Dosing of immunocytokines for cancer treatment

Abstract

The invention provides an immunocytokine for use in the treatment of cancer, wherein the immunocytokine administered at a dose of 0.02 to 0.6 mg / kg. The immunocytokine comprises: an antibody or antigen binding fragment thereof binding to PD1 and an interleukin- 15 (IL-15) conjugate, wherein said IL- 15 conjugate is located N-terminal of the antibody and comprises an IL- 15 polypeptide and an IL-15Rα sushi domain polypeptide. The invention further provides a method of treating cancer comprising administration of the immunocytokine to a subject in need thereof.

Description

[0001] Attorney Docket No. 0183-0350-00PCT-KAD-ZSP ZSP ref.: 589-395 PCT

[0002] DOSING OF IMMUNOCYTOKINES FOR CANCER TREATMENT

[0003] CROSS-REFERENCE TO RELATED APPLICATIONS

[0004] This application claims priority to U.S. Provisional Application No. 63 / 807,001, filed May 16, 2025, and U.S. Provisional Application No. 63 / 749,956, filed January 27, 2025, the entire contents of which are incorporated by reference herein for all purposes.

[0005] SEQUENCE LISTING

[0006] The present application contains a Sequence Listing which has been submitted electronically in XML format. Said XML copy, created on January 22, 2026, is named “01183-0350-00PCT-KAD-ZSP” and is 31,128 bytes in size. The information in the electronic format of the sequence listing is incorporated herein by reference in its entirety.

[0007] The present invention relates generally to methods for treating cancer using an interleukin-15-based immunocytokine. More particularly, the present invention relates to advantageous doses and regimens for administration of the interleukin- 15-based immunocytokine.

[0008] BACKGROUND OF THE INVENTION

[0009] Immunocytokines comprising interleukin- 15 (IL- 15) and an antibody have emerged as promising candidates for cancer immunotherapy. IL- 15 has the ability to stimulate the proliferation and cytotoxic functions of immune effector cells, particularly T cells and natural killer (NK) cells, via binding to IL- 15 receptor a (IL-15Ra). The therapeutic effect of IL- 15 can be improved if IL-15 is provided in a complex with the sushi domain of the IL-15Ra (IL-15Ra sushi).

[0010] In an immunocytokine, a cytokine is combined with an antibody, usually in the form of a fusion protein. The combination of IL- 15 with an immunostimulatory antibody that acts on immune effector cells, such as anti-PD-1, has been found to be particularly advantageous. Depending on the format of the immunocytokine, the IL- 15 and the anti-PD-1 may act on different cells (in trans) or on the same cell (in cis).

[0011] The optimal dosing regimen for immunocytokines is a critical challenge in maximizing therapeutic efficacy while minimizing toxicity. There is thus a need for optimizing the dosage and timing of the administration of immunocytokines to achieve the best possible therapeutic outcomeAttorney Docket No. 0183-0350-00PCT-KAD-ZSP ZSP ref.: 589-395 PCT while reducing the risk for severe side effects. This patent application presents novel dosing strategies for IL- 15-based immunocytokines in cancer treatment. The inventors provide advantageous dosing regimens for an immunocytokine comprising IL-15, IL-15Ra sushi and anti-PD-1, resulting in efficient anti-cancer responses while exhibiting acceptable side effects. Thus, a solution to the problem stated above is provided by the subject-matter of the independent claims.

[0012] SUMMARY OF THE INVENTION

[0013] In a first aspect, the present invention relates to an immunocytokine for use in the treatment of cancer, wherein the immunocytokine comprises:

[0014] a) an antibody or antigen binding fragment thereof binding to PD1 comprising a heavy chain variable domain (VH) comprising complementarity determining regions HCDR1 to HCDR3 as set forth in SEQ ID NO: 3 to 5, respectively, and a light chain variable domain (VL) comprising complementarity determining regions LCDR1 to LCDR3 as set forth in SEQ ID NO: 6 to 8, respectively; and

[0015] b) an interleukin- 15 (IL- 15) conjugate, wherein said IL- 15 conjugate is located N-terminal of VH and comprises: (i) an IL-15 polypeptide comprising an amino acid sequence as set forth in SEQ ID NO: 9 or an amino acid sequence having at least 80 % identity to SEQ ID NO: 9; and (ii) an IL-15Ra sushi domain polypeptide comprising an amino acid sequence as set forth in SEQ ID NO: 11; and (iii) optionally a polypeptide linker LI linking (i) and (ii); and c) optionally a polypeptide linker L2 linking a) and b);

[0016] characterized in that the immunocytokine is administered at a dose of 0.02 to 0.6 mg / kg.

[0017] In a second aspect, the present invention relates to method of treating cancer, comprising administration of an immunocytokine to a human subject in need thereof, wherein the immunocytokine comprises:

[0018] d) an antibody or antigen binding fragment thereof binding to PD1 comprising a heavy chain variable domain (VH) comprising complementarity determining regions HCDR1 to HCDR3 as set forth in SEQ ID NO: 3 to 5, respectively, and a light chain variable domain (VL) comprising complementarity determining regions LCDR1 to LCDR3 as set forth in SEQ ID NO: 6 to 8, respectively; and

[0019] e) an interleukin- 15 (IL- 15) conjugate, wherein said IL- 15 conjugate is located N-terminal of VH and comprises: (i) an IL-15 polypeptide comprising an amino acid sequence as set forth in SEQ ID NO: 9 or an amino acid sequence having at least 80 % identity to SEQ ID NO: 9;Attorney Docket No. 0183-0350-00PCT-KAD-ZSP ZSP ref.: 589-395 PCT and (ii) an IL-15Ra sushi domain polypeptide comprising an amino acid sequence as set forth in SEQ ID NO: 11; and (iii) optionally a polypeptide linker LI linking (i) and (ii); and f) optionally a polypeptide linker L2 linking a) and b);

[0020] characterized in that the immunocytokine is administered at a dose of 0.02 to 0.6 mg / kg.

[0021] DETAILED DESCRIPTION

[0022] Before the present invention is described in detail below, it is to be understood that this invention is not limited to the particular methodology, protocols and reagents described herein as these may vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to limit the scope of the present invention, which will be limited only by the appended claims. Unless defined otherwise, all technical and scientific terms used herein have the same meanings as commonly understood by one of ordinary skill in the art. Preferably, the terms used herein are defined as described in “A multilingual glossary of biotechnological terms: (IUPAC Recommendations)”, Leuenberger, H.G.W, Nagel, B. and Kolbl, H. eds. (1995), Helvetica Chimica Acta, CH-4010 Basel, Switzerland).

[0023] Several documents are cited throughout the text of this specification. Each of the documents cited herein (including all patents, patent applications, scientific publications, manufacturer's specifications, instructions, etc.), whether supra or infra, are hereby incorporated by reference in their entirety. Nothing herein is to be construed as an admission that the invention is not entitled to antedate such disclosure by virtue of prior invention. Some of the documents cited herein are characterized as being “incorporated by reference". In the event of a conflict between the definitions or teachings of such incorporated references and definitions or teachings recited in the present specification, the text of the present specification takes precedence.

[0024] To practice the present invention, unless otherwise indicated, conventional methods of chemistry, biochemistry, and recombinant DNA techniques are employed which are explained in the literature in the field (cf., e.g., Molecular Cloning: A Laboratory Manual, 2ndEdition, J. Sambrook et al. eds., Cold Spring Harbor Laboratory Press, Cold Spring Harbor 1989).

[0025] Throughout this specification and the claims which follow, unless the context requires otherwise, the words “comprising”, “including”, “having” or grammatical variants thereof, will be understood to imply the inclusion of a stated integer or step or group of integers or steps but not the exclusion of any other integer or step or group of integers or steps. The terms “comprising” / “including” / “having” encompass the preferred embodiments “consisting of’ orAttorney Docket No. 0183-0350-00PCT-KAD-ZSP ZSP ref.: 589-395 PCT “consisting essentially of’. Thus, whenever the terms “comprising’7“including’7“having” are used herein, they can be replaced by “consisting essentially of’ or, more preferably, by “consisting of’.

[0026] As used in this specification and the appended claims, the singular forms “a”, “an”, and “the” include plural referents, unless the content clearly dictates otherwise.

[0027] The term “about” when used in connection with a numerical value is meant to encompass numerical values within a range having a lower limit that is 10% smaller than the indicated numerical value and having an upper limit that is 10% larger than the indicated numerical value (each limit preferably 5%, 4%, 3%, 2% or 1% instead of 10%).

[0028] In the following, the elements of the present invention will be described. These elements are listed with specific embodiments, however, it should be understood that they may be combined in any manner and in any number to create additional embodiments. The variously described examples and preferred embodiments should not be construed to limit the present invention to only the explicitly described embodiments. This description should be understood to support and encompass embodiments, which combine the explicitly described embodiments with any number of the disclosed and / or preferred elements. Furthermore, any permutations and combinations of all described elements in this application should be considered disclosed by the description of the present application unless the context indicates otherwise.

[0029] In a first aspect, the present invention relates to an immunocytokine for use in the treatment of cancer, wherein the immunocytokine comprises:

[0030] a) an antibody or antigen binding fragment thereof binding to PD-1 comprising a heavy chain variable domain (VH) comprising complementarity determining regions HCDR1 to HCDR3 as set forth in SEQ ID NO: 3 to 5, respectively, and a light chain variable domain (VL) comprising complementarity determining regions LCDR1 to LCDR3 as set forth in SEQ ID NO: 6 to 8, respectively; and

[0031] b) an interleukin- 15 (IL- 15) conjugate, wherein said IL- 15 conjugate is located N-terminal of VH and comprises (i) an IL- 15 polypeptide comprising an amino acid sequence as set forth in SEQ ID NO: 9 or an amino acid sequence having at least 80 % identity to SEQ ID NO: 9; and (ii) an IL-15Ra sushi domain polypeptide comprising an amino acid sequence as set forth in SEQ ID NO: 11; and (iii) optionally a polypeptide linker LI linking (i) and (ii); and c) optionally a polypeptide linker L2 linking a) and b);

[0032] characterized in that the immunocytokine is administered at a dose of 0.02 to 0.6 mg / kg.Attorney Docket No. 0183-0350-00PCT-KAD-ZSP ZSP ref.: 589-395 PCT Interleukin- 15 (IL- 15)

[0033] IL-15 is a 14-15 kDa cytokine that is secreted by monocytes, dendritic cells (DCs), epithelial cells, bone marrow stromal cells and fibroblasts. IL- 15 regulates T and NK cell activation and proliferation, and the number of CD8+ memory cells is affected by a balance between IL- 15 and IL-2. As used herein, the term “IL- 15 polypeptide” includes IL- 15, preferably mammalian IL-15, more preferably primate IL-15, most preferably human IL-15, and derivatives thereof having at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, or at least 90% of the activity of human IL- 15, and preferably not more than 100% of the activity of human IL-15. In some preferred embodiments, IL- 15 polypeptide has a reduced activity compared to the activity of human IL-15. Such derivatives can be obtained through substitutions of amino acids and the reduced activity checked by means known in the art. Techniques for making such derivatives or variants are known in the art. The human mature IL-15 amino acids sequence is provided as SEQ ID NO: 9.

[0034] Thus, in some embodiments, the IL- 15 polypeptide comprises an amino acid sequence as set forth in SEQ ID NO: 9. In some embodiments, the IL-15 polypeptide, in particular having a reduced activity as described above, comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 98% or at least 99% identity to SEQ ID NO: 9. It is envisioned that the IL-15 polypeptide may comprise an amino acid sequence comprising one or more, in particular 1-5, preferably 1, 2, or 3 amino acid mutations compared to SEQ ID NO: 9. An amino acid mutation may be selected from an insertion, a deletion and a substitution, preferably a substitution. In some embodiments, the amino acid substitutions are located at positions 1, 4, 8, 30, 45, 61, 64, 65, and / or 108 of SEQ ID NO: 9. In some embodiments, the amino acid substitutions are located at position 45, 65, and / or 108 of SEQ ID NO: 9. In some embodiments the amino acid substitutions in SEQ ID NO: 9 are N1D, N4D, D8N, D30N, D61N, E64Q, L45A, N65S / A / D / K, and / or Q108S / E. In some embodiments, the amino acid substitutions in SEQ ID NO: 9 are L45A, N65S / A / D / K, and / or Q108S. In some embodiments, the IL-15 polypeptide comprises an amino acid sequence comprising a substitution at position 65 of SEQ ID NO: 9, in particular N65 A or N65S, preferably N65S. In some embodiments, theIL-15 polypeptide comprises an amino acid sequence comprising a substitution at position 65 and at position 78 and / or 79 of SEQ ID NO: 9. The IL-15 polypeptide may comprise an amino acid sequence comprising substitutions at positions 65, 78 and 79 of SEQ ID NO: 9, in particular N65S, G78A and N79Q.Attorney Docket No. 0183-0350-00PCT-KAD-ZSP ZSP ref.: 589-395 PCT The IL-15 polypeptide may comprise an amino acid sequence as set forth in SEQ ID NO: 10 or 31. Preferably, the IL- 15 polypeptide comprises an amino acid sequence as set forth in SEQ ID NO: 10.

[0035] IL- 15 conjugate

[0036] The immunocytokine for use of the present invention comprises an IL- 15 conjugate, comprising the IL- 15 polypeptide and an IL-15Ra sushi domain polypeptide.

[0037] The IL-15 receptor complex specifically binds IL-15 with high affinity and consists of a unique interleukin 15 receptor a subunit (IL-15Ra), IL-2 / IL-15RP, and the common y-chain / IL-2Ry subunit. IL-15Ra is expressed by mitogen-activated macrophages, NK cells, and CD4+ and CD8+ T cells. As used herein, the term “sushi domain” of IL-15Ra refers to a domain of IL-15Ra beginning at the first cysteine residue (Cl) after the signal peptide of IL-15Ra and ending at the fourth cysteine residue (C4) after said signal peptide. The sushi domain corresponds to a portion of the extracellular region of IL-15Ra and is involved in binding to IL-15.

[0038] The IL-15Ra sushi domain polypeptide of the immunocytokine for use of the present invention comprises an amino acid sequence as set forth in SEQ ID NO: 11. It is also envisioned that the IL-15Ra sushi domain polypeptide may comprise an amino acid sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 98% or at least 99% identity to SEQ ID NO: 11, wherein the IL-15Ra sushi domain polypeptide has at least 95%, 96%, 97%, 98%, 99% or 100% of the binding activity to IL- 15 compared to the IL-15Ra sushi domain polypeptide comprising the amino acid sequence as set forth in SEQ ID NO: 11. It is envisioned that the IL-15Ra sushi domain polypeptide may comprise an amino acid sequence comprising one or more, in particular 1-5, preferably 1, 2, or 3 amino acid mutations compared to SEQ ID NO: 11. In some embodiments, the IL-15Ra sushi domain polypeptide further comprises an amino acid sequence as set forth in SEQ ID NO: 26. Preferably, the IL-15Ra sushi domain polypeptide comprises or consists of the amino acid sequence set forth in SEQ ID NO 27 or 28.

[0039] Within the IL-15 conjugate, the IL-15Ra sushi domain polypeptide may be located N-terminal or C-terminal of the IL-15 polypeptide. Preferably, the IL-15Ra sushi domain polypeptide is located N-terminal of the IL- 15 polypeptide.

[0040] In some embodiments, the IL- 15 conjugate comprises a polypeptide linker LI. It is envisioned that LI is a serine-glycine linker, i.e. a polypeptide linker comprising predominantly serines and glycines. In some embodiments, LI comprises at least 80%, at least 85%, at least 95% or 100% serines and glycines. In some embodiments, LI has a length of 1-40 amino acids,Attorney Docket No. 0183-0350-00PCT-KAD-ZSP ZSP ref.: 589-395 PCT preferably 5-30 amino acids, more preferably 15-25 amino acids. LI may comprise or consist of an amino acid as set forth in SEQ ID NO: 19-25, preferably SEQ ID NO: 19 or 20.

[0041] In some embodiments, the immunocytokine for use of the present invention comprises an IL-15 conjugate comprising an amino acid sequence as set forth in SEQ ID NO: 12 or 29, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 98% or at least 99% identity to SEQ ID NO: 12 or 29. Preferably, the immunocytokine for use of the present invention comprises an IL- 15 conjugate comprising an amino acid sequence as set forth in SEQ ID NO: 12.

[0042] Anti-PD-1 antibody

[0043] The immunocytokine for use of the present invention comprises an antibody or antigen binding fragment thereof binding to PD-1, comprising a VH comprising HCDR1 to HCDR3 as set forth in SEQ ID NO: 3 to 5, respectively, and a VL comprising LCDR1 to LCDR3 as set forth in SEQ ID NO: 6 to 8, respectively.

[0044] The term “antibody” is used herein to refer to an immunoglobulin molecule comprising an antigen-binding site capable of specifically binding to an antigen. An antibody is a Y-shaped molecule comprising four polypeptide chains: two heavy chains (HC) and two light chains (LC), wherein the heavy chains are linked to each other by disulfide bonds and each heavy chain is linked to a light chain by a disulfide bond. There are five main heavy chain classes (or isotypes) which determine the functional activity of an antibody molecule: IgM, IgD, IgG, IgA and IgE. When referring to IgG in general, IgGl, IgG2, IgG3 and IgG4 are included, unless defined otherwise. Each light chain consists of two domains, the N-terminal domain being known as the variable or VL domain and the C-terminal domain being known as the constant (or CL) domain (constant kappa (CK) or constant lambda (Ck) domain). Each heavy chain includes four or five domains depending on the antibody isotype: a variable domain (VH) followed by the first constant domain (CHI), the hinge region, and then constant domains CH2 and CH3, and in some isotypes CH4. The constant domains confer important biological properties such as antibody chain association, secretion, trans-placental mobility, complement binding, and binding to Fcreceptors (FCR). In an assembled antibody, the VL and VH domains associate to form an antigen binding domain. Also, the CL and CHI domains associate to keep one heavy chain associated with one light chain. The two heavy-light chains associate by interaction of the CH2 and CH3 domains and interaction between the hinge regions of the two heavy chains.

[0045] The term “antigen-binding fragment” of an antibody as used herein, refers to a fragment of an antibody that retains the ability of the antibody to specifically bind to an antigen. As used herein,Attorney Docket No. 0183-0350-00PCT-KAD-ZSP ZSP ref.: 589-395 PCT the term “antigen-binding fragment” includes Fv, Fab, F(ab')2, Fab', dsFv, (dsFv)2, scFv and sc(Fv)2.

[0046] By “specific binding” is meant that the binding is selective for the antigen and can be discriminated from unwanted or non-specific interactions. In certain embodiments, an antibody is said to specifically bind an antigen when it preferentially recognizes its target antigen in a complex mixture of proteins and / or macromolecules. The ability of an antigen binding molecule to bind to a specific antigen can be measured either through an enzyme-linked immunosorbent assay (ELISA) or other techniques familiar to one of skill in the art, e.g. Surface Plasmon Resonance (SPR) technique. As such, the specified ligand binds to its target molecule and does not bind in a substantial amount to other molecules present. Generally, an antibody that “specifically binds” a target molecule has an equilibrium dissociation constant smaller than about IO’5(e.g., 10'6, 10'7, 10-8, IQ-9, JO-10, 10'11, or 10'12) M for that target molecule.

[0047] In some embodiments, the antibody or antigen binding fragment thereof comprises a VH as set forth in SEQ ID NO: 1 and a VL as set forth in SEQ ID NO: 2.

[0048] As used herein, VH and VL refer to the antibody variable domains of the heavy chain (HC) and light chain (LC), respectively. A “variable domain” or “antibody variable domain” as used herein refers to each of a pair of domains forming an antigen binding domain of an antibody. Each variable domain comprises four framework (FR) regions whose sequences are widely conserved, connected by three hypervariable regions (or complementarity determining regions, CDRs). The framework regions form a “core” P-sheet structure displaying the CDRs on the surface of the variable domain.

[0049] As used herein, the term “Complementarity Determining Regions” (CDRs) refers to portions of an antibody variable domain that are (typically) involved in antigen binding. The six CDRs present in each antigen-binding domain are short sequences of amino acids that are specifically positioned to form the antigen-binding domain. The length and composition of the CDR sequences are variable, especially in the CDR3. The remainder of the amino acids located in the antigen-binding domains or the “framework” regions, show less variability. The antigenbinding domain formed by the positioned CDRs defines a surface complementary to the epitope on the immunoreactive antigen. This complementary surface promotes the noncovalent binding of the antibody to its cognate epitope.

[0050] Herein, the CDRs of the antibody or antigen binding fragment thereof binding to PD-1 are defined according to Kabat nomenclature (Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md. (1987, 1991)).Attorney Docket No. 0183-0350-00PCT-KAD-ZSP ZSP ref.: 589-395 PCT The anti-PD-1 antibody or antigen binding-fragment thereof comprised in the immunocytokine for use of the invention binds specifically to PD-1. The term “PD-1” refers to programmed cell death 1 protein 1 (also known as CD279), a cell surface receptor on T cells and B cells that has a role in regulating the immune system's response to cells of the human body by down-regulating the immune system and promoting self-tolerance by suppressing T cell inflammatory activity. The amino acid sequence of full-length PD-1 is provided in GenBank as accession number NP_ 005009.2.

[0051] In a particular embodiment, the immunocytokine for use of the present invention comprises an anti-PD-1 antibody (i.e. a 4-chain antibody). The anti-PD-1 antibody may be any class of antibody, such as IgG, IgA, or IgM (or a subclass thereof), and the antibody need not be of any particular class, and any of the immunoglobulin molecules comprising the antigen recognition site of the required specificity, other modified configurations (including glycosylation variants of antibodies, amino acid sequence variants of antibodies, and covalently modified antibodies) can be encompassed. Modified versions of each of these classes and isotypes are known to a person skilled in the art.

[0052] Preferably, the anti-PD-1 antibody belongs to the IgG class of immunoglobulin molecules. A standard IgG immunoglobulin molecule comprises two identical light chain polypeptides, and two identical heavy chain polypeptides. The anti-PD-1 antibody may be an IgGl, IgG2, IgG3 or IgG4 antibody, preferably an IgGl antibody.

[0053] The HC of an IgG antibody comprises a variable domain (VH) and three constant domains (CHI, CH2, CH3). CH2 and CH3 together are also referred to as “Fc domain” herein. CHI, CH2 and CH3 together are also referred to as “antibody constant region” herein.

[0054] In some embodiments, the antibody comprises a first heavy chain (HC1), a second heavy chain (HC2), and two identical light chains (LC), wherein HC1 and HC2 are not identical.

[0055] The Fc domain of the anti-PD-1 antibody comprised in the immunocytokine may be a Fc domain variant comprising one or more of the amino acid substitutions described herein below.

[0056] In some embodiments, the anti-PD-1 antibody, in particular HC1 and HC2, comprise LALA mutations L234A and L235A. These substitutions reduce binding to the IgG Fc receptors FcyRI, FcyRII and FcyRIII as well as to complement component Clq. Such substitutions are useful where binding and activation of Fc receptors is undesirable.

[0057] In some embodiments, the anti-PD-1 antibody in particular HC1 andHC2, comprise “knob-into-hole (kiH)” mutations. The “knob-into-hole” technology refers to mutations in the CH3 interface of a dimeric Fc domain to create a “knob” on one CH3 domain and a “hole” on the other CH3 domain to promote heterodimer formation. By way of non-limiting example, particularAttorney Docket No. 0183-0350-00PCT-KAD-ZSP ZSP ref.: 589-395 PCT “knob-into-hole” mutations are Y349C, T366S, L368A, Y407V, S354C, T366W, and Y407T. Preferably, the anti-PD-1 antibody comprises at least one, in particular all, of the “knob-into-hole” Y349C, T366S, L368A, Y407V, S354C and T366W mutations. It is envisioned that HC1 comprises S354C and T366W mutations and HC2 comprises Y349C, T366S, L368A and Y407V mutations.

[0058] In some cases artificially introduced cysteine bonds may (further) improve the stability of the heterodimer, optimally without interfering with the binding characteristics of the antibody. Such cysteine bonds can further improve heterodimerization.

[0059] The skilled person is readily capable of determining whether the above-described mutations / substitutions are present in the disclosed sequences.

[0060] The anti-PD-1 antibody may comprise polypeptide chains comprising a constant region as set forth in SEQ ID NO: 13, 14 and / or 18, in particular LC constant regions as set forth in SEQ ID NO: 18 and HC constant regions as set forth in SEQ ID NO: 13 and / or 14.

[0061] Preferably, the antibody comprises a HC1 comprising a constant region as set forth in SEQ ID NO: 13, a HC2 comprising a constant region as set forth in SEQ ID NO: 14, and two LC each comprising a constant region as set forth in SEQ ID NO: 18.

[0062] Immunocytokine

[0063] In the immunocytokine for use of the present invention, the IL-15 conjugate and the anti-PD-1 antibody or antigen binding fragment thereof are covalently linked. The covalent linkage may be either directly via a peptide bond or indirectly via a polypeptide linker.

[0064] If not specified otherwise, the binding proteins of the present disclosure are oriented with the amino terminus (“N-terminal end” or “N-term”) on the left-hand side and the carboxylterminus (“C-terminal end” or “C-term”) on the right-hand side, in accordance with standard usage and convention.

[0065] In the immunocytokine for use of the present invention, the IL-15 conjugate is located N-terminal of the VH of the anti-PD-1 antibody or antigen binding fragment thereof. Thus, the IL-15 conjugate is covalently linked to the N-terminus of the heavy chain variable domain of the anti-PD-1 antibody or antigen binding fragment thereof.

[0066] It is preferred that the immunocytokine comprises only one IL-15 conjugate. Thus, the IL-15 conjugate is linked to the N-terminus of one and only one heavy chain of the anti-PD-1 antibody.

[0067] In some embodiments, the immunocytokine comprises a polypeptide linker L2. It is envisioned that L2 is a serine-glycine linker, i.e. a polypeptide linker comprising predominantlyAttorney Docket No. 0183-0350-00PCT-KAD-ZSP ZSP ref.: 589-395 PCT serines and glycines. In some embodiments, L2 comprises at least 80%, at least 85%, at least 95% or 100% serines and glycines. In some embodiments, L2 has a length of 1-40 amino acids, preferably 10-40 amino acids, more preferably 25-35 amino acids. LI may comprise or consist of an amino acid as set forth in SEQ ID NO: 19-25, preferably SEQ ID NO: 22.

[0068] The immunocytokine for use of the present invention may comprise polypeptide chains comprising or consisting of SEQ ID NO: 15, 16 and / or 17. In some embodiments, the immunocytokine for use of the present invention consists of two LC each consisting of an amino acid sequence as set forth in SEQ ID NO: 17, HC1 consisting of an amino acid sequence as set forth in SEQ ID NO: 15 and HC2 consisting of an amino acid sequence as set forth in SEQ ID NO: 16. The amino acid sequence of SEQ ID NO: 16 comprises the second heavy chain of the anti-PD-1 antibody N-terminally linked via polypeptide linker L2 to the IL- 15 conjugate.

[0069] Dosing and administration

[0070] In some embodiments of the immunocytokine for use, the use comprises administering the immunocytokine at a dose of at least 0.02, 0.036, 0.07, 0.1 or 0.14 mg / kg. In some embodiments, the use comprises administering the immunocytokine at a dose of no more than 0.57, 0.43 or 0.28 mg / kg or no more than 0.43 or 0.28 mg / kg. In some embodiments, the use comprises administering the immunocytokine at a dose of at least 0.02, 0.036, 0.07, 0.1 or 0.14 mg / kg, and at a dose of no more than 0.57, 0.43 or 0.28 mg / kg or no more than 0.43 or 0.28 mg / kg. It is preferred that the immunocytokine is administered at a dose between about 0.1 mg / kg and about 0.5 mg / kg, preferably at a dose from 0.14 mg / kg to 0.43 mg / kg, or preferably from 0.14 mg / kg to about 0.28 mg / kg or from 0.28 mg / kg to 0.43 mg / kg.

[0071] It is envisioned that the immunocytokine is administered at a dose of about 0.1 mg / kg. It is envisioned that the immunocytokine is administered at a dose of about 0.14 mg / kg. It is envisioned that the immunocytokine is administered at a dose of about 0.28 mg / kg. It is envisioned that the immunocytokine is administered at a dose of about 0.43 mg / kg. It is envisioned that the immunocytokine is administered at a dose of about 0.5 mg / kg. Preferably, the immunocytokine is administered at a dose of 0.14, 0.28 or 0.43 mg / kg.

[0072] In some embodiments, the immunocytokine is administered at a fixed dose. In particular, a fixed dose can be determined assuming an average body weight of about 70 kg.

[0073] Thus, in some embodiments, the use comprises administering the immunocytokine at a dose of 1.4 to 40 mg. In some embodiments, the use comprises administering the immunocytokine at a dose from 5 to 30 or 35 mg. The immunocytokine may be administered at a dose of at least 1.4, 2.5, 5, 7.5 or 10 mg. The immunocytokine may be administered at a dose of no more than 30Attorney Docket No. 0183-0350-00PCT-KAD-ZSP ZSP ref.: 589-395 PCT or 20 mg. The immunocytokine may be administered at a dose of at least 1.4, 2.5, 5, 7.5 or 10 mg and no more than 30 or 20 mg. It is preferred that the immunocytokine is administered at a dose from about 10 mg to about 30 mg, preferably at a dose from about 10 mg to about 20 mg or preferably at a dose from about 20 mg to about 30 mg.

[0074] It is envisioned that the immunocytokine is administered at a dose of about 7.5 mg. It is envisioned that the immunocytokine is administered at a dose of about 10 mg. It is envisioned that the immunocytokine is administered at a dose of about 20 mg. It is envisioned that the immunocytokine is administered at a dose of about 30 mg. It is envisioned that the immunocytokine is administered at a dose of about 35 mg. Preferably, the immunocytokine is administered at a dose of about 10, 20 or 30 mg.

[0075] The immunocytokine for use of the present invention may be administered weekly (referred to as QW herein) or bi-weekly (referred to as Q2W herein). The preferred administration is biweekly.

[0076] In some embodiments, the immunocytokine for use of the present invention is administered bi-weekly at a dose of at least 0.02, 0.036, 0.07, 0.1 or 0.14 mg / kg, and at a dose of no more than 0.57, 0.43 or 0.28 mg / kg or no more than 0.43 or 0.28 mg / kg. In some embodiments, the immunocytokine is administered bi-weekly at a dose between about 0.1 mg / kg and about 0.5 mg / kg, preferably at a dose from 0.14 mg / kg to 0.43 mg / kg, or preferably from 0.14 mg / kg to about 0.28 mg / kg or from 0.28 mg / kg to 0.43 mg / kg. In some embodiments, the immunocytokine for use of the present invention is administered bi-weekly at a dose of 0.02 mg / kg. In some embodiments, the immunocytokine for use of the present invention is administered bi-weekly at a dose of 0.07 mg / kg. In some embodiments, the immunocytokine for use of the present invention is administered bi-weekly at a dose of 0.14 mg / kg. In some embodiments, the immunocytokine for use of the present invention is administered bi-weekly at a dose of 0.28 mg / kg. In some embodiments, the immunocytokine for use of the present invention is administered bi-weekly at a dose of 0.43 mg / kg. In some embodiments, the immunocytokine for use of the present invention is administered bi-weekly at a dose of 0.5 mg / kg. In some embodiments, the immunocytokine for use of the present invention is administered bi-weekly at a dose of 0.57 mg / kg.

[0077] In some embodiments, the immunocytokine for use of the present invention is administered weekly at a dose of 0.036 mg / kg. In some embodiments, the immunocytokine for use of the present invention is administered weekly at a dose of 0.07 mg / kg. In some embodiments, the immunocytokine for use of the present invention is administered weekly at a dose of 0.1 mg / kg.

[0078] In some embodiments, the immunocytokine for use of the present invention is administered bi-weekly at a dose of at least 1.4, 2.5, 5, 7.5 or 10 mg and no more than 30 or 20 mg. In someAttorney Docket No. 0183-0350-00PCT-KAD-ZSP ZSP ref.: 589-395 PCT embodiments, the immunocytokine for use of the present invention is administered bi-weekly at a dose from about 10 mg to about 30 mg, preferably at a dose from about 10 mg to about 20 mg or preferably at a dose from about 20 mg to about 30 mg. In some embodiments, the immunocytokine for use of the present invention is administered bi-weekly at a dose of 1.4 mg. In some embodiments, the immunocytokine for use of the present invention is administered bi-weekly at a dose of 5 mg. In some embodiments, the immunocytokine for use of the present invention is administered bi-weekly at a dose of 10 mg. In some embodiments, the immunocytokine for use of the present invention is administered bi-weekly at a dose of 20 mg. In some embodiments, the immunocytokine for use of the present invention is administered bi-weekly at a dose of 30 mg. In some embodiments, the immunocytokine for use of the present invention is administered biweekly at a dose of 35 mg. In some embodiments, the immunocytokine for use of the present invention is administered bi-weekly at a dose of 40 mg.

[0079] In some embodiments, the immunocytokine for use of the present invention is administered weekly at a dose of 2.5 mg. In some embodiments, the immunocytokine for use of the present invention is administered weekly at a dose of 5 mg. In some embodiments, the immunocytokine for use of the present invention is administered weekly at a dose of 7.5 mg.

[0080] All doses described herein may be used in the above schemes and regimens.

[0081] The immunocytokine for use of the present invention is administered intravenously.

[0082] Indications

[0083] The immunocytokine for use of the present invention is provided for the treatment of cancer.

[0084] In some embodiments, the cancer is selected from the group consisting of non-small cell lung cancer, hepatocellular carcinoma, gastric cancer, gastroesophageal junction adenocarcinoma, sigmoid adenocarcinoma, cutaneous squamous cell carcinoma, colorectal cancer, rectal adenocarcinoma, ascending colon cancer, penile squamous cell carcinoma, sarcomatoid lung cancer, urothelial carcinoma, uveal melanoma, squamous cell carcinoma of the head and neck, squamous cell carcinoma of the scalp, gastric adenocarcinoma, myxofibrosarcoma, salivary gland acinic cell carcinoma, and soft tissue sarcoma.

[0085] In preferred embodiments, the cancer is non-small cell lung cancer (NSCLC) or colorectal cancer (CRC), in particular metastatic NSCLC or metastatic CRC. In most preferred embodiments, the cancer is CRC, in particular metastatic CRC.

[0086] In some embodiments, the gastric cancer and the gastroesophageal junction adenocarcinoma has non-MSI-H status, proficient MMR status or HER2 / neu negative status.Attorney Docket No. 0183-0350-00PCT-KAD-ZSP ZSP ref.: 589-395 PCT In some embodiments, the cancer is unresectable. In some embodiments, the cancer is metastatic.

[0087] It is envisioned that the use comprises administering the immunocytokine to a subject that is refractory to standard approved therapy. It is also envisioned that the use comprises administering the immunocytokine to a subject that is not a candidate for standard approved therapy.

[0088] Combination therapy

[0089] In some embodiments the immunocytokine for use according to the invention, the use further comprises administration of a second anti-cancer drug.

[0090] In some embodiments the immunocytokine for use according to the invention, the use further comprises administration of a chemotherapeutic agent.

[0091] As used herein, the term “chemotherapeutic agent” refers to a compound that is capable of inhibiting, reducing, preventing, or treating the growth, proliferation, survival, or spread of cancerous or pre-cancerous cells in a subject. Chemotherapeutic agents include, without limitation, small-molecule cytotoxic or cytostatic compounds, alkylating agents, antimetabolites, microtubule inhibitors, topoisomerase inhibitors, hormonal agents, targeted therapies, and biological agents. In certain embodiments, the chemotherapeutic agent comprises a therapeutic antibody, including monoclonal, chimeric, humanized, or fully human antibodies, or antigen-binding fragments thereof, that specifically bind to a tumor-associated antigen or an immune regulatory target. Preferably, the chemotherapeutic agent is a therapeutic antibody.

[0092] In some embodiments, the chemotherapeutic agent is an inhibitor of epidermal growth factor receptor (EGFR), preferably an antagonistic EGFR antibody. It is envisioned that the antagonistic EGFR antibody is selected from cetuximab, matuzumab, panitumumab and necitumumab. Preferably, the antagonistic EGFR antibody is cetuximab. It is preferred that the EGFR antibody, in particular cetuximab, is administered intravenously, most preferably infused over 120 minutes. Preferably, the EGFR antibody, in particular cetuximab, is administered at a concentration of 500mg / m2. Preferably, the EGFR antibody, in particular cetuximab, is administered biweekly. In particularly preferred embodiments, the EGFR antibody, in particular cetuximab, is administered biweekly intravenously at a concentration of 500mg / m2infused over 120 minutes.

[0093] It is preferred that the second anti-cancer drug is a therapeutic antibody. The therapeutic antibody is preferably a monoclonal antibody. In some embodiments, the therapeutic antibody is directed against EGFR, cytotoxic T-lymphocyte-associated protein 4 (anti CTLA-4), vascularAttorney Docket No. 0183-0350-00PCT-KAD-ZSP ZSP ref.: 589-395 PCT endothelial growth factor (VEGF) or programmed cell death 1 (PD-1). Preferably, the therapeutic antibody is an antagonistic antibody. The therapeutic antibody is preferably administered intravenously.

[0094] It is envisioned that the antagonistic VEGF antibody is selected from bevacizumab, ranibizumab, aflibercept and ramucirumab. Preferably, the antagonistic VEGF antibody is bevacizumab. It is envisioned that the antagonistic PD-1 antibody is selected from nivolumab, pembrolizumab, cemiplimab and dostarlimab. Preferably, the antagonistic PD-1 antibody is nivolumab. It is envisioned that the antagonistic CTLA-4 antibody is selected from ADG126 (also referred to as Muzastotug), ipilimumab and tremelimumab. Preferably, the antagonistic CTLA-4 antibody is ipilimumab or ADG126, more preferably ADG126. Accordingly, the invention relates to the herein described immunocytokine for use in the treatment of a cancer, wherein said immunocytokine is administered in combination with a chemotherapeutic agent. In other words, the invention relates to the described immunocytokine for use in the treatment of cancer, wherein a chemotherapeutic agent is administered before, concomitantly, simultaneously or after administration of the immunocytokine. More preferably, the invention relates to the herein described immunocytokine for use in the treatment of a cancer, wherein said immunocytokine is administered in combination with a therapeutic antibody. In other words, the invention relates to the described immunocytokine for use in the treatment of cancer, wherein a therapeutic antibody is administered before, concomitantly, simultaneously or after administration of the immunocytokine.

[0095] “Co-administration”, “administered in combination”, “administration of a combination”, “co-administration of a combination”, “combined therapy” and / or “combined treatment regimen” is used herein in the broadest sense and refers to at least two therapeutically active drugs or compositions which may be administered or co-administered, simultaneously, in either separate or combined formulations, or sequentially at different times separated by minutes, hours or days, but in some way act together to provide the desired therapeutic response. Accordingly, the described immunocytokine and the chemotherapeutic agent or therapeutic antibody may be administered or co-administered, simultaneously, in either separate or combined formulations, or sequentially at different times separated by minutes, hours or days.

[0096] Simultaneous administration means that the therapeutically active agents can be administered at the same time. This can occur either by combining them into a single formulation / composition or by administering them separately but concurrently or concomitantly.

[0097] The described immunocytokine and the chemotherapeutic agent or therapeutic antibody may be administered in separate formulations which means that the active agents may be providedAttorney Docket No. 0183-0350-00PCT-KAD-ZSP ZSP ref.: 589-395 PCT in different formulations / compositions, meaning each drug or composition retains its own distinct form. Even though they are administered together or in close succession, they are not physically combined into one dosage form (e.g. pharmaceutical composition).

[0098] Sequential administration means that the described immunocytokine and the chemotherapeutic agent or therapeutic antibody be administered at different times. This could mean administering them minutes, hours or even days apart. Despite this staggered timing, the agents are considered part of a combined therapy because they interact or work in concert to achieve the desired therapeutic effect.

[0099] The skilled person knows that the key aspect of these combined approaches is that, regardless of the timing or method of delivery, the active agents may act in synergy or complement each other in a way that contributes to the intended therapeutic outcome.

[0100] As mentioned above it is envisaged that the e.g. described immunocytokine and the chemotherapeutic agent or therapeutic antibody may be provided in different formulations / compositions. Accordingly, the invention relates to the described immunocytokine for use in the treatment of cancer, wherein the immunocytokine is administered in combination with a chemotherapeutic agent or therapeutic antibody and wherein the immunocytokine is formulated in a first pharmaceutical composition and the chemotherapeutic agent or therapeutic antibody is formulated in a second pharmaceutical composition.

[0101] It is envisaged that the chemotherapeutic agent or therapeutic antibody is administered every two weeks. It is envisaged that the chemotherapeutic agent or therapeutic antibody is administered every three weeks. It is envisaged that the chemotherapeutic agent or therapeutic antibody is administered every four weeks. It is envisaged that the chemotherapeutic agent or therapeutic antibody is administered every five weeks. It is also envisaged that the chemotherapeutic agent is or therapeutic antibody administered every six weeks.

[0102] In some embodiments the immunocytokine for use according to the invention, the use further comprises administration of a premedication. The premedication may be selected from the group consisting of dexamethasone, acetaminophen, diphenhydramine, cetirizine, promethazine, dexchlorpheniramine, meperidine and montelukast.

[0103] Premedication with glucocorticoids, such as dexamethasone or prednisone, is commonly employed in cancer treatment to mitigate various adverse reactions associated with chemotherapy, radiation therapy, or biological agents. These medications are administered to prevent or alleviate side effects such as nausea, vomiting, allergic reactions, inflammation, edema, hypersensitivity reactions, fatigue, pain, neutropenia and appetite suppression. Glucocorticoids function by suppressing immune responses and reducing inflammation, making them effective in managing aAttorney Docket No. 0183-0350-00PCT-KAD-ZSP ZSP ref.: 589-395 PCT spectrum of treatment-related complications and symptoms. Accordingly, it is envisaged that in the herein described uses and methods are combined with glucocorticoid treatment. In other words, glucocorticoids may be administered before, concomitantly, simultaneously or after administration of the immunocytokine in the herein described uses.

[0104] The invention also relates to the use of the described immunocytokine in the manufacture of a medicament for the treatment of cancer.

[0105] In an alternative aspect, the invention provides an immunocytokine for use in the treatment of cancer, wherein the immunocytokine comprises:

[0106] a) an antibody or antigen binding fragment thereof binding to PD-1 comprising a heavy chain variable domain (VH) comprising complementarity determining regions HCDR1 to HCDR3, and a light chain variable domain (VL) comprising complementarity determining regions LCDR1 to LCDR3; and

[0107] b) an interleukin- 15 (IL-15) conjugate, wherein said IL-15 comprises: (i) an IL-15 polypeptide comprising an amino acid sequence as set forth in SEQ ID NO: 9 or an amino acid sequence having at least 80 % identity to SEQ ID NO: 9; and (ii) an IL-15Ra sushi domain polypeptide comprising an amino acid sequence as set forth in SEQ ID NO: 11; and (iii) optionally a polypeptide linker LI linking (i) and (ii); and

[0108] c) optionally a polypeptide linker L2 linking a) and b);

[0109] characterized in that the immunocytokine is administered at a dose of 0.02 to 0.6 mg / kg.

[0110] The immunocytokine provided in this alternative aspect differs from the immunocytokine described in the first aspect in that the an antibody or antigen binding fragment thereof binding to PD-1 may comprise complementarity determining regions that are different from the ones specified for the immunocytokine described in the first aspect, and in that the IL- 15 conjugate may be located N-terminal or C-terminal of the antibody or antigen binding fragment thereof, preferably C-terminal. If the IL-15 conjugate is located N-terminal of the antibody or antigen binding fragment thereof, it may be linked to the N-terminus of the LC or the HC. If the IL-15 conjugate is located C-terminal of the antibody or antigen binding fragment thereof, it may be linked to the C-terminus of one or two heavy chains, preferably to the C-terminus of only one heavy chain.

[0111] All embodiments described for the first aspect of the invention are also disclosed for the alternative aspect, where applicable.

[0112] In some preferred embodiments, the immunocytokine of the alternative aspect comprises an IL-15 polypeptide comprising an amino acid sequence as set forth in SEQ ID NO: 31 and an IL-15Ra sushi domain polypeptide as set forth in SEQ ID NO: 30.Attorney Docket No. 0183-0350-00PCT-KAD-ZSP ZSP ref.: 589-395 PCT

[0113] Method of treatment

[0114] In a second aspect, the present invention relates to method of treating cancer, comprising administration of an immunocytokine to a human subject in need thereof, wherein the immunocytokine comprises:

[0115] a) an antibody or antigen binding fragment thereof binding to PD1 comprising a heavy chain variable domain (VH) comprising complementarity determining regions HCDR1 to HCDR3 as set forth in SEQ ID NO: 3 to 5, respectively, and a light chain variable domain (VL) comprising complementarity determining regions LCDR1 to LCDR3 as set forth in SEQ ID NO: 6 to 8, respectively; and

[0116] b) an interleukin- 15 (IL- 15) conjugate, wherein said IL- 15 conjugate is located N-terminal of VH and comprises (i) an IL- 15 polypeptide comprising an amino acid sequence as set forth in SEQ ID NO: 9 or an amino acid sequence having at least 80 % identity to SEQ ID NO: 9; and (ii) an IL-15Ra sushi domain polypeptide comprising an amino acid sequence as set forth in SEQ ID NO: 11; and (iii) optionally a polypeptide linker LI linking i and ii; and

[0117] c) optionally a polypeptide linker L2 linking a) and b);

[0118] characterized in that the immunocytokine is administered at a dose of 0.02 to 0.6 mg / kg.

[0119] The invention also relates to a method of treating cancer. All uses of the immunocytokine described herein are also disclosed as corresponding methods. In particular, all doses, schemes and regimens disclosed in context of uses are also disclosed in context of methods of treatment. All embodiments relating to the use of the immunocytokine described herein also apply to embodiments of the methods of treatment, where applicable.

[0120] In the context of the invention, the terms “treating” or “treatment”, as used herein include all types of preventive and / or therapeutic interventions medically allowed for the purpose of cure, temporary remission, prevention, delaying or stopping the progress of a disease, making a lesion regress or disappear, or inhibiting recurrence. Thus, the term “treatment of a disease” includes curing, shortening the duration, ameliorating, preventing, slowing down or inhibiting progression or worsening, or preventing or delaying the onset of a disease or the symptoms thereof. In some embodiments, the treatment comprises or consists of the killing of cancer cells. In some embodiments, the treatment comprises or consists of the prevention of metastatic spread. The term “in need of treatment” refers to a subject having a given disease or being at risk of developing said disease.Attorney Docket No. 0183-0350-00PCT-KAD-ZSP ZSP ref.: 589-395 PCT It is preferred that the described uses and methods are for the treatment of humans. Accordingly, the herein described uses and methods preferably relate to the treatment of cancer in humans. Accordingly, the patients referred to herein are preferably humans.

[0121] Pharmaceutical composition

[0122] The invention also relates to the use of the immunocytokine as described herein as a pharmaceutical composition. The invention relates to a pharmaceutical composition for use in the treatment of cancer comprising an immunocytokine, wherein the immunocytokine comprises: a) an antibody or antigen binding fragment thereof binding to PD-1 comprising a heavy chain variable domain (VH) comprising complementarity determining regions HCDR1 to HCDR3 as set forth in SEQ ID NO: 3 to 5, respectively, and a light chain variable domain (VL) comprising complementarity determining regions LCDR1 to LCDR3 as set forth in SEQ ID NO: 6 to 8, respectively; and

[0123] b) an interleukin- 15 (IL- 15) conjugate, wherein said IL- 15 conjugate is located N-terminal of VH and comprises (i) an IL- 15 polypeptide comprising an amino acid sequence as set forth in SEQ ID NO: 9 or an amino acid sequence having at least 80 % identity to SEQ ID NO: 9; and (ii) an IL-15Ra sushi domain polypeptide comprising an amino acid sequence as set forth in SEQ ID NO: 11; and (iii) optionally a polypeptide linker LI linking (i) and (ii); and c) optionally a polypeptide linker L2 linking a) and b);

[0124] characterized in that the pharmaceutical composition is administered at a dose of 0.02 to 0.6 mg / kg.

[0125] All embodiments relating to the immunocytokine for use described herein also apply to embodiments of the pharmaceutical composition, where applicable. This applies in particular to the described immunocytokine, doses and indications.

[0126] The terms “pharmaceutical composition” or “therapeutic composition” as used herein refer to a compound or composition capable of inducing a desired therapeutic effect when properly administered to a subject. The subject may also be referred to as patient.

[0127] Such therapeutic or pharmaceutical compositions may comprise a therapeutically effective amount of the immunocytokine, in admixture with a pharmaceutically or physiologically acceptable formulation agent, carrier or aqueous medium selected for suitability with the mode of administration.

[0128] The immunocytokine will usually be supplied as part of a sterile, pharmaceutical composition which may include a pharmaceutically acceptable carrier.

[0129] “Pharmaceutically acceptable” means approved by a regulatory agency of the Federal or a state government or listed in the U.S. Pharmacopeia or other generally recognized pharmacopeiaAttorney Docket No. 0183-0350-00PCT-KAD-ZSP ZSP ref.: 589-395 PCT for use in animals, and more particularly in humans. “Pharmaceutically acceptable” refers to molecular entities and compositions that do not produce an adverse, allergic or other untoward reaction when administered to a mammal, especially a human, as appropriate. A pharmaceutically acceptable carrier or excipient refers to a non-toxic solid, semi-solid or liquid filler, diluent, encapsulating material or formulation auxiliary of any type. A “pharmaceutically acceptable carrier” may also be referred to as “pharmaceutically acceptable diluent” or “pharmaceutically acceptable vehicle” and may include solvents, bulking agents, stabilizing agents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like which are physiologically compatible. The carrier may be an aqueous carrier or an aqueous medium.

[0130] The term “carrier”, as used herein, refers to a pharmacologically inactive substance such as but not limited to diluents, surfactants, stabilizers, physiological buffer solutions or vehicles with which the pharmaceutically active substance is administered. Such pharmaceutical carriers can be liquid or solid. Liquid carriers include but are not limited to sterile liquids, such as saline solutions in water and oils, including but not limited to those of petroleum, animal, vegetable or synthetic origin, such as peanut oil, soybean oil, mineral oil, sesame oil and the like. Saline solutions and aqueous dextrose and glycerol solutions can also be employed as liquid carriers, particularly for injectable solutions. A saline solution is a preferred carrier when the pharmaceutical composition is administered intravenously. Examples of suitable pharmaceutical carriers are described in “Remington's Pharmaceutical Sciences” by E. W. Martin.

[0131] Suitable pharmaceutical “excipients” include starch, glucose, lactose, sucrose, gelatine, malt, rice, flour, chalk, silica gel, sodium stearate, glycerol monostearate, talc, sodium chloride, dried skim milk, glycerol, propylene, glycol, water, ethanol and the like.

[0132] “Surfactants” include anionic, cationic, and non-ionic surfactants such as but not limited to sodium deoxy cholate, sodium dodecyl sulfate, Triton X-100, and polysorbates such as polysorbate 20, polysorbate 40, polysorbate 60, polysorbate 65 and polysorbate 80.

[0133] “Stabilizers” include but are not limited to mannitol, sucrose, trehalose, albumin, as well as protease and / or nuclease antagonists.

[0134] “Physiological buffer solution” include but are not limited to sodium chloride solution, demineralized water, as well as suitable organic or inorganic buffer solutions such as but not limited to phosphate buffer, citrate buffer, tris buffer (tris(hydroxymethyl)aminomethane), HEPES buffer ([4 (2 hydroxyethyl)piperazino]ethanesulphonic acid) or MOPS buffer (3 morpholino-1 propanesulphonic acid). The choice of the respective buffer in general depends on the desired buffer molarity. Phosphate buffers are suitable, for example, for injection and infusion solutions.Attorney Docket No. 0183-0350-00PCT-KAD-ZSP ZSP ref.: 589-395 PCT The pharmaceutical composition may further comprise a cryoprotectant to enable freezing of the isolated targeted delivery system. The cryoprotectant may comprise dimethyl sulfoxide (DMSO), glucose, dextrose, mannitol, trehalose, salts, glutamine, glycerol, propylene glycol, sodium pyruvate, sodium bicarbonate and / or human serum albumin.

[0135] In particular, the pharmaceutical compositions may contain vehicles, which are pharmaceutically acceptable for a formulation suitable for injection. These may be in particular isotonic, sterile, saline solutions (monosodium or disodium phosphate, sodium, potassium, calcium or magnesium chloride and the like or mixtures of such salts), or dry, especially freeze-dried compositions which upon addition, depending on the case, of sterilized water or physiological saline, permit the constitution of injectable solutions.

[0136] The pharmaceutical forms suitable for injectable use include sterile aqueous solutions or dispersions; formulations including sesame oil, peanut oil or aqueous propylene glycol; and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions. In all cases, the form must be sterile and must be fluid to the extent that administration via a syringe is possible. It must be stable under the conditions of manufacture and storage and must be preserved against the contaminating action of microorganisms, such as bacteria and fungi.

[0137] Solutions of the active compounds as free base or pharmacologically acceptable salts can be prepared in water suitably mixed with a surfactant, such as hydroxypropylcellulose. Dispersions can also be prepared in glycerol, liquid polyethylene glycols, and mixtures thereof and in oils. Under ordinary conditions of storage and use, these preparations may contain a preservative to prevent the growth of microorganisms.

[0138] An immunocytokine described herein may be formulated into a composition in a neutral or salt form using pharmaceutically acceptable salts.

[0139] Sterile injectable solutions are prepared by incorporating the active compounds (immunocytokine) in the required amount in the appropriate solvent with several of the other ingredients enumerated above, as required, followed by filtered sterilization. Generally, dispersions are prepared by incorporating the various sterilized active ingredients into a sterile vehicle which contains the basic dispersion medium and the required other ingredients from those enumerated above. In the case of sterile powders for the preparation of sterile injectable solutions, the preferred methods of preparation are vacuum-drying and freeze-drying techniques which yield a powder of the active ingredient plus any additional desired ingredient from a previously sterile-filtered solution thereof.Attorney Docket No. 0183-0350-00PCT-KAD-ZSP ZSP ref.: 589-395 PCT The preparation of more, or highly concentrated solutions for direct injection is also contemplated, where the use of DMSO as solvent is envisioned to result in extremely rapid penetration, delivering high concentrations of the active agents to a small tumor area.

[0140] The pharmaceutical compositions may be provided in a sealed container and may be provided as part of a kit. Such a kit may comprise instructions for use, e.g. a leaflet. Furthermore, the pharmaceutical composition or the kit may comprise a further medicament as herein described for co-therapy.

[0141] To prepare pharmaceutical compositions, an effective amount of immunocytokine may be dissolved or dispersed in a pharmaceutically acceptable carrier or aqueous medium. Accordingly, the invention relates to a pharmaceutical composition, wherein the herein described immunocytokine is dissolved or dispersed in a pharmaceutically acceptable carrier or aqueous medium.

[0142] Definitions

[0143] The specification uses a variety of terms and phrases, which have certain meanings as defined below. Preferred meanings are to be construed as preferred embodiments of the aspects of the invention described herein. As such, they and also further embodiments described in the following can be combined with any embodiment of the aspects of the invention and in particular any preferred embodiment of the aspects of the invention described above.

[0144] The terms “peptide” or “polypeptide” are used interchangeably in the context of the present invention to refer to a chain of at least two amino acids linked by peptide bonds. Thus, the term “polypeptide” in the context of the present invention is also used to refer to amino acid chains with more than 50, more than 100 or more than 150 amino acids.

[0145] In context of the present application, it is preferred that CDR / FR definition in an immunoglobulin light or heavy chain are determined according to Kabat numbering (Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md. (1987, 1991)). Accordingly, amino acid sequences of the CDR1, CDR2 and CDR3 of a given variable chain of an antibody or antibody fragment are indicated according to said Kabat definition.

[0146] Knowing the amino acid sequence of the CDRs of an antibody, one skilled in the art can easily determine the framework regions. However, in cases where the CDRs are not indicated, the person skilled in the art can first determine the CDR amino acid sequences and then determine the amino acid sequences of the framework regions.Attorney Docket No. 0183-0350-00PCT-KAD-ZSP ZSP ref.: 589-395 PCT The term “amino acid” encompasses naturally occurring amino acids as well as amino acid derivatives. In the context of the present specification, amino acids are identified using the 1 -letter code (Hausman RE, Cooper GM (2004). The cell: a molecular approach. Washington, D.C: ASM Press, p. 51. ISBN 978-0-87893-214-6). An amino acid identified with the letter X corresponds to any amino acid. An amino acid identified with the letter B corresponds to either D (asparagine) or N (aspartic acid). An amino acid identified with the letter Z corresponds to either E (glutamine) or Q (glutamic acid).

[0147] In the description of the invention, in some instances amino acid numbers are used with respect to antibodies or antibody domains, which do not indicate a SEQ ID NO. These numbers refer to amino acid positions in antibodies according to the UniProtKB database (www.uniprot.org / uniprot) in the version as disclosed on August 26, 2016. Unless specified otherwise, the number(s) correspond to a position in human IgG, in particular in human IgGl. The UniProtKB sequences (in the version as disclosed on August 26, 2016) of antibody domains referred to herein, including those of human IgGl are incorporated by reference as particular embodiments of the domains described herein, as well as variants thereof as defined below. When it is referred herein to amino acid positions in the Fc domain, these amino acid positions or residues are indicated according to the EU numbering system as described, for example in Edelman et al., Proc. Natl. Acad. USA, 1969, 63, 78-85.

[0148] The term “antigen” is used to refer to a substance, preferably an immunogenic polypeptide that comprises at least one epitope, preferably an epitope that elicits a B cell response.

[0149] “Fused” or “fusion” in the context of the invention means that two polypeptides are covalently linked, in particular via a peptide bond.

[0150] The term “cytokine” in the context of the invention refers to small proteins (about 5-25 kDa) which are involved in autocrine, paracrine and endocrine signaling as immunomodulating agents. Cytokines cannot cross the lipid bilayer of cells to enter the cytoplasm and act through cell surface receptors. The term “cytokine” as used herein includes chemokines, interferons, interleukins, lymphokines, and tumor necrosis factors.

[0151] The term “immunocytokine” in the context of the invention refers to a molecule comprising a fusion protein comprising a cytokine and at least an immunoglobulin variable domain.

[0152] The determination of percent identity between two sequences is accomplished using the mathematical algorithm of Karlin and Altschul, Proc. Natl. Acad. Sci. USA 90, 5873-5877, 1993. Such an algorithm is incorporated into the BLASTN and BLASTP programs of Altschul et al. (1990) J. Mol. Biol. 215, 403-410. To obtain gapped alignments for comparative purposes, Gapped BLAST is utilized as described in Altschul et al. (1997) Nucleic Acids Res. 25, 3389-3402. WhenAttorney Docket No. 0183-0350-00PCT-KAD-ZSP ZSP ref.: 589-395 PCT utilizing BLAST and Gapped BLAST programs, the default parameters of the respective programs are used. Alternatively, a variant can also be defined as having up to 20, 15, 10, 5, 4, 3, 2, or 1 amino acid substitutions, in particular conservative amino acid substitutions. Conservative substitutions are well known in the art (see for example Creighton (1984) Proteins. W.H. Freeman and Company). Families of amino acid residues having similar side chains are known in the art, and include amino acids with basic side chains (e.g., lysine, arginine, histidine), acidic side chains (e.g., aspartic acid, glutamic acid), uncharged polar side chains (e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine), nonpolar side chains (e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine), and aromatic side chains (e.g., tyrosine, phenylalanine, tryptophan).

[0153] The term “variant” refers, with respect to a polypeptide, generally to a modified version of the polypeptide, e.g., a mutation, so one or more amino acids of the polypeptide may be deleted, inserted, modified and / or substituted. Generally, the variant is a functional variant, meaning that it preserves the function of the parent polypeptide. Characteristics of functional variants, e.g., binding characteristics, can be analysed by known methods, e.g., using an ELISA-assay. More specific functions are defined herein and have precedence over the general definition. A “mutation” or “amino acid mutation” can be an amino acid substitution, deletion and / or insertion (“and” may apply if there is more than one mutation). Preferably, it is a substitution (i.e., a conservative or non-conservative amino acid substitution), more preferably a conservative amino acid substitution. In some embodiments, a substitution also includes the exchange of a naturally occurring amino acid with a not naturally occurring amino acid. A conservative substitution comprises the substitution of an amino acid with another amino acid having a chemical property similar to the amino acid that is substituted. Preferably, the conservative substitution is a substitution selected from the group consisting of:

[0154] (i) a substitution of a basic amino acid with another, different basic amino acid;

[0155] (ii) a substitution of an acidic amino acid with another, different acidic amino acid; (iii) a substitution of an aromatic amino acid with another, different aromatic amino acid;

[0156] (iv) a substitution of a non-polar, aliphatic amino acid with another, different non-polar, aliphatic amino acid; and

[0157] (v) a substitution of a polar, uncharged amino acid with another, different polar, uncharged amino acid.

[0158] A basic amino acid is preferably selected from the group consisting of arginine, histidine, and lysine. An acidic amino acid is preferably aspartate or glutamate. An aromatic amino acid isAttorney Docket No. 0183-0350-00PCT-KAD-ZSP ZSP ref.: 589-395 PCT preferably selected from the group consisting of phenylalanine, tyrosine and tryptophane. An aliphatic uncharged amino acid is preferably selected from the group consisting of glycine, alanine, valine, leucine, and isoleucine. A hydrophilic uncharged amino acid is preferably selected from the group consisting of serine, threonine, asparagine, and glutamine. A non-polar uncharged amino acid is preferably selected from the group consisting of cysteine, methionine, proline. In contrast to a conservative amino acid substitution, a non-conservative amino acid substitution is the exchange of one amino acid with any amino acid that does not fall under the above-outlined conservative substitutions (i) through (v).

[0159] Amino acids of a protein may also be modified, e.g., chemically modified. For example, the side chain or a free amino- or carboxy -terminus of an amino acid of the protein or polypeptide may be modified by e.g., glycosylation, amidation, phosphorylation, ubiquitination, etc. The chemical modification can also take place in vivo, e.g., in a host-cell, as is well known in the art. For example, a suitable chemical modification motif, e.g., glycosylation sequence motif present in the amino acid sequence of the protein will cause the protein to be glycosylated. Unless a modification leads to a change in identity of a modified amino acid (e.g., a substitution or deletion), a modified polypeptide is within the scope of polypeptide as mentioned with respect to a certain SEQ ID NO, i.e., it is not a variant as defined herein.

[0160] Preferably the degree of identity between a given amino acid sequence and an amino acid sequence which is a variant of said given amino acid sequence will be at least about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%. The degree of identity is given for an amino acid region which is 100% of the entire length of the reference amino acid sequence.

[0161] The amino acid sequence variants described herein may readily be prepared by the skilled person, for example, by recombinant DNA manipulation or with the aid of known peptide synthesis techniques such as, for example, by solid phase synthesis and similar methods. The manipulation of DNA sequences for preparing peptides or proteins having substitutions, additions, insertions or deletions, is described in detail in Sambrook et al. (1989), for example.

[0162] BRIEF DESCRIPTION OF DRAWINGS FIG. 1 is a line graph showing anti-tumor activity of the study drug administered at different doses and different regimens. Each line represents the tumor evolution in one patient.Attorney Docket No. 0183-0350-00PCT-KAD-ZSP ZSP ref.: 589-395 PCT FIG. 2 is a line graph showing PK profiles for the study drug at biweekly administration at doses of 1.4, 5, 10, 20, 30 and 40 mg at cycle 1 (A) and cycle 3 (B).

[0163] FIG. 3 is a line graph showing peripheral PD-1 Receptor Occupancy at biweekly administration (Q2W) at doses of 5, 10, 20, 30 and 40 mg, and at weekly administration (Q1W) at doses of 2.5 and 7.5 mg. Abbreviations: C1D1 = cyclel day 1; C1D4 = cycle 1 day 4; C3D1 = cycle 3 day 1; C3D4 = cycle 3 day 4; SOI = start of infusion; EOI = end of infusion; 8H = 8 hours; 72H = 72 hours.

[0164] FIG. 4 is a line graph showing NK cell (A) and CD8+ T Cell (B) Proliferation at biweekly administration (Q2W) at doses of 5, 10, 20, 30 and 40 mg (expressed as mean fold change over baseline C1D1 of fluorescence intensity of Ki67 staining).

[0165] FIG. 5 is a line graph showing CD3+ T cell, CD8+ T cell, lymphocyte and NK cell Counts at biweekly administration (Q2W) at doses of 5, 10, 20, 30 and 40 mg.

[0166] FIG. 6 is a line graph showing activated peripheral CD8 T Cell Counts at biweekly administration (Q2W) at doses of 5, 10, 20, 30 and 40 mg.

[0167] FIG. 7 is a line graph showing the mean value expressed in ng / L (A, B, D, E) and the mean fold change over baseline C1D1 (C) of cytokines IFNy, IL10, IL6 and TNFa. Figures 7A and 7B show biweekly administration (Q2W) at doses of 1.4, 5, 10 and 20 mg (A) and 1.4, 5, 10, 20 and 40 mg (B); values at 1.4, 5, 10 and 20 mg in Figures 7A and 7B are the same, but at different scale. Figure 7C is a line graph showing the mean fold change over baseline C1D1 of IFNy, IL10, IL6 and TNFa at biweekly administration (Q2W) at doses of 1.4, 5, 10, 20 and 40mg. Figure 7D shows biweekly administration (Q2W) at doses of 1.4 mg (DL1), 5 mg (DL2), 10 mg (DL3), 20 mg (DL4), 40 mg (DL5) and 30 mg (DL6). Figure 7E shows weekly administration (QW) at doses of 2.5 mg (DLla), 5 mg (DL2) and 7.5 mg (DL3). Abbreviations: IFN = Interferon; IL = Interleukin; TNF = Tumor Necrosis Factor; CxDy = Cycle XDay Y (e.g., C1D1 = Cycle 1 Day 1); SOI = Start of Infusion; EOI = End of Infusion, EOI+8H = 8-hours post EOI; EOI+24H = 24-hours post EOI; EOI+48H = 48-hours post EOI; EOI+168H = 168-hours post EOI. In Figures 7D and 7E the following applies: the values show the mean abundance of the cytokines across visits and time points (exposed population). For y-axis, log(2) value is presented in the plot but original unit is displayed as the label. The error bar is derived as Mean + Standard Error.Attorney Docket No. 0183-0350-00PCT-KAD-ZSP ZSP ref.: 589-395 PCT

[0168] FIG. 8 is a line graph showing the mean value expressed in ng / L (A, C, D) and the mean fold change over baseline C1D1 (B) of chemokines MCP-1, IP-10, MIP-la and MIP-ip. Figures A and B show biweekly administration (Q2W) at doses of 1.4, 5, 10, 20 and 40 mg. Figure 8C shows biweekly administration (Q2W) at doses of 1.4 mg (DL1), 5 mg (DL2), 10 mg (DL3), 20 mg (DL4), 40 mg (DL5) and 30 mg (DL6). Figure 8D shows weekly administration (QW) at doses of 2.5 mg (DLla), 5 mg (DL2) and 7.5 mg (DL3). Abbreviations :CCL2 = Chemokine (C-C Motif) Ligand 2; CXCL10 = Chemokine (C-X-C Motif) Ligand 10; MIP 1 a = Macrophage Inflammatory Protein 1 Alpha; MIP 1 13 = Macrophage Inflammatory Protein 1 Beta; CxDy = Cycle X Day Y (e.g., C1D1 = Cycle 1 Day 1); SOI = Start of Infusion; EOI = End of Infusion, EOI+8H = 8-hours post EOI; EOI+24H = 24-hours post EOI; EOI+48H = 48-hours post EOI; EOI+168H = 168-hours post EOI. In Figures 8C and 8D the following applies: the values show the mean abundance of the chemokines across visits and time points (exposed population). For y-axis, log(2) value is presented in the plot but original unit is displayed as the label. The error bar is derived as Mean + Standard Error.

[0169] FIG. 9 is a line graph showing PK profiles for the study drug at weekly administration.

[0170] FIG. 10 shows an analysis of circulating tumor DNA(ctDNA) in patient peripheral blood samples. Abbreviations: C4D1 : cycle 4 day 1; BOR: best overall response; SD, stable disease (with less and equal 6 months treatment duration); PD, progressive disease; CB, clinical benefit including confirmed partial response, unconfirmed partial response, and long stable disease (> 6 months).

[0171] EXAMPLE SECTION

[0172] Example 1: Clinical study of a IL15-based immunocytokine in Participants with Advanced Solid Tumors

[0173] This Example outlines the protocol for a phase 1 / 2, open label, first-in-human, dose escalation and expansion study for the evaluation of safety, pharmacokinetics, pharmacodynamics, and anti-tumor activity of a IL15-based immunocytokine, composed of an anti-PDl antibody, a mutated IL 15 and a IL15Ra sushi (herein after the study drug), administered as monotherapy or in combination with other anticancer therapies in adults with advanced solid tumors.Attorney Docket No. 0183-0350-00PCT-KAD-ZSP ZSP ref.: 589-395 PCT 1.1 Study design

[0174] The study includes 2 parts:

[0175] A dose escalation Part 1: for finding the therapeutic dose(s) of the study drug in a monotherapy given every 2 weeks (Q2W) or weekly (QW) and in combination with other anticancer therapies when applicable.

[0176] A multicohort dose expansion / dose optimization Part 2: for the assessment of safety and efficacy of the study drug in monotherapy and in combination with cetuximab or with next generation aCTLA4 (ADG126) or with bevacizumab (2 recommended doses for expansion / optimization (RDE / O) of the study drug identified from dose escalation Part 1 are be tested in different indications monotherapy and in combination with other anticancer therapies as applicable).

[0177] The study population for Part 1 is participants with advanced unresectable or metastatic solid tumors with no alternative therapeutic options or for those who decline alternative therapies. For combination with next generation anti-CTLA4: only participants with tumors for which current generation anti-CTLA4 as single agent or in combination treatments are approved. For combination with bevacizumab: participants with metastatic colorectal cancer (CRC).

[0178] The study population for Part 2A are: Cohorts Al and A2, participants with metastatic nonsmall cell lung cancer (NSCLC) not amenable to available standard of care (SOC), Cohort B, participants with advanced unresectable or metastatic hepatocellular carcinoma (HCC) who relapsed on prior PD1 / PD-L1 -based regimens, Cohorts Cl and C2, participants with gastric cancer / gastro esophageal junction adenocarcinoma (GC / GEJ) who relapsed to at least 1 prior regimen which may or may not include an anti-PDl / PD-Ll -based treatment depending on local standard of care, regardless combined positivity score (CPS), Cohort D: biomarkers cohort in infiltrated tumor type(s) and Cohorts El and E2: participants with metastatic CRC.

[0179] The study population for Part 2B is Cohort E3, participants with metastatic CRC regardless RAS mutation status.

[0180] The study population for Part 2C is Cohorts Gl, G2 and G3, participants with metastatic melanoma who relapsed to at least 1 prior line of therapy.Attorney Docket No. 0183-0350-00PCT-KAD-ZSP ZSP ref.: 589-395 PCT The study population for Part 2D is Cohorts Hl and H2, participants with metastatic CRC.

[0181] In Part 1A monotherapy, the recommended starting dose of 1.4 mg (approximately 0.02 mg / kg [assuming an average body weight is 70 kg]) was selected based on minimal anticipated biological effect level (MABEL) approach,. The incidence of dose limiting toxi cities (DLTs) will be tested across the dose levels.

[0182] In Part IB combination with ADG126, the incidence of DLTs occurring within Cycles 1 to 3 is tested in several pairs of doses of study drug and ADG126.

[0183] In Part 2A, 15 to 30 participants per cohort are be enrolled and treated in each of the following independent cohorts to assess the antitumor activity and for dose optimization in some indications.

[0184] Cohort 1) (= cohort Al and A2): up to 30 participants assess the study drug as a monotherapy in participants with metastatic non-small cell lung cancer (NSCLC) not amenable to available standard of care (SOC) or who have progressed on prior regimens.

[0185] Cohort 2) (= cohort Cl and C2): up to 30 participants assess the study drug as a monotherapy in participants with advanced unresectable or metastatic gastric cancer (GC) or Siewert Type 2 and 3 gastro esophageal junction adenocarcinoma (GEJ), and for whom receiving the SOC is not in his or her best interest, or where no SOC is established. Human epidermal growth factor receptor 2 (HER2) positive cases are not be eligible unless they have progressed on a HER2 targeted therapy. Those participants should have received at least 1 prior line of anti-cancer treatment which may or may not include ananti-PDl / PD-Ll based treatment depending on local standard of care. Metastatic microsatellite instability-high (MSLH) or mismatch repair deficient (dMMR) cases are not eligible.

[0186] Cohort 3) (cohort El and E2): up to 30 participants assess the study drug as monotherapy in participants with advanced unresectable or metastatic CRC who have progressed on prior regimens. Participants must have non-MSI-H disease to be eligible.

[0187] Enrolment continues with the optimal dose to accrue approximately 15 participants in up to 2 selected indications.

[0188] In Part 2B, up to 45 participants are enrolled and treated in the following independent cohort to assess the antitumor activity:Attorney Docket No. 0183-0350-00PCT-KAD-ZSP ZSP ref.: 589-395 PCT Cohort E3: up to 45 participants assess the study drug at one of the RDE / O used in Part 2A in combination with cetuximab in participants with advanced unresectable or metastatic CRC any RAS who have progressed on at least one prior regimen. Participants must have non-MSI-H disease to be eligible.

[0189] For Part 2A, 2B, 2C and 2D, different two-stage designs are implemented to conduct dose expansion / optimization for each indication with monotherapy and for combination with Cetuximab, ADG126 or bevacizumab to select the optimal dose for those indications with activity as applicable.

[0190] In Part 2C, up to 120 participants are enrolled and treated in the following independent cohorts to assess the antitumor activity and for dose optimization.

[0191] Cohorts G1 and G2: up to 80 participants assess the study drug at one of the RDE / O used in Part 2A in combination with aCTLA4 ADG126 versus Cohort G3: up to 40 participants treated in combination with nivolumab plus ipilimumab in participants with metastatic melanoma.

[0192] In Part 2D, up to 80 participants are enrolled and treated in the following independent cohorts to assess the antitumor activity and for dose optimization.

[0193] Cohorts Hl and H2: up to 80 participants assess the study drug at one of the RDE from study drug monotherapy dose escalation in participants with advanced unresectable or metastatic CRC who have progressed on two prior lines of therapy.

[0194] The study population for Japan Cohort F is Japan specific participants with advanced unresectable or metastatic solid tumors with no alternative therapeutic options. In Japan Cohort F, up to 9 Japanese participants assess the study drug as a monotherapy at 10 or 30 mg Q2W.

[0195] Additional cohorts with different regimens, indications or combinations may be added to this protocol at a later stage of development.

[0196] Number of participants:

[0197] Approximately 473 participants are exposed to the study intervention:

[0198] • For Part 1: approximately 99 participants are exposed in the escalation part.

[0199] • For Part 2: up to 365 participants are exposed in expansion / dose optimization parts.Attorney Docket No. 0183-0350-00PCT-KAD-ZSP ZSP ref.: 589-395 PCT • For Japan Cohort F: up to 9 Japanese participants are exposed in the Japan specific cohort to obtain at least 3 DLT-evaluable Japanese participants in each dose.

[0200] 1.2. Study arms and duration:

[0201] There are 7 intervention groups:

[0202] 1. Part 1A (Q2W or QW): study drug single agent dose escalation in participants with advanced unresectable or metastatic solid tumor

[0203] 2. Part IB: Study drug in combination with ADG 126 in participants with advanced unresectable or metastatic melanoma, NSCLC; renal cell carcinoma (RCC), HCC, colorectal cancer (MSI- FI / dMMR), malignant pleural mesothelioma or esophageal squamous cell carcinoma (ESCC).

[0204] 3. Part 2A: study drug single agent expansion

[0205] a. Cohorts Al and A2: in metastatic NSCLC not amenable to available SOC

[0206] b. Cohorts Cl and C2: in advanced unresectable or metastatic GC / GEJ, regardless CPS and c. Cohorts El and E2: in advanced unresectable or metastatic CRC who have progressed on at least 2 prior regimens. Participants must have non-MSI-H disease to be eligible.

[0207] 4. Japan Cohort F: Study drug single agent dose escalation in Japanese participants with advanced unresectable or metastatic solid tumor

[0208] 5. Part 2B: Study drug in combination with cetuximab expansion

[0209] a. Cohort E3 : in advanced unresectable or metastatic CRC who have progressed on at least one prior regimen. Participants must have non-MSI-H disease to be eligible

[0210] 6. Part 2C: Study drug in combination with ADG126 expansion

[0211] a. Cohorts Gl, G2 and G3 : in metastatic melanoma who have progressed on at least one prior regimen

[0212] 7. Part 2D: Study drug in combination with bevacizumab expansion

[0213] a. Cohorts Hl and H2: in advanced unresectable or metastatic CRC who have progressed on 2 prior lines of therapy

[0214] The duration of each cycle is 14 days and the DLT observation period consists of the first 2-3 cycles of treatment, as applicable.

[0215] The duration of the study for a participant includes:

[0216] Screening Period: up to 28 daysAttorney Docket No. 0183-0350-00PCT-KAD-ZSP ZSP ref.: 589-395 PCT Treatment Period: enrolled and exposed participants receive continuous treatment until progressive disease (PD), or an occurrence of an unacceptable adverse event (AE), a withdrawal of consent, or until other permanent discontinuation criteria described in the protocol are met.

[0217] The end of treatment (EOT) visit occurs 30 days ±7 days from the last investigational medicinal product (IMP) administration or prior to the initiation of further therapy, whichever occurs first.

[0218] The follow-up period will occur until, death, withdrawal of participant’s consent, or study cut-off date, whichever comes first.

[0219] 1.3. Study intervention(s)

[0220] Escalation Part 1 A, Part IB and Expansion Part 2A - Cohort Al, A2, Cl, C2, El, E2 and Japan cohort F (study drug monotherapy) Part 2B - Cohort E3: study drug + Cetuximab Part 2C - Cohorts G1 and G2: study drug + ADG126 Cohort G3:Nivolumab + Ipilimumab Part 2D - Cohorts Hl and H2: study drug + bevacizumab

[0221] General dosing sequence: premedication for the study drug (30 to 60 minutes prior to start of the study drug infusion), and the study drug.

[0222] Dosing sequence for combination with cetuximab: premedication for cetuximab (30 to 60 minutes prior to the first dose of cetuximab), cetuximab, premedication for study drug (30 to 60 minutes prior to start of study drug infusion), and study drug (start of study drug infusion should be at least 60 minutes after completion of cetuximab infusion).

[0223] Dosing sequence for combination with bevacizumab: premedication for study drug (30 to 60 minutes prior to start of study drug infusion), start of study drug IV infusion followed by bevacizumab.

[0224] Investigational medicinal product(s):

[0225] IL-15 based immunocytokine

[0226] • The IL 15-based immunocytokine tested in this clinical trial is composed of an anti-PDl antibody, a mutated IL15 and a IL15Ra sushi. This IL15-based immunocytokine is composed of two light chains (LC) each consisting of an amino acid sequence as set forth in SEQ ID NO: 17, a first heavy chain (HC1) consisting of an amino acid sequence as set forth in SEQ ID NO: 15 and a second heavy chain (HC2) consisting of an amino acid sequence as set forth in SEQ ID NO: 16 (“study drug”)

[0227] • Route(s) of administration: intravenous (IV) infusionAttorney Docket No. 0183-0350-00PCT-KAD-ZSP ZSP ref.: 589-395 PCT • Dose regimen: the starting dose is 1.4 mg per administration Q2W or 2.5 mg per administration QW.

[0228] • Dose for Part IB: the starting dose is 10 mg per administration Q2W

[0229] • Doses for Part 2 A: 30 mg as a higher recommended dose and 10 mg as a lower recommended dose both given Q2W to be randomized in Part 2A dose expansion / optimization for dose ranging in monotherapy in Cohorts Al and A2, Cohorts Cl and C2, Cohorts El and E2 • Doses for Part 2C & 2D: 20 mg as a higher recommended dose and 10 mg as a lower recommended dose both given Q2W to be randomized in Part 2C & 2D dose expansion / optimization for dose ranging in combination with the other anti-tumor agent in Cohorts Gl, G2, and Cohorts Hl and H2

[0230] • 30 mg Q2W will be given in combination with cetuximab 500 mg / m2Q2W in Part 2B safety run in of Cohort E3

[0231] • Japan Cohort F: The starting dose is 10 mg Q2W in monotherapy.

[0232] Cetuximab

[0233] • Formulation:

[0234] - 200 mg / 100 mL (2 mg / mL) (US only)

[0235] - 500 mg / 100 mL (5 mg / mL)

[0236] - any other cetuximab formulation approved locally.

[0237] • Route of administration: IV infusion

[0238] • Dose regimen: Cetuximab will be given on Cycle 1 Day 1 500 mg / m2infused over 120 minutes Q2W.

[0239] ADG126

[0240] Bevacizumab

[0241] • Formulation:

[0242] - 400 mg / vial

[0243] - 100 mg / vial

[0244] • Route of administration: IV infusion

[0245] • Dose regimen: bevacizumab will be given on DI 5 mg / kg Q2WAttorney Docket No. 0183-0350-00PCT-KAD-ZSP ZSP ref.: 589-395 PCT - The first infusion will be given over 90 minutes. If this is well tolerated the second infusion may be given over 60 minutes. If the 60-minute infusion is well tolerated, all subsequent infusions may be administered over 30 minutes.

[0246] Nivolumab

[0247] • Formulation: 10 mg / mL in a single dose vial

[0248] • Route of administration: IV infusion

[0249] • Dose regimen: 1 mg / kg every 3 weeks for 4 doses, then 240 mg every 2 weeks or 480 mg every 4 weeks over 30 minutes

[0250] • Complete dosing information can be found in approved label

[0251] Ipilimumab

[0252] • Formulation: 5 mg / mL in a single-dose vial

[0253] • Route of administration: IV infusion

[0254] • Dose regimen: 3 mg / kg every 3 weeks for 4 doses over 30 minutes.

[0255] Non-investigational medicinal product(s) / Auxiliary medicinal product(s):

[0256] Non-investigational medicinal products include the premedication administered for the study drug.

[0257] Premedication for the study drug

[0258] All participants receive the following premedication to prevent or reduce the severity of potential acute AEs, approximately 30 to 60 minutes prior to the study drug infusion for the first 4 administrations:

[0259] • Dexamethasone 15 mg IV or equivalent.

[0260] • Acetaminophen (paracetamol) 650 to 1000 mg oral route (PO) or equivalent, and then optionally thereafter as needed.

[0261] • Diphenhydramine 25 to 50 mg IV (preferred but PO is also acceptable when IV formulation is not available) or equivalent (e.g., cetirizine, promethazine, dexchlorpheniramine, according to local approval and availability), and then optionally thereafter as needed.

[0262] • Additional premedication (e.g., meperidine or montelukast) may be considered at the Investigator’s discretion.Attorney Docket No. 0183-0350-00PCT-KAD-ZSP ZSP ref.: 589-395 PCT The study drug premedication may be optional after 4 infusions as described below:

[0263] • For a participant who has no CRS or infusion-related reaction (IRR) during the first 4 infusions: premedication for the subsequent infusions is optional at the Investigator’s discretion. However, if during the subsequent infusions without premedication the participant experiences a CRS / IRR (any grade), premedication must be restarted for all subsequent infusions.

[0264] • If a participant develops a CRS / IRR Grade <3 during their first cycle only and then experiences no further CRS / IRRs during their next 2 infusions: the Investigator may consider omitting premedication for the next cycle. If no CRS / IRR is observed during the next infusion without premedication, premedication is optional for the subsequent infusions at the Investigator’ s discretion. However, if during the next infusion without premedication the participant experiences a CRS / IRR (any grade), premedication must be restarted for all subsequent infusions.

[0265] Premedication for cetuximab:

[0266] All participants who will receive cetuximab should be premedicated with diphenhydramine 25 to 50 mg IV (or equivalent) prior to the first dose of cetuximab or any other recommended premedication as per local requirements. Premedication for subsequent doses of cetuximab should be given per medical judgment and history of prior IRRs.

[0267] Each dose of cetuximab is be followed by a 1-2 hours observation period. If there are no reported IRRs during this observation period, then the study drug dose may be administered at applicable dosing visits.

[0268] • When study drug and cetuximab are given on the same day, participants who have received diphenhydramine as cetuximab premedication may skip the diphenhydramine as study drug premedication.

[0269] 1.4. Objectives

[0270] a. Primary

[0271] Dose escalation: To determine the maximum tolerated dose (MTD) or maximum administered dose (MAD), recommended dose(s), and the overall safety and tolerability profile of the study drug when administered as monotherapy or in combination

[0272] Japan Cohort F: To confirm the tolerability of study drug monotherapy in Japanese participantsAttorney Docket No. 0183-0350-00PCT-KAD-ZSP ZSP ref.: 589-395 PCT Dose expansion / optimization: To determine the objective response rate (ORR) of the study drug administered as monotherapy or in combination at the recommended dose(s)

[0273] b. Secondary

[0274] Dose escalation and Japan Cohort F: To assess clinical activity of the study drug monotherapy or in combination at the recommended dose(s)

[0275] Dose escalation and expansion / optimization and Japan Cohort F: To characterize the pharmacokinetic (PK) profile of the study drug when administered as monotherapy or in combination and the PK of combined therapies

[0276] Dose escalation and expansion / optimization and Japan Cohort F: To assess the potential immunogenicity of the study drug and ADG126

[0277] Dose expansion / optimization: To assess other indicators of antitumor activity Dose expansion / optimization: To characterize the safety profile of the study drug monotherapy or in combination

[0278] c. Tertiary / Exploratory

[0279] Dose escalation and expansion / optimization:

[0280] - To monitor immune cell activation, expansion and kinetics in blood induced by the study drug in monotherapy or combined therapies

[0281] - To document the immune contexture and the modulation of immune responses in the tumor microenvironment by the study drug in monotherapy or combined therapies

[0282] - To measure PD-1 receptor occupancy in blood

[0283] - To identify additional biomarkers that maybe related to safety and efficacy in response to the study drug treatment at different dose levels in monotherapy or combined therapies

[0284] - To identify potential predictive biomarkers for the study drug treatment in monotherapy or combined therapies

[0285] - To explore Exposure / Response analysis for efficacy and safety, as data permits Dose expansion / optimization:

[0286] - To explore other indicators of antitumor activity

[0287] - To explore patient-reported experience of treatmentAttorney Docket No. 0183-0350-00PCT-KAD-ZSP ZSP ref.: 589-395 PCT 1.5.

[0288] a. Primary endpoints:

[0289] • Presence of dose limiting toxi cities (DLTs) in Cycles 1 and 2 or Cycles 1-3 as applicable • Presence of treatment-emergent adverse events (TEAEs), serious adverse events (SAEs), and lab abnormalities, according to the National Cancer Institute (NCI) Common Terminology Criteria for Adverse Events (CTCAE) Version 5.0 and American Society for Transplantation and Cellular Therapy (ASTCT) consensus grading

[0290] • Objective response rate, which is defined as the proportion of participants who have a confirmed complete response (CR) or a partial response (PR), as the best overall response determined by the Investigator as per the Response Evaluation Criteria in Solid Tumors (RECIST) v 1.1

[0291] b. Secondary endpoints:

[0292] • Objective response rate (ORR) based on RECIST 1.1 criteria

[0293] • Duration of response (DoR), defined as the time from the first documented evidence of confirmed CR or PR until progressive disease (PD) determined by Investigator per RECIST 1.1 or death from any cause, whichever occurs first

[0294] • Study drug PK parameters (Cmax, Tmax, AUCrau) and at least Ctrough for combined therapies • Proportion of participants with presence of ADA against the study drug

[0295] • Percentage of participants with presence of ADA against study drug and ADG126

[0296] • Time to response, defined as the time from the first administration of investigational medicinal product (IMP) to the first documented evidence of confirmed PR or CR determined by Investigator per RECIST 1.1

[0297] • Duration of response (DoR), defined as the time from first documented evidence of confirmed CR or PR until progressive disease (PD) determined by Investigator per RECIST 1.1 or death from any cause, whichever occurs first

[0298] • Clinical benefit rate including confirmed CR or PR at any time or stable disease (SD) of at least 6 months determined by Investigator per RECIST 1.1

[0299] • Progression-free survival (PFS), defined as the time from the date of first administration of IMP to the date of the first documented disease progression determined by Investigator as per RECIST 1.1 or death from any cause, whichever occurs first

[0300] • Overall survival (OS), defined as the time from the first dose of IMP to the date of death due to any causeAttorney Docket No. 0183-0350-00PCT-KAD-ZSP ZSP ref.: 589-395 PCT • Presence of TEAEs, SAEs, and lab abnormalities, according to the NCI-CTCAE Version 5.0 and the ASTCT consensus grading

[0301] c. Tertiary / Exploratory

[0302] • Activation and expansion of CD8+ T cells (particularly effector memory subpopulation), T regulatory (Treg) cells, natural killer (NK) cells, and CD8 / Treg ratio in blood.

[0303] • Infiltration and proliferation of CD8+ T cells and CD8 / Treg ratio

[0304] • Determine the drug-free PD-1 on T cells by flow cytometry

[0305] • Assessment of change in cytokine levels in peripheral blood

[0306] • Assessment of dynamic change of transcription profile of immune cells from tumor biopsies

[0307] • Potential decrease of detectable ctDNA in peripheral blood

[0308] • The change of soluble PD1 in peripheral blood

[0309] • Baseline immune contexture and programmed cell death-ligand 1 (PD-L1) expression from tumor biopsy

[0310] • Next-Generation Sequencing to detect tumor mutational burden (TMB), microsatellite instability (MSI), deficient mismatch repair (dMMR), interferon (IFN) related gene signature, etc from tumor and blood

[0311] • To analyze PK parameters and efficacy / safety endpoints of interest

[0312] • ORR defined as the proportion of participants who have a confirmed complete response (CR) or partial response (PR) determined by Investigator per immune Response Evaluation Criteria in Solid Tumors (iRECIST) for immune-based therapies or modified Response Evaluation Criteria in Solid Tumors (mRECIST) for HCC

[0313] • Patient-reported benefits and disadvantages of treatment will be measured by the Patient’s Qualitative Assessment of Treatment version 2 (PQATv2)

[0314] 1.6. Study population

[0315] a. Inclusion criteria

[0316] Participants are eligible to be included in the study only if all of the following criteria apply:

[0317] Age

[0318] Participant must be at least 18 years of age inclusive (or country’s legal age of majority if >18 years), at the time of signing the informed consent.

[0319] Type of participant and disease characteristicsAttorney Docket No. 0183-0350-00PCT-KAD-ZSP ZSP ref.: 589-395 PCT Dose escalation Part 1A and Japan Cohort F:

[0320] Participants with advanced unresectable or metastatic solid tumors for which, in the judgement of the investigator, no standard alternative therapy is available or is not in the best interest of the participant.

[0321] Dose expansion / Optimization Part 2:

[0322] Cancer diagnosis:

[0323] Participants in Cohorts Al and A2: Histologically or cytologically confirmed diagnosis of metastatic NSCLC.

[0324] Participants in Cohort B: Histologically or cytologically confirmed diagnosis of advanced unresectable or metastatic HCC, or clinically by American Association for the Study of Liver Diseases (AASLD) criteria in cirrhotic patients (participants without cirrhosis must have had histological confirmation of diagnosis).

[0325] Participants in Cohort Cl and C2: Histologically or cytologically confirmed diagnosis of advanced unresectable or metastatic GC or Siewert Type 2 & 3 GEJ.

[0326] For participants in Cohort Cl and C2: Disease with any CPS scoring. No need for CPS determination at local laboratory.

[0327] For participants in Cohort Cl and C2: Participants must have MSI or MMR status known or determined locally and must have non-MSI-H or proficient MMR (pMMR) disease to be eligible.

[0328] For participants in Cohort Cl and C2: Participants with unknown HER2 / neu status must have their HER2 / neu status determined locally. Participants with HER2 / neu negative are eligible. Participants with HER2 / neu positive tumors must have documentation of disease progression on treatment containing an approved HER2 targeted therapy to be eligible.

[0329] Prior anticancer therapy (for dose expansion / optimization Part 2 only):

[0330] Participants in Cohort Al and A2: Participants must have received at least 1 systemic therapy for the metastatic setting and must not be amenable to the available SOC.

[0331] Participants in Cohort B: Participants who have received at least 1 prior anticancer therapy, including an anti-PDl / PD-Ll containing regimen, and for whom have progressed after a primary or secondary resistance to an anti-PDl / PD-Ll.

[0332] Primary resistance: participant must have experienced PD or SD lasting <6 months since the initiation of the anti-PDl / PD-Ll inhibitor-based treatment and the participant must have received PD1 / PD-L1 for at least 6 weeks. Radiographic confirmation of the PD must be documented after a minimum of 4 weeks after the initial identification of progression, unless: 1)Attorney Docket No. 0183-0350-00PCT-KAD-ZSP ZSP ref.: 589-395 PCT investigator confirms clinical progression / deterioration attributed to PD, or 2) the first radiographic assessment indicated critical tumor growth by imaging (size or location).

[0333] Secondary resistance: participants must have experienced PD, either during or within 3 months of discontinuing treatment with an anti-PDl based therapy, occurring after previous clear benefit (any complete [CR] or partial response [PR]), or after previous SD of >6 months. There is no requirement for radiographic confirmation of the progression.

[0334] Participants in Cohorts Cl and C2: Participants should have failed or relapsed after at least 1 prior line of treatment which may or may not include an anti-PDl / PD-L1 -based treatment depending on local standard of care.

[0335] Measurable Disease:

[0336] At least 1 measurable lesion per RECIST 1.1 criteria. Target lesions may be located in a previously irradiated field if there is a documented radiographic disease progression in that site.

[0337] Weight

[0338] Body weight within 40-150 kg (inclusive).

[0339] Sex, contraceptive / barrier method and pregnancy testing requirements / breastfeeding All:

[0340] Contraceptive use by men and women should be consistent with local regulations regarding the methods of contraception for those participating in clinical studies.

[0341] Male participants:

[0342] • Male participants are eligible to participate if they agree to the following during the intervention period and for at least 5 half-lives of the study drug after the last dose of the study drug administration:

[0343] - Refrain from donating sperm, plus either:

[0344] - Be abstinent from heterosexual intercourse as their preferred and usual lifestyle (being abstinent on a long term and persistent basis) and agree to remain abstinent, or

[0345] - Must agree to the use of a contraception / barrier (a male condom). Also, the participant should be advised of the benefit for the female partner to use a highly effective method of contraception (as described in Section 10.4) as a condom may break or leak when having sexual intercourse with a woman of childbearing potential (WOCBP), who is not currently pregnant.Attorney Docket No. 0183-0350-00PCT-KAD-ZSP ZSP ref.: 589-395 PCT Female participants:

[0346] • A female participant is eligible to participate if she is not pregnant or breastfeeding, and if at least one of the following conditions applies:

[0347] - She is not a WOCBP as defined in Appendix 4 Contraceptive and barrier guidance (Section 10.4).

[0348] OR

[0349] - She is a WOCBP and agrees to use highly effective contraceptive methods, with a failure rate of <1% per year, starting from the time of the informed consent, and through the duration of the treatment period, to at least 60 days after the last dose of the study intervention administration.

[0350] • A WOCBP must have a negative result in a highly sensitive pregnancy test (as required by local regulations) within 72 hours before the first administration of study intervention.

[0351] - The Investigator is responsible for reviewing the medical history, menstrual history, and the participants’ recent sexual activity to decrease the risk for inclusion of a woman with an early undetected pregnancy.

[0352] Informed Consent

[0353] Capable of giving signed informed consent, which includes compliance with the requirements and restrictions listed in the informed consent form (ICF) and in this protocol.

[0354] Capable of and willing to comply with scheduled visits, treatment plans, laboratory tests and other study related tests and procedures.

[0355] For participants in Cohort D: Participants with tumors that are usually considered immunogenic "hot" tumors infiltrated by T cells as any of the following: histologically or cytologically confirmed diagnosis of advanced unresectable or metastatic melanoma, non-small cell lung cancer, bladder, head and neck, kidney, and liver cancer or other indications known to be immunogenic at the discretion of the investigator with agreement of the study medical monitor.

[0356] Participants in Cohort D: Participants must have received at least 1 systemic therapy for their advanced / metastatic setting and must not be amenable to the available SOC.

[0357] Further criteria (introduced with amendments)

[0358] Participants in Part 2A Cohorts El and E2, Part 2B Cohort E3, Parti C and Part 2D Cohorts Hl and H2: Histologically or cytologically confirmed diagnosis of advanced unresectable or metastatic colorectal cancer.Attorney Docket No. 0183-0350-00PCT-KAD-ZSP ZSP ref.: 589-395 PCT Participants in Part 2A Cohorts El, and E2 and Part 2B Cohort E3 MSI status: Participants must have MSI status known or determined locally and must have non-MSI-H disease to be eligible.

[0359] Participants in Part 2A Cohorts El, E2, Part 2B Cohort E3, and Part 2D Cohorts Hl and H2: Participantswith RAS-mutant and BRAF-mutant colorectal cancer are eligible for enrollment.

[0360] Prior anticancer therapy: Participants in Part 2A Cohorts El and E2 and Part 2D Cohorts Hl and H2 should have failed or relapsed on at least 2 prior regimens.

[0361] Participants in cohort E3 should have failed or relapsed on at least 1 prior regimen. Participants who have received cetuximab or other anti-EGFR therapy as part of their prior line of treatment are eligible.

[0362] Further criteria (introduced with amendments):

[0363] Dose escalation Part IB: Participants with advanced unresectable or metastatic melanoma, NSCLC; renal cell carcinoma (RCC); HCC, colorectal cancer (MSI-H / dMMR), malignant pleural mesothelioma or esophageal squamous cell carcinoma (ESCC). and for who, in the judgement of the investigator, no standard alternative therapy is available or is not in the best interest of the participant.

[0364] Dose expansi on / optimizati on Part 2C Cohorts Gl, G2 and G3: Participants with histologically confirmed unresectable locally advanced or metastatic melanoma

[0365] Dose expansion / optimization Part 2C Cohorts Gl, G2 and G3: Participants must have received at least one prior line of therapy for advanced / metastatic melanoma and / or does not have any standard of care (SoC) treatment option or decline or is intolerant to be treated with SoC treatment.

[0366] Part 2D: Adequate coagulation function for all participants. For participants receiving anticoagulant therapy (except platelet anti-aggregates) the adequate therapeutic levels of INR should be confirmed.

[0367] b. Exclusion criteria

[0368] Participants are excluded from the study if any of the following criteria apply:

[0369] Medical conditions

[0370] Eastern Cooperative Oncology Group (ECOG) performance status of >2.

[0371] Predicted life expectancy <3 months.Attorney Docket No. 0183-0350-00PCT-KAD-ZSP ZSP ref.: 589-395 PCT For participants with HCC- Cohort B (Part 2): Child Pugh Class B or C liver score. Participants with Child Pugh Class B-7 score are allowed for Part 1.

[0372] Diagnosed of any other malignancies, either progressing or requiring active treatments, within 2 years prior to enrollment, except for basal cell carcinoma or squamous cell carcinoma of the skin, which are in-situ malignancies, as well as superficial bladder carcinoma, or low risk prostate cancer, and any tumors that have been deemed as effectively treated with definitive local control.

[0373] Active brain metastases or leptomeningeal metastases:

[0374] • Participants with previously treated brain metastases are eligible, provided they are clinically stable for at least 4 weeks with no evidence of new or enlarging brain metastases and have not received corticosteroids for at least 2 weeks prior to the first IMP administration.

[0375] • Participants with asymptomatic brain metastases (i.e., no neurological symptoms, no requirements for corticosteroids, and no lesion >1.5 cm) are eligible but require regular imaging of the brain at the site of the disease.

[0376] History of treatment-related immune-mediated (or immune-related) AEs from immunomodulatory agents (including but not limited to anti-PDl / PD-Ll agents and anti -cytotoxic T lymphocyte associated protein 4 monoclonal antibodies) that caused permanent discontinuation of the agent, or that were Grade 4 in severity, or have not resolved to Grade <1.

[0377] Has any condition requiring ongoing / continuous corticosteroid therapy (>10 mg prednisone / day or an anti-inflammatory equivalent) within 1 week prior to the first dose of the study medicine. Physiologic replacement doses are allowed even if they are >10 mg of prednisone / day or equivalent, as long as they are not being administered for immunosuppressive intent. Inhaled or topical steroids are permitted, provided that they are not for treatment of an autoimmune disorder. Note: Participants who require a brief course of steroids (up to 2 days in the week before enrollment) or physiologic replacement are eligible to be enrolled in the study.

[0378] Any clinically significant cardiac (including valvular) or vascular (thromboembolic disorders) disease, within 6 months prior to the first IMP administration, such as:

[0379] • Congestive heart failure (New York Heart Association Class II to IV, or left ventricular ejection fraction <50%).

[0380] • Increased troponin (higher than upper limit of normal [ULN] per local laboratory) at screening. Participants with a higher troponin may be allowed upon cardiologist consultation and discussion with the Study Medical Monitor on a case-by-case basis.Attorney Docket No. 0183-0350-00PCT-KAD-ZSP ZSP ref.: 589-395 PCT • Unstable or poorly controlled angina, myocardial infarction, coronary artery revascularization procedure (e.g., coronary artery bypass graft, percutaneous coronary intervention).

[0381] • Transient ischemic attack, cerebral infarction (stroke), symptomatic pulmonary embolism, and peripheral artery revascularization procedure.

[0382] • Uncontrolled cardiac arrhythmia requiring medication (> Grade 2, according to the NCICTCAE V5.0).

[0383] History of congenital long QT syndrome, torsades de pointes or prolonged QTc interval >480 msec using Fridericia’s formula (unless a pacemaker is in place).

[0384] Ongoing AEs >Grade 2 (NCI CTCAE V5.0) caused by any prior anti-cancer therapy with the exception of Grade 2 alopecia, vitiligo, fatigue, and active hypothyroidism, and stable neuropathy.

[0385] Ongoing or recent (within 2 years) evidence of significant autoimmune disease that required treatment with systemic immunosuppressive treatments, which may suggest risk for immune-related AEs (irAEs). Participants with the following conditions are eligible: vitiligo, childhood asthma that has resolved, residual hypothyroidism that required only hormone replacement or psoriasis that does not require systemic treatment.

[0386] Has a known history or any evidence of interstitial lung disease or active, non-infectious pneumonitis within 3 years prior to the first dose of the study drug. A history of radiation pneumonitis in the radiation field is permitted.

[0387] Organ transplant requiring immunosuppressive treatment.

[0388] Uncontrolled or active infection with human immunodeficiency virus (HIV), hepatitis B, or hepatitis C infection, or has a diagnosis of immunodeficiency:

[0389] • Participants are tested for hepatitis C virus (HCV) and hepatitis B virus (HBV) at screening.

[0390] • Participants with a known HIV infection, if controlled (undetectable viral load [HIV RNA PCR] and CD4 count above 350, either spontaneously or on a stable antiviral regimen), are permitted. For participants with a controlled HIV infection, monitoring will be performed per local standards.

[0391] • Participants with a controlled hepatitis B (HBsAg+) infection (serum HBV DNA PCR that is below the limit of detection and receiving anti-viral therapy for hepatitis B) are permitted. Participants with controlled infections must undergo periodic monitoring of the HBV DNA. Participants must remain on anti-viral therapy throughout the study treatment.Attorney Docket No. 0183-0350-00PCT-KAD-ZSP ZSP ref.: 589-395 PCT • Participants who are hepatitis C virus antibody positive (HCV Ab+), and whose infection is controlled (undetectable HCV RNA by PCR either spontaneously or in response to a successful prior course of anti-HCV therapy), may be enrolled into the study.

[0392] Clinically not controlled chronic or ongoing infectious disease requiring hospitalization or treatment within the 14 days prior first IMP administration.

[0393] Known severe hypersensitivity (> Grade 3) to or contraindication for the use of any study intervention or components thereof, including premedication that may be administered in this study. Otherwise, sensitivity of any grade or allergies that, in the opinion of the investigator contraindicates participation in the study. For Cohort E3 (receiving cetuximab) Participant has a history of allergic reactions attributed to compounds of chemical or biologic composition similar to those of cetuximab, or if the participant had red meat allergy / tick bite history.

[0394] Prior / concomitant therapy

[0395] Is unable or unwilling to take the premedication that may be used.

[0396] Last administration of prior antitumor therapy within 21 days or less than 5 times the halflife, whichever is shorter; major surgery or local therapy within 21 days prior to first dose of IMP. Participants who received anti-PD-1 therapy require an interval of 90 days prior to first dose.

[0397] Washout period of less than 2 weeks prior to radiotherapy. A 1-week washout is permitted for palliative radiation for a non-central nervous system disease.

[0398] Receipt of a live-virus vaccination within 28 days of the planned treatment start date.

[0399] Prior / concurrent clinical study experience

[0400] Participation in any other clinical study in which the last investigational study treatment administration was within 5 half-lives from this study’s intervention initiation ie, prior to the first IMP administration.

[0401] Diagnostic assessments

[0402] Inadequate organ and bone marrow functions as evidenced by:

[0403] • Absolute neutrophil count (ANC) <1500 cells / pL (1.5 x 109 / L). No granulocyte colony stimulating factor (G-CSF) is permitted in order to reach this value.

[0404] • Platelets <100 x 103 / pL or <100 x 109 / L. Platelet transfusion is not allowed within 3 days prior to the screening hematological test.

[0405] • Hemoglobin <9 g / dL (transfusion support within prior 2 weeks is not permitted). Participants can be on stable dose of erythropoietin (> approximately 3 months).Attorney Docket No. 0183-0350-00PCT-KAD-ZSP ZSP ref.: 589-395 PCT • Total bilirubin >1.5 x ULN unless direct bilirubin < ULN (Participants with known Gilbert disease who have serum bilirubin level <3 x ULN may be enrolled).

[0406] • Aspartate aminotransferase (AST / SGOT) and / or alanine aminotransferase (ALT / SGPT) >2.5 x ULN (or >5 x ULN for participants with liver metastases). Alkaline phosphatase (ALP) >2.5 x ULN (>5.0 x ULN if bone or liver metastases).

[0407] • Estimated glomerular filtration rate (eGFR) <50 mL / min / 1.73 m2(Modification of Diet in Renal Disease [MDRD] Formula).

[0408] • International Normalized Ratio (INR) or Prothrombin Time (PT) (or Activated Partial Thromboplastin Time [aPTT]) >1.5 x ULN, unless the participant is receiving anticoagulant therapy, and as long as PT or aPTT is within therapeutic range of intended use of anticoagulants.

[0409] Other exclusions

[0410] Individuals accommodated in an institution because of regulatory or legal order, such as prisoners or participants who are legally institutionalized.

[0411] Participant not suitable for participation, whatever the reason, as judged by the Investigator, including medical or clinical conditions, or if the participants are potentially at risk of noncompliance to the study procedures.

[0412] Participants are employees of the clinical study site or other individuals directly involved in the conduct of the study, or immediate family members of such individuals (in conjunction with section 1.61 of the ICH-Good Clinical Practice [GCP] Ordinance E6).

[0413] Any country-related specific regulation that would prevent the participant from entering the study.

[0414] Example 2: Clinical results, Safety and Efficacy Data

[0415] A total of 49 patients have received the study drug intravenously (IV) in dose escalation monotherapy, 32 in Q2W and 17 in QW respectively. A summary of patient demographics and baseline characteristics (dose escalation study) is shown in Tables la and lb.Attorney Docket No. 0183-0350-00PCT-KAD-ZSP ZSP ref.: 589-395 PCT Table 1a - Dose Escalation by Dose Level

[0416]

[0417] Table 1b - Dose Escalation by Dose Level (detailed)

[0418]

[0419] 2A. Safety Profile

[0420] The study drug was generally well tolerated. In a preliminary analysis and confirmed in the complete analysis, 6 Dose limiting toxicities (DLTs) have been reported, 1 at 30mg Q2W, 3 at 40mg Q2W and 2 at 5mg QW out of 49 participants exposed. A brief description of the six cases follows:

[0421] At 40ms Q2W:Attorney Docket No. 0183-0350-00PCT-KAD-ZSP ZSP ref.: 589-395 PCT A 72-year-old male with adenocarcinoma of the lung experienced Grade 4 metabolic acidosis on cycle 2 day 7 (C2D7) of dose level 5 IV Q2W presenting with dyspnea (Grade 3). Evaluation noted elevated lactate level and positive urine ketones. Concomitant medications included oral antidiabetic medication. Treatments received were inhalation therapy, prednisone, hydrocortisone, sodium bicarbonate, tocilizumab and furosemide. Participant recovered from event 11 days after onset. Study drug was withdrawn. The event was assessed as related to study treatment.

[0422] A 69-y ear-old male with urothelial neoplasm experienced Grade 4 pneumonia on C1D6 of dose level 5 IV Q2W presenting with fever (Grade 2), dyspnea (Grade 3), and tachycardia (Grade 1). Radiologic evaluation demonstrated findings consistent with pneumonia. Infectious disease evaluation was negative. Treatments received were cefepime and amikacin. Participant recovered and was discharged from hospital on C1D13. No action was taken with study drug. The event was assessed as not related to study treatment.

[0423] A 52-year-old male with stage 4 squamous cell carcinoma of the esophagus experienced Grade 3 immune-related hyperbilirubinemia on C1D4 of dose level 5 IV Q2W presenting with oral thrush (Grade 2), dysgeusia (Grade 1), nausea (Grade 1), anorexia (Grade 1), arthralgia (Grade 1) and fatigue (Grade 1). Laboratory evaluation noted total bilirubin of 4.4 mg / dL and direct bilirubin of 3.1 mg / dL. Treatment received was prednisone at 1 mg / kg. Event was resolving. Study drug was withdrawn. The event was assessed as related to study treatment.

[0424] At 30ms Q2W:

[0425] A 28-year-old male with intraabdominal desmoplastic small round cell tumor experienced Grade 3 gastrointestinal bleeding on C2D4 of dose level 6 IV Q2W. Etiology of bleeding was not determined. Treatments received were dipyrone, packed red blood cell transfusions, omeprazole, and pantoprazole. Participant recovered from event on C2D11. No action was taken with study drug. The event was assessed as related to study treatment.

[0426] At 5mg OW:

[0427] A 70-year-old male with stage 4 mucinous adenocarcinoma of the stomach experienced Grade 2 cytokine release syndrome (CRS) on CID 1 of dose level 1 IV QW presenting with fever (Grade 1) and hypotension (Grade 1). Treatments were IV fluids and tocilizumab. On C1D2 the participant experienced Grade 3 CRS with hypotension (blood pressure [BP] = 83 / 58). Treatments received were IV fluids, tocilizumab and norepinephrine. Participant recovered 2 days after Grade 3 CRS event. No action was taken with the study drug. The event was assessed as related to study treatment.Attorney Docket No. 0183-0350-00PCT-KAD-ZSP ZSP ref.: 589-395 PCT A 60-year-old female with stage 4 colon adenocarcinoma experienced Grade 1 CRS on C1D1 of dose level 1 IV QW presenting with fever (Grade 1) and diarrhea (Grade 1). On C1D3 the participant progressed to Grade 4 CRS with fever (Grade 4), hypoxia (Grade 4), diarrhea (Grade 4) and hypotension (Grade 4). Gastrointestinal infection evaluation noted positive results for norovirus and salmonella. Treatments were IV fluids, antibiotics, tocilizumab, oxygen therapy, and methylprednisolone. Participant was recovering from event. Study drug was withdrawn. The event was assessed as related to study treatment.

[0428] All participants experienced at least one treatment emergent adverse effect (TEAE), and 28 participants (57%) experienced a Grade >3 TEAE. Treatment related TEAE was observed in 46 participants (94%) with 15 participants experiencing a Grade >3 treatment related TEAE. The most common treatment related TEAEs were cytokine release syndrome (CRS) (n=31; 63%), pyrexia (n=16; 33%), fatigue (n=12; 25%), and nausea (n=ll; 22%).

[0429] CRS was the most common treatment related TEAE observed in 33 participants (67%) with 13 participants (27%) experiencing treatment emergent SAE.

[0430] The majority of episodes were Grade 1 or 2. There were two Grade 3 and one Grade 4 CRS episodes related to study drug. All episodes recovered with supportive care.

[0431] Based on the available data (Table 2a), for monotherapy with the study drug a dose of 30mg given Q2W is the Maximum tolerated dose (MTD), as 6 participants were treated by 30 mg Q2W and targeted probability of toxicity at 30 mg was highest (data not shown).

[0432] Table 2a - Summary of Treated Patients Who Had DLT by Dose Level

[0433]

[0434] Table 2b shows the complete (updated)detailed summary of all adverse events and lists TEAEs, treatment-related adverse events (TRAEs) and treatment-emergent serious adverse events (TESAE).Attorney Docket No. 0183-0350-00PCT-KAD-ZSP ZSP ref.: 589-395 PCT Table 2b - Adverse events

[0435] > >

[0436]

[0437] § Cause of death was progressive disease during post-treatment period.

[0438] Table 2c shows TEAEs of special interest by system organ class (SOC) and preferred term (PT) (all grade).

[0439] Table 2c - Adverse events

[0440]

[0441] Attorney Docket No. 0183-0350-00PCT-KAD-ZSP ZSP ref.: 589-395 PCT

[0442]

[0443] AESI, adverse event of special interest; SAE, serious adverse events

[0444] Maximum administered dose (MAD) for monotherapy with the study drug is a dose of 40mg given Q2W, as a total of 4 patients were treated by 40mg Q2W and DLT evaluable. 3 DLTs were observed.

[0445] Q2W schedule is safe, shows efficacy, and is patient-friendly (in terms of both patient preferences and compliance). At the highest dose tested (7.5mg) in QW schedule a total of 4 patients were treated and DLT evaluable. No DLT was observed.

[0446] 2B. Clinical Efficacy

[0447] Assessment of tumor response is conducted using RECIST vl.l (Figure 1, Table 3 and 4 represent results from the preliminary analysis).

[0448] Table 3 - Data summary clinical response and safety by dose (Q2W)

[0449]

[0450] Attorney Docket No. 0183-0350-00PCT-KAD-ZSP ZSP ref.: 589-395 PCT Table 4 - Data summary clinical response and safety by dose (QW)

[0451]

[0452] cPR: confirmed Partial Response ; uPR: unconfirmed Partial Response ; SD: stable disease ; DLT: dose limiting toxicity

[0453] Table 5 - A Total of 6 Partial Response (PR) and 3 Long-Stable Disease (SD) > 6 Months or 1 year Occurred in Q2W Cohorts (complete and updated analysis)

[0454] >

[0455] >

[0456] > >

[0457] >

[0458]

[0459] cPR = confirmed partial response; uPR = unconfirmed partial response; IO = immuno-oncology; CRC = colorectal cancer; MSS = microsatellite stable; SCO = squamous cell carcinoma.Attorney Docket No. 0183-0350-00PCT-KAD-ZSP ZSP ref.: 589-395 PCT Table 6 - A Total of 3 cPR and 3 Long SD > 6 Months or 1 year Occurred in QW Cohorts (complete and updated analysis)

[0460] > >

[0461] > >

[0462] >

[0463]

[0464] cPR = confirmed partial response; SCCHN = squamous cell carcinoma of the head and neck; SCC = squamous cell carcinoma.

[0465] Clinical activity was seen at different doses including the lowest ones in both schedules.

[0466] 2C. Pharmacokinetic (PK) and Pharmacodynamic (PD) Data

[0467] Study drug PK has Target-Mediated Drug Disposition (TMDD) shaped kinetics, exhibiting both dose and time dependency (Figure 2). After first dosing (cycle 1), study drug is quantifiable up to 72h or 1 week depending on the dose level. Study drug exposure increases with dose increase. Then, study drug exposure decreases over cycles (not in all patients). Mean elimination half-life ranges from 13 to 32 hours. At Cycle 3, exposure was lower in most of participants and the study drug was quantifiable up to 3 days.

[0468] In addition, most of the participants developed ADA (Anti-Drug Antibody) during treatment but no apparent impact on clinical responses was observed.

[0469] 2D. Receptor occupancy

[0470] Binding of study drug to PD-1 resulted in reduced drug-free PD-1 on CD8 T cells from 8 to 72 hours post-infusion. Very little free PD1 was detected at C1D4 except for the samples from 2.5 mg dose level, suggesting that administered study drug at 5 mg was sufficient to saturate PD-1 binding until 72 hours post treatment in cycle 1. Compared with C1D4, faster recovery of free PD-1 at C3D4 was detected, which was correlated with lower study drug exposure in cycle 3. Data indicated that samples from 2.5 mg, 5 mg, and 7.5 mg dose levels showed faster free PD-1 recovery at C3D4 compared with those from higher dose levels. In summary, 10 mg would be the minimal dose to maintain longer PD-1 occupancy in repeat dosing of study drug. PD-1 Receptor occupancy (RO) and T cell proliferation data suggest 10 mg as suitable minimal dose to maintain long PD-1Attorney Docket No. 0183-0350-00PCT-KAD-ZSP ZSP ref.: 589-395 PCT receptor occupancy and to have good target engagement during repeat dosing of study drug (Figure 3).

[0471] 2E. Cell proliferation

[0472] Cell proliferation was studied and summarized descriptively using change from baseline and over time following administration of the study drug

[0473] T cell proliferation measured by Ki67 indicated that study drug induced NK and CD8+ T cell proliferation. Samples from 5 mg dose level (cohort 2) showed the lowest NK and CD8+ T cell proliferation in cycle 3 (only one sample from cohort 2). NK and CD8+ T cell Ki67 expressions level were relatively close. The data suggest that 10 mg (cohort 3) would be the minimal dose level for activate NK and CD8 T cell proliferation after repeat dosing of study drug (Figure 4).

[0474] Study drug also showed an increase in CD3+ and CD8+ T cell, lymphocyte and NK cell counts in peripheral blood, whatever the administered dose (Figure 5).

[0475] Finally, the drug increased activated Peripheral CD8 T Cell Counts (Figure 6).

[0476] 2F. Cytokine and chemokine analysis

[0477] Change in cytokine and chemokine levels following treatment administration from baseline and over time was analysed and summarized descriptively, and their relationship with serum exposure of the study drug was assessed.

[0478] Blood levels of cytokines and chemokines were assessed at different time points. Cytokine analysis was performed in a central laboratory. Results were correlated with participants’ clinical parameters. IL-6, IL-8, TNFa, and INFy levels were used for Cytokine Release Syndrome (CRS) evaluation.

[0479] Cytokine release, measured by both mean concentration (ng / L) and mean fold change, is observed at all the administered doses and as low as 1.4 mg; 40 mg cohort is significantly higher than the rest of cohorts (Figure 7).

[0480] Chemokine release, measured by both mean concentration (ng / L) and mean fold change, show a trend to a dose dependent increase (Figure 8).

[0481] In conclusion, treatment with the study drug resulted in increased:

[0482] - NK and CD8+ T cell proliferation (Figure 4),

[0483] - CD3+ T cell, CD8+ T cell, lymphocyte and NK cell counts in periphery blood (Figure 5), - activated Peripheral CD8 T Cell Counts (Figure 6),Attorney Docket No. 0183-0350-00PCT-KAD-ZSP ZSP ref.: 589-395 PCT - serum cytokine release (Figure 7), wherein cytokine release is observed at all the administered doses; and

[0484] - serum chemokine release (Figure 8), wherein a dose dependent increase of chemokine release is measured by both mean concentration and mean fold change.

[0485] Taken together, biomarker data indicate that the tested compound is active at a dose as low as 1.4 mg.

[0486] Circulating tumor DNA (ctDNA) was analysed in patient peripheral blood samples (Figure 10). For ctDNA analysis, patient peripheral blood samples were collected in Streck Cell-Free DNA BCT® tube for extraction of plasma at the central lab. Batched plasma samples were then shipped to Guardant Health and analyzed by Guardant Infinity Platform. Molecular response scores were calculated as mean variant allele frequencies (mVAF) for patients with paired samples at screening and during the treatment (at C4D1). Results were transferred per data transfer specifications as mutually agreed upon.

[0487] The data from Figure 10 shows a correlation between the best overall response [PR and long SD (> 6 months)] measured by RECIST vl.l and the molecular responders measured by ctDNA MR score.

[0488] Conclusion

[0489] Clinical and biomarker data show an efficacy of the tested compound in all the tested range, i.e., from 1.4 mg to 40 mg [corresponding to a concentration of 0.02 mg / kg to 0.57 mg / kg for a mean patient weight of 70kg],

[0490] Safety data show a high frequency of dose limiting toxicities (DLT) at 40 mg, suggesting that the tested compound should be administered at a dose of less than 40mg. In particular, for a Q2W administration, a dose of 30 mg [concentration of 0.43 mg / kg] has been shown to be the Maximum tolerated dose (MTD).

[0491] Example 3: Clinical results from Dose Expansion / Dose Optimization Study

[0492] Efficacy Data

[0493] Part 1 A stage 1 in mCRC

[0494] Table 7 presents efficacy data from the Dose Expansion Phase (Stage 1) of the clinical trial evaluating the investigational molecule (study drug) across two cohorts (El receiving 10 mg andAttorney Docket No. 0183-0350-00PCT-KAD-ZSP ZSP ref.: 589-395 PCT E2 receiving 30 mg, n=15 each; total N=30). Tumor response was assessed using Response Evaluation Criteria in Solid Tumors version 1.1 (RECIST vl.l).

[0495] Table 7 - Summary of Best Overall Response in mCRC Stage 1 Dose Optimization

[0496]

[0497] Abbreviations: Cl = confidence interval; RECIST = Response Evaluation Criteria in Solid Tumors.

[0498] Notes:

[0499] Objective Response Rate (ORR) is based on RECIST 1.1 criteria, EUROPEAN JOURNAL OF CANCER 45 (2009) 228 — 247: Table 2. ORR is based on confirmed Best Overall Response (BOR), which is obtained combining target and non-target lesion assessment results. Exact binomial (Clopper-Pearson) 90% confidence intervals are presented.

[0500] Key Findings During Dose Optimization in CRC Patients:

[0501] The study drug demonstrated measurable anti-tumor activity in the exposed population, with an overall Objective Response Rate (ORR) of 16.7% (90% CI: 6.81-31.90). Cohort E2 exhibited higher response rates (26.7%; 90% CI: 9.67-51.08) compared to Cohort El (6.7%; 90% CI: 0.34-27.94).

[0502] Disease Control: 30.0% of patients achieved stable disease (SD), indicating the molecule's potential to halt tumor progression.

[0503] Response Distribution: Five patients (16.7%) achieved partial response (PR); no complete responses (CR) were observed in this early-stage cohort.

[0504] Progressive Disease: 46.7% of patients experienced disease progression, which is consistent with the advanced / refractory nature of the study population.

[0505] This data supports the molecule's biological activity and provides evidence of therapeutic benefit, which is relevant to patent claims regarding therapeutic efficacy and clinical utility in the specified indication.Attorney Docket No. 0183-0350-00PCT-KAD-ZSP ZSP ref.: 589-395 PCT NSCLC

[0506] Table 8 presents efficacy data from the Dose Expansion Phase (Stage 1) of the clinical trial evaluating the investigational molecule (study drug) across two cohorts (Al receiving 10 mg and A2 receiving 30 mg, n=15 (Al), 14 (A2); total N=29). Tumor response was assessed using Response Evaluation Criteria in Solid Tumors version 1.1 (RECIST vl.l).

[0507] Table 8 - Summary of Best Overall Response in NSCLC Stage 1 Dose Optimization

[0508]

[0509] Abbreviations: Cl = confidence interval; RECIST = Response Evaluation Criteria in Solid Tumors.

[0510] Notes:

[0511] Objective Response Rate (ORR) is based on RECIST 1.1 criteria, EUROPEAN JOURNAL OF CANCER 45 (2009) 228 — 247: Table 2. ORR is based on confirmed Best Overall Response (BOR), which is obtained combining target and non-target lesion assessment results. Exact binomial (Clopper-Pearson) 90% confidence intervals are presented.

[0512] Key Findings During Dose Optimization in NSCLC Patients:

[0513] The study drug demonstrated measurable anti-tumor activity in the exposed population, with an overall Objective Response Rate (ORR) of 20.7% (90% CI: 9.42-36.80). Response rate in Cohort Al was 20.0% (90% CI: 5.68-43.98). Response rate in Cohort A2 was 21.4% (90% CI: 5.68-43.98).

[0514] Disease Control: 31.0% of patients achieved stable disease (SD), indicating the molecule's potential to halt tumor progression.

[0515] Response Distribution: Six patients (20.7%) achieved partial response (PR); no complete responses (CR) were observed in this early-stage cohort.

[0516] Progressive Disease: 37.9% of patients experienced disease progression, which is consistent with the advanced / refractory nature of the study population.

[0517] This data supports the molecule's biological activity and provides evidence of therapeutic benefit, which is relevant to patent claims regarding therapeutic efficacy and clinical utility in the specified indication.Attorney Docket No. 0183-0350-00PCT-KAD-ZSP ZSP ref.: 589-395 PCT

[0518] Safety Data

[0519] Table 9 provides a summary of major TEAEs observed by dose in all tumor types (excluding lab abnormalities).

[0520] Table 9 - Number (%) of participants with TEAE(s) with a PT incidence > 20% in at least one dose level by primary SOC and PT (worst grade by participant) - Exposed population &

[0521]

[0522] Table lOpresents an overview of participants with TEAE(s) leading to permanent study intervention discontinuation (lung cancer).Attorney Docket No. 0183-0350-00PCT-KAD-ZSP ZSP ref.: 589-395 PCT Table 10 - Number (%) of Participants with TEAE(s) Leading to Permanent Study Intervention Discontinuation by Primary SOC and PT (Exposed Population) for Lung Cancer

[0523] > > >

[0524]

[0525] Attorney Docket No. 0183-0350-00PCT-KAD-ZSP ZSP ref.: 589-395 PCT Table 11 presents an overview of participants with TEAE(s) leading to permanent study intervention discontinuation (mCRC).

[0526] Table 11 - Number (%) of Participants with TEAE(s) Leading to Permanent Study Intervention Discontinuation by Primary SOC and PT (Exposed Population) for mCRC

[0527] > > >

[0528]

[0529] Attorney Docket No. 0183-0350-00PCT-KAD-ZSP ZSP ref.: 589-395 PCT Table 12 - Summary of TEAEs leading to permanent discontinuation

[0530]

[0531] Abbreviations: CTCAE = Common Terminology Criteria for Adverse Events; MedDRA = Medical Dictionary for Regulatory Activities; PT = Preferred Term; SOC = System Organ Class; TEAE = treatment-emergent adverse event.

[0532] Notes:

[0533] At each level of summarization within each column (any event and preferred term), participants reporting more than one adverse event are counted only once using the highest CTCAE grade.

[0534] CTCAE grades are scored as: Grade 1 = Mild; Grade 2 = Moderate; Grade 3 = Severe or medically significant; Grade 4 = Life-threatening; Grade 5 = Death related to AE.

[0535] Adverse events are coded to system organ class and preferred term using MedDRA, Version 28.1

[0536] All adverse events were coded using MedDRA Version 28.1 and graded according to CTCAE criteria. Participants reporting multiple adverse events within each category were counted only once using the highest CTCAE grade, ensuring conservative safety estimates.

[0537] Discontinuation rates are comparable to or more favorable than approved oncology therapeutics, with no unexpected or novel safety signals identified. The safety profile is consistent with mechanism of action expectations.

[0538] The discontinuation rates caused by treatment-emergent adverse events, regardless of relationship to the molecule, were comparable between cohorts, with Cohort Al (10 mg) demonstrating 13.3% and Cohort A2 (30 mg) showing 14.3% overall discontinuation rates. This consistency across cohorts indicates reproducible safety characteristics of the study drug. Notably,Attorney Docket No. 0183-0350-00PCT-KAD-ZSP ZSP ref: 589-395 PCT the majority of discontinuations (75%, 3 of 4 participants) were associated with severe or lifethreatening events (Grade >3).

[0539] SEQUENCES

[0540] Table 13. Sequences according to the invention

[0541]

[0542] Attorney Docket No. 0183-0350-00PCT-KAD-ZSP ZSP ref.: 589-395 PCT

[0543]

[0544] Attorney Docket No. 0183-0350-00PCT-KAD-ZSP ZSP ref.: 589-395 PCT

[0545]

Claims

1. Attorney Docket No. 0183-0350-00PCT-KAD-ZSP ZSP ref.: 589-395 PCT CLAIMS1. An immunocytokine for use in the treatment of cancer, wherein the immunocytokine comprises:a) an antibody or antigen binding fragment thereof binding to PD1 comprising a heavy chain variable domain (VH) comprising complementarity determining regions HCDR1 to HCDR3 as set forth in SEQ ID NO: 3 to 5, respectively, and a light chain variable domain (VL) comprising complementarity determining regions LCDR1 to LCDR3 as set forth in SEQ ID NO: 6 to 8, respectively; andb) an interleukin- 15 (IL-15) conjugate, wherein said IL-15 conjugate is located N-terminal of VH and comprises:i. an IL- 15 polypeptide comprising an amino acid sequence as set forth in SEQ ID NO: 9 or an amino acid sequence having at least 80% identity to SEQ ID NO: 9; andii. an IL-15Ra sushi domain polypeptide comprising an amino acid sequence as set forth in SEQ ID NO: 11; andiii. optionally a polypeptide linker LI linking i and ii; andc) optionally a polypeptide linker L2 linking a) and b);characterized in that the immunocytokine is administered at a dose of 0.02 to 0.6 mg / kg.

2. The immunocytokine for use of claim 1, wherein the antibody or antigen binding fragment thereof comprises a VH as set forth in SEQ ID NO: 1 and a VL as set forth in SEQ ID NO: 1.

3. The immunocytokine for use of claim 1 or 2, wherein the IL-15 polypeptide comprises an amino acid sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 9, in particular an amino acid sequence as set forth in SEQ ID NO: 10.

4. The immunocytokine for use of any of claims 1-3, wherein the IL-15Ra sushi domain polypeptide further comprises an amino acid sequence as set forth in SEQ ID NO: 26.

5. The immunocytokine for use of any of claims 1-4, wherein the IL-15Ra sushi domain polypeptide comprises or consists of an amino acid sequence as set forth in SEQ ID NO: 27 or 28.Attorney Docket No. 0183-0350-00PCT-KAD-ZSP ZSP ref.: 589-395 PCT 6. The immunocytokine for use of any of claims 1-5, wherein the IL-15Ra sushi domain polypeptide is located N-terminal of the IL- 15 polypeptide.

7. The immunocytokine for use of any of claims 1-6, wherein the IL- 15 conjugate comprises a polypeptide linker LI, wherein LI is a serine-glycine linker of 1-40 amino acids, preferably comprising an amino acid sequence as set forth in SEQ ID NO: 19 or 20.

8. The immunocytokine for use of any of claims 1-7, wherein the IL-15 conjugate comprises an amino acid sequence as set forth in SEQ ID NO: 12 or 29.

9. The immunocytokine for use of any of claims 1-8, wherein the antibody is an IgGl antibody.

10. The immunocytokine for use of any of claims 1-9, wherein the antibody comprises LALA mutations L234A and L235A.

11. The immunocytokine for use of any of claims 1-10, wherein the antibody comprises knob- into-hole (kiH) mutations, preferably at least one, in particular all, of the Y349C, T366S, L368A, Y407V, S354C and T366W mutations.

12. The immunocytokine for use of any of claims 1-11, wherein the antibody comprises a first heavy chain (HC1) comprising a constant region as set forth in SEQ ID NO: 13, a second heavy chain (HC2) comprising a constant region as set forth in SEQ ID NO: 14, and two light chains (LC) each comprising a constant region as set forth in SEQ ID NO: 18.

13. The immunocytokine for use of any of claims 1-12, wherein the immunocytokine comprises a polypeptide linker L2, wherein L2 is a serine-glycine linker of 1-40 amino acids, preferably comprising an amino acid sequence as set forth in SEQ ID NO: 22.

14. The immunocytokine for use of any of claims 1-13, wherein the immunocytokine consists of two LC each consisting of an amino acid sequence as set forth in SEQ ID NO: 17, HC1 consisting of an amino acid sequence as set forth in SEQ ID NO: 15 and HC2 consisting of an amino acid sequence as set forth in SEQ ID NO: 16.Attorney Docket No. 0183-0350-00PCT-KAD-ZSP ZSP ref.: 589-395 PCT 15. The immunocytokine for use of any of claims 1-14, wherein the use comprises administering the immunocytokine at a dose of at least 0.02, 0.036, 0.07, 0.1 or 0.14 mg / kg and no more than 0.57, 0.43 or 0.28 mg / kg, preferably at a dose between 0.1 mg / kg and about 0.5 mg / kg.

16. The immunocytokine for use of any of claims 1-15, wherein the use comprises administering the immunocytokine at a dose of at least 0.02, 0.036, 0.07, 0.1 or 0.14 mg / kg and no more than 0.43 or 0.28 mg / kg, preferably at a dose between 0.14 mg / kg to 0.43 mg / kg or from 0.14 mg / kg to about 0.28 mg / kg.

17. The immunocytokine for use of any of claims 1-16, wherein the use comprises administering the immunocytokine at a dose of about 0.1, 0.14, 0.28, 0.43 or 0.5 mg / kg, preferably 0.14, 0.28 or 0.43 mg / kg.

18. The immunocytokine for use of any of claims 1-17, wherein the use comprises administering the immunocytokine at a dose of 1.4 to 40 mg.

19. The immunocytokine for use of any of claims 1-18, wherein the use comprises administering the immunocytokine at a dose of at least 1.4, 2.5, 5, 7.5 or 10 mg and no more than 40, 30 or 20 mg, preferably at a dose between 7.5 mg and about 35 mg.

20. The immunocytokine for use of any of claims 1-19, wherein the use comprises administering the immunocytokine at a dose of at least 1.4, 2.5, 5, 7.5 or 10 mg and no more than 30 or 20 mg, preferably at a dose between 10 to 30 mg or from 10 to 20 mg.

21. The immunocytokine for use of any of claims 20, wherein the use comprises administering the immunocytokine at a dose of about 7.5, 10, 20, 30 or 35 mg, preferably 10, 20 or 30 mg.

22. The immunocytokine for use of any of claims 1-21, wherein the use comprises biweekly administration of the immunocytokine.

23. The immunocytokine for use of any of claims 1-22, wherein the use comprises intravenous administration of the immunocytokine.Attorney Docket No. 0183-0350-00PCT-KAD-ZSP ZSP ref.: 589-395 PCT 24. The immunocytokine for use of any of claims 1-23, wherein the cancer is selected from the group consisting of non-small cell lung cancer, hepatocellular carcinoma, gastric cancer, gastroesophageal junction adenocarcinoma, sigmoid adenocarcinoma, cutaneous squamous cell carcinoma, colorectal cancer, rectal adenocarcinoma, ascending colon cancer, penile squamous cell carcinoma, sarcomatoid lung cancer, urothelial carcinoma, uveal melanoma, squamous cell carcinoma of the head and neck, squamous cell carcinoma of the scalp, gastric adenocarcinoma, myxofibrosarcoma, salivary gland acinic cell carcinoma, and soft tissue sarcoma.

25. The immunocytokine for use of claim 24, wherein the cancer is non-small cell lung cancer (NSCLC).

26. The immunocytokine for use of claim 24, wherein the cancer is colorectal cancer (CRC).

27. The immunocytokine for use of claim 24, wherein the gastric cancer and the gastroesophageal junction adenocarcinoma has non-MSI-H status, proficient MMR status or HER2 / neu negative status.

28. The immunocytokine for use of any of claims 1-27, whereina. the cancer is unresectable;b. the cancer is metastatic;c. use comprises administering the immunocytokine to a subject that is refractory to standard approved therapy;d. use comprises administering the immunocytokine to a subj ect that is not a candidate for standard approved therapy; ore. any combination of a)-d).

29. The immunocytokine for use of any of claims 1-26, wherein the use further comprises administration of a second anti-cancer drug.

30. The immunocytokine for use of claim 27, wherein the second anti-cancer drug is a therapeutic antibody.Attorney Docket No. 0183-0350-00PCT-KAD-ZSP ZSP ref.: 589-395 PCT 31. The immunocytokine for use of any of claims 1-28, wherein the use further comprises administration of a premedication.

32. The immunocytokine for use of claim 29, wherein the premedication is selected from the group consisting of dexamethasone, acetaminophen, diphenhydramine, cetirizine, promethazine, dexchlorpheniramine, meperidine and montelukast.

33. A method of treating cancer, comprising administration of an immunocytokine to a human subject in need thereof, wherein the immunocytokine comprises:a) an antibody or antigen binding fragment thereof binding to PD1 comprising a heavy chain variable domain (VH) comprising complementarity determining regions HCDR1 to HCDR3 as set forth in SEQ ID NO: 3 to 5, respectively, and a light chain variable domain (VL) comprising complementarity determining regions LCDR1 to LCDR3 as set forth in SEQ ID NO: 6 to 8, respectively; andb) an interleukin- 15 (IL-15) conjugate, wherein said IL-15 conjugate is located N-terminal of VH and comprises:i. IL- 15 or an IL- 15 derivative, wherein the IL- 15 comprises an amino acid sequence as set forth in SEQ ID NO: 9 and the IL-15 derivative comprises an amino acid sequence having at least 80 % identity to SEQ ID NO: 9; andii. a sushi domain of IL- 15 receptor alpha (IL-15Ra) as defined in SEQ ID NO: 11;andiii. optionally a polypeptide linker LI linking i and ii; andc) optionally a polypeptide linker L2 linking a) and b);characterized in that the immunocytokine is administered at a dose of 0.02 to 0.6 mg / kg.

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