inventors developed the following

- 101 041360 general analysis.

Colo-205 cells were seeded in culture medium (RPMI1640/Glutamax, Gibco 61870-010; plus 10% FCS, Gibco 26140). After resting overnight at 37°C and 5% CO ₂ , the cells were incubated for 24 h with 50 μl of various dilutions of antibodies or binding molecules at desired concentrations. Cell viability was then assessed using the CellTiter-Glo Luminescent Cell Viability Assay (Promega G7571) according to the instructions provided by the manufacturer. Finally, luminescence was recorded using a Perkin Elmer VICTOR X4 2030 multilabile plate reader.

The effect of bispecific TRAILR2/CDH17 binding molecules on cell viability is shown in FIG. 3. Colo-205 cells were incubated for 24 hours with (i) the bispecific molecule, (ii) anti-TRAILR2 antibody alone, (iii) anti-CDH17 antibody alone, or (iv) an equivalent free combination of TRAILR2 and CDH17 antibodies.

As shown in FIG. 3, the anti-CDH17 antibody had no effect on the viability of Colo-205 cells when used in the concentration range from 7 x 10' ⁴ to 70 nM. The TRAILR2 antibody was able to significantly reduce cell viability when used between 7 and 70 nM and the addition of anti-CDH17 in free combination did not alter the effect seen with anti-TRAILR2 antibody alone. Finally, incubation of Colo-205 cells with a bispecific TRAIL-R2/CDH17 binding molecule as described in the present invention (CDH17v1/TR2v1 containing the heavy chain sequence of SEQ ID NO: 145 and the light chain sequence of SEQ ID NO: 146), resulting in more than 1000-fold improved performance compared to both an anti-TRAIL-R2 antibody (containing a TRv1 antigen-binding site with the sequence VH and VL of SEQ ID NO: 96 and SEQ ID NO: 97, respectively) alone, and anti-TRAIL-R2 an antibody in combination with an anti-CDH17 antibody (comprising the CDH17v1 antigen-binding site with the sequence VH and VL of SEQ ID NO: 118 and SEQ ID NO: 119, respectively).

TRAILR2/CDH17 molecules induce apoptosis in Colo-205 cells.

It was then examined whether there was a decrease in cell viability as shown in FIG. 3 is caused by the induction of apoptosis. TRAIL-induced apoptosis is mediated by recruitment and activation of caspase-8, which in turn activates effector caspase-3, leading to apoptosis. Therefore, in order to determine whether the antibodies and binding molecules produced in the present invention are capable of specifically activating the apoptotic pathway, the inventors measured the activities of both caspase-8 and -3 in target cell lines.

Caspase-3 and -8 activity was measured using Promega Apo-ONE Homogeneous Caspase-3/7 Assay (Cat. No. G7790) and Promega Caspase-Glo 8 Assay (Cat. No. G8201), respectively. After keeping overnight at 37°C and 5% CO ₂ , the cells were incubated at different time points with 50 μl of various dilutions of antibodies or binding molecules to achieve the desired concentrations. Caspase-3 and -8 activity was then assessed according to the instructions provided by the manufacturer. The luminescence of each sample was then measured using a Perkin Elmer VICTOR X4 2030 multilabile plate reader.

The results of this analysis are shown in FIG. 4. Here it can be seen that both anti-TRAILR2 antibodies (containing the TRv1 antigen-binding site with the sequence VH and VL of SEQ ID NO: 96 and SEQ ID NO: 97, respectively) alone and the TRAILR2/CDH17 bispecific binding molecules (CDH17v1/TR2v1 containing the heavy chain sequence of SEQ ID NO: 145 and the light chain sequence of SEQ ID NO: 146) were able to effectively activate both caspase 8 (A) and 3 (B). No effect was observed after incubation of an anti-CDH17 antibody (containing the CDH17v1 antigen binding site with the VH and VL sequence of SEQ ID NO: 118 and SEQ ID NO: 119, respectively) when compared to untreated controls. These data demonstrate that the decrease in cell viability seen in FIG. 3 is not associated with any non-specific mechanism, and that both molecules are able to effectively and specifically induce apoptosis in target cells. Consistent with cell viability data, the inventors found that incubation of Colo-205 cells with bispecific TRAILR2/CDH17 binding molecules clearly resulted in superior activation of caspase-8 and -3 (more than 3 log shift in activity) compared to only anti-TRAILR2 antibody.

Apoptosis in Colo-205 cells is specifically induced by the TRAILR2/CDH17 bispecific molecules.

The inventors also wished to confirm that the increase in apoptosis modulated by the bispecific TRAILR2/CDH17 binding molecules according to the invention is specifically mediated by CDH17 present on the surface of Colo-205 cells. To demonstrate this, the inventors created an additional bispecific, tetravalent molecule recognizing human TRAILR2 and trinitrophenol (TNP), where the IgG primary anti-CDH1 7 antibody was replaced with a single TNP recognition. TNP binding region acts as non-specific

- 102 041360 control for the molecule because the TNP antigen cannot be easily detected on the cell surface.

Colo-205 cells were treated for 24 hours with different concentrations of (i) anti-TNP antibody only, (ii) anti-CDH17 antibody only, (iii) anti-TRAILR2 antibody only, (iv) TRAILR2/TNP bispecific binding molecule ( v) TRAILR2/CDH17 bispecific molecules and cell viability was measured. The data obtained is shown in Fig. 5.

As shown in FIG. 5, the TRAILR2/CDH17 bispecific binding molecule (CDH17v1/TR2v1 containing the heavy chain sequence of SEQ ID NO: 145 and the light chain sequence of SEQ ID NO: 146) is able to induce a clear superior effect in reducing cell viability compared to an anti-TRAILR2 antibody (containing an antigen binding the TRv1 region with the VH and VL sequence of SEQ ID NO: 96 and SEQ ID NO: 97, respectively). This effect is not observed with the bispecific TRAILR2/TNP binding molecule. Therefore, replacing CDH17 for TNP with TNP abolishes the superior apoptotic effect of the TRAILR2/CDH17 bispecific binding molecule, and the TRAILR2/TNP bispecific binding molecule has a comparable effect with only one anti-TRAILR2 antibody.

Effect of TRAILR2/CDH17 bispecific molecules on HepG2 cells.

To study the effect of bispecific TRAILR2/CDH17 binding molecules on liver-derived CDH17-negative and TRAILR2-responsive cells, a similar experiment as described above was performed using HepG2 cells. This cell line was originally derived from hepatocellular carcinoma of the liver of a 15 year old Caucasian male.

HepG2 cells were cultured at 37°C and 5% CO ₂ in DMEM (Lonza, BE12-604F) plus 10% FCS (Gibco 26140).

As described for Colo-205, surface protein expression and cell viability were analyzed as described in Example 3. FIG. 6A shows surface protein expression of TRAILR2 and CDH17 in HepG2 cells, with only TRAILR2 showing significant expression.

HepG2 cells were incubated with (i) anti-TRAILR2 (anti-TR2v1 containing a TRv1 antigen-binding site with only VH and VL sequences of SEQ ID NO: 96 and SEQ ID NO: 97, respectively), (ii) TRAILR2/CDH17 bispecific binding molecule ( CDH17v1/TR2v1 containing the heavy chain sequence of SEQ ID NO: 145 and the light chain sequence of SEQ ID NO: 146).

In addition, as a control for maximal pro-apoptotic signaling, antiTRAILR2 antibodies as described in the literature for various human TRAILR1 and TRAILR2 agonist antibodies (Ichikawa et al., 2001. Nat Med. Aug;7(8):954-60; Li et al., 2008 BMC Cancer. Nov 7;8:325; Natoni et al., 2007, British journal of haematology 2007 Nov;139(4):568-77; Pukac et al., 2005 Br J Cancer. 2005 Apr 25 ;92(8):1430-41 Yada et al. -57), HepG2 cells were also incubated with (iii) antiTRAILR2 (containing the TRv1 antigen-binding site of VH and VL sequences of SEQ ID NO: 96 and SEQ ID NO: 97, respectively) in the presence of a cross-linker, goat anti-human IgG (anti -TR2v1 (Fc crosslinking)). As shown in FIG. 6B, both the anti-TRAILR2 antibody alone and the TRAILR2/CDH17 bispecific molecule do not affect viability. Only artificial cross-linking of the Fc of the TRAILR2 antibody can significantly reduce cell viability.

Effect of TRAILR2/CDH17 bispecific molecules on additional CRC cells.

To explore whether the observed effect in Colo-205 could be applied to other CRC cell lines, an experiment similar to that described above was performed using CL-34 (KRAS wild-type) and SK-CO-1 (KRAS p.G12V) cells, two additional CRC cell lines.

CL-34 and SK-CO-1 cells were cultured at 37°C and 5% CO ₂ in DMEM/F-12 (ATCC 30-2006) plus 20% FCS (Gibco 26140) and EMEM (Lonza BE12-662F) plus 10 % FCS (Gibco 26140), respectively.

As described in Example 6, surface protein expression and cell viability were analyzed. In FIG. 11A and 11B show surface expression of TRAILR2 and CDH17 protein in CL34 (A) and SK-CO-1 (B) cells, both proteins showing significant expression.

CL-34 and SK-CO-1 cells were incubated with (i) a TRAILR2/CDH17 bispecific binding molecule (CDH17v1/TR2v1 containing SEQ ID NO: 145 heavy chain and SEQ ID NO: 146 heavy chain), (ii) anti- TRAILR2 antibody (comprising the TRv1 antigen-binding region of VH and VL sequence of SEQ ID NO: 96 and SEQ ID NO: 97, respectively), or (iii) an anti-CDH17 antibody alone (containing the CDH17v1 antigen-negative region of VH and VL sequence of SEQ ID NO: 118 and SEQ ID NO: 119, respectively).

As shown in FIG. 11, the anti-CDH17 antibody does not affect cell viability, while the TRAILR2 antibody can significantly reduce cell viability when used between 7 and

- 103 041360 nM. As described for Colo-205 cells, incubation of both CL-34 and SK-CO-1 cells with bispecific TRAILR2/CDH17 binding molecules resulted in a clear shift of more than 3 log activity in cell viability compared to anti- TRAILR2 antibody.

Consistent with a similar response observed in CL-34 and SK-CO-1 cells, no significant difference in susceptibility was found between KRASmut and KRASwt cell lines in an expanded panel of 19 CRC cell lines (Wilcoxon-Rank-Sum-Test, data not shown).

Discussion.

The inventors have demonstrated above that a bispecific TRAILR2/CDH17 binding molecule induces apoptosis in CRC cell lines. This apoptotic effect is achieved using a concentration that is more than 3 orders of magnitude less than the anti-TRAILR2 antibody alone and is at therapeutically acceptable dosage levels. It has also been demonstrated that this effect is specifically triggered by a portion of the CDH17 bispecific molecule, and that no significant effect on cell viability of liver-derived CDH17-negative and TRAILR2-responsive HepG2 cells was found at maximum effect concentration.

Example 7 Use of different IgG scaffolds in the TRAILR2/CDH17 bispecific molecule.

The inventors also investigated whether altering the IgG framework used in the core antibody element of the TRAILR2/CDH17 bispecific molecules alters the induction of apoptosis.

Colo-205 cells were treated for 24 hours with various concentrations of anti-TRAILR2 antibody alone and TRAILR2/CDH17 bispecific binding molecules containing IgG1 (A and C) or IgG4Pro(B) constant region. After antibody incubation, cell viability was measured (FIG. 7).

As shown in FIG. 7, replacement of the previously used IgG1 framework with a second variant (IgG1 KO; SEQ ID NO: 121) with 2 additional mutations (L234A and L235A) or a third variant (FcRnmut) with 1 additional mutation (H310A), SEQ ID NO: 270) does not change the effect on the decrease in the viability of Colo-205 cells.

Like the wild-type variant, the newly designed bispecific binding molecules (TRv1/CDH17v1 IgG1 (KO) containing the heavy chain of SEQ ID NO: 147 and the light chain of SEQ ID NO: 148) were able to significantly reduce cell viability when used at concentrations as low as 10 ^-2 nM, while the parental TRAILR2 antibody (containing the TRv1 antigen binding site with the VH and VL sequences of SEQ ID NO: 96 and SEQ ID NO: 97, respectively) showed no effect. This is also true for the second bispecific binding molecule where the TRAILR2 binding domain has been replaced with a slightly modified sequence (TRAILR2v2 containing SEQ ID NO: 149 heavy chain and SEQ ID NO: 150 light chain compared to anti-TR2v2 with VH and VL sequences of SEQ ID NO: 98 and SEQ ID NO: 99, respectively)). In addition, various bispecific molecules have been generated in which the IgG scaffold used in the core antibody element of the bispecific scaffold has been completely replaced by an IgG4 Pro isoform (containing the heavy chain sequence of SEQ ID NO: 151 and the light chain sequence of SEQ ID NO: 152) or IgG1 FcRnmut (CDH17H2/TR2v2) containing the heavy chain sequence of SEQ ID NO: 271 and the light chain sequence of SEQ ID NO: 218). As shown in FIG. 7, this change in the molecule did not affect its ability to reduce the viability of Colo-205 cells.

Example 8 Effect of scFv Variants on Decreased Cell Viability.

It was then investigated whether altering the second component of the scFv antibody affects the induction of apoptosis by the molecule according to the invention.

In the molecules used in Example 6, the scFv is oriented so that the VL forms the N-terminus of the binding fragment, and thus connects via a linker to the C-terminus of the main antibody heavy chain, while the VH of scFv forms the C-terminus of the entire heavy chain molecule. .

Consequently, several different bispecific TRAILR2/CDH17 binding molecules have been generated containing binding sites defined by identical variable domain sequences but with different orientation and/or other modifications of said domains (scFv versions 1 to 4).

1. The TRAILR2/CDH17 scFv1 bispecific molecule has a VH-VL orientation, i. The VH scFv is fused via a linker to the C-terminus of the antibody, and the VL scFv forms the C-terminus of the heavy chain of the whole molecule (CDH17v1/TRv1 with scFv sequence SEQ ID NO: 131).

2. The TRAILR2/CDH17 scFv2 bispecific molecule also has a VH-VL orientation, i. The scFv VH is fused via a linker to the C-terminus of the antibody, and the scFv VL forms the C-terminus of the heavy chain of the whole molecule, and has an additional disulfide bond to stabilize the scFv binding domain (CDH17v1/TRv1 with scFv sequence SEQ ID NO: 132).

3. The TRAILR2/CDH17 scFv3 bispecific molecule used in FIG. 7 and thus has a VL-VH orientation, i.e. The VL scFv is fused via a linker to the C-terminus of the antibody, and the VH scFv forms the C-terminus of the heavy chain of the whole molecule (CDH17v1/TRv1 with scFv sequence SEQ ID NO: 124).

4. The bispecific TRAILR2/CDH17 scFv4 molecule also has a VL-VH orientation, i. The VL scFv is fused via a linker to the C-terminus of the antibody, and the VH scFv forms the C-terminus of the heavy chain of the whole molecule,

- 104 041360 and has an additional disulfide bond to stabilize the scFv binding domain (CDH17v1/TRv1 with scFv sequence SEQ ID NO: 133).

The function of these TRAILR2/CDH17-binding variants of the bispecific molecules was then examined in the Colo-205 cell apoptosis assay described in Example 6. After incubation with the binding molecules, cell viability was measured.

The data shown in FIG. 8 show that the TRAILR2/CDH17 bispecific binding molecule can induce apoptosis in Colo-205 cells regardless of the orientation of the VH and VL domains within the scFv antibody domain, however, VL-VH has a stronger apoptotic effect than VH-VL orientation. Molecules bearing an additional disulfide bond do not have a significant effect compared to the corresponding parent molecules without these additional bonds.

Therefore, changes in the structure of the bispecific antibody format described in Example 1 can be tolerated, although VL-VH has a surprisingly large apoptotic effect.

Example 9 Effect of bispecific TRAILR2 and CDH17 bi-scFv molecules using alternative binding sequences in reducing the viability of Colo-205 cells.

Variations of TRAILR2/CDH17 bispecific molecules with different binding domains have been generated. Details of the specific binding molecules used are shown in Table 1. in Example 5. The effect of varying the binding domains used in the Colo-205 cell apoptosis assay was then examined as described in Example 6.

Colo-205 cells were treated for 24 h with various concentrations of various bispecific molecules containing various newly identified CDH17 (A) and TRAILR2 (B and C) binding sequences. After antibody incubation, cell viability was measured.

The data shown in FIG. 9 (A-C) show that bispecific binding molecules with different TRAILR2 and CDH17 binding sequences (CDH17A6/TR2v2 containing HC SEQ ID NO: 182 and LC SEQ ID NO: 188; CDH17E2/TR2v2 containing HC SEQ OD NO: 197 and LC SEQ ID NO: 203 CDH17E9/TR2v2 containing HC SEQ ID NO: 167 and LC SEQ ID NO: 173 CDH17v1/TR2#1 containing HC SEQ ID NO: 219 and LC SEQ ID NO: 233, CDH17v1 /TR2#2 containing HC SEQ ID NO: 220 and LC SEQ ID NO: 233 CDH17v1/TR2#3 containing HC SEQ ID NO: 221 and LC SEQ ID NO: 233 CDH17v1/TR2#8 containing HC SEQ ID NO: 223 and LC SEQ ID NO: 233 CDH17v1/TR2#10 containing HC SEQ ID NO: 224 and LC SEQ ID NO: 233 CDH17H2/TR2v2 containing HC SEQ ID NO: 212 and LC SEQ ID NO: 218, CDH17H2/TR2v3 containing HC SEQ ID NO: 213 and LC SEQ ID NO: 218 CDH17H2/TR2v4 containing HC SEQ ID NO: 214 and LC SEQ ID NO: 218 CDH17H2/TR2v5 containing HC SEQ ID NO: 215 and LC SEQ ID NO: 218 CDH17H2/TR2v6 containing HC SEQ ID NO: 216 and LC SEQ ID NO: 218 CDH17H2/TR2v7 containing HC SEQ ID NO: 217 and LC SEQ ID NO: 218) are able to significantly reduce cell viability when used at concentrations as low as 10 ^-2 nM, while the parent antibody TRAILR2 (TR2v2 containing VH SEQ ID NO: 98 and VL SEQ ID NO: 99) showed no effect at the same concentration. In FIG. 9D, some of these CDH17H2/TR2v2 molecules containing HC SEQ ID NO: 212 and LC SEQ ID NO: 218, CDH17H2/TR2v3 containing HC SEQ ID NO: 213 and LC SEQ ID NO: 218; CDH17H2/TR2v4 containing HC SEQ ID NO: 214 and LC SEQ ID NO: 218; CDH17H2/TR2v5 containing HC SEQ ID NO: 215 and LC SEQ ID NO: 218; CDH17H2/TR2v6 containing HC SEQ ID NO: 216 and LC SEQ ID NO: 218; CDH17H2/TR2v7 containing HC SEQ ID NO: 217 and LC SEQ ID NO: 218) were tested in the aforementioned liver-derived CDH17-negative and TRAILR2-responsive HepG2 cells. As shown in the figure, concentrations up to 1 nM did not significantly affect HepG2 cell viability for any of the molecules tested. In FIG. 9C-D also show that the anti-CDH17 parental antibody (CDH17H2 containing VH SEQ ID NO: 116 and VL SEQ ID NO: 117) showed no cell viability-reducing effect either on Colo-205 or within the concentration range tested.

Example 10 Bispecific molecules recognizing TRAILR2 and CD44v6 have no additional effect on reducing the viability of Colo-205 cells.

It was also investigated whether an alternative anchor target could be used instead of CDH17. Therefore, a bispecific molecule recognizing TRAILR2 and CD44v6 was generated.

As in example 6, the inventors first measured the surface expression of the TRAILR2 protein and CD44v6 using FACS. For detection of CD44v6, the BIWA1 antibody was used, followed by a goat anti-mouse IgG PE secondary antibody (Dako, R0480). Mouse IgG1 isotype control (AbDserotec, MCA928EL) was used as a control.

In FIG. 10A shows surface expression of TRAILR2 and CD44v6 protein in Colo-205 cells, both proteins showing significant expression.

Cell viability was then measured using the CellTiter-Glo luminescent cell viability assay. Colo-205 cells were incubated with either the TRAILR2/CD44v6 bispecific binding molecule, anti-TRAILR2 antibody alone, or anti-CD44v6 antibody alone. The data is shown in Fig. 10V.

Incubation of anti-CD44v6 antibody in the concentration range from 10 ^-2 ng/ml to 10 mg/ml nM does not affect

- 105 041360 on the viability of Colo-205 cells. Incubation with aHTu-TRAILR2 antibody (TR2v1 containing VH

SEQ ID NO: 96 and VL SEQ ID NO: 97) was able to significantly reduce cell viability when used between 0.1 and 10 μg/mL, and incubation of Colo-205 cells with an equivalent amount of a bispecific, tetravalent molecule recognizing human TRAILR2 and human CD44v6 did not differ from the effect seen with only one anti-TRAILR2 antibody.

Thus, CD44v6 cannot be used as an anchor target for modulating TRAILR2-induced apoptosis in Colo-205 cells.

Example 11 In vivo efficacy of bispecific molecules in xenograft models of CRC.

The in vivo efficacy of binding molecules recognizing TRAILR2 and CDH17 was also investigated. For this purpose, Colo205 (KRAS wild-type) and Gp2d (KRAS p.G12D), human CRC cancer cell lines expressing both TRAILR2 and CDH17, were implanted in immunodeficient mice, and the effect of administering a molecule according to the invention was measured in tumor volume.

BomTac:NMRI-Foxnlnu-homozygous female mice are inoculated with 5.0x106 Colo205 cells (0.1 ml PBS + 5% FCS) or 5.0x106 Gp2d cells (0.1 ml with cells: matrigel ratio 1:1 (v/v .)) and tumor growth was controlled until reaching 180-200 mm ³ . Mice were randomized into two groups and given one injection of a vehicle control or binding molecule according to the invention - 5 mg/kg for Colo205 and 1.67 mg/kg for Gp2d (CDH17H2/TR2v2-(Ig G1 KO) containing the heavy chain sequence of SEQ ID NO : 212 and light chain sequence SEQ ID NO: 218); 5 mg/kg for Colo205 and Gp2d (CDH17H2/TR2v2-(Ig G1 FcRnmut) containing the heavy chain sequence of SEQ ID NO: 271 and the light chain sequence of SEQ ID NO: 218).

The data shown in FIG. 12 demonstrate that the binding molecules according to the invention are capable of producing a significant and lasting reduction in tumor volume compared to the control group.

Example 12 Detection of Surface Expression of CDH17 Using Newly Generated Anti-CDH17 Antibodies.

In FIG. 13 shows how the newly generated anti-CDH17H2 antibody (comprising VH SEQ ID NO: 116 and VL SEQ ID NO: 117 and the sequence of IgG1SEQ ID NO: 121) is able to detect surface expression of the CDH17 protein. FACS analysis was performed as described in Example 6. A newly generated anti-CDH17H2 antibody was used as the primary detection antibody, followed by a secondary rabbit anti-mouse IgG PE antibody (Dako, R0439). Human IgG1 APC isotype control (Biolegend, 409306) was used as a control.

Example 13. Pharmaceutical composition for intravenous administration.

Any of the above binding molecules in accordance with the invention can be selected for the manufacture of a pharmaceutical composition for iv. introductions. An example of a suitable pharmaceutical formulation for an antibody of the invention is as follows:

ml vial contains 10 mg/ml of the binding molecule of the invention in a buffer consisting of 10 mM citrate, pH 5.5, 207 mM sucrose, 25 mM lysine HCl and 0.02% polysorbate 20, and water for injection (WFI ).

Example 14 ProtA Affinity Chromatography and Monomer Content Determination by Analytical Size Exclusion Chromatography (aSEC).

The various binding molecules described in the present invention were sampled from the collected cell culture fluid by recombinant protein-A affinity chromatography using MabSelect SuRe resin (GE Healthcare). Samples were eluted isocratically using 30 mM sodium acetate, pH 3.5 and eluted samples were neutralized to pH 5.0 using 1% 3M sodium acetate, pH 9.0.

aSEC was performed on a Waters BEH200 column on a Waters UHPLC system. The mobile phase buffer contained 50 mM sodium phosphate, pH 6.8, 200 mM arginine, 0.05% sodium azide. For each cycle, 5-10 µg of sample was injected at a current flow rate of 0.5 ml/min.

Was determined by the percentage of monomers, and the results for various binding molecules are shown in table. 4.

- 106 041360

Binding molecule % monomers after protA CDH17H2/TRv2 (SEQ IDNOs 212/218) 98% CDH17H2/TRv3 (SEQ IDNOs 213/218) 99% CDH17H2/TRv4 (SEQ IDNOs 214/218) 96% CDH17H2/TRv5 (SEQ IDNOs 215/218) 96% CDH17H2/TRv6 (SEQ IDNOs 216/218) 97.7% CDH17H2/TRv7 (SEQ IDNOs 217/218) 98.3%

CLAIM

Claims (26)

CLAIM 1. A bispecific and tetravalent binding molecule that is a modified immunoglobulin (Ig) molecule and contains two antigen-binding sites that specifically bind to TNF-associated apoptosis-inducing receptor ligand 2 (TRAILR2) and two antigen-binding sites that specifically bind to cadherin- 17 (CDH17), where the two antigen-binding sites that specifically bind to cadherin-17 (CDH17) are in the variable region (Fv) of the Ig molecule and the two antigen-binding sites that specifically bind to TRAILR2 are scFv(s), with scFv( s) fused to the C-terminus of the heavy chain of the Ig molecule. 2. The binding molecule of claim 1, wherein the scFv(s) have a VL-VH orientation from N- to C-terminus. 3. The binding molecule according to any one of claims 1 or 2, wherein the Ig molecule is IgG. 4. A binding molecule according to any one of claims 1 to 3, wherein the two scFv(s) are fused to the Ig molecule via a peptide linker, preferably a peptide linker having a length of about 4 to 20 amino acids. 5. A binding molecule according to any one of claims 1-4, wherein the antigen-binding site that specifically binds to TRAILR2 is selected from the group consisting of antigen-binding sites i)-xi): i) an antigen-binding site comprising heavy chain CDRs containing the amino acid sequences of SEQ ID NO: 1 (CDR1), SEQ ID NO: 2 (CDR2) and SEQ ID NO: 3 (CDR3), and light chain CDRs containing the amino acid sequences of SEQ ID NO: 4 (CDR1), SEQ ID NO: 5 (CDR2) and SEQ ID NO: 6 (CDR3); ii) an antigen-binding site comprising heavy chain CDRs containing the amino acid sequences of SEQ ID NO: 7 (CDR1), SEQ ID NO: 8 (CDR2) and SEQ ID: NO: 9 (CDR3), and light chain CDRs containing the amino acid sequences of SEQ ID NO: 10 (CDR1), SEQ ID NO: 11 (CDR2) and SEQ ID NO: 12 (CDR3); iii) an antigen-binding site comprising heavy chain CDRs containing the amino acid sequences of SEQ ID NO: 13 (CDR1), SEQ ID NO: 14 (CDR2) and SEQ ID NO: 15 (CDR3), and light chain CDRs containing the amino acid sequences of SEQ ID NO: 16 (CDR1), SEQ ID NO: 17 (CDR2) and SEQ ID NO: 18 (CDR3); iv) an antigen-binding site comprising heavy chain CDRs containing the amino acid sequences of SEQ ID NO: 25 (CDR1), SEQ ID NO: 26 (CDR2) and SEQ ID NO: 27 (CDR3), and light chain CDRs containing the amino acid sequences of SEQ ID NO: 28 (CDR1), SEQ ID NO: 29 (CDR2) and SEQ ID NO: 30 (CDR3); v) an antigen-binding site comprising heavy chain CDRs containing the amino acid sequences of SEQ ID NO: 31 (CDR1), SEQ ID NO: 32 (CDR2) and SEQ ID NO: 33 (CDR3), and light chain CDRs containing the amino acid sequences of SEQ ID NO: 34 (CDR1), SEQ ID NO: 35 (CDR2) and SEQ ID NO: 36 (CDR3); vi) an antigen-binding site comprising a heavy chain CDR containing SEQ ID NO: 43 (CDR1), SEQ ID NO: 44 (CDR2) and SEQ ID NO: 48 (CDR3), and a light chain CDR containing the amino acid sequences of SEQ ID NO: 49 (CDR1), SEQ ID NO: 50 (CDR2) and SEQ ID NO: 54 (CDR3); vii) an antigen-binding site comprising heavy chain CDRs comprising SEQ ID NO: 43 - 107 041360 (CDR1), SEQ ID NO: 45 (CDR2) and SEQ ID NO: 48 (CDR3), and light chain CDRs containing the amino acid sequences of SEQ ID NO: 49 (CDR1), SEQ ID NO: 51 (CDR2) and SEQ ID NO: 55 (CDR3); viii) an antigen-binding site comprising a heavy chain CDR comprising SEQ ID NO: 43 (CDR1), SEQ ID NO: 45 (CDR2) and SEQ ID NO: 48 (CDR3), and a light chain CDR comprising the amino acid sequences of SEQ ID NO: 49 (CDR1), SEQ ID NO: 52 (CDR2) and SEQ ID NO: 56 (CDR3); ix) an antigen-binding site comprising a heavy chain CDR comprising SEQ ID NO: 43 (CDR1), SEQ ID NO: 45 (CDR2) and SEQ ID NO: 48 (CDR3), and a light chain CDR comprising the amino acid sequences of SEQ ID NO: 49 (CDR1), SEQ ID NO: 50 (CDR2) and SEQ ID NO: 56 (CDR3); x) an antigen-binding site comprising a heavy chain CDR containing SEQ ID NO: 43 (CDR1), SEQ ID NO: 46 (CDR2) and SEQ ID NO: 48 (CDR3), and a light chain CDR containing the amino acid sequences of SEQ ID NO: 49 (CDR1), SEQ ID NO: 51 (CDR2) and SEQ ID NO: 57 (CDR3); and xi) an antigen-binding site comprising a heavy chain CDR comprising SEQ ID NO: 43 (CDR1), SEQ ID NO: 47 (CDR2), and SEQ ID NO: 48 (CDR3), and a light chain CDR comprising the amino acid sequences of SEQ ID NO : 49 (CDR1), SEQ ID NO: 53 (CDR2) and SEQ ID NO: 57 (CDR3). 6. The binding molecule according to any one of claims 1-5, wherein the antigen-binding site that specifically binds to TRAILR2 is selected from the group consisting of antigen-binding sites i)-xii): i) an antigen binding site comprising a heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 82 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 83; ii) an antigen binding site comprising a heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 84 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 85; iii) an antigen binding site comprising a heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 86 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 87; iv) an antigen binding site comprising a heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 90 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 91; v) an antigen binding site comprising a heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 92 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 93; vi) an antigen binding site comprising a heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 96 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 97; vii) an antigen binding site comprising a heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 98 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 99; viii) an antigen binding site comprising a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 100 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 101; ix) an antigen binding site comprising a heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 102 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 103; x) an antigen binding site comprising a heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 104 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 105; xi) an antigen binding site comprising a heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 106 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 107; and xii) an antigen binding site comprising a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 108 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 109. 7. A binding molecule according to any one of claims 1-6, wherein the antigen-binding site that specifically binds to CDH17 is selected from the group consisting of antigen-binding sites i)-iv): i) an antigen-binding site comprising heavy chain CDRs containing the amino acid sequences of SEQ ID NO: 58 (CDR1), SEQ ID NO: 59 (CDR2) and SEQ ID NO: 60 (CDR3), and light chain CDRs containing the amino acid sequences of SEQ ID NO: 61 (CDR1), SEQ ID NO: 62 (CDR2) and SEQ ID NO: 63 (CDR3); ii) an antigen-binding site comprising a heavy chain CDR containing the amino acid sequences of SEQ ID NO: 64 (CDR1), SEQ ID NO: 65 (CDR2) and SEQ ID NO: 66 (CDR3), and CDR - 108 041360 light chain containing the amino acid sequences of SEQ ID NO: 67 (CDR1), SEQ ID NO: 68 (CDR2) and SEQ ID NO: 69 (CDR3); iii) an antigen-binding site comprising heavy chain CDRs containing the amino acid sequences of SEQ ID NO: 70 (CDR1), SEQ ID NO: 71 (CDR2) and SEQ ID NO: 72 (CDR3), and light chain CDRs containing the amino acid sequences of SEQ ID NO: 73 (CDR1), SEQ ID NO: 74 (CDR2) and SEQ ID NO: 75 (CDR3); and iv) an antigen-binding site comprising heavy chain CDRs comprising the amino acid sequences of SEQ ID NO: 76 (CDR1), SEQ ID NO: 77 (CDR2), and SEQ ID NO: 78 (CDR3), and light chain CDRs comprising the amino acid sequences of SEQ ID NO: 79 (CDR1), SEQ ID NO: 80 (CDR2) and SEQ ID NO: 81 (CDR3). 8. The binding molecule according to any one of claims 1-7, wherein the antigen-binding site that specifically binds to CDH17 is selected from the group consisting of antigen-binding sites i)-v): i) an antigen binding site comprising a heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 110 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 111; ii) an antigen binding site comprising a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 112 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 113; iii) an antigen binding site comprising a heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 114 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 115; iv) an antigen binding site comprising a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 116 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 117; v) an antigen binding site comprising a heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 118 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 119. 9. A binding molecule according to any one of claims 1 to 8, containing: (i) a heavy chain containing the amino acid sequence of SEQ ID NO: 159, or a heavy chain containing the amino acid sequence of SEQ ID NO: 160, or a heavy chain containing the amino acid sequence of SEQ ID NO: 161, or a heavy chain containing the amino acid sequence of SEQ ID NO: 163, or a heavy chain containing the amino acid sequence of SEQ ID NO: 164, or a heavy chain containing the amino acid sequence of SEQ ID NO: 166, or a heavy chain containing the amino acid sequence of SEQ ID NO: 167, or a heavy chain containing the amino acid sequence SEQ ID NO: 168, or a heavy chain containing the amino acid sequence of SEQ ID NO: 169, or a heavy chain containing the amino acid sequence of SEQ ID NO: 170, or a heavy chain containing the amino acid sequence of SEQ ID NO: 171, or a heavy chain containing the amino acid sequence of SEQ ID NO: 172; (ii) a light chain containing the amino acid sequence of SEQ ID NO: 173. 10. A binding molecule according to any one of claims 1 to 8, containing: (i) a heavy chain containing the amino acid sequence of SEQ ID NO: 174, or a heavy chain containing the amino acid sequence of SEQ ID NO: 175, or a heavy chain containing the amino acid sequence of SEQ ID NO: 176, or a heavy chain containing the amino acid sequence of SEQ ID NO: 178, or a heavy chain containing the amino acid sequence of SEQ ID NO: 179, or a heavy chain containing the amino acid sequence of SEQ ID NO: 181, or a heavy chain containing the amino acid sequence of SEQ ID NO: 183, or a heavy chain containing the amino acid sequence SEQ ID NO: 184, or a heavy chain containing the amino acid sequence of SEQ ID NO: 185, or a heavy chain containing the amino acid sequence of SEQ ID NO: 186, or a heavy chain containing the amino acid sequence of SEQ ID NO: 187; (ii) a light chain containing the amino acid sequence of SEQ ID NO: 188. 11. A binding molecule according to any one of claims 1 to 8, containing: (i) a heavy chain containing the amino acid sequence of SEQ ID NO: 189, or a heavy chain containing the amino acid sequence of SEQ ID NO: 190, or a heavy chain containing the amino acid sequence of SEQ ID NO: 191, or a heavy chain containing the amino acid sequence of SEQ ID NO: 193, or a heavy chain containing the amino acid sequence of SEQ ID NO: 194, or a heavy chain containing the amino acid sequence of SEQ ID NO: 196, or a heavy chain containing the amino acid sequence of SEQ ID NO: 197, or a heavy chain containing the amino acid sequence SEQ ID NO: 198, or a heavy chain containing the amino acid sequence of SEQ ID NO: 199, or a heavy chain containing - 109 041360 amino acid sequence of SEQ ID NO: 200, or a heavy chain containing the amino acid sequence of SEQ ID NO: 201, or a heavy chain containing the amino acid sequence of SEQ ID NO: 202; (ii) a light chain containing the amino acid sequence of SEQ ID NO: 203. 12. The binding molecule according to any one of claims 1 to 8, containing: (i) a heavy chain containing the amino acid sequence of SEQ ID NO: 204, or a heavy chain containing the amino acid sequence of SEQ ID NO: 205, or a heavy chain containing the amino acid sequence of SEQ ID NO: 206, or a heavy chain containing the amino acid sequence of SEQ ID NO: 208, or a heavy chain containing the amino acid sequence of SEQ ID NO: 209, or a heavy chain containing the amino acid sequence of SEQ ID NO: 211, or a heavy chain containing the amino acid sequence of SEQ ID NO: 212, or a heavy chain containing the amino acid sequence SEQ ID NO: 213, or a heavy chain containing the amino acid sequence of SEQ ID NO: 214, or a heavy chain containing the amino acid sequence of SEQ ID NO: 215, or a heavy chain containing the amino acid sequence of SEQ ID NO: 216, or a heavy chain containing the amino acid sequence of SEQ ID NO: 217; (ii) a light chain containing the amino acid sequence of SEQ ID NO: 218. 13. A binding molecule according to any one of claims 1 to 8, containing: (i) a heavy chain containing the amino acid sequence of SEQ ID NO: 145, or a heavy chain containing the amino acid sequence of SEQ ID NO: 147, or a heavy chain containing the amino acid sequence of SEQ ID NO: 149, or a heavy chain containing the amino acid sequence of SEQ ID NO: 151, or a heavy chain containing the amino acid sequence of SEQ ID NO: 153, or a heavy chain containing the amino acid sequence of SEQ ID NO: 155, or a heavy chain containing the amino acid sequence of SEQ ID NO: 157, or a heavy chain containing the amino acid sequence SEQ ID NO: 219, or a heavy chain containing the amino acid sequence of SEQ ID NO: 220, or a heavy chain containing the amino acid sequence of SEQ ID NO: 221, or a heavy chain containing the amino acid sequence of SEQ ID NO: 223, or a heavy chain containing amino acid sequence of SEQ ID NO: 224, or a heavy chain containing the amino acid sequence activity of SEQ ID NO: 226, or a heavy chain containing the amino acid sequence of SEQ ID NO: 227, or a heavy chain containing the amino acid sequence of SEQ ID NO: 228, or a heavy chain containing the amino acid sequence of SEQ ID NO: 229, or a heavy chain, containing the amino acid sequence of SEQ ID NO: 230, or a heavy chain containing the amino acid sequence of SEQ ID NO: 231, or a heavy chain containing the amino acid sequence of SEQ ID NO: 232; (ii) a light chain containing the amino acid sequence of SEQ ID NO: 233. 14. An isolated nucleic acid molecule encoding a heavy chain variable region and a light chain variable region of a bispecific and tetravalent binding molecule according to any one of claims 5-8. 15. An isolated nucleic acid molecule encoding the heavy chain and light chain of a bispecific and tetravalent binding molecule according to any one of claims 9-13. 16. An expression vector containing a nucleic acid molecule according to claim 14 or 15. 17. A host cell designed to obtain a bispecific and tetravalent binding molecule according to claims 1 to 13, transfected with an expression vector according to claim 16. 18. A method of obtaining a bispecific and tetravalent binding molecule according to any one of claims 1 to 13, including (a) culturing the host cell according to claim 17 under conditions that allow the expression of the molecule; and (b) isolating the molecule. 19. The method of claim 18, wherein further purification of the molecule is carried out. 20. A pharmaceutical composition containing a bispecific and tetravalent binding molecule according to any one of claims 1 to 13 and a pharmaceutically acceptable carrier. 21. A method of treating cancer expressing both TRAILR2 and CDH17, comprising administering to a patient in need thereof a therapeutically effective amount of a bispecific and tetravalent binding molecule according to any one of claims 1-13. 22. The method of claim 21, wherein the cancer is colorectal cancer (CRC), gastric cancer (GC), pancreatic cancer (PC), esophageal cancer, or a neuroendocrine tumor. 23. The use of a bispecific and tetravalent binding molecule according to any one of claims 1 to 13 for the preparation of a pharmaceutical composition for the treatment of cancer expressing both TRAILR2 and CDH17. 24. Use according to claim 23, wherein the cancer is colorectal cancer (CRC), gastric cancer (GC), pancreatic cancer (PC), esophageal cancer, or a neuroendocrine tumor. 25. Use of a bispecific and tetravalent binding molecule according to any one of claims 1-13 - 110 041360 for the treatment of cancer expressing both TRAILR2 and CDH17. 26. Use according to claim 25, wherein the cancer is colorectal cancer (CRC), gastric cancer (GC), pancreatic cancer (PC), esophageal cancer, or neuroendocrine tumor.