Anticancer combination therapy

Abstract

To provide a method for treating cancer.SOLUTION: The present invention provides a polypeptide capable of specifically binding to LRP5 and LRP6 for use in a method for treating and / or preventing a hyperproliferative disease, preferably cancer, wherein the method comprises administering the polypeptide capable of specifically binding to LRP5 and LRP6 in combination with a PD-1 antibody to a patient in need thereof.SELECTED DRAWING: None

Description

Technical Field

[0001] Field of the Invention The present invention relates to combination therapies in the treatment of cancer and compounds for use in such combination therapies. The compounds for combination are LRP5 / 6 antagonists and PD-1 antagonists.

[0002] Background of the Invention Activation of the Wnt signaling pathway requires binding of extracellular Wnt ligands to the Frizzled receptor and the co-receptor LRP5 (accession number: UniProtKB - O75197 / LRP5_HUMAN) or its closely related homolog LRP6 (accession number: UniProtKB - O75581 / LRP6_HUMAN). Mammalian cells contain 19 Wnt proteins and 10 Frizzled receptors. In the absence of Wnt ligands, cytoplasmic beta-catenin is phosphorylated by a protein complex consisting of the scaffold proteins Axin and APC and the kinases GSK3 beta and CK1a. Subsequently, when recognized by the ubiquitin ligase beta-TrCP, ubiquitin-mediated degradation of beta-catenin occurs. In the presence of Wnt ligands, when Wnt binds to Frizzled and LRP5 or LRP6, the cytoplasmic effector protein Dvl is recruited, and the cytoplasmic tail of LRP5 or LRP6 is phosphorylated, which provides a docking site for Axin. Isolation of Axin by LRP5 or LRP6 inactivates the Axin-APC-GSK3 beta complex, and thus, intracellular beta-catenin is stabilized and accumulates. Consequently, the cytoplasmic level of beta-catenin increases, beta-catenin translocates to the nucleus, and forms a complex with members of the transcription factor family of T cell factor (TCF) / lymphoid enhancer-binding factor (LEF). Subsequently, the basal transcription apparatus and transcription co-activator containing cAMP response element-binding protein (CREB)-binding protein (CBP) or its homolog p300 are recruited, and various target genes including Axin2, Cyclin D1, Naked1, Notum, and c-Myc are expressed.

[0003] Higher levels of ligand-dependent Wnt pathway regulation are mediated by the E3 ligase RNF43 and its closely related homolog ZNRF3, as well as secreted R-spondin proteins (de Lau et al., "The R-spondin / Lgr5 / Rnf43 module: regulator of Wnt signal strength," Genes Dev. 2014; 28(4):305-16). RNF43 mediates the ubiquitination of the Frizzled / LRP5 or LRP6 receptor complex at the cell surface, leading to its degradation, thereby inhibiting ligand-dependent Wnt pathway activity. The activity of RNF43 is counteracted by R-spondin family members (R-spondin 1-4 ligands). In the presence of R-spondin ligands, the R-spondin ligands remove RNF43 from the cell surface, allowing for the accumulation of the Frizzled / LRP5 or LRP6 complex and enhancement of Wnt signaling in the presence of Wnt ligands.

[0004] Hyperactivation of Wnt signaling is involved in the etiology of various types of cancer in at least two different ways, although not in all. In some types of cancer, frequent mutations in downstream signaling molecules contribute to a constitutively activated Wnt pathway (e.g., APC mutations in colorectal cancer, beta-catenin activating mutations in hepatocellular carcinoma). On the other hand, in other types of cancer, such as triple-negative breast cancer (TNBC), non-small cell lung cancer (NSCLC), pancreatic adenocarcinoma, etc., as well as in subsets of colorectal cancer (CRC) and endometrial cancer, activation of Wnt signaling is driven by a ligand-dependent mechanism (i.e., by autocrine / paracrine Wnt activation) such that it is detected by intracellular accumulation of beta-catenin.In NSCLC, TNBC, and pancreatic adenocarcinoma, ligand-dependent Wnt activation is mediated by multiple mechanisms, including increased expression of Wnt ligands and / or LRP5 and LRP6 receptors or silencing of DKK1, a negative regulator of LRP5 and LRP6 (TNBC: Liu et al., "LRP6 overexpression defines a class of breast cancer subtype and is a target for therapy", Proc Natl Acad Sci U S A 2010; 107(11):5136-41; Khramtsov et al., "Wnt / beta-catenin pathway activation is enriched in basal-like breast cancers and predicts poor outcome", Am J Pathol. 2010; 176(6): 2911-20; NSCLC: Nakashima et al., "Wnt1 overexpression associated with tumor proliferation and a poor prognosis in non-small cell lung cancer patients", Oncol Rep. 2008; 19(1):203-9; pancreatic cancer: Zhang et al., "Canonical wnt signaling is required for pancreatic carcinogenesis", Cancer Res. 2013; 73(15):4909-22). In particular, published data show that in healthy tissues (e.g., breast and lung epithelium), beta-catenin is localized only to the cell membrane. In contrast, in the majority of primary clinical samples of TNBC, NSCLC, and pancreatic adenocarcinoma, beta-catenin intracellular accumulation due to abnormal Wnt signaling (i.e., cytoplasmic / nuclear, a biomarker of Wnt signaling activation) was shown.In recent publications, ligand-dependent activation of Wnt signaling has been shown to be mediated by activation of mutant / inactivated RNF43 (Giannakis et al., "RNF43 is frequently mutated in colorectal and endometrial cancers", Nat Genet. 2014; 46(12):1264-6) or R-spondin fusion transcripts (encoding R-spondin 2 or R-spondin 3 proteins driven by a constitutively active and strong promoter; Seshagiri et al., "Recurrent R-spondin fusions in colon cancer", Nature 2012; 488(7413):660-4) in subsets of CRC and endometrial cancer. Both inactivating RNF43 mutations and R-spondin fusion transcripts have been shown to enhance ligand-dependent Wnt signaling in vitro by increasing the abundance of Frizzled on the cell surface. Ligand-dependent Wnt activation in tumors has been shown to drive tumor growth and resistance to chemotherapy or immunotherapy and is associated with recurrence in preclinical models.

[0005] Since LRP5 and LRP6 function as gatekeepers for ligand-dependent activation of Wnt signaling, they can be considered as targets to achieve complete blockade of the pathway mediated by all 19 Wnt ligands and 10 Frizzled receptors.

[0006] An alternative approach to the above-described methods that directly target cancer / cancer cells is cancer immunotherapy. Cancer immunotherapy is a field of oncology in which the immune system is used to treat cancer and is in stark contrast to existing common treatment methods that directly excise or treat tumors. This treatment concept is based on the identification of a number of proteins on the surface of T cells that act to inhibit the immune function of these cells. Among these proteins, PD-1 (programmed cell death 1) is listed.

[0007] PD-1 is a cell surface receptor protein expressed on T cells. PD-1 has two ligands: PD-L1 and PD-L2. These ligands interact with the cell surface receptor. Upon ligand binding, PD-1 recruits intracellular signals that negatively regulate T cell responses. Thus, typically, this protein functions as an "immune checkpoint" inhibitor, i.e., it acts to modulate the activity of cells of the immune system so as to regulate and limit autoimmune diseases. It has been understood in recent years that many cancers can protect themselves from the immune system and thus avoid detection by modifying "immune checkpoint" inhibitors.

[0008] Therefore, it has also been shown in a range of different cancer settings that antagonist PD-1 antibody molecules, such as nivolumab and pembrolizumab, etc., can stimulate the immune system and thereby be used to treat cancer.

[0009] Despite the above-mentioned progress in the treatment of cancer, there is still a need for new treatment concepts for the treatment of cancer that show advantages over standard therapies. These advantages can include efficacy in vivo (e.g., improvement of clinical response, prolongation of response, increase in response rate, duration of response, disease stabilization rate, duration of stabilization, time to disease progression, progression-free survival (PFS) and / or overall survival (OS), later onset of resistance, etc.), safe and well-tolerated administration, and a reduction in the frequency and severity of adverse events. Specifically, there is a need for additional treatment options for patients with cancers such as lung cancer (e.g., NSCLC), melanoma, bladder cancer, and gastrointestinal cancer, etc.

[0010] For this reason, the object of the present invention is to provide a treatment for cancer that is more advantageous than currently used and / or known treatments / treatment methods in the prior art and is novel.

[0011] Summary of the Invention The present invention provides a method for treating a patient having a proliferative disorder by using an antibody specific for programmed cell death 1 (PD-1) (this term is used interchangeably herein with the terms "anti-PD-1 antibody", "PD-1 antibody" or "PD-1 antagonist") together with an LRP5 / LRP6 antagonist (this term is used interchangeably herein with the terms "polypeptide that specifically binds to LRP5 and LRP6" or "polypeptide that can specifically bind to LRP5 and LRP6"), thereby antagonizing the PD-1 signaling pathway. Accordingly, the present invention provides a combination therapy comprising an LRP5 / LRP6 antagonist and an anti-PD-1 antibody.

[0012] In one aspect, the present invention is a polypeptide that can specifically bind to LRP5 and LRP6 for use in a method for treating and / or preventing a proliferative disorder, preferably cancer, wherein the method comprises administering a polypeptide that can specifically bind to LRP5 and LRP6 in combination with a PD-1 antibody to a patient in need thereof, wherein the polypeptide that can specifically bind to LRP5 and LRP6 (i) a first immunoglobulin single variable domain (ISVD) (a) comprising the following CDR sequences: CDR1: TYTVG (= SEQ ID NO: 40) CDR2: AIRRRGSSTYYADSVKG (= SEQ ID NO: 41) CDR3: DTRTVALLQYRYDY (= SEQ ID NO: 42) and a second ISVD (b) comprising the following CDR sequences: CDR1: SYAMG (= SEQ ID NO: 49) CDR2: AISWSGGSTYYADSVKG (= SEQ ID NO: 50) CDR3: SPIPYGSLLRRRNNYDY (= SEQ ID NO: 51) ; (ii) the following CDR sequences: CDR1: SYAMG (= SEQ ID NO: 43) CDR2: AIRRSGRRTYYADSVKG (= Sequence number: 44) CDR3: ARRVRSSTRYNTGTWWWEY (= Sequence number: 45) A first ISVD (a) comprising The following CDR sequences: CDR1: SYAMG (= Sequence number: 49) CDR2: AISWSGGSTYYADSVKG (= Sequence number: 50) CDR3: SPIPYGSLLRRRNNYDY (= Sequence number: 51) A polypeptide comprising a second ISVD (b); (iii) The following CDR sequences: CDR1: RYTMG (= Sequence number: 46) CDR2: AIVRSGGSTYYADSVKG (= Sequence number: 47) CDR3: DRRGRGENYILLYSSGRYEY (= Sequence number: 48) A first ISVD (a) comprising The following CDR sequences: CDR1: SYAMG (= Sequence number: 49) CDR2: AISWSGGSTYYADSVKG (= Sequence number: 50) CDR3: SPIPYGSLLRRRNNYDY (= Sequence number: 51) A polypeptide comprising a second ISVD (b); (iv) The following CDR sequences: CDR1: TYTVG (= Sequence number: 40) CDR2: AIRRRGSSTYYADSVKG (= Sequence number: 41) CDR3: DTRTVALLQYRYDY (= Sequence number: 42) A first ISVD (a) comprising The following CDR sequences: CDR1: SYAMG (= Sequence number: 52) CDR2: AISWRSGSTYYADSVKG (= Sequence number: 53) CDR3: DPRGYGVAYVSAYYEY (= Sequence number: 54) A polypeptide comprising a second ISVD (b); (v) The following CDR sequences: CDR1: SYAMG (= Sequence No.: 43) CDR2: AIRRSGRRTYYADSVKG (= Sequence No.: 44) CDR3: ARRVRSSTRYNTGTWWWEY (= Sequence No.: 45) A first ISVD (a) comprising: (v) The following CDR sequences: CDR1: SYAMG (= Sequence No.: 52) CDR2: AISWRSGSTYYADSVKG (= Sequence No.: 53) CDR3: DPRGYGVAYVSAYYEY (= Sequence No.: 54) A polypeptide comprising a second ISVD (b) comprising: (vi) The following CDR sequences: CDR1: RYTMG (= Sequence No.: 46) CDR2: AIVRSGGSTYYADSVKG (= Sequence No.: 47) CDR3: DRRGRGENYILLYSSGRYEY (= Sequence No.: 48) A first ISVD (a) comprising: (v) The following CDR sequences: CDR1: SYAMG (= Sequence No.: 52) CDR2: AISWRSGSTYYADSVKG (= Sequence No.: 53) CDR3: DPRGYGVAYVSAYYEY (= Sequence No.: 54) A polypeptide comprising a second ISVD (b) comprising: Selected from the group consisting of: wherein the PD-1 antibody is (i) An anti-PD1 antibody comprising a heavy chain CDR comprising the amino acid sequences of SEQ ID NO: 1 (HCDR1), SEQ ID NO: 2 (HCDR2) and SEQ ID NO: 3 (HCDR3) and a light chain CDR comprising the amino acid sequences of SEQ ID NO: 4 (LCDR1), SEQ ID NO: 5 (LCDR2) and SEQ ID NO: 6 (LCDR3); (ii) An anti-PD1 antibody comprising a heavy chain CDR comprising the amino acid sequences of SEQ ID NO: 7 (HCDR1), SEQ ID NO: 8 (HCDR2) and SEQ ID NO: 9 (HCDR3) and a light chain CDR comprising the amino acid sequences of SEQ ID NO: 10 (LCDR1), SEQ ID NO: 11 (LCDR2) and SEQ ID NO: 12 (LCDR3), and (iii) An anti-PD1 antibody comprising a heavy chain CDR comprising the amino acid sequences of SEQ ID NO: 13 (HCDR1), SEQ ID NO: 14 (HCDR2) and SEQ ID NO: 15 (HCDR3) and a light chain CDR comprising the amino acid sequences of SEQ ID NO: 16 (LCDR1), SEQ ID NO: 17 (LCDR2) and SEQ ID NO: 18 (LCDR3) A polypeptide selected from the group consisting of

[0013] In another aspect, the present invention is a method of treating and / or preventing a proliferative disorder, preferably cancer, comprising administering to a patient in need thereof a therapeutically effective amount of a polypeptide capable of specifically binding to LRP5 and LRP6 and a therapeutically effective amount of a PD-1 antibody, wherein the polypeptide capable of specifically binding to LRP5 and LRP6 is (i) The following CDR sequences: CDR1: TYTVG (= SEQ ID NO: 40) CDR2: AIRRRGSSTYYADSVKG (= SEQ ID NO: 41) CDR3: DTRTVALLQYRYDY (= SEQ ID NO: 42) comprising a first immunoglobulin single variable domain (ISVD) (a) and the following CDR sequences: CDR1: SYAMG (= SEQ ID NO: 49) CDR2: AISWSGGSTYYADSVKG (= SEQ ID NO: 50) CDR3: SPIPYGSLLRRRNNYDY (= SEQ ID NO: 51) comprising a second ISVD (b); (ii) The following CDR sequences: CDR1: SYAMG (= SEQ ID NO: 43) CDR2: AIRRSGRRTYYADSVKG (= Sequence number: 44) CDR3: ARRVRSSTRYNTGTWWWEY (= Sequence number: 45) a first ISVD (a) comprising the following CDR sequences: CDR1: SYAMG (= Sequence number: 49) CDR2: AISWSGGSTYYADSVKG (= Sequence number: 50) CDR3: SPIPYGSLLRRRNNYDY (= Sequence number: 51) a polypeptide comprising a second ISVD (b); (iii) the following CDR sequences: CDR1: RYTMG (= Sequence number: 46) CDR2: AIVRSGGSTYYADSVKG (= Sequence number: 47) CDR3: DRRGRGENYILLYSSGRYEY (= Sequence number: 48) a first ISVD (a) comprising the following CDR sequences: CDR1: SYAMG (= Sequence number: 49) CDR2: AISWSGGSTYYADSVKG (= Sequence number: 50) CDR3: SPIPYGSLLRRRNNYDY (= Sequence number: 51) a polypeptide comprising a second ISVD (b); (iv) the following CDR sequences: CDR1: TYTVG (= Sequence number: 40) CDR2: AIRRRGSSTYYADSVKG (= Sequence number: 41) CDR3: DTRTVALLQYRYDY (= Sequence number: 42) a first ISVD (a) comprising the following CDR sequences: CDR1: SYAMG (= Sequence number: 52) CDR2: AISWRSGSTYYADSVKG (= Sequence number: 53) CDR3: DPRGYGVAYVSAYYEY (= Sequence number: 54) a polypeptide comprising a second ISVD (b); (v) The following CDR sequences: CDR1: SYAMG (= Sequence No.: 43) CDR2: AIRRSGRRTYYADSVKG (= Sequence No.: 44) CDR3: ARRVRSSTRYNTGTWWWEY (= Sequence No.: 45) A first ISVD (a) comprising: (v) The following CDR sequences: CDR1: SYAMG (= Sequence No.: 52) CDR2: AISWRSGSTYYADSVKG (= Sequence No.: 53) CDR3: DPRGYGVAYVSAYYEY (= Sequence No.: 54) A polypeptide comprising a second ISVD (b) comprising: (vi) The following CDR sequences: CDR1: RYTMG (= Sequence No.: 46) CDR2: AIVRSGGSTYYADSVKG (= Sequence No.: 47) CDR3: DRRGRGENYILLYSSGRYEY (= Sequence No.: 48) A first ISVD (a) comprising: (vi) The following CDR sequences: CDR1: SYAMG (= Sequence No.: 52) CDR2: AISWRSGSTYYADSVKG (= Sequence No.: 53) CDR3: DPRGYGVAYVSAYYEY (= Sequence No.: 54) A polypeptide comprising a second ISVD (b) comprising: Selected from the group consisting of: wherein the PD-1 antibody is (i) An anti-PD1 antibody comprising a heavy chain CDR comprising the amino acid sequences of SEQ ID NO: 1 (HCDR1), SEQ ID NO: 2 (HCDR2) and SEQ ID NO: 3 (HCDR3) and a light chain CDR comprising the amino acid sequences of SEQ ID NO: 4 (LCDR1), SEQ ID NO: 5 (LCDR2) and SEQ ID NO: 6 (LCDR3); (ii) an anti-PD1 antibody comprising a heavy chain CDR comprising the amino acid sequences of SEQ ID NO: 7 (HCDR1), SEQ ID NO: 8 (HCDR2), and SEQ ID NO: 9 (HCDR3) and a light chain CDR comprising the amino acid sequences of SEQ ID NO: 10 (LCDR1), SEQ ID NO: 11 (LCDR2), and SEQ ID NO: 12 (LCDR3), and (iii) an anti-PD1 antibody comprising a heavy chain CDR comprising the amino acid sequences of SEQ ID NO: 13 (HCDR1), SEQ ID NO: 14 (HCDR2), and SEQ ID NO: 15 (HCDR3) and a light chain CDR comprising the amino acid sequences of SEQ ID NO: 16 (LCDR1), SEQ ID NO: 17 (LCDR2), and SEQ ID NO: 18 (LCDR3) selected from the group consisting of.

[0014] In another aspect, the present invention is a PD-1 antibody for use in a method of treating and / or preventing a proliferative disorder, preferably cancer, wherein the method comprises administering to a patient in need thereof a combination of a PD-1 antibody with a polypeptide capable of specifically binding to LRP5 and LRP6, wherein the polypeptide capable of specifically binding to LRP5 and LRP6 is (i) the following CDR sequences: CDR1: TYTVG (= SEQ ID NO: 40) CDR2: AIRRRGSSTYYADSVKG (= SEQ ID NO: 41) CDR3: DTRTVALLQYRYDY (= SEQ ID NO: 42) comprising a first immunoglobulin single variable domain (ISVD) (a) and the following CDR sequences: CDR1: SYAMG (= SEQ ID NO: 49) CDR2: AISWSGGSTYYADSVKG (= SEQ ID NO: 50) CDR3: SPIPYGSLLRRRNNYDY (= SEQ ID NO: 51) comprising a second ISVD (b); (ii) the following CDR sequences: CDR1: SYAMG (= SEQ ID NO: 43) CDR2: AIRRSGRRTYYADSVKG (= Sequence number: 44) CDR3: ARRVRSSTRYNTGTWWWEY (= Sequence number: 45) a first ISVD (a) comprising the following CDR sequences: CDR1: SYAMG (= Sequence number: 49) CDR2: AISWSGGSTYYADSVKG (= Sequence number: 50) CDR3: SPIPYGSLLRRRNNYDY (= Sequence number: 51) a polypeptide comprising a second ISVD (b); (iii) the following CDR sequences: CDR1: RYTMG (= Sequence number: 46) CDR2: AIVRSGGSTYYADSVKG (= Sequence number: 47) CDR3: DRRGRGENYILLYSSGRYEY (= Sequence number: 48) a first ISVD (a) comprising the following CDR sequences: CDR1: SYAMG (= Sequence number: 49) CDR2: AISWSGGSTYYADSVKG (= Sequence number: 50) CDR3: SPIPYGSLLRRRNNYDY (= Sequence number: 51) a polypeptide comprising a second ISVD (b); (iv) the following CDR sequences: CDR1: TYTVG (= Sequence number: 40) CDR2: AIRRRGSSTYYADSVKG (= Sequence number: 41) CDR3: DTRTVALLQYRYDY (= Sequence number: 42) a first ISVD (a) comprising the following CDR sequences: CDR1: SYAMG (= Sequence number: 52) CDR2: AISWRSGSTYYADSVKG (= Sequence number: 53) CDR3: DPRGYGVAYVSAYYEY (= Sequence number: 54) a polypeptide comprising a second ISVD (b); (v) The following CDR sequences: CDR1: SYAMG (= Sequence number: 43) CDR2: AIRRSGRRTYYADSVKG (= Sequence number: 44) CDR3: ARRVRSSTRYNTGTWWWEY (= Sequence number: 45) A first ISVD (a) comprising and The following CDR sequences: CDR1: SYAMG (= Sequence number: 52) CDR2: AISWRSGSTYYADSVKG (= Sequence number: 53) CDR3: DPRGYGVAYVSAYYEY (= Sequence number: 54) A polypeptide comprising a second ISVD (b) comprising and (vi) The following CDR sequences: CDR1: RYTMG (= Sequence number: 46) CDR2: AIVRSGGSTYYADSVKG (= Sequence number: 47) CDR3: DRRGRGENYILLYSSGRYEY (= Sequence number: 48) A first ISVD (a) comprising and The following CDR sequences: CDR1: SYAMG (= Sequence number: 52) CDR2: AISWRSGSTYYADSVKG (= Sequence number: 53) CDR3: DPRGYGVAYVSAYYEY (= Sequence number: 54) A polypeptide comprising a second ISVD (b) comprising selected from the group consisting of wherein the PD-1 antibody is (i) An anti-PD1 antibody comprising a heavy chain CDR comprising the amino acid sequences of SEQ ID NO: 1 (HCDR1), SEQ ID NO: 2 (HCDR2) and SEQ ID NO: 3 (HCDR3) and a light chain CDR comprising the amino acid sequences of SEQ ID NO: 4 (LCDR1), SEQ ID NO: 5 (LCDR2) and SEQ ID NO: 6 (LCDR3) (ii) An anti-PD1 antibody comprising a heavy chain CDR comprising the amino acid sequences of SEQ ID NO: 7 (HCDR1), SEQ ID NO: 8 (HCDR2), and SEQ ID NO: 9 (HCDR3) and a light chain CDR comprising the amino acid sequences of SEQ ID NO: 10 (LCDR1), SEQ ID NO: 11 (LCDR2), and SEQ ID NO: 12 (LCDR3), and (iii) An anti-PD1 antibody comprising a heavy chain CDR comprising the amino acid sequences of SEQ ID NO: 13 (HCDR1), SEQ ID NO: 14 (HCDR2), and SEQ ID NO: 15 (HCDR3) and a light chain CDR comprising the amino acid sequences of SEQ ID NO: 16 (LCDR1), SEQ ID NO: 17 (LCDR2), and SEQ ID NO: 18 (LCDR3) provided, which is selected from the group consisting of PD-1 antibodies.

[0015] In another aspect, the present invention is the use of a polypeptide capable of specifically binding to LRP5 and LRP6 for the manufacture of a pharmaceutical composition for use in treating and / or preventing a proliferative disorder, preferably cancer, wherein the polypeptide capable of specifically binding to LRP5 and LRP6 is used in combination with a PD-1 antibody, wherein the polypeptide capable of specifically binding to LRP5 and LRP6 is (i) The following CDR sequences: CDR1: TYTVG (= SEQ ID NO: 40) CDR2: AIRRRGSSTYYADSVKG (= SEQ ID NO: 41) CDR3: DTRTVALLQYRYDY (= SEQ ID NO: 42) comprising a first immunoglobulin single variable domain (ISVD) (a) and the following CDR sequences: CDR1: SYAMG (= SEQ ID NO: 49) CDR2: AISWSGGSTYYADSVKG (= SEQ ID NO: 50) CDR3: SPIPYGSLLRRRNNYDY (= SEQ ID NO: 51) comprising a second ISVD (b); (ii) The following CDR sequences: CDR1: SYAMG (= SEQ ID NO: 43) CDR2: AIRRSGRRTYYADSVKG (= Sequence number: 44) CDR3: ARRVRSSTRYNTGTWWWEY (= Sequence number: 45) A first ISVD (a) comprising The following CDR sequences: CDR1: SYAMG (= Sequence number: 49) CDR2: AISWSGGSTYYADSVKG (= Sequence number: 50) CDR3: SPIPYGSLLRRRNNYDY (= Sequence number: 51) A polypeptide comprising a second ISVD (b); (iii) The following CDR sequences: CDR1: RYTMG (= Sequence number: 46) CDR2: AIVRSGGSTYYADSVKG (= Sequence number: 47) CDR3: DRRGRGENYILLYSSGRYEY (= Sequence number: 48) A first ISVD (a) comprising The following CDR sequences: CDR1: SYAMG (= Sequence number: 49) CDR2: AISWSGGSTYYADSVKG (= Sequence number: 50) CDR3: SPIPYGSLLRRRNNYDY (= Sequence number: 51) A polypeptide comprising a second ISVD (b); (iv) The following CDR sequences: CDR1: TYTVG (= Sequence number: 40) CDR2: AIRRRGSSTYYADSVKG (= Sequence number: 41) CDR3: DTRTVALLQYRYDY (= Sequence number: 42) A first ISVD (a) comprising The following CDR sequences: CDR1: SYAMG (= Sequence number: 52) CDR2: AISWRSGSTYYADSVKG (= Sequence number: 53) CDR3: DPRGYGVAYVSAYYEY (= Sequence number: 54) A polypeptide comprising a second ISVD (b); (v) The following CDR sequences: CDR1: SYAMG (= Sequence No.: 43) CDR2: AIRRSGRRTYYADSVKG (= Sequence No.: 44) CDR3: ARRVRSSTRYNTGTWWWEY (= Sequence No.: 45) A first ISVD (a) comprising and The following CDR sequences: CDR1: SYAMG (= Sequence No.: 52) CDR2: AISWRSGSTYYADSVKG (= Sequence No.: 53) CDR3: DPRGYGVAYVSAYYEY (= Sequence No.: 54) A polypeptide comprising a second ISVD (b) and (vi) The following CDR sequences: CDR1: RYTMG (= Sequence No.: 46) CDR2: AIVRSGGSTYYADSVKG (= Sequence No.: 47) CDR3: DRRGRGENYILLYSSGRYEY (= Sequence No.: 48) A first ISVD (a) comprising and The following CDR sequences: CDR1: SYAMG (= Sequence No.: 52) CDR2: AISWRSGSTYYADSVKG (= Sequence No.: 53) CDR3: DPRGYGVAYVSAYYEY (= Sequence No.: 54) A polypeptide comprising a second ISVD (b) selected from the group consisting of wherein the PD-1 antibody is (i) An anti-PD1 antibody comprising a heavy chain CDR containing the amino acid sequences of SEQ ID NO: 1 (HCDR1), SEQ ID NO: 2 (HCDR2) and SEQ ID NO: 3 (HCDR3) and a light chain CDR containing the amino acid sequences of SEQ ID NO: 4 (LCDR1), SEQ ID NO: 5 (LCDR2) and SEQ ID NO: 6 (LCDR3), (ii) An anti-PD1 antibody comprising a heavy chain CDR comprising the amino acid sequences of SEQ ID NO: 7 (HCDR1), SEQ ID NO: 8 (HCDR2) and SEQ ID NO: 9 (HCDR3) and a light chain CDR comprising the amino acid sequences of SEQ ID NO: 10 (LCDR1), SEQ ID NO: 11 (LCDR2) and SEQ ID NO: 12 (LCDR3), and (iii) An anti-PD1 antibody comprising a heavy chain CDR comprising the amino acid sequences of SEQ ID NO: 13 (HCDR1), SEQ ID NO: 14 (HCDR2) and SEQ ID NO: 15 (HCDR3) and a light chain CDR comprising the amino acid sequences of SEQ ID NO: 16 (LCDR1), SEQ ID NO: 17 (LCDR2) and SEQ ID NO: 18 (LCDR3) selected from the group consisting of, is provided for use.

[0016] In another aspect, the present invention is the use of a PD-1 antibody for the manufacture of a pharmaceutical composition for use in treating and / or preventing a proliferative disorder, preferably cancer, wherein the PD-1 antibody is used in combination with a polypeptide capable of specifically binding to LRP5 and LRP6, and the polypeptide capable of specifically binding to LRP5 and LRP6 is (i) The following CDR sequences: CDR1: TYTVG (= SEQ ID NO: 40) CDR2: AIRRRGSSTYYADSVKG (= SEQ ID NO: 41) CDR3: DTRTVALLQYRYDY (= SEQ ID NO: 42) comprising a first immunoglobulin single variable domain (ISVD) (a) and the following CDR sequences: CDR1: SYAMG (= SEQ ID NO: 49) CDR2: AISWSGGSTYYADSVKG (= SEQ ID NO: 50) CDR3: SPIPYGSLLRRRNNYDY (= SEQ ID NO: 51) comprising a second ISVD (b); (ii) The following CDR sequences: CDR1: SYAMG (= SEQ ID NO: 43) CDR2: AIRRSGRRTYYADSVKG (= Sequence number: 44) CDR3: ARRVRSSTRYNTGTWWWEY (= Sequence number: 45) a first ISVD (a) comprising the following CDR sequences: CDR1: SYAMG (= Sequence number: 49) CDR2: AISWSGGSTYYADSVKG (= Sequence number: 50) CDR3: SPIPYGSLLRRRNNYDY (= Sequence number: 51) a polypeptide comprising a second ISVD (b); (iii) the following CDR sequences: CDR1: RYTMG (= Sequence number: 46) CDR2: AIVRSGGSTYYADSVKG (= Sequence number: 47) CDR3: DRRGRGENYILLYSSGRYEY (= Sequence number: 48) a first ISVD (a) comprising the following CDR sequences: CDR1: SYAMG (= Sequence number: 49) CDR2: AISWSGGSTYYADSVKG (= Sequence number: 50) CDR3: SPIPYGSLLRRRNNYDY (= Sequence number: 51) a polypeptide comprising a second ISVD (b); (iv) the following CDR sequences: CDR1: TYTVG (= Sequence number: 40) CDR2: AIRRRGSSTYYADSVKG (= Sequence number: 41) CDR3: DTRTVALLQYRYDY (= Sequence number: 42) a first ISVD (a) comprising the following CDR sequences: CDR1: SYAMG (= Sequence number: 52) CDR2: AISWRSGSTYYADSVKG (= Sequence number: 53) CDR3: DPRGYGVAYVSAYYEY (= Sequence number: 54) a polypeptide comprising a second ISVD (b); (v) The following CDR sequences: CDR1: SYAMG (= Sequence No.: 43) CDR2: AIRRSGRRTYYADSVKG (= Sequence No.: 44) CDR3: ARRVRSSTRYNTGTWWWEY (= Sequence No.: 45) A first ISVD (a) comprising: and The following CDR sequences: CDR1: SYAMG (= Sequence No.: 52) CDR2: AISWRSGSTYYADSVKG (= Sequence No.: 53) CDR3: DPRGYGVAYVSAYYEY (= Sequence No.: 54) A polypeptide comprising a second ISVD (b) comprising: and (vi) The following CDR sequences: CDR1: RYTMG (= Sequence No.: 46) CDR2: AIVRSGGSTYYADSVKG (= Sequence No.: 47) CDR3: DRRGRGENYILLYSSGRYEY (= Sequence No.: 48) A first ISVD (a) comprising: and The following CDR sequences: CDR1: SYAMG (= Sequence No.: 52) CDR2: AISWRSGSTYYADSVKG (= Sequence No.: 53) CDR3: DPRGYGVAYVSAYYEY (= Sequence No.: 54) A polypeptide comprising a second ISVD (b) comprising: Selected from the group consisting of wherein the PD-1 antibody is (i) An anti-PD1 antibody comprising a heavy chain CDR comprising the amino acid sequences of SEQ ID NO: 1 (HCDR1), SEQ ID NO: 2 (HCDR2) and SEQ ID NO: 3 (HCDR3) and a light chain CDR comprising the amino acid sequences of SEQ ID NO: 4 (LCDR1), SEQ ID NO: 5 (LCDR2) and SEQ ID NO: 6 (LCDR3); (ii) An anti-PD1 antibody comprising a heavy chain CDR comprising the amino acid sequences of SEQ ID NO: 7 (HCDR1), SEQ ID NO: 8 (HCDR2), and SEQ ID NO: 9 (HCDR3) and a light chain CDR comprising the amino acid sequences of SEQ ID NO: 10 (LCDR1), SEQ ID NO: 11 (LCDR2), and SEQ ID NO: 12 (LCDR3), and (iii) An anti-PD1 antibody comprising a heavy chain CDR comprising the amino acid sequences of SEQ ID NO: 13 (HCDR1), SEQ ID NO: 14 (HCDR2), and SEQ ID NO: 15 (HCDR3) and a light chain CDR comprising the amino acid sequences of SEQ ID NO: 16 (LCDR1), SEQ ID NO: 17 (LCDR2), and SEQ ID NO: 18 (LCDR3) selected from the group consisting of, is provided for use.

[0017] In another aspect, the present invention · A polypeptide capable of specifically binding to LRP5 and LRP6, · A PD-1 antibody, · Optionally, one or more pharmaceutically acceptable carriers, excipients, and / or vehicles, wherein the polypeptide capable of specifically binding to LRP5 and LRP6 (i) The following CDR sequences: CDR1: TYTVG (= SEQ ID NO: 40) CDR2: AIRRRGSSTYYADSVKG (= SEQ ID NO: 41) CDR3: DTRTVALLQYRYDY (= SEQ ID NO: 42) comprising a first immunoglobulin single variable domain (ISVD) (a) and the following CDR sequences: CDR1: SYAMG (= SEQ ID NO: 49) CDR2: AISWSGGSTYYADSVKG (= SEQ ID NO: 50) CDR3: SPIPYGSLLRRRNNYDY (= SEQ ID NO: 51) comprising a second ISVD (b); (ii) The following CDR sequences: CDR1: SYAMG (= SEQ ID NO: 43) CDR2: AIRRSGRRTYYADSVKG (= Sequence number: 44) CDR3: ARRVRSSTRYNTGTWWWEY (= Sequence number: 45) A first ISVD (a) containing The following CDR sequences: CDR1: SYAMG (= Sequence number: 49) CDR2: AISWSGGSTYYADSVKG (= Sequence number: 50) CDR3: SPIPYGSLLRRRNNYDY (= Sequence number: 51) A polypeptide containing a second ISVD (b); (iii) The following CDR sequences: CDR1: RYTMG (= Sequence number: 46) CDR2: AIVRSGGSTYYADSVKG (= Sequence number: 47) CDR3: DRRGRGENYILLYSSGRYEY (= Sequence number: 48) A first ISVD (a) containing The following CDR sequences: CDR1: SYAMG (= Sequence number: 49) CDR2: AISWSGGSTYYADSVKG (= Sequence number: 50) CDR3: SPIPYGSLLRRRNNYDY (= Sequence number: 51) A polypeptide containing a second ISVD (b); (iv) The following CDR sequences: CDR1: TYTVG (= Sequence number: 40) CDR2: AIRRRGSSTYYADSVKG (= Sequence number: 41) CDR3: DTRTVALLQYRYDY (= Sequence number: 42) A first ISVD (a) containing The following CDR sequences: CDR1: SYAMG (= Sequence number: 52) CDR2: AISWRSGSTYYADSVKG (= Sequence number: 53) CDR3: DPRGYGVAYVSAYYEY (= Sequence number: 54) A polypeptide containing a second ISVD (b); (v) The following CDR sequences: CDR1: SYAMG (= Sequence No.: 43) CDR2: AIRRSGRRTYYADSVKG (= Sequence No.: 44) CDR3: ARRVRSSTRYNTGTWWWEY (= Sequence No.: 45) A first ISVD (a) comprising and The following CDR sequences: CDR1: SYAMG (= Sequence No.: 52) CDR2: AISWRSGSTYYADSVKG (= Sequence No.: 53) CDR3: DPRGYGVAYVSAYYEY (= Sequence No.: 54) A polypeptide comprising a second ISVD (b) comprising and (vi) The following CDR sequences: CDR1: RYTMG (= Sequence No.: 46) CDR2: AIVRSGGSTYYADSVKG (= Sequence No.: 47) CDR3: DRRGRGENYILLYSSGRYEY (= Sequence No.: 48) A first ISVD (a) comprising and The following CDR sequences: CDR1: SYAMG (= Sequence No.: 52) CDR2: AISWRSGSTYYADSVKG (= Sequence No.: 53) CDR3: DPRGYGVAYVSAYYEY (= Sequence No.: 54) A polypeptide comprising a second ISVD (b) comprising selected from the group consisting of wherein the PD-1 antibody is (i) An anti-PD1 antibody comprising a heavy chain CDR comprising the amino acid sequences of SEQ ID NO: 1 (HCDR1), SEQ ID NO: 2 (HCDR2) and SEQ ID NO: 3 (HCDR3) and a light chain CDR comprising the amino acid sequences of SEQ ID NO: 4 (LCDR1), SEQ ID NO: 5 (LCDR2) and SEQ ID NO: 6 (LCDR3) (ii) An anti-PD1 antibody comprising a heavy chain CDR comprising the amino acid sequences of SEQ ID NO: 7 (HCDR1), SEQ ID NO: 8 (HCDR2) and SEQ ID NO: 9 (HCDR3) and a light chain CDR comprising the amino acid sequences of SEQ ID NO: 10 (LCDR1), SEQ ID NO: 11 (LCDR2) and SEQ ID NO: 12 (LCDR3), and (iii) An anti-PD1 antibody comprising a heavy chain CDR comprising the amino acid sequences of SEQ ID NO: 13 (HCDR1), SEQ ID NO: 14 (HCDR2) and SEQ ID NO: 15 (HCDR3) and a light chain CDR comprising the amino acid sequences of SEQ ID NO: 16 (LCDR1), SEQ ID NO: 17 (LCDR2) and SEQ ID NO: 18 (LCDR3) A pharmaceutical composition selected from the group consisting of

[0018] In some embodiments, the pharmaceutical composition is for use in a method of treating and / or preventing a proliferative disorder, preferably cancer.

[0019] In another aspect, the present invention provides in one or more containers · A first pharmaceutical composition or dosage form comprising a polypeptide capable of specifically binding to LRP5 and LRP6, and optionally one or more pharmaceutically acceptable carriers, excipients and / or vehicles, · A second pharmaceutical composition or dosage form comprising a PD-1 antibody and optionally one or more pharmaceutically acceptable carriers, excipients and / or vehicles, and · An accompanying document comprising, optionally, printed instructions, wherein the polypeptide capable of specifically binding to LRP5 and LRP6 is (i) The following CDR sequences: CDR1: TYTVG (= SEQ ID NO: 40) CDR2: AIRRRGSSTYYADSVKG (= SEQ ID NO: 41) CDR3: DTRTVALLQYRYDY (= SEQ ID NO: 42) Comprising a first immunoglobulin single variable domain (ISVD) (a) and The following CDR sequences: CDR1: SYAMG (= SEQ ID NO: 49) CDR2: AISWSGGSTYYADSVKG (= Sequence number: 50) CDR3: SPIPYGSLLRRRNNYDY (= Sequence number: 51) A polypeptide comprising a second ISVD(b) containing the same; (ii) The following CDR sequences: CDR1: SYAMG (= Sequence number: 43) CDR2: AIRRSGRRTYYADSVKG (= Sequence number: 44) CDR3: ARRVRSSTRYNTGTWWWEY (= Sequence number: 45) A first ISVD(a) containing the same and The following CDR sequences: CDR1: SYAMG (= Sequence number: 49) CDR2: AISWSGGSTYYADSVKG (= Sequence number: 50) CDR3: SPIPYGSLLRRRNNYDY (= Sequence number: 51) A polypeptide comprising a second ISVD(b) containing the same; (iii) The following CDR sequences: CDR1: RYTMG (= Sequence number: 46) CDR2: AIVRSGGSTYYADSVKG (= Sequence number: 47) CDR3: DRRGRGENYILLYSSGRYEY (= Sequence number: 48) A first ISVD(a) containing the same and The following CDR sequences: CDR1: SYAMG (= Sequence number: 49) CDR2: AISWSGGSTYYADSVKG (= Sequence number: 50) CDR3: SPIPYGSLLRRRNNYDY (= Sequence number: 51) A polypeptide comprising a second ISVD(b) containing the same; (iv) The following CDR sequences: CDR1: TYTVG (= Sequence number: 40) CDR2: AIRRRGSSTYYADSVKG (= Sequence number: 41) CDR3: DTRTVALLQYRYDY (= Sequence number: 42) A first ISVD (a) comprising and The following CDR sequences: CDR1: SYAMG (= SEQ ID NO: 52) CDR2: AISWRSGSTYYADSVKG (= SEQ ID NO: 53) CDR3: DPRGYGVAYVSAYYEY (= SEQ ID NO: 54) A polypeptide comprising a second ISVD (b) comprising; (v) The following CDR sequences: CDR1: SYAMG (= SEQ ID NO: 43) CDR2: AIRRSGRRTYYADSVKG (= SEQ ID NO: 44) CDR3: ARRVRSSTRYNTGTWWWEY (= SEQ ID NO: 45) A first ISVD (a) comprising and The following CDR sequences: CDR1: SYAMG (= SEQ ID NO: 52) CDR2: AISWRSGSTYYADSVKG (= SEQ ID NO: 53) CDR3: DPRGYGVAYVSAYYEY (= SEQ ID NO: 54) A polypeptide comprising a second ISVD (b) comprising and (vi) The following CDR sequences: CDR1: RYTMG (= SEQ ID NO: 46) CDR2: AIVRSGGSTYYADSVKG (= SEQ ID NO: 47) CDR3: DRRGRGENYILLYSSGRYEY (= SEQ ID NO: 48) A first ISVD (a) comprising and The following CDR sequences: CDR1: SYAMG (= SEQ ID NO: 52) CDR2: AISWRSGSTYYADSVKG (= SEQ ID NO: 53) CDR3: DPRGYGVAYVSAYYEY (= SEQ ID NO: 54) A polypeptide comprising a second ISVD (b) comprising Selected from the group consisting of Wherein the PD-1 antibody is (i) An anti-PD1 antibody comprising a heavy chain CDR containing the amino acid sequences of SEQ ID NO: 1 (HCDR1), SEQ ID NO: 2 (HCDR2), and SEQ ID NO: 3 (HCDR3) and a light chain CDR containing the amino acid sequences of SEQ ID NO: 4 (LCDR1), SEQ ID NO: 5 (LCDR2), and SEQ ID NO: 6 (LCDR3), (ii) An anti-PD1 antibody comprising a heavy chain CDR containing the amino acid sequences of SEQ ID NO: 7 (HCDR1), SEQ ID NO: 8 (HCDR2), and SEQ ID NO: 9 (HCDR3) and a light chain CDR containing the amino acid sequences of SEQ ID NO: 10 (LCDR1), SEQ ID NO: 11 (LCDR2), and SEQ ID NO: 12 (LCDR3), and (iii) An anti-PD1 antibody comprising a heavy chain CDR containing the amino acid sequences of SEQ ID NO: 13 (HCDR1), SEQ ID NO: 14 (HCDR2), and SEQ ID NO: 15 (HCDR3) and a light chain CDR containing the amino acid sequences of SEQ ID NO: 16 (LCDR1), SEQ ID NO: 17 (LCDR2), and SEQ ID NO: 18 (LCDR3) A kit is provided, which is selected from the group consisting of.

[0020] In some embodiments, the kit of the present invention is for use in a method of treating and / or preventing a proliferative disorder, preferably cancer.

[0021] In a preferred embodiment of the present invention, the polypeptide capable of specifically binding to LRP5 and LRP6 is (i) A polypeptide comprising a first ISVD containing the amino acid sequence of SEQ ID NO: 58 and a second ISVD containing the sequence of SEQ ID NO: 61, (ii) A polypeptide comprising a first ISVD containing the amino acid sequence of SEQ ID NO: 59 and a second ISVD containing the sequence of SEQ ID NO: 61, (iii) A polypeptide comprising a first ISVD containing the sequence of SEQ ID NO: 60 and a second ISVD containing the sequence of SEQ ID NO: 61, (iv) A polypeptide comprising a first ISVD containing the amino acid sequence of SEQ ID NO: 58 and a second ISVD containing the sequence of SEQ ID NO: 62, (v) A polypeptide comprising a first ISVD comprising the amino acid sequence of SEQ ID NO: 59 and a second ISVD comprising the sequence of SEQ ID NO: 62, and (vi) A polypeptide comprising a first ISVD comprising the amino acid sequence of SEQ ID NO: 60 and a second ISVD comprising the sequence of SEQ ID NO: 62 selected from the group consisting of, preferably, wherein the polypeptide capable of specifically binding to LRP5 and LRP6 further comprises an Alb11 domain comprising the amino acid sequence of SEQ ID NO: 63.

[0022] In a particularly preferred embodiment, the polypeptide capable of specifically binding to LRP5 and LRP6 comprises a polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 64, SEQ ID NO: 65, and SEQ ID NO: 66.

[0023] In a preferred embodiment of the present invention, the PD-1 antibody is (i) An antibody having a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 19 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 20, (ii) An antibody having a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 21 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 22, (iii) An antibody having a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 23 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 24, (iv) An antibody having a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 25 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 26, and (v) An antibody having a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 27 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 28 selected from the group consisting of.

[0024] In a particularly preferred embodiment of the present invention, the PD-1 antibody is (i) An antibody comprising a heavy chain comprising the amino acid sequence of SEQ ID NO: 29 and a light chain comprising the amino acid sequence of SEQ ID NO: 30, (ii) An antibody comprising a heavy chain comprising the amino acid sequence of SEQ ID NO: 31 and a light chain comprising the amino acid sequence of SEQ ID NO: 32, (iii) An antibody comprising a heavy chain comprising the amino acid sequence of SEQ ID NO: 33 and a light chain comprising the amino acid sequence of SEQ ID NO: 34, (iv) An antibody comprising a heavy chain comprising the amino acid sequence of SEQ ID NO: 35 and a light chain comprising the amino acid sequence of SEQ ID NO: 36 and (v) An antibody comprising a heavy chain comprising the amino acid sequence of SEQ ID NO: 37 and a light chain comprising the amino acid sequence of SEQ ID NO: 38 selected from the group consisting of.

[0025] In some embodiments of the present invention, the PD-1 antibody is administered simultaneously, contemporaneously, sequentially, continuously, alternately or separately with a polypeptide capable of specifically binding to LRP5 and LRP6.

[0026] In a preferred embodiment, a polypeptide capable of specifically binding to LRP5 and LRP6 and a PD-1 antibody are administered according to the following treatment schedule: (i) A first treatment period in which a polypeptide capable of specifically binding to LRP5 and LRP6 and a PD-1 antibody are administered simultaneously or contemporaneously, preferably every 3 or 4 weeks, and (ii) A second treatment period in which only the PD-1 antibody is administered and a polypeptide capable of specifically binding to LRP5 and LRP6 is not administered, preferably where the PD-1 antibody is administered every 3 or 4 weeks according to.

[0027] In a preferred embodiment of the present invention, the proliferative disease to be treated is a cancer selected from the group consisting of gastrointestinal cancer, melanoma tumor, bladder cancer and lung cancer (e.g., NSCLC), and even more preferably, the cancer is an immune therapy-resistant gastrointestinal cancer (esophageal cancer (e.g., gastroesophageal junction cancer), gastric (stomach) cancer, hepatocellular cancer, cholangiocarcinoma (e.g., bile duct cancer), gallbladder cancer, pancreatic cancer or colorectal cancer (CRC), including but not limited to these), immune therapy-resistant melanoma, immune therapy-resistant bladder cancer or immune therapy-resistant lung cancer.

[0028] In an alternative preferred embodiment of the present invention, the hyperproliferative disease to be treated is an immune therapy-resistant solid tumor.

Brief Description of the Drawings

[0029]

Figure 1

Figure 2

Figure 3

[0030] Detailed Description of the Invention Definitions The above and other aspects and embodiments of the present invention will become apparent from further description herein.

[0031] Unless otherwise specified, all terms used have their ordinary meaning in the art, which will be apparent to those skilled in the art to which the present invention pertains. In case of conflict, the patent specification including the definitions shall prevail. For example, standard handbooks such as Sambrook et al, "Molecular Cloning: A Laboratory Manual" (2nd Ed.), VoIs. 1-3, Cold Spring Harbor Laboratory Press (1989); Lewin, "Genes IV", Oxford University Press, New York, (1990) and Roitt et al., "Immunology" (2 nd Ed.), Gower Medical Publishing, London, New York (1989) and the general background art cited herein are referred to. Further, unless otherwise specified, all methods, procedures, techniques and operations not specifically described in detail can be carried out in a manner known per se, as will be apparent to those skilled in the art. Again, for example, standard handbooks, the general background art mentioned above and further references cited therein are referred to.

[0032] The term "antibody" encompasses antibodies, antibody fragments, antibody-like molecules, and conjugates with any of the foregoing. Antibodies include, but are not limited to, polyclonal antibodies, monoclonal antibodies, chimeric antibodies, humanized antibodies, human antibodies, monospecific antibodies, bispecific antibodies, or multispecific antibodies. The term "antibody" refers to molecules produced by lymphocytes, such as intact immunoglobulins present in serum, monoclonal antibodies secreted by hybridoma cell lines, polypeptides produced by recombinant expression that have the binding specificity of an immunoglobulin or monoclonal antibody, and molecules obtained from such immunoglobulins, monoclonal antibodies, or polypeptides by further processing while maintaining the binding specificity. In particular, the term "antibody" includes intact immunoglobulins containing two heavy chains and two light chains. In another embodiment, the term encompasses fragments of immunoglobulins, such as Fab fragments. In another embodiment, the term "antibody" includes polypeptides having one or more variable domains obtained from immunoglobulins, such as single-chain antibodies (scFv), single-domain antibodies, and the like.

[0033] A "human antibody" is an antibody having an amino acid sequence corresponding to the amino acid sequence of an antibody produced by human cells or obtained from a non-human source that utilizes a human antibody repertoire or other human antibody coding sequences. This definition of human antibody specifically excludes humanized antibodies that contain non-human antigen-binding residues.

[0034] As used herein, the term "recombinant human antibody" is intended to include all human antibodies prepared, expressed, produced or isolated by recombinant means, such as antibodies isolated from a host cell, such as an NS0 or CHO cell, or an animal transgenic for a human immunoglobulin gene (e.g., a mouse), or antibodies expressed using a recombinant expression vector transfected into a host cell. Such recombinant human antibodies have variable and constant regions in rearranged forms. The recombinant human antibodies of the invention have been subjected to in vivo somatic hypermutation. As a result, the amino acid sequences of the VH and VL regions of the recombinant antibodies are derived from and related to human germline VH and VL sequences, but are sequences that may not naturally occur within the human antibody germline repertoire in vivo.

[0035] A "humanized" antibody refers to a chimeric antibody that contains amino acid residues from non-human hypervariable regions (HVRs) and amino acid residues from human framework regions (FRs). In certain embodiments, a humanized antibody will comprise substantially all of at least one, typically two, variable domains. In the same variable domains, all or substantially all of the HVRs (e.g., complementarity determining regions (CDRs)) correspond to those of a non-human antibody, and all or substantially all of the entire framework regions (FRs) correspond to those of a human antibody. A humanized antibody may optionally include at least a portion of an antibody constant region obtained from a human antibody. The "humanized form" of an antibody, e.g., a non-human antibody, refers to the antibody that has been humanized.

[0036] As used herein, the terms "variable domain" or "variable region" or "Fv" refer to each of the pair of light and heavy chains that are directly involved in the binding of an antibody to an antigen. The variable domain of the light chain is abbreviated as "VL" and the variable domain of the heavy chain is abbreviated as "VH". The variable light chain domain and the variable heavy chain domain have the same overall structure, and each domain contains four framework (FR) regions, the sequences of which are widely conserved and are linked by three hypervariable regions (HVRs) or complementarity-determining regions (CDRs). The framework regions adopt a beta-sheet conformation, and the CDRs can form loops that connect the beta-sheet structures. The CDRs in each chain are held in their three-dimensional structure by the framework regions and, together with the CDRs from the other chain, form the antigen-binding site. The heavy and light chain CDR regions of an antibody play a particularly important role in the binding specificity / affinity of the antibodies of the present invention and thus provide a further object of the present invention.

[0037] In the context of the present invention, references to CDRs related to an antibody (e.g., a PD1 antibody) are based on the definitions of Kabat (E.A. Kabat, T.T. Wu, H. Bilofsky, M. Reid-Miller and H. Perry, Sequence of Proteins of Immunological Interest, National Institutes of Health, Bethesda (1983)) together with Chothia (Chothia and Lesk, J. Mol. Biol. 1987, 196: 901-917).

[0038] Unless otherwise indicated, the terms "immunoglobulin" and "immunoglobulin sequence" are used herein as general terms that include both full-length antibodies, their individual chains and all of their parts, domains or fragments (including, but not limited to, antigen-binding domains or fragments, e.g., VHH domains or VH / VL domains, respectively), whether used to refer to heavy-chain antibodies or conventional four-chain antibodies. In addition, as used herein, the term "sequence" (e.g., as used in terms such as "immunoglobulin sequence", "antibody sequence", "(single) variable domain sequence", "VHH sequence" or "protein sequence") is generally understood to include both the relevant amino acid sequence and the nucleic acid or nucleotide sequence encoding it, unless the context requires a more limited interpretation.

[0039] As used herein, the term "(domain of a polypeptide or protein)" refers to a folded protein structure that has the ability to maintain its tertiary structure independently of the rest of the protein. Generally, a domain confers distinct functional properties on a protein and can often be added to, removed from or transferred to other proteins without impairing the function of the protein and / or the rest of the domain.

[0040] As used herein, the term "immunoglobulin domain" refers to a globular region of an antibody chain (e.g., of a conventional four-chain antibody or a chain of a heavy-chain antibody) or a polypeptide consisting essentially of such a globular region. Immunoglobulin domains are characterized by their retention of the immunoglobulin-fold properties of the antibody molecule, which consists of a two-layer sandwich of approximately seven antiparallel beta-strands arranged in two beta-sheets, optionally stabilized by conserved disulfide bonds.

[0041] As used herein, the term "immunoglobulin variable domain" means an immunoglobulin domain that consists essentially of four "framework regions" that are referred to in the art and herein as "framework region 1" or "FR1", "framework region 2" or "FR2", "framework region 3" or "FR3", and "framework region 4" or "FR4", respectively. These framework regions are interrupted in the art and herein by three "complementary determining regions" or "CDRs" that are referred to as "complementary determining region 1" or "CDR1", "complementary determining region 2" or "CDR2", and "complementary determining region 3" or "CDR3", respectively. Thus, the general structure or sequence of an immunoglobulin variable domain can be shown as follows: FR1-CDR1-FR2-CDR2-FR3-CDR3-FR4. An immunoglobulin variable domain confers specificity for an antigen by having an antigen-binding site.

[0042] As used herein, the term "immunoglobulin single variable domain" (or "ISVD") means an immunoglobulin variable domain that can specifically bind to an epitope of an antigen without pairing with an additional variable immunoglobulin domain. An example of an ISVD in the context of the present invention is a "domain antibody", e.g., an ISVD VH and VL (VH domain and VL domain). Another important example of an ISVD is the "VHH domain" (or simply "VHH") from camel, as defined below.

[0043] In view of the above definition, the antigen-binding domains of conventional four-chain antibodies (e.g., IgG, IgM, IgA, IgD or IgE molecules; known in the art) or Fab fragments, F(ab’)2 fragments, Fv fragments, e.g., disulfide-bonded Fv or scFv fragments or diabodies obtained from such conventional four-chain antibodies (all known in the art) will generally not be regarded as ISVDs. In these cases, binding to each epitope of an antigen usually does not occur by one (single) immunoglobulin domain, but by a pair of (associating) immunoglobulin domains, e.g., the light and heavy chain variable domains, i.e., by the VH-VL pair of immunoglobulin domains that bind cooperatively to the epitope of each antigen.

[0044] The “VHH domain” (also known as VHH, VH domain, VHH antibody fragment and VHH antibody) has originally been described as the antigen-binding immunoglobulin (variable) domain of “heavy-chain antibodies” (i.e., “antibodies lacking light chains”; Hamers-Casterman C, Atarhouch T, Muyldermans S, Robinson G, Hamers C, Songa EB, Bendahman N, Hamers R.: “Naturally occurring antibodies devoid of light chains”; Nature 363, 446-448 (1993)). The term “VHH domain” differentiates these variable domains from the heavy-chain variable domains (referred to herein as “V H domain” or “VH domain”) present in conventional four-chain antibodies and the light-chain variable domains (referred to herein as “V H domain”) present in conventional four-chain antibodies. LIt has been selected to distinguish it from the so-called "domain" or "VL domain". The VHH domain can specifically bind to an epitope without an additional antigen-binding domain (in contrast to the VH domain or VL domain in a conventional four-chain antibody, where in this case the epitope is recognized by the VL domain together with the VH domain). The VHH domain is a small, robust, and efficient antigen recognition unit formed by a single immunoglobulin domain.

[0045] In the context of the present invention, the terms VHH domain, VHH, VH domain, VHH antibody fragment, VHH antibody, and "Nanobody (registered trademark)" and "Nanobody (registered trademark) domain" (where "Nanobody" is a trademark of Ablynx N.V.; Ghent; Belgium) are used interchangeably and are representative examples of ISVDs (having a structure of FR1-CDR1-FR2-CDR2-FR3-CDR3-FR4 and specifically binding to an epitope without the need for the presence of a second immunoglobulin variable domain), and can also be distinguished from the VH domain by so-called "hallmark residues", as defined in FIG. 1 of WO 2009 / 109635, for example.

[0046] The amino acid residues of the VHH domain are numbered according to the general numbering for the V domain given by Kabat et al. ("Sequence of proteins of immunological interest", US Public Health Services, NIH Bethesda, MD, Publication No. 91) as applied to the VHH domain from camel as shown in FIG. 2 of Riechmann and Muyldermans, J. Immunol. Methods 231, 25-38 (1999). According to this numbering, H -FR1 contains amino acid residues at positions 1 to 30, -CDR1 contains amino acid residues at positions 31 to 35, ​​ - FR2 contains the amino acids at positions 36 to 49, - CDR2 contains the amino acid residues at positions 50 to 65, - FR3 contains the amino acid residues at positions 66 to 94, - CDR3 contains the amino acid residues at positions 95 to 102 - FR4 contains the amino acid residues at positions 103 to 113.

[0047] However, as is well known in the art for V H domains and VHH domains, the total number of amino acid residues in each CDR may vary, and as a result, it should be noted that it may not correspond to the total number of amino acid residues indicated by Kabat numbering (i.e., one or more positions according to Kabat numbering may not be occupied in the actual sequence or the actual sequence may contain more amino acid residues than the number allowed by Kabat numbering). This means that although the numbering of the amino acid residues in the VHH domain is based on Kabat numbering, the actual numbering of the amino acid residues in the actual sequence may be different. Since this type of change is well known in the art, the numbering and assignment of each framework region and CDR within such sequences can be determined by those skilled in the art without further effort.

[0048] V H Alternative methods for numbering the amino acid residues of the domain (which can also be applied to the VHH domain in a similar manner) are known in the art. However, in this specification, the claims, and the drawings related to the ISVD described herein, unless otherwise specified, shall follow the Kabat-based numbering applied to the VHH domain as described above.

[0049] The total number of amino acid residues in a VHH domain will typically be in the range of 110 - 120, and often in the range of 112 - 115. However, it should be noted that for the purposes described herein, shorter and longer sequences may also be appropriate.

[0050] Methods for obtaining VHH domains that bind to a specific antigen or epitope have been previously described, for example, in WO 2006 / 040153 and WO 2006 / 122786. VHH domains obtained from camels can be "humanized" by replacing one or more amino acid residues in the amino acid sequence of the original VHH sequence with one or more amino acid residues present at the corresponding positions in the VH domain from a conventional four-chain antibody derived from humans. A humanized VHH domain can contain one or more complete human framework region sequences, and in an even more specific embodiment, can optionally contain a human framework region sequence obtained from DP-29, DP-47, DP-51, or a portion thereof, in combination with a JH sequence, for example, JH5.

[0051] The terms "epitope" and "antigenic determinant" can be used interchangeably and refer to a macromolecule, such as a portion of a polypeptide, recognized by an antigen-binding molecule, such as a conventional antibody or a polypeptide of the present invention, particularly by the antigen-binding site of said molecule. An epitope defines the minimal binding site for an immunoglobulin and thus represents the target of immunoglobulin specificity.

[0052] The portion of an antigen-binding molecule (e.g., a conventional antibody or a polypeptide described herein) that recognizes an epitope is called a paratope.

[0053] As used herein, the term "biparatopic" (antigen) binding molecule or "biparatopic" polypeptide means a polypeptide comprising a first ISVD and a second ISVD as defined herein, where these two variable domains are capable of binding to two different epitopes of one antigen, and these epitopes are not usually simultaneously bound by one monospecific immunoglobulin, such as a conventional antibody or one ISVD, etc. The biparatopic polypeptides of the present invention are composed of variable domains having different epitope specificities and do not contain pairs of variable domains that are complementary to each other and bind to the same epitope. Thus, they do not compete with each other for binding to LRP5 or LRP6.

[0054] A polypeptide (e.g., an immunoglobulin, antibody, ISVD or generally an antigen-binding molecule or a fragment thereof) that can "bind to", "bind", "specifically bind to", "be specifically bindable to" or "specifically bind" to a particular epitope, antigen or protein (or at least a part, fragment or epitope thereof), and has "affinity" and / or "specificity" for these, is a molecule "directed against" or "directed towards" said epitope, antigen or protein or a "binding" molecule for such an epitope, antigen or protein.

[0055] Generally, the term "specificity" refers to the number of different types of antigens or epitopes to which a particular antigen-binding molecule or antigen-binding protein (e.g., an immunoglobulin, antibody, ISVD) can bind. The specificity of an antigen-binding protein can be determined based on its affinity and / or binding strength. The affinity, represented by the equilibrium constant (K D ) for the dissociation of the antigen from the antigen-binding protein, is a measure of the binding strength between the epitope and the antigen-binding site on the antigen-binding protein, and the smaller the value of K D , the higher the binding strength between the epitope and the antigen-binding molecule (alternatively, the affinity is the affinity constant (K D ) which is 1 / K A)). As will be clear to the skilled artisan (e.g., based on the further disclosure herein), affinity can be determined in a manner known per se depending on the specific antigen of interest. Avidity is a measure of the binding strength between an antigen-binding molecule (e.g., immunoglobulin, antibody, ISVD) and the relevant antigen. Affinity is related to both the affinity between the epitope and its antigen-binding site on the antigen-binding molecule and the number of relevant binding sites present on the antigen-binding molecule.

[0056] Typically, an antigen-binding protein (e.g., a polypeptide capable of specifically binding to LRP5 and LRP6) has a dissociation constant (K) of 10E-5 to 10E-14 moles / liter (M) or less, preferably 10E-7 to 10E-14 moles / liter (M) or less, more preferably 10E-8 to 10E-14 moles / liter, and even more preferably 10E-11 to 10E-13 (e.g., as measured by Kinexa assay; known in the art). D ) and / or an association constant (K) of at least 10E7 ME-1, preferably at least 10E8 ME-1, more preferably at least 10E9 ME-1, e.g., at least 10E11 ME-1. A ) will bind with any K greater than 10E-4M. D Preferably, an antigen binding protein (e.g., a polypeptide capable of specifically binding to LRP5 and LRP6) has a K of less than 500 nM, preferably less than 200 nM, more preferably less than 10 nM, e.g., less than 500 pM. D and will bind to the desired antigen. Specific binding of an antigen-binding protein to an antigen or epitope can be determined in any suitable manner known per se, including, for example, the assays described herein, Scatchard analysis and / or competitive binding assays known per se in the art, such as radioimmunoassays (RIA), enzyme immunoassays (EIA) and sandwich competition assays and their various modifications.

[0057] The term "cross-reactivity" ("LRP5 / LRP6 cross-reactivity") as related to binding molecules capable of binding to LRP5 and LRP6 is intended to mean that such binding molecules can specifically bind to epitopes contained in LRP5 molecules and, alternatively, can also specifically bind to epitopes contained in LRP6 molecules. Usually, such cross-reactivity occurs when the epitopes of different proteins bound by such binding molecules have similar structures and / or sequences, for example, represent conserved epitopes, for example, are shared by proteins belonging to the same protein family (such as LRP5 and LRP6 belonging to the LRP protein family).

[0058] The polypeptides specifically bindable to LRP5 and LRP6 described herein (also referred to herein as LRP5 / LRP6 antagonists) are specific for LRP5 and LRP6 in that they contain an immunoglobulin single variable domain (LRP5 / LRP6 cross-reactive binding molecule) that specifically binds to epitopes contained in both of these molecules. They do not cross-react or essentially do not cross-react with epitopes having a structure similar to the epitopes of LRP5 and LRP6 or epitopes having an unrelated structure.

[0059] As used herein, the term "comprising" and variations thereof, for example, "comprises" and "compris", can be replaced by the terms "containing" or "including" or "having". Further, the term "comprising" also explicitly encompasses "consisting of" embodiments of the recited elements.

[0060] Combination therapy An object of the present invention is to provide a novel treatment method for treating or controlling various hyperproliferative diseases, particularly various malignant tumors.

[0061] The inventors of the present application have surprisingly discovered that the use of an LRP5 / LRP6 antagonist in combination with an anti-PD-1 (programmed cell death 1) antibody has the potential to improve clinical outcomes as compared to the use of the LRP5 / LRP6 antagonist or the anti-PD-1 antibody alone.

[0062] Specifically, in preclinical studies, the inventors tested the immunomodulatory function and antitumor activity of the LRP5 / LRP6 antagonist alone or in combination with an anti-PD-1 antibody (see Example 1 below). Complete responses and abundant T cell tumor infiltration determined by histopathological analysis were observed only for the combination of the LRP5 / LRP6 antagonist and the anti-PD-1 antibody. FACS analysis of tumor draining lymph nodes further showed that this combination treatment resulted in an increase in the number of activated dendritic cells (DCs) in the draining lymph nodes. As further shown in Example 3 below, treatment with the Wnt3a ligand in co-cultures of tumor spheroids and activated human PBMCs resulted in a significant blockade of PBMC-mediated inhibition of tumor cell viability. Treatment with the LRP5 / LRP6 antagonist in co-cultures of tumor spheroids and activated human PBMCs in the presence of Wnt3a restored PBMC-mediated inhibition of tumor cell viability. The combination treatment of the LRP5 / LRP6 antagonist of the present invention and the anti-human PD1 antibody results in enhanced PBMC-mediated tumor cell killing as compared to monotherapy with the LRP5 / LRP6 antagonist alone.

[0063] Without wishing to be bound by theory, these findings suggest that the combination treatment of the LRP5 / LRP6 antagonist and the anti-PD-1 antibody results in inhibition of the Wnt signaling pathway in DCs, subsequently leading to upregulation of pro-inflammatory cytokines, restoration of cross-priming, and promotion of tumor T cell infiltration and antitumor activity.

[0064] A variety of combination therapies are known in the art and are currently under investigation (e.g., in preclinical or clinical trials), but still lack the ability to satisfy treatment concepts for the treatment of cancer diseases, particularly solid tumors such as lung cancer (e.g., NSCLC), melanoma, bladder cancer, and gastrointestinal cancer. Therefore, any treatment showing advantages over standard treatments, such as better treatment outcomes, beneficial effects, superior efficacy, and / or improved tolerability, e.g., reduced side effects, etc., would represent an important development.

[0065] From the surprising results shown in the following examples, it is shown that a combination of an LRP5 / LRP6 antagonist that has no therapeutic effect by itself in a tumor model and an anti-PD-1 antibody that has only a limited therapeutic effect provides excellent results in that it results in a synergistic (i.e., more than additive) interaction of these two compounds and a complete response is obtained.

[0066] For this reason, the present invention relates to methods for the treatment and / or prevention of proliferative diseases, particularly cancer, comprising the combined administration of an LRP5 / LRP6 antagonist and an anti-PD-1 antibody as described herein, respectively, as well as to medical uses, uses, pharmaceutical compositions, or combinations and kits comprising such therapeutic agents.

[0067] Furthermore, the present invention relates to an anti-cancer therapy comprising the use of an LRP5 / LRP6 antagonist and an anti-PD-1 antibody as described herein in combination.

[0068] Such a combined treatment can be provided as a non-fixed (e.g., free) combination of substances or in the form of a fixed combination comprising a kit of parts.

[0069] For the treatment of diseases of oncological nature, numerous anti-cancer agents (including target-specific and non-target-specific anti-cancer agents) have already been proposed, which can be used as monotherapy or in combination therapy including two or more agents (for example, combination therapy of two or three agents) and / or can be combined with radiotherapy (for example, irradiation treatment), radioimmunotherapy and / or surgery. Thus, the combination treatments described herein can be provided in addition to further therapeutic agents and / or treatments, for example, radiotherapy, radioimmunotherapy and surgery.

[0070] LRP5 / LRP6 antagonist In the meaning of the present invention and all its embodiments, a polypeptide capable of specifically binding to LRP5 and LRP6 (also referred to herein as an LRP5 / LRP6 antagonist) is an LRP5 / LRP6 cross-reactive bivalent polypeptide comprising two or more immunoglobulin single variable domains that bind to LRP5 and / or LRP6 at different epitopes. As described above, "cross-reactive" and "bivalent" can be defined as molecules in which an LRP5 / LRP6 cross-reactive bivalent molecule can bind to LRP5 at two different epitopes contained in the LRP5 protein and can also bind to LRP6 at the corresponding two epitopes contained in the LRP6 protein.

[0071] More specifically, the polypeptide capable of specifically binding to LRP5 and LRP6 is - a first immunoglobulin single variable domain (LRP5 / LRP6 cross-reactive) that can specifically bind to LRP5 and LRP6 via an epitope in such a way as to result in inhibition of the Wnt1 signaling pathway and, as a result, inhibition of the transcription of Wnt1-driven target genes, and - a second immunoglobulin single variable domain (LRP5 / LRP6 cross-reactive) that can specifically bind to LRP5 and LRP6 via an epitope in such a way as to result in inhibition of the Wnt3a signaling pathway and, as a result, inhibition of the transcription of Wnt3a-driven target genes. comprises

[0072] Due to the presence of two immunoglobulin single variable domains in such polypeptides, where two domains bind to different epitopes (related to Wnt1 / Wnt3a signaling), these molecules are bivalent binding molecules. It should be noted that in this context, it is contemplated that the LRP5 / LRP6 antagonists described herein may bind to one single LRP5 or LRP6 molecule via both of its LRP5 / LRP6 binding domains. However, other binding modes may occur as well.

[0073] In some embodiments of the invention, the polypeptide capable of specifically binding to LRP5 and LRP6 is - the following CDR sequences: CDR1: TYTVG (= SEQ ID NO: 40) CDR2: AIRRRGSSTYYADSVKG (= SEQ ID NO: 41) CDR3: DTRTVALLQYRYDY (= SEQ ID NO: 42) comprising a first ISVD (a) and - the following CDR sequences: CDR1: SYAMG (= SEQ ID NO: 49) CDR2: AISWSGGSTYYADSVKG (= SEQ ID NO: 50) CDR3: SPIPYGSLLRRRNNYDY (= SEQ ID NO: 51) comprising a second ISVD (b) comprises.

[0074] This particular combination of CDR sequences is contained, for example, in the LRP5 / LRP6 antagonist referred to herein below as LRP5 / LRP6#1.

[0075] In some embodiments of the invention, the polypeptide capable of specifically binding to LRP5 and LRP6 is - the following CDR sequences: CDR1: SYAMG (= SEQ ID NO: 43) CDR2: AIRRSGRRTYYADSVKG (= Sequence number: 44) CDR3: ARRVRSSTRYNTGTWWWEY (= Sequence number: 45) A first ISVD (a) comprising - The following CDR sequences: CDR1: SYAMG (= Sequence number: 49) CDR2: AISWSGGSTYYADSVKG (= Sequence number: 50) CDR3: SPIPYGSLLRRRNNYDY (= Sequence number: 51) A second ISVD (b) comprising is included.

[0076] This specific combination of CDR sequences is contained, for example, in the LRP5 / LRP6 antagonist called LRP5 / LRP6#2 hereinafter in this specification.

[0077] In some embodiments of the present invention, a polypeptide capable of specifically binding to LRP5 and LRP6 is - The following CDR sequences: CDR1: RYTMG (= Sequence number: 46) CDR2: AIVRSGGSTYYADSVKG (= Sequence number: 47) CDR3: DRRGRGENYILLYSSGRYEY (= Sequence number: 48) A first ISVD (a) comprising - The following CDR sequences: CDR1: SYAMG (= Sequence number: 49) CDR2: AISWSGGSTYYADSVKG (= Sequence number: 50) CDR3: SPIPYGSLLRRRNNYDY (= Sequence number: 51) A second ISVD (b) comprising is included.

[0078] This specific combination of CDR sequences is contained, for example, in the LRP5 / LRP6 antagonist called LRP5 / LRP6#3 hereinafter in this specification.

[0079] In some embodiments of the present invention, the polypeptide capable of specifically binding to LRP5 and LRP6 is - the following CDR sequences: CDR1: TYTVG (= SEQ ID NO: 40) CDR2: AIRRRGSSTYYADSVKG (= SEQ ID NO: 41) CDR3: DTRTVALLQYRYDY (= SEQ ID NO: 42) a first ISVD (a) comprising - the following CDR sequences: CDR1: SYAMG (= SEQ ID NO: 52) CDR2: AISWRSGSTYYADSVKG (= SEQ ID NO: 53) CDR3: DPRGYGVAYVSAYYEY (= SEQ ID NO: 54) a second ISVD (b) comprising and

[0080] This particular combination of CDR sequences is contained, for example, in the LRP5 / LRP6 antagonist, hereinafter referred to as LRP5 / LRP6#4, herein.

[0081] In some embodiments of the present invention, the polypeptide capable of specifically binding to LRP5 and LRP6 is - the following CDR sequences: CDR1: SYAMG (= SEQ ID NO: 43) CDR2: AIRRSGRRTYYADSVKG (= SEQ ID NO: 44) CDR3: ARRVRSSTRYNTGTWWWEY (= SEQ ID NO: 45) a first ISVD (a) comprising - the following CDR sequences: CDR1: SYAMG (= SEQ ID NO: 52) CDR2: AISWRSGSTYYADSVKG (= SEQ ID NO: 53) CDR3: DPRGYGVAYVSAYYEY (= SEQ ID NO: 54) a second ISVD (b) comprising and

[0082] This particular combination of CDR sequences is contained, for example, in an LRP5 / LRP6 antagonist called LRP5 / LRP6#5 hereinafter in this specification.

[0083] In some embodiments of the invention, the polypeptide capable of specifically binding to LRP5 and LRP6 is - the following CDR sequences: CDR1: RYTMG (= SEQ ID NO: 46) CDR2: AIVRSGGSTYYADSVKG (= SEQ ID NO: 47) CDR3: DRRGRGENYILLYSSGRYEY (= SEQ ID NO: 48) a first ISVD (a) comprising - the following CDR sequences: CDR1: SYAMG (= SEQ ID NO: 52) CDR2: AISWRSGSTYYADSVKG (= SEQ ID NO: 53) CDR3: DPRGYGVAYVSAYYEY (= SEQ ID NO: 54) a second ISVD (b) comprising and

[0084] This particular combination of CDR sequences is contained, for example, in an LRP5 / LRP6 antagonist called LRP5 / LRP6#6 hereinafter in this specification.

[0085] Generally, as used herein, the terms "first" and "second" with respect to such ISVDs or domains are merely intended to indicate that these domains are two different domains (since they contain at least different CDR sequences). For this reason, these terms should not be understood to refer to the exact order or arrangement of the domains within such a polypeptide chain. That is, the above ISVD (a) and (b) can be arranged in either order (a)-(b) or order (b)-(a) within the polypeptides described herein.

[0086] The terms "capable of specifically binding to LRP5 and LRP6" and "specifically binds to LRP5 or LRP6" are intended to mean that the immunoglobulin single variable domains (a) and (b) are cross-reactive with LRP5 and LRP6. Of course, the binding properties of such molecules are determined by their (CDR) sequences, and as a result, the features of "capable of specifically binding to LRP5 and LRP6" and "specifically binds to LRP5 or LRP6" described above and in the claims are only intended to illustrate the usefulness of the present invention and are not intended to limit the scope of the present invention.

[0087] Specifically, the ISVDs of the polypeptides described herein (e.g., ISVDs containing the CDR sequences defined above) are VHH domains, preferably humanized VHH domains.

[0088] In some embodiments of the present invention, the polypeptide capable of specifically binding to LRP5 and LRP6 includes a polypeptide having a first ISVD (a) and a second ISVD (b), wherein the first ISVD includes a VHH domain having a sequence selected from the group consisting of SEQ ID NO: 58, SEQ ID NO: 59, and SEQ ID NO: 60, and the second ISVD includes a VHH domain having a sequence selected from the group consisting of SEQ ID NO: 61 and SEQ ID NO: 62, where these sequences are as follows.

[0089]

Chemical formula

Chemical formula

Chemical formula

Chemical formula

Chemical formula

[0090] In some embodiments, the first ISVD comprises the sequence of SEQ ID NO: 58, and the second ISVD comprises the sequence of SEQ ID NO: 61 (LRP5 / LRP6#1).

[0091] In some embodiments of the present invention, the first ISVD comprises the sequence of SEQ ID NO: 59, and the second ISVD comprises the sequence of SEQ ID NO: 61 (LRP5 / LRP6#2).

[0092] In some embodiments, the first ISVD comprises the sequence of SEQ ID NO: 60, and the second ISVD comprises the sequence of SEQ ID NO: 61 (LRP5 / LRP6#3).

[0093] In some embodiments, the first ISVD comprises the sequence of SEQ ID NO: 58, and the second ISVD comprises the sequence of SEQ ID NO: 62 (LRP5 / LRP6#4).

[0094] In some embodiments, the first ISVD comprises the sequence of SEQ ID NO: 59, and the second ISVD comprises the sequence of SEQ ID NO: 62 (LRP5 / LRP6#5).

[0095] In some embodiments, the first ISVD comprises the sequence of SEQ ID NO: 60, and the second ISVD comprises the sequence of SEQ ID NO: 62 (LRP5 / LRP6#6).

[0096] In a preferred embodiment of the present invention, the LRP5 / LRP6 antagonist is any one of LRP5 / LRP6#1, LRP5 / LRP6#5 or LRP5 / LRP6#6 defined by the above CDR and / or VHH sequence.

[0097] According to a preferred embodiment of the present invention, the LRP5 / LRP6 antagonist comprises a polypeptide having a first (a) LRP5 / LRP6-binding ISVD, a second (b) LRP5 / LRP6-binding ISVD, and a third (c) ISVD. Preferably, the LRP5 / LRP6 antagonist comprises the first and second ISVDs defined by the above CDR sequences and a third ISVD that directly or indirectly binds the first and second ISVDs. In some embodiments, the first ISVD is covalently bound via a peptide linker to a third ISVD that is covalently bound via a peptide linker to the second ISVD. The two linkers can be the same or different linkers. Also included is the case where only one linker is present. The terms "first" and "second" do not indicate their positions within the polypeptide as described above. Thus, the ISVD sequences within the polypeptide can be arranged in any of the orders of ISVD (a)-(c)-(b), (a)-[linker]-(c)-[linker]-(b), (b)-(c)-(a), (b)-[linker]-(c)-[linker]-(a), (a)-[linker]-(c)-(b), (a)-(c)-[linker]-(b), (b)-[linker]-(c)-(a), (b)-(c)-[linker]-(a) from the N-terminus to the C-terminus.

[0098] Preferably, the third ISVD (c) is an albumin-binding ISVD. Non-limiting examples of such albumin-binding ISVDs are the following CDRs: CDR(Alb11)1: SFGMS (=SEQ ID NO: 55) CDR(Alb11)2: SISGSGSDTLYADSVKG (=SEQ ID NO: 56) CDR(Alb11)3: GGSLSR (=SEQ ID NO: 57) which is the Alb11 domain containing the same.

[0099] Thereby, a preferred group of LRP5 / LRP6 antagonists having the following structure is obtained.

[0100] FR(a)1-CDR(a)1-FR(a)2-CDR(a)2-FR(a)3-CDR(a)3-FR(a)4-[Any linker peptide]-FR(Alb11)1-CDR(Alb11)1-FR(Alb11)2-CDR(Alb11)2-FR(Alb11)3-CDR(Alb11)3-FR(Alb11)4-[Any linker peptide]-FR(b)1-CDR(b)1-FR(b)2-CDR(b)2-FR(b)3-CDR(b)3-FR(b)4, preferably, where the CDRs include the sequences described above.

[0101] Again, the order of the three ISVDs (a), (b), and Alb11 is not fixed, and the above domains are as follows: (b)-Alb11-(a) Polypeptides arranged in the order of are also included. Furthermore, polypeptides having an Alb11 domain at the N-terminus or C-terminus of the polypeptide (e.g., Alb11-(a)-(b), Alb11-(b)-(a), (a)-(b)-Alb11, or (b)-(a)-Alb11) are also included in the present invention.

[0102] In some embodiments of the present invention, the polypeptide capable of specifically binding to LRP5 and LRP6 is - The following CDR sequences: CDR1: TYTVG (= SEQ ID NO: 40) CDR2: AIRRRGSSTYYADSVKG (= SEQ ID NO: 41) CDR3: DTRTVALLQYRYDY (= SEQ ID NO: 42) The first ISVD containing - The following CDR sequences: CDR1: SYAMG (= SEQ ID NO: 49) CDR2: AISWSGGSTYYADSVKG (= SEQ ID NO: 50) CDR3: SPIPYGSLLRRRNNYDY (= SEQ ID NO: 51) The second ISVD containing - The following CDR sequences: CDR1: SFGMS (= SEQ ID NO: 55) CDR2: SISGSGSDTLYADSVKG (=SEQ ID NO: 56) CDR3: GGSLSR (=SEQ ID NO: 57) An albumin-binding ISVD containing the same (the third ISVD) is included.

[0103] This specific combination of CDR sequences is contained, for example, in the LRP5 / LRP6 antagonist called LRP5 / LRP6#1 hereinafter in this specification.

[0104] In some embodiments of the present invention, the polypeptide capable of specifically binding to LRP5 and LRP6 is - the following CDR sequences: CDR1: SYAMG (=SEQ ID NO: 43) CDR2: AIRRSGRRTYYADSVKG (=SEQ ID NO: 44) CDR3: ARRVRSSTRYNTGTWWWEY (=SEQ ID NO: 45) a first ISVD containing the same - the following CDR sequences: CDR1: SYAMG (=SEQ ID NO: 49) CDR2: AISWSGGSTYYADSVKG (=SEQ ID NO: 50) CDR3: SPIPYGSLLRRRNNYDY (=SEQ ID NO: 51) a second ISVD containing the same and - the following CDR sequences: CDR1: SFGMS (=SEQ ID NO: 55) CDR2: SISGSGSDTLYADSVKG (=SEQ ID NO: 56) CDR3: GGSLSR (=SEQ ID NO: 57) an albumin-binding ISVD containing the same is included.

[0105] This specific combination of CDR sequences is contained, for example, in the LRP5 / LRP6 antagonist called LRP5 / LRP6#2 hereinafter in this specification.

[0106] In some embodiments of the present invention, the polypeptide capable of specifically binding to LRP5 and LRP6 is - the following CDR sequences: CDR1: RYTMG (= SEQ ID NO: 46) CDR2: AIVRSGGSTYYADSVKG (= SEQ ID NO: 47) CDR3: DRRGRGENYILLYSSGRYEY (= SEQ ID NO: 48) a first ISVD comprising - the following CDR sequences: CDR1: SYAMG (= SEQ ID NO: 49) CDR2: AISWSGGSTYYADSVKG (= SEQ ID NO: 50) CDR3: SPIPYGSLLRRRNNYDY (= SEQ ID NO: 51) a second ISVD having - the following CDR sequences: CDR1: SFGMS (= SEQ ID NO: 55) CDR2: SISGSGSDTLYADSVKG (= SEQ ID NO: 56) CDR3: GGSLSR (= SEQ ID NO: 57) an albumin-binding ISVD comprising and

[0107] This specific combination of CDR sequences is contained, for example, in the LRP5 / LRP6 antagonist, hereinafter referred to as LRP5 / LRP6#3, herein.

[0108] In some embodiments of the present invention, the polypeptide capable of specifically binding to LRP5 and LRP6 is - the following CDR sequences: CDR1: TYTVG (= SEQ ID NO: 40) CDR2: AIRRRGSSTYYADSVKG (= SEQ ID NO: 41) CDR3: DTRTVALLQYRYDY (= SEQ ID NO: 42) a first ISVD comprising - the following CDR sequences: CDR1: SYAMG (= SEQ ID NO: 52) CDR2: AISWRSGSTYYADSVKG (=SEQ ID NO: 53) CDR3: DPRGYGVAYVSAYYEY (=SEQ ID NO: 54) A second ISVD comprising - The following CDR sequences: CDR1: SFGMS (=SEQ ID NO: 55) CDR2: SISGSGSDTLYADSVKG (=SEQ ID NO: 56) CDR3: GGSLSR (=SEQ ID NO: 57) An albumin-binding ISVD comprising is included.

[0109] This particular combination of CDR sequences is contained, for example, in an LRP5 / LRP6 antagonist called LRP5 / LRP6#4 hereinafter in this specification.

[0110] In some embodiments of the invention, a polypeptide capable of specifically binding to LRP5 and LRP6 is - The following CDR sequences: CDR1: SYAMG (=SEQ ID NO: 43) CDR2: AIRRSGRRTYYADSVKG (=SEQ ID NO: 44) CDR3: ARRVRSSTRYNTGTWWWEY (=SEQ ID NO: 45) A first ISVD comprising - The following CDR sequences: CDR1: SYAMG (=SEQ ID NO: 52) CDR2: AISWRSGSTYYADSVKG (=SEQ ID NO: 53) CDR3: DPRGYGVAYVSAYYEY (=SEQ ID NO: 54) A second ISVD comprising - The following CDR sequences: CDR1: SFGMS (=SEQ ID NO: 55) CDR2: SISGSGSDTLYADSVKG (=SEQ ID NO: 56) CDR3: GGSLSR (=SEQ ID NO: 57) An albumin-binding ISVD comprising is included.

[0111] This particular combination of CDR sequences is contained, for example, in an LRP5 / LRP6 antagonist called LRP5 / LRP6#5 hereinafter in this specification.

[0112] In some embodiments of the invention, the polypeptide capable of specifically binding to LRP5 and LRP6 is - the following CDR sequences: CDR1: RYTMG (= SEQ ID NO: 46) CDR2: AIVRSGGSTYYADSVKG (= SEQ ID NO: 47) CDR3: DRRGRGENYILLYSSGRYEY (= SEQ ID NO: 48) a first ISVD comprising - the following CDR sequences: CDR1: SYAMG (= SEQ ID NO: 52) CDR2: AISWRSGSTYYADSVKG (= SEQ ID NO: 53) CDR3: DPRGYGVAYVSAYYEY (= SEQ ID NO: 54) a second ISVD comprising - the following CDR sequences: CDR1: SFGMS (= SEQ ID NO: 55) CDR2: SISGSGSDTLYADSVKG (= SEQ ID NO: 56) CDR3: GGSLSR (= SEQ ID NO: 57) an albumin-binding ISVD comprising and

[0113] This particular combination of CDR sequences is contained, for example, in an LRP5 / LRP6 antagonist called LRP5 / LRP6#6 hereinafter in this specification.

[0114] In some embodiments, the ISVDs defined by those CDR sequences in the polypeptide capable of specifically binding to LRP5 and LRP6 are arranged such that the albumin-binding ISVD binds the first and second ISVDs directly or indirectly (e.g., (a) via a linker peptide).

[0115] The sequence of the above-mentioned Alb11 immunoglobulin single variable domain is as follows.

[0116]

Chemical Formula

[0117] The above-mentioned CDR sequences are summarized in Table 1A, Table 1B, and Table 1C.

[0118]

Table 1

[0119]

Table 2

[0120]

Table 3

[0121] The three preferred LRP5 / LRP6 antagonists described in this specification are as follows.

[0122] The first preferred LRP5 / LRP6 antagonist: the following - The first (LRP5 / LRP6 binding) ISVD containing the amino acid sequence shown in SEQ ID NO: 58 - The albumin-binding ISVD containing the amino acid sequence shown in SEQ ID NO: 63 - The second (LRP5 / LRP6 binding) ISVD containing the amino acid sequence shown in SEQ ID NO: 61 A polypeptide containing them in this order or in an order in which the above three domains are changed.

[0123] The second preferred LRP5 / LRP6 antagonist: the following - The first (LRP5 / LRP6 binding) ISVD containing the amino acid sequence shown in SEQ ID NO: 59 - An albumin-binding ISVD comprising the amino acid sequence shown in SEQ ID NO: 63 - A second (LRP5 / LRP6-binding) ISVD comprising the amino acid sequence shown in SEQ ID NO: 62 A polypeptide comprising, in this order or in an order in which the three domains are changed, the above.

[0124] A third preferred LRP5 / LRP6 antagonist: the following - A first (LRP5 / LRP6-binding) ISVD comprising the amino acid sequence shown in SEQ ID NO: 60 - An albumin-binding ISVD comprising the amino acid sequence shown in SEQ ID NO: 63 - A second (LRP5 / LRP6-binding) ISVD comprising the amino acid sequence shown in SEQ ID NO: 62 A polypeptide comprising, in this order or in an order in which the three domains are changed, the above.

[0125] In an even more specifically preferred embodiment, the albumin-binding ISVD is located between two LRP5 / LRP6-binding ISVDs.

[0126] The sequences of the above-mentioned VHHs are summarized in Tables 2A, 2B and 2C.

[0127]

Table 4

[0128]

Table 5

[0129]

Table 6

[0130] In a preferred embodiment of the present invention, the LRP5 / LRP6 antagonist comprises a sequence selected from SEQ ID NOs: 64, 65 and 66 (these preferred polypeptides capable of specifically binding to LRP5 and LRP6 are herein also referred to as LRP5 / LRP6#1, LRP5 / LRP6#5 and LRP5 / LRP6#6, respectively), where the exact amino acid sequence can be obtained from Table 2D below.

[0131]

Table 7

[0132] The production and therapeutic use of the aforementioned polypeptides capable of specifically binding to LRP5 and LRP6 are disclosed in WO 2017 / 093478. In particular, this document provides a sufficient disclosure of methods for preparing polypeptides capable of specifically binding to LRP5 and LRP6 for use in the present invention.

[0133] Anti-PD-1 antibody An anti-PD-1 antibody (also referred to herein as a "PD-1 antibody") is a compound that inhibits the interaction between PD-1 and its ligand in the present invention and all its embodiments. Preferably, the anti-PD-1 antibody is a humanized or fully human anti-PD-1 antibody. Any one of these antibodies may be a recombinant human antibody.

[0134] The PD-1 gene encodes a 55 kDa type I transmembrane protein that is part of the Ig gene superfamily (Agata et al. (1996) Int Immunol. 8:765-72). The complete PD-1 sequence can be found in GenBank accession number U64863. Although structurally similar to CTLA-4, PD-1 lacks the MYPPY motif (SEQ ID NO: 39) important for B7-1 and B7-2 binding.

[0135] PD-1 is an inhibitory member of the extended CD28 / CTLA-4 family of T cell regulators. Other members of the CD28 family include CD28, CTLA-4, ICOS and BTLA. PD-1 is suggested to exist as a monomer lacking the asymmetric cysteine residues characteristic of other CD28 family members. PD-1 is expressed on activated B cells, T cells and monocytes (Okazaki et al. (2002) Curr Opin Immunol 14:391779-82; Bennett et al. (2003) J. Immunol. 170:711-8). Two ligands for PD-1, PD-L1 (B7-H1) and PD-L2 (B7-DC), have been identified, and binding to PD-1 has been shown to down-regulate T cell activation (Freeman et al. (2000) J. Exp. Med. 192:1027-3; Carter et al. (2002) Eur. J. Immunol. 32:634-43). Both PD-L1 and PD-L2 are B7 homologs that bind to PD-1. PD-L1 is abundantly present in various human cancers (Dong et al. (2002) Nat. Med. 8:787-9).

[0136] PD-1 is known as an immune inhibitory protein that negatively regulates TCR signals (Ishida, Y. et al. (1992) EMBO J. 11:3887-3895; Blank, C. et al. (2006) Immunol. Immunother. 56(6):739-745). The interaction between PD-1 and PD-L1 can act as an immune checkpoint that can, for example, result in a decrease in tumor-infiltrating lymphocytes, a decrease in T cell receptor-mediated proliferation, and / or immune evasion by cancerous cells (Dong et al. (2003) J. Mol. Med. 81:281-7; Blank et al. (2005) Cancer Immunol. Immunother. 54:307-314; Konishi et al. (2004) Clin. Cancer Res. 10:5094-100). Immune suppression can be reversed by inhibiting the local interaction between PD-1 and PD-L1 or PD-L2. When the interaction between PD-1 and both PD-L1 and PD-L2 is blocked, the effect is additive (Iwai et al. (2002) Proc. Nat’l. Acad. Sci USA 99:12293-7; Brown et al. (2003) J. Immunol. 170:1257-66).

[0137] In one aspect of the invention, the anti-PD-1 antibody is any one of antibodies PD1-1, PD1-2, PD1-3, PD1-4, and PD1-5 defined by the sequences shown in Table 3 by SEQ ID NO., where VH refers to the heavy chain variable domain, VL refers to the light chain variable domain, HC refers to the (full-length) heavy chain, and LC refers to the (full-length) light chain.

[0138] [Table 8]

[0139] Also, here, the amino acid sequences (and sequence names) of SEQ ID NO. are as shown in Table 4.

[0140]

Table 9

[0141] Specifically, the anti-PD-1 antibody molecule described in this specification includes a heavy-chain CDR containing the amino acid sequences of SEQ ID NO: 1 (HCDR1), SEQ ID NO: 2 (HCDR2), and SEQ ID NO: 3 (HCDR3), and a light-chain CDR containing the amino acid sequences of SEQ ID NO: 4 (LCDR1), SEQ ID NO: 5 (LCDR2), and SEQ ID NO: 6 (LCDR3), or (b) a heavy-chain CDR containing the amino acid sequences of SEQ ID NO: 7 (HCDR1), SEQ ID NO: 8 (HCDR2), and SEQ ID NO: 9 (HCDR3), and a light-chain CDR containing the amino acid sequences of SEQ ID NO: 10 (LCDR1), SEQ ID NO: 11 (LCDR2), and SEQ ID NO: 12 (LCDR3), or (c) a heavy-chain CDR containing the amino acid sequences of SEQ ID NO: 13 (HCDR1), SEQ ID NO: 14 (HCDDR2), and SEQ ID NO: 15 (HCDR3), and a light-chain CDR containing the amino acid sequences of SEQ ID NO: 16 (LCDR1), SEQ ID NO: 17 (LCDR2), and SEQ ID NO: 18 (LCDR3).

[0142] In some embodiments, the anti-PD-1 antibody molecule includes a heavy-chain variable domain containing an amino acid sequence selected from SEQ ID NO: 19, 21, 23, 25, and 27.

[0143] In some embodiments, the anti-PD-1 antibody molecule includes a light-chain variable domain containing an amino acid sequence selected from SEQ ID NO: 20, 22, 24, 26, and 28.

[0144] In some embodiments, the anti-PD-1 antibody molecule comprises (a) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 19 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 20, (b) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 21 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 22, (c) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 23 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 24, (d) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 25 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 26, or (e) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 27 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 28.

[0145] In some embodiments, the anti-PD-1 antibody comprises (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 29 and a light chain comprising the amino acid sequence of SEQ ID NO: 30, (b) a heavy chain comprising the amino acid sequence of SEQ ID NO: 31 and a light chain comprising the amino acid sequence of SEQ ID NO: 32, (c) a heavy chain comprising the amino acid sequence of SEQ ID NO: 33 and a light chain comprising the amino acid sequence of SEQ ID NO: 34, (d) a heavy chain comprising the amino acid sequence of SEQ ID NO: 35 and a light chain comprising the amino acid sequence of SEQ ID NO: 36, or (e) a heavy chain comprising the amino acid sequence of SEQ ID NO: 37 and a light chain comprising the amino acid sequence of SEQ ID NO: 38.

[0146] In a preferred embodiment, the anti-PD-1 antibody is PD1-1.

[0147] In a preferred embodiment, the anti-PD-1 antibody is PD1-2.

[0148] In a preferred embodiment, the anti-PD-1 antibody is PD1-3.

[0149] In a preferred embodiment, the anti-PD-1 antibody is PD1-4.

[0150] In a preferred embodiment, the anti-PD-1 antibody is PD1-5.

[0151] In one aspect, the present invention provides a method for treating and / or preventing a proliferative disorder, preferably cancer, the method comprising administering to a patient in need thereof a therapeutically effective amount of an LRP5 / LRP6 antagonist (e.g., any one of LRP5 / LRP6#1, LRP5 / LRP6#2, LRP5 / LRP6#3, LRP5 / LRP6#4, LRP5 / LRP6#5, LRP5 / LRP6#6 defined by the CDR and / or VHH sequences of Table 1a, Table 1b, Table 1c, Table 2a, Table 2b, Table 2c) and a therapeutically effective amount of an anti-PD-1 antibody (e.g., any one of PD1-1, PD1-2, PD1-3, PD1-4, PD1-5 defined by the CDR and / or VH / VL sequences of Table 3 and Table 4). In a preferred embodiment, the LRP5 / LRP6 antagonist comprises the amino acid sequence of SEQ ID NO: 64, SEQ ID NO: 65 or SEQ ID NO: 66, and the PD-1 antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 29 and a light chain comprising the amino acid sequence of SEQ ID NO: 30. In a preferred embodiment, the LRP5 / LRP6 antagonist comprises the amino acid sequence of SEQ ID NO: 64, SEQ ID NO: 65 or SEQ ID NO: 66, and the PD-1 antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 31 and a light chain comprising the amino acid sequence of SEQ ID NO: 32. In a preferred embodiment, the LRP5 / LRP6 antagonist comprises the amino acid sequence of SEQ ID NO: 64, SEQ ID NO: 65 or SEQ ID NO: 66, and the PD-1 antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 33 and a light chain comprising the amino acid sequence of SEQ ID NO: 34.

[0152] In another aspect, the present invention provides a combination of an LRP5 / LRP6 antagonist (e.g., any one of LRP5 / LRP6#1, LRP5 / LRP6#2, LRP5 / LRP6#3, LRP5 / LRP6#4, LRP5 / LRP6#5, LRP5 / LRP6#6 defined by the CDR and / or VHH sequences in Table 1a, Table 1b, Table 1c, Table 2a, Table 2b, Table 2c) and an anti-PD-1 antibody (e.g., any one of PD1-1, PD1-2, PD1-3, PD1-4, PD1-5 defined by the CDR and / or VH / VL sequences in Table 3 and Table 4) for use in a method of treating and / or preventing a proliferative disorder, preferably cancer. Here, the method includes administering a therapeutically effective amount of the combination to a patient in need thereof. In a preferred embodiment, the LRP5 / LRP6 antagonist comprises the amino acid sequence of SEQ ID NO: 64, SEQ ID NO: 65 or SEQ ID NO: 66, and the PD-1 antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 29 and a light chain comprising the amino acid sequence of SEQ ID NO: 30. In a preferred embodiment, the LRP5 / LRP6 antagonist comprises the amino acid sequence of SEQ ID NO: 64, SEQ ID NO: 65 or SEQ ID NO: 66, and the PD-1 antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 31 and a light chain comprising the amino acid sequence of SEQ ID NO: 32. In a preferred embodiment, the LRP5 / LRP6 antagonist comprises the amino acid sequence of SEQ ID NO: 64, SEQ ID NO: 65 or SEQ ID NO: 66, and the PD-1 antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 33 and a light chain comprising the amino acid sequence of SEQ ID NO: 34.

[0153] In another aspect, the present invention relates to an LRP5 / LRP6 antagonist (e.g., any one of LRP5 / LRP6#1, LRP5 / LRP6#2, LRP5 / LRP6#3, LRP5 / LRP6#4, LRP5 / LRP6#5, LRP5 / LRP6#6 defined by the CDR and / or VHH sequences in Table 1a, Table 1b, Table 1c, Table 2a, Table 2b, Table 2c) for use in a method of treating and / or preventing a proliferative disorder, preferably cancer. Here, the method comprises administering a therapeutically effective amount of the LRP5 / LRP6 antagonist in combination with an anti-PD-1 antibody (e.g., any one of PD1-1, PD1-2, PD1-3, PD1-4, PD1-5 defined by the CDR and / or VH / VL sequences in Table 3 and Table 4) to a patient in need thereof. In a preferred embodiment, the LRP5 / LRP6 antagonist comprises the amino acid sequence of SEQ ID NO: 64, SEQ ID NO: 65 or SEQ ID NO: 66, and the PD-1 antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 29 and a light chain comprising the amino acid sequence of SEQ ID NO: 30. In a preferred embodiment, the LRP5 / LRP6 antagonist comprises the amino acid sequence of SEQ ID NO: 64, SEQ ID NO: 65 or SEQ ID NO: 66, and the PD-1 antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 31 and a light chain comprising the amino acid sequence of SEQ ID NO: 32. In a preferred embodiment, the LRP5 / LRP6 antagonist comprises the amino acid sequence of SEQ ID NO: 64, SEQ ID NO: 65 or SEQ ID NO: 66, and the PD-1 antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 33 and a light chain comprising the amino acid sequence of SEQ ID NO: 34.

[0154] In another aspect, the present invention refers to an anti-PD-1 antibody described herein (e.g., any one of PD1-1, PD1-2, PD1-3, PD1-4, PD1-5 defined by the CDR and / or VH / VL sequences in Tables 3 and 4) for use in a method of treating and / or preventing a proliferative disorder, preferably cancer. The method includes administering to a patient in need thereof a therapeutically effective amount of the anti-PD-1 antibody in combination with an LRP5 / LRP6 antagonist (e.g., any one of LRP5 / LRP6#1, LRP5 / LRP6#2, LRP5 / LRP6#3, LRP5 / LRP6#4, LRP5 / LRP6#5, LRP5 / LRP6#6 defined by the CDR and / or VHH sequences in Tables 1a, 1b, 1c, 2a, 2b, 2c). In a preferred embodiment, the LRP5 / LRP6 antagonist includes the amino acid sequence of SEQ ID NO: 64, SEQ ID NO: 65 or SEQ ID NO: 66, and the PD-1 antibody includes a heavy chain including the amino acid sequence of SEQ ID NO: 29 and a light chain including the amino acid sequence of SEQ ID NO: 30. In a preferred embodiment, the LRP5 / LRP6 antagonist includes the amino acid sequence of SEQ ID NO: 64, SEQ ID NO: 65 or SEQ ID NO: 66, and the PD-1 antibody includes a heavy chain including the amino acid sequence of SEQ ID NO: 31 and a light chain including the amino acid sequence of SEQ ID NO: 32. In a preferred embodiment, the LRP5 / LRP6 antagonist includes the amino acid sequence of SEQ ID NO: 64, SEQ ID NO: 65 or SEQ ID NO: 66, and the PD-1 antibody includes a heavy chain including the amino acid sequence of SEQ ID NO: 33 and a light chain including the amino acid sequence of SEQ ID NO: 34.

[0155] In another aspect, the present invention provides in one or more containers · a first pharmaceutical composition or dosage form comprising an LRP5 / LRP6 antagonist (e.g., any one of LRP5 / LRP6#1, LRP5 / LRP6#2, LRP5 / LRP6#3, LRP5 / LRP6#4, LRP5 / LRP6#5, LRP5 / LRP6#6 defined by the CDR and / or VHH sequences in Tables 1a, 1b, 1c, 2a, 2b, 2c) and optionally one or more pharmaceutically acceptable carriers, excipients and / or vehicles · A second pharmaceutical composition or dosage form comprising an anti-PD-1 antibody described herein (e.g., any one of PD1-1, PD1-2, PD1-3, PD1-4, PD1-5 defined by the CDR and / or VH / VL sequences in Tables 3 and 4), and optionally one or more pharmaceutically acceptable carriers, excipients and / or vehicles, and · Refers to a kit, optionally including an accompanying document containing printed instructions.

[0156] In a preferred embodiment of the kit of the present invention, the LRP5 / LRP6 antagonist comprises the amino acid sequence of SEQ ID NO: 64, SEQ ID NO: 65 or SEQ ID NO: 66, and the PD-1 antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 29 and a light chain comprising the amino acid sequence of SEQ ID NO: 30. In a preferred embodiment of the kit of the present invention, the LRP5 / LRP6 antagonist comprises the amino acid sequence of SEQ ID NO: 64, SEQ ID NO: 65 or SEQ ID NO: 66, and the PD-1 antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 31 and a light chain comprising the amino acid sequence of SEQ ID NO: 32. In a preferred embodiment of the kit of the present invention, the LRP5 / LRP6 antagonist comprises the amino acid sequence of SEQ ID NO: 64, SEQ ID NO: 65 or SEQ ID NO: 66, and the PD-1 antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 33 and a light chain comprising the amino acid sequence of SEQ ID NO: 34.

[0157] Preferably, the accompanying document includes printed instructions for the simultaneous, concurrent, sequential, continuous, alternating or separate use in the treatment and / or prevention of hyperproliferative diseases, particularly cancers described herein, in a patient in need thereof.

[0158] In another aspect, the present invention relates to the aforementioned kit for use in a method of treating and / or preventing a hyperproliferative disease, preferably a cancer described herein.

[0159] In another aspect, the present invention is · An LRP5 / LRP6 antagonist (for example, any one of LRP5 / LRP6#1, LRP5 / LRP6#2, LRP5 / LRP6#3, LRP5 / LRP6#4, LRP5 / LRP6#5, LRP5 / LRP6#6 defined by the CDR and / or VHH sequences in Table 1a, Table 1b, Table 1c, Table 2a, Table 2b, Table 2c), and · An anti-PD-1 antibody described herein (for example, any one of PD1-1, PD1-2, PD1-3, PD1-4, PD1-5 defined by the CDR and / or VH / VL sequences in Table 3 and Table 4), and · Optionally, one or more pharmaceutically acceptable carriers, excipients, and / or media A pharmaceutical composition comprising the same.

[0160] In a preferred embodiment of the pharmaceutical composition of the present invention, the LRP5 / LRP6 antagonist comprises the amino acid sequence of SEQ ID NO: 64, SEQ ID NO: 65, or SEQ ID NO: 66, and the anti-PD-1 antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 29 and a light chain comprising the amino acid sequence of SEQ ID NO: 30. In a preferred embodiment of the pharmaceutical composition of the present invention, the LRP5 / LRP6 antagonist comprises the amino acid sequence of SEQ ID NO: 64, SEQ ID NO: 65, or SEQ ID NO: 66, and the anti-PD-1 antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 31 and a light chain comprising the amino acid sequence of SEQ ID NO: 32. In a preferred embodiment of the pharmaceutical composition of the present invention, the LRP5 / LRP6 antagonist comprises the amino acid sequence of SEQ ID NO: 64, SEQ ID NO: 65, or SEQ ID NO: 66, and the anti-PD-1 antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 33 and a light chain comprising the amino acid sequence of SEQ ID NO: 34.

[0161] In another aspect, the present invention relates to the use of an LRP5 / LRP6 antagonist (e.g., any one of LRP5 / LRP6#1, LRP5 / LRP6#2, LRP5 / LRP6#3, LRP5 / LRP6#4, LRP5 / LRP6#5, LRP5 / LRP6#6 defined by the CDR and / or VHH sequences of Table 1a, Table 1b, Table 1c, Table 2a, Table 2b, Table 2c) for the manufacture of a pharmaceutical composition for use in treating and / or preventing a proliferative disorder, preferably a cancer as described herein. Here, the LRP5 / LRP6 antagonist is used in combination with a PD-1 antibody (e.g., any one of PD1-1, PD1-2, PD1-3, PD1-4, PD1-5 defined by the CDR and / or VH / VL sequences of Table 3 and Table 4) as described herein. In a preferred embodiment, the LRP5 / LRP6 antagonist comprises the amino acid sequence of SEQ ID NO: 64, SEQ ID NO: 65 or SEQ ID NO: 66, and the PD-1 antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 29 and a light chain comprising the amino acid sequence of SEQ ID NO: 30. In a preferred embodiment, the LRP5 / LRP6 antagonist comprises the amino acid sequence of SEQ ID NO: 64, SEQ ID NO: 65 or SEQ ID NO: 66, and the PD-1 antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 31 and a light chain comprising the amino acid sequence of SEQ ID NO: 32. In a preferred embodiment, the LRP5 / LRP6 antagonist comprises the amino acid sequence of SEQ ID NO: 64, SEQ ID NO: 65 or SEQ ID NO: 66, and the PD-1 antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 33 and a light chain comprising the amino acid sequence of SEQ ID NO: 34.

[0162] In another aspect, the present invention refers to the use of a PD-1 antibody described herein (e.g., any one of PD1-1, PD1-2, PD1-3, PD1-4, PD1-5 defined by the CDR and / or VH / VL sequences in Tables 3 and 4) for manufacturing a pharmaceutical composition for use in treating and / or preventing a proliferative disorder, preferably a cancer described herein. Here, the PD-1 antibody is used in combination with an LRP5 / LRP6 antagonist (e.g., any one of LRP5 / LRP6#1, LRP5 / LRP6#2, LRP5 / LRP6#3, LRP5 / LRP6#4, LRP5 / LRP6#5, LRP5 / LRP6#6 defined by the CDR and / or VHH sequences in Tables 1a, 1b, 1c, 2a, 2b, 2c). In a preferred embodiment, the LRP5 / LRP6 antagonist comprises the amino acid sequence of SEQ ID NO: 64, SEQ ID NO: 65 or SEQ ID NO: 66, and the PD-1 antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 29 and a light chain comprising the amino acid sequence of SEQ ID NO: 30. In a preferred embodiment, the LRP5 / LRP6 antagonist comprises the amino acid sequence of SEQ ID NO: 64, SEQ ID NO: 65 or SEQ ID NO: 66, and the PD-1 antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 31 and a light chain comprising the amino acid sequence of SEQ ID NO: 32. In a preferred embodiment, the LRP5 / LRP6 antagonist comprises the amino acid sequence of SEQ ID NO: 64, SEQ ID NO: 65 or SEQ ID NO: 66, and the PD-1 antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 33 and a light chain comprising the amino acid sequence of SEQ ID NO: 34.

[0163] In another aspect, the present invention relates to the use of an LRP5 / LRP6 antagonist (e.g., any one of LRP5 / LRP6#1, LRP5 / LRP6#2, LRP5 / LRP6#3, LRP5 / LRP6#4, LRP5 / LRP6#5, LRP5 / LRP6#6 defined by the CDR and / or VHH sequences of Table 1a, Table 1b, Table 1c, Table 2a, Table 2b, Table 2c) and a PD-1 antibody (e.g., any one of PD1-1, PD1-2, PD1-3, PD1-4, PD1-5 defined by the CDR and / or VH / VL sequences of Table 3 and Table 4) for manufacturing a pharmaceutical composition for use in treating and / or preventing a hyperproliferative disease, preferably a cancer as described herein. In a preferred embodiment, the LRP5 / LRP6 antagonist comprises the amino acid sequence of SEQ ID NO: 64, SEQ ID NO: 65 or SEQ ID NO: 66, and the PD-1 antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 29 and a light chain comprising the amino acid sequence of SEQ ID NO: 30. In a preferred embodiment, the LRP5 / LRP6 antagonist comprises the amino acid sequence of SEQ ID NO: 64, SEQ ID NO: 65 or SEQ ID NO: 66, and the PD-1 antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 31 and a light chain comprising the amino acid sequence of SEQ ID NO: 32. In a preferred embodiment, the LRP5 / LRP6 antagonist comprises the amino acid sequence of SEQ ID NO: 64, SEQ ID NO: 65 or SEQ ID NO: 66, and the PD-1 antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 33 and a light chain comprising the amino acid sequence of SEQ ID NO: 34.

[0164] In another aspect, the present invention relates to an LRP5 / LRP6 antagonist (e.g., any one of LRP5 / LRP6#1, LRP5 / LRP6#2, LRP5 / LRP6#3, LRP5 / LRP6#4, LRP5 / LRP6#5, LRP5 / LRP6#6 defined by the CDR and / or VHH sequences of Table 1a, Table 1b, Table 1c, Table 2a, Table 2b, Table 2c) and an anti-PD-1 antibody (e.g., any one of PD1-1, PD1-2, PD1-3, PD1-4, PD1-5 defined by the CDR and / or VH / VL sequences of Table 3 and Table 4) for use in a method of treating and / or preventing a proliferative disorder, preferably a cancer as described herein, and combinations, pharmaceutical compositions or kits of the invention as described herein, respectively, comprising, consisting of or essentially consisting of the same. In a preferred embodiment, the LRP5 / LRP6 antagonist comprises the amino acid sequence of SEQ ID NO: 64, SEQ ID NO: 65 or SEQ ID NO: 66, and the PD-1 antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 29 and a light chain comprising the amino acid sequence of SEQ ID NO: 30. In a preferred embodiment, the LRP5 / LRP6 antagonist comprises the amino acid sequence of SEQ ID NO: 64, SEQ ID NO: 65 or SEQ ID NO: 66, and the PD-1 antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 31 and a light chain comprising the amino acid sequence of SEQ ID NO: 32. In a preferred embodiment, the LRP5 / LRP6 antagonist comprises the amino acid sequence of SEQ ID NO: 64, SEQ ID NO: 65 or SEQ ID NO: 66, and the PD-1 antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 33 and a light chain comprising the amino acid sequence of SEQ ID NO: 34.

[0165] Embodiments permutations related to LRP5 / LRP6 antagonists (e.g., any one of LRP5 / LRP6#1, LRP5 / LRP6#2, LRP5 / LRP6#3, LRP5 / LRP6#4, LRP5 / LRP6#5, LRP5 / LRP6#6) together with permutations related to PD-1 antagonists PD1-1, PD1-2, PD1-3, PD1-4, PD1-5 are considered to be specifically disclosed in their entirety and are considered to be embodiments of the present invention, including specific combinations and methods of applying specific dosing / administration regimens detailed below, resulting in all of their combinations, compositions, kits, methods, uses, and compounds for the treatment of specific cancers detailed below.

[0166] The routes of administration for the LRP5 / LRP6 antagonists and / or anti-PD1 antibodies described herein include, but are not limited to, parenteral (e.g., intramuscular, intraperitoneal, intravenous, transdermal, or subcutaneous injection or implant), oral, enteral, nasal, vaginal, rectal, or topical administration. In a preferred embodiment, the route of administration is intravenous administration, particularly intravenous infusion or injection. The compounds of the present invention can be formulated, alone or together, into suitable pharmaceutical dosage forms containing conventional non-toxic pharmaceutically acceptable carriers, excipients, and / or media suitable for each route of administration. More preferably, the formulations include solid, semi-solid, or liquid dosage forms, such as lyophilized products, solutions (e.g., injectable and infusible solutions), dispersions, or suspensions, liposomes, and suppositories. The preferred mode is determined by the intended mode of administration and therapeutic application. Particularly preferred embodiments include liquid formulations and lyophilization. In the case of lyophilization, the lyophilized product can be reconstituted in a liquid, preferably water.

[0167] Administration of the anti-PD-1 antibodies described herein can be by injection (e.g., subcutaneous or intravenous) at a dose of, for example, about 0.1 to 30 mg / kg patient body weight, such as about 0.5 to 25 mg / kg patient body weight, about 1 to 20 mg / kg patient body weight, about 2 to 5 mg / kg patient body weight, or about 3 mg / kg patient body weight.

[0168] In some embodiments, the anti-PD-1 antibody is administered at a dose of about 10-20 mg / kg of patient body weight every two weeks. The antibody molecule is administered by intravenous infusion at a rate of more than 20 mg / min, for example, 20-40 mg / min, typically 40 mg / min or more, to reach a dose of about 35-440 mg / m 2 , typically about 70-310 mg / m 2 , more typically about 110-130 mg / m 2 . In some embodiments, an infusion rate of about 110-130 mg / m 2 achieves a level of about 3 mg / kg of patient body weight. In other embodiments, the antibody molecule is administered by intravenous infusion at a rate of less than 10 mg / min, for example, 5 mg / min or less, to reach a dose of about 1-100 mg / m 2 , for example, about 5-50 mg / m 2 , about 7-25 mg / m 2 or about 10 mg / m 2 . In some embodiments, the antibody is infused over a period of about 30 minutes.

[0169] Preferred dosing regimens for the anti-PD-1 antibodies described herein include 1 mg / kg of patient body weight by intravenous administration or alternatively 3 mg / kg of patient body weight, where the antibody is given every three or four weeks.

[0170] The LRP5 / LRP6 antagonists or compositions containing them described herein can be administered in an effective amount or dose, for example, intravenously (i.v.), subcutaneously (s.c.), intramuscularly (i.m.), intraperitoneally (i.p.), transdermally, orally, sublingually (e.g., in the form of a sublingual tablet, placed sublingually and adsorbed through the mucosa into the sublingual capillary network as a spray or droplet), nasally (intranasally) (e.g., in the form of a nasal spray and / or as an aerosol), topically, as an agent, by inhalation or in any other suitable manner.

[0171] The LRP5 / LRP6 antagonists described herein will generally be administered in an amount of 0.005 to 20.0 mg, preferably 0.05 to 10.0 mg / kg / dose, more preferably 0.5 to 10 mg / kg / dose, per kilogram of patient body weight per dose, but may vary specifically depending on the particular disease, disorder or condition being treated, the potency of the specific LRP5 / LRP6 antagonist being used, the specific route of administration and the particular pharmaceutical formulation or composition being used. For this reason, in some cases, it may be sufficient to use less than the minimum dose given above, while in other cases, the upper limit may have to be exceeded. When administering in large amounts, it may be desirable to divide it into small amounts and disperse it over a day.

[0172] It should be noted that the dosage value may vary depending on the type and severity of the condition being alleviated. Furthermore, it should be understood that for any particular subject, a specific dosage regimen should be adjusted over time according to the expert judgment of the person managing or supervising the administration of the individual needs and composition.

[0173] The LRP5 / LRP6 antagonists and anti-PD1 antibodies described herein can be administered in a therapeutically effective amount as a single dose or as divided doses at appropriate time intervals. A therapeutically effective amount refers to an amount effective in the dosage and period required to achieve the desired therapeutic result and is the minimum amount necessary to prevent, ameliorate or treat a disease or disorder. The therapeutically effective amount of the compounds described herein may vary according to factors such as the individual's disease state, age, gender and body weight and the ability of the compound to induce the desired response in the individual. Also, a therapeutically effective amount is also an amount in which any toxic or adverse effects of the compound are less than the therapeutically beneficial effects. The therapeutically effective dose preferably inhibits a measurable parameter, such as tumor growth rate, by at least about 20%, more preferably at least about 40%, even more preferably at least about 60%, even more preferably at least about 80%, relative to an untreated subject or relative to a previous untreated period of the same subject to be treated.

[0174] The active compound can be administered at a therapeutically effective dose in monotherapy or at a dose lower or higher than the dose used in monotherapy, but when combined, a desired (synergistic) therapeutically effective amount is provided. This can be useful, for example, to avoid, limit or reduce any undesirable side effects associated with the use of one or more substances or principles while still obtaining the desired pharmacological or therapeutic effect when they are used in normal amounts.

[0175] The amount of the compounds described herein required for use in treatment can be adapted to the particular compound selected, the route of administration, the nature of the condition being treated and the age and condition of the patient and will ultimately be at the discretion of the attending physician or clinician. Also, the dosage of the compounds described herein can be adapted according to the target cells, tumors, tissues, grafts or organs.

[0176] The desired dosages of the LRP5 / LRP6 antagonist or anti-PD-1 antibody described herein can both be administered as a fixed amount per administration or as a bolus so as to reach a set blood concentration in the patient.

[0177] It should be understood that in the present invention, the LRP5 / LRP6 antagonist and anti-PD1 antibody can be formulated and administered either subordinately (i.e., mixed together in one composition) or independently (i.e., as separate compositions). Here, such administration provides therapeutically effective levels of the two compounds in the patient's body. Also, the latter is also applicable to cocktail therapies, for example, the administration of three or more active agents. That is, the LRP5 / LRP6 antagonist and anti-PD1 antibody can be administered as part of the same pharmaceutical composition / dosage form or preferably, in separate pharmaceutical compositions / dosage forms. As long as the administration is in separate pharmaceutical compositions / dosage forms, according to the present invention, it should be understood that the administration contemplates simultaneous, concurrent, sequential or alternating administration of the active agents or components.

[0178] The term "simultaneous" (also referred to herein as "concomitant") refers to administering both compounds / compositions substantially simultaneously.

[0179] Simultaneous administration includes administering the active agents within the same general period, e.g., on the same day, but not necessarily simultaneously.

[0180] Sequential administration involves administering one agent over a first period (e.g., over several hours, days, or a week) using one or more doses, and subsequently administering the other agent over a second period (e.g., over several hours, days, or a week) using one or more doses. A repeated schedule can also be used, which includes administering the active agents on different days over the treatment period and does not necessarily follow a regular order. Alternatively, sequential administration is also contemplated where the second administration step is carried out immediately upon completion of the administration of the first compound. One of ordinary skill in the art knows how to determine the end of the first administration step and thereby identify a suitable time point to initiate the second administration step.

[0181] Alternating administration involves administering one agent over a period, e.g., over several hours, days, or a week, and subsequently administering the other agent over a subsequent period, e.g., over several hours, days, or a week, and then repeating this pattern over one or more cycles, where the total number of repetitions is determined by the selected dosing regimen.

[0182] Variations of these general guidelines can also be utilized, e.g., according to the agents used and the condition of the subject.

[0183] In a preferred embodiment of the present invention, in the method of the present invention, the LRP5 / LRP6 antagonist and the anti-PD1 antibody described herein are administered simultaneously or contemporaneously (e.g., by intravenous injection or subcutaneously) during a first period, and subsequently, during a second period, the anti-PD1 antibody is administered (e.g., by intravenous injection or subcutaneously), and the LRP5 / LRP6 antagonist is not administered. In some embodiments, the first period is 3 weeks or 6 weeks when the polypeptide specifically bindable to LRP5 and LRP6 and the PD1 antibody are administered every 3 weeks. In some embodiments, the first period is 4 weeks or 8 weeks when the polypeptide specifically bindable to LRP5 and LRP6 and the PD1 antibody are administered every 4 weeks. This dosing schedule is particularly preferably utilized when the LRP5 / LRP6 antagonist is LRP5 / LRP6#1 and the anti-PD-1 antibody is PD1-1. Even more preferably, this dosing schedule is utilized for the treatment of gastrointestinal cancer, melanoma, bladder cancer or lung cancer (including gastrointestinal cancer, melanoma, bladder cancer and lung cancer refractory or resistant to checkpoint inhibitor therapy) or any solid tumor refractory or resistant to checkpoint inhibitor therapy. This dosing schedule is particularly preferably utilized when the LRP5 / LRP6 antagonist is LRP5 / LRP6#5 and the anti-PD-1 antibody is PD1-1. Even more preferably, this dosing schedule is utilized for the treatment of gastrointestinal cancer, melanoma, bladder cancer or lung cancer (including gastrointestinal cancer, melanoma, bladder cancer and lung cancer refractory or resistant to checkpoint inhibitor therapy) or any solid tumor refractory or resistant to checkpoint inhibitor therapy. This dosing schedule is particularly preferably utilized when the LRP5 / LRP6 antagonist is LRP5 / LRP6#6 and the anti-PD-1 antibody is PD1-1. Even more preferably, this dosing schedule is utilized for the treatment of gastrointestinal cancer, melanoma, bladder cancer or lung cancer (including gastrointestinal cancer, melanoma, bladder cancer and lung cancer refractory or resistant to checkpoint inhibitor therapy) or any solid tumor refractory or resistant to checkpoint inhibitor therapy.This dosing schedule is particularly preferably used when the LRP5 / LRP6 antagonist is LRP5 / LRP6#1 and the anti-PD-1 antibody is PD1-2. Even more preferably, this dosing schedule is used for the treatment of gastrointestinal cancer, melanoma, bladder cancer or lung cancer (including gastrointestinal cancer, melanoma, bladder cancer and lung cancer refractory or resistant to checkpoint inhibitor therapy) or any solid tumor refractory or resistant to checkpoint inhibitor therapy. This dosing schedule is particularly preferably used when the LRP5 / LRP6 antagonist is LRP5 / LRP6#5 and the anti-PD-1 antibody is PD1-2. Even more preferably, this dosing schedule is used for the treatment of gastrointestinal cancer, melanoma, bladder cancer or lung cancer (including gastrointestinal cancer, melanoma, bladder cancer and lung cancer refractory or resistant to checkpoint inhibitor therapy) or any solid tumor refractory or resistant to checkpoint inhibitor therapy. This dosing schedule is particularly preferably used when the LRP5 / LRP6 antagonist is LRP5 / LRP6#6 and the anti-PD-1 antibody is PD1-2. Even more preferably, this dosing schedule is used for the treatment of gastrointestinal cancer, melanoma, bladder cancer or lung cancer (including gastrointestinal cancer, melanoma, bladder cancer and lung cancer refractory or resistant to checkpoint inhibitor therapy) or any solid tumor refractory or resistant to checkpoint inhibitor therapy. This dosing schedule is particularly preferably used when the LRP5 / LRP6 antagonist is LRP5 / LRP6#1 and the anti-PD-1 antibody is PD1-3. Even more preferably, this dosing schedule is used for the treatment of gastrointestinal cancer, melanoma, bladder cancer or lung cancer (including gastrointestinal cancer, melanoma, bladder cancer and lung cancer refractory or resistant to checkpoint inhibitor therapy) or any solid tumor refractory or resistant to checkpoint inhibitor therapy. This dosing schedule is particularly preferably used when the LRP5 / LRP6 antagonist is LRP5 / LRP6#5 and the anti-PD-1 antibody is PD1-3.Even more preferably, this dosing schedule is utilized for the treatment of gastrointestinal cancer, melanoma, bladder cancer or lung cancer (including gastrointestinal cancer, melanoma, bladder cancer and lung cancer refractory or resistant to checkpoint inhibitor therapy) or any solid tumor refractory or resistant to checkpoint inhibitor therapy. This dosing schedule is particularly preferably utilized when the LRP5 / LRP6 antagonist is LRP5 / LRP6#6 and the anti-PD-1 antibody is PD1-3. Even more preferably, this dosing schedule is utilized for the treatment of gastrointestinal cancer, melanoma, bladder cancer or lung cancer (including gastrointestinal cancer, melanoma, bladder cancer and lung cancer refractory or resistant to checkpoint inhibitor therapy) or any solid tumor refractory or resistant to checkpoint inhibitor therapy.

[0184] In another preferred embodiment of the present invention, both the LRP5 / LRP6 antagonist and the anti-PD1 antibody described herein are administered every three weeks (by intravenous injection or subcutaneously simultaneously or contemporaneously) for a first period (e.g., three weeks or six weeks), and then the anti-PD1 antibody is administered every three weeks (e.g., by intravenous injection or subcutaneously) for, e.g., a second period. For example, the LRP5 / LRP6 antagonist and the anti-PD1 antibody are administered (i) in the first week or (ii) in the first and fourth weeks simultaneously or contemporaneously (e.g., by intravenous injection or subcutaneously), and then the PD1 antibody is administered, e.g., in the seventh week, the tenth week and any subsequent third week (the thirteenth week, the sixteenth week, etc.) until the treatment is completed. In the case of option (i), the PD1 antibody is administered alone in advance in the fourth week (i.e., instead of co-administration with the LRP5 antagonist as in option (ii)).

[0185] This dosing schedule is particularly preferably used when the LRP5 / LRP6 antagonist is LRP5 / LRP6#1 and the anti-PD-1 antibody is PD1-1. Even more preferably, this dosing schedule is used for the treatment of gastrointestinal cancer, melanoma, bladder cancer or lung cancer (including gastrointestinal cancer, melanoma, bladder cancer and lung cancer refractory or resistant to checkpoint inhibitor therapy) or any solid tumor refractory or resistant to checkpoint inhibitor therapy. This dosing schedule is particularly preferably used when the LRP5 / LRP6 antagonist is LRP5 / LRP6#5 and the anti-PD-1 antibody is PD1-1. Even more preferably, this dosing schedule is used for the treatment of gastrointestinal cancer, melanoma, bladder cancer or lung cancer (including gastrointestinal cancer, melanoma, bladder cancer and lung cancer refractory or resistant to checkpoint inhibitor therapy) or any solid tumor refractory or resistant to checkpoint inhibitor therapy. This dosing schedule is particularly preferably used when the LRP5 / LRP6 antagonist is LRP5 / LRP6#6 and the anti-PD-1 antibody is PD1-1. Even more preferably, this dosing schedule is used for the treatment of gastrointestinal cancer, melanoma, bladder cancer or lung cancer (including gastrointestinal cancer, melanoma, bladder cancer and lung cancer refractory or resistant to checkpoint inhibitor therapy) or any solid tumor refractory or resistant to checkpoint inhibitor therapy. This dosing schedule is particularly preferably used when the LRP5 / LRP6 antagonist is LRP5 / LRP6#1 and the anti-PD-1 antibody is PD1-2. Even more preferably, this dosing schedule is used for the treatment of gastrointestinal cancer, melanoma, bladder cancer or lung cancer (including gastrointestinal cancer, melanoma, bladder cancer and lung cancer refractory or resistant to checkpoint inhibitor therapy) or any solid tumor refractory or resistant to checkpoint inhibitor therapy. This dosing schedule is particularly preferably used when the LRP5 / LRP6 antagonist is LRP5 / LRP6#5 and the anti-PD-1 antibody is PD1-2.Even more preferably, this dosing schedule is utilized for the treatment of gastrointestinal cancer, melanoma, bladder cancer or lung cancer (including gastrointestinal cancer, melanoma, bladder cancer and lung cancer that are refractory or resistant to checkpoint inhibitor therapy) or any solid tumor that is refractory or resistant to checkpoint inhibitor therapy. This dosing schedule is particularly preferably utilized when the LRP5 / LRP6 antagonist is LRP5 / LRP6#6 and the anti-PD-1 antibody is PD1-2. Even more preferably, this dosing schedule is utilized for the treatment of gastrointestinal cancer, melanoma, bladder cancer or lung cancer (including gastrointestinal cancer, melanoma, bladder cancer and lung cancer that are refractory or resistant to checkpoint inhibitor therapy) or any solid tumor that is refractory or resistant to checkpoint inhibitor therapy. This dosing schedule is particularly preferably utilized when the LRP5 / LRP6 antagonist is LRP5 / LRP6#1 and the anti-PD-1 antibody is PD1-3. Even more preferably, this dosing schedule is utilized for the treatment of gastrointestinal cancer, melanoma, bladder cancer or lung cancer (including gastrointestinal cancer, melanoma, bladder cancer and lung cancer that are refractory or resistant to checkpoint inhibitor therapy) or any solid tumor that is refractory or resistant to checkpoint inhibitor therapy. This dosing schedule is particularly preferably utilized when the LRP5 / LRP6 antagonist is LRP5 / LRP6#5 and the anti-PD-1 antibody is PD1-3. Even more preferably, this dosing schedule is utilized for the treatment of gastrointestinal cancer, melanoma, bladder cancer or lung cancer (including gastrointestinal cancer, melanoma, bladder cancer and lung cancer that are refractory or resistant to checkpoint inhibitor therapy) or any solid tumor that is refractory or resistant to checkpoint inhibitor therapy. This dosing schedule is particularly preferably utilized when the LRP5 / LRP6 antagonist is LRP5 / LRP6#6 and the anti-PD-1 antibody is PD1-3.Even more preferably, this dosing schedule is utilized for the treatment of gastrointestinal cancer, melanoma, bladder cancer or lung cancer (including gastrointestinal cancer, melanoma, bladder cancer and lung cancer refractory or resistant to checkpoint inhibitor therapy) or any solid tumor refractory or resistant to checkpoint inhibitor therapy.

[0186] In another preferred embodiment of the invention, both the LRP5 / LRP6 antagonist and the anti-PD-1 antibody described herein are administered every 4 weeks (by intravenous infusion or subcutaneously simultaneously or contemporaneously) for a first period (e.g., 4 weeks or 8 weeks), and then the anti-PD1 antibody is administered every 4 weeks (e.g., by intravenous infusion or subcutaneously) for, e.g., a second period. For example, the LRP5 / LRP6 antagonist and the anti-PD1 antibody are administered (i) simultaneously or contemporaneously (e.g., by intravenous infusion or subcutaneously) in the first week or (ii) in the first and fifth weeks, and then the PD1 antibody is administered, e.g., in the ninth week, the thirteenth week and any subsequent 4-week period (the seventeenth week, the twenty-first week, etc.) until the treatment is completed. In the case of option (i), the PD1 antibody is administered alone in advance in the fifth week (i.e., instead of co-administration with the LRP5 antagonist as in option (ii)).

[0187] This dosing schedule is particularly preferably used when the LRP5 / LRP6 antagonist is LRP5 / LRP6#1 and the anti-PD-1 antibody is PD1-1. Even more preferably, this dosing schedule is used for the treatment of gastrointestinal cancer, melanoma, bladder cancer or lung cancer (including gastrointestinal cancer, melanoma, bladder cancer and lung cancer refractory or resistant to checkpoint inhibitor therapy) or any solid tumor refractory or resistant to checkpoint inhibitor therapy. This dosing schedule is particularly preferably used when the LRP5 / LRP6 antagonist is LRP5 / LRP6#5 and the anti-PD-1 antibody is PD1-1. Even more preferably, this dosing schedule is used for the treatment of gastrointestinal cancer, melanoma, bladder cancer or lung cancer (including gastrointestinal cancer, melanoma, bladder cancer and lung cancer refractory or resistant to checkpoint inhibitor therapy) or any solid tumor refractory or resistant to checkpoint inhibitor therapy. This dosing schedule is particularly preferably used when the LRP5 / LRP6 antagonist is LRP5 / LRP6#6 and the anti-PD-1 antibody is PD1-1. Even more preferably, this dosing schedule is used for the treatment of gastrointestinal cancer, melanoma, bladder cancer or lung cancer (including gastrointestinal cancer, melanoma, bladder cancer and lung cancer refractory or resistant to checkpoint inhibitor therapy) or any solid tumor refractory or resistant to checkpoint inhibitor therapy. This dosing schedule is particularly preferably used when the LRP5 / LRP6 antagonist is LRP5 / LRP6#1 and the anti-PD-1 antibody is PD1-2. Even more preferably, this dosing schedule is used for the treatment of gastrointestinal cancer, melanoma, bladder cancer or lung cancer (including gastrointestinal cancer, melanoma, bladder cancer and lung cancer refractory or resistant to checkpoint inhibitor therapy) or any solid tumor refractory or resistant to checkpoint inhibitor therapy. This dosing schedule is particularly preferably used when the LRP5 / LRP6 antagonist is LRP5 / LRP6#5 and the anti-PD-1 antibody is PD1-2.Even more preferably, this dosing schedule is utilized for the treatment of gastrointestinal cancer, melanoma, bladder cancer or lung cancer (including gastrointestinal cancer, melanoma, bladder cancer and lung cancer that are refractory or resistant to checkpoint inhibitor therapy) or any solid tumor that is refractory or resistant to checkpoint inhibitor therapy. This dosing schedule is particularly preferably utilized when the LRP5 / LRP6 antagonist is LRP5 / LRP6#6 and the anti-PD-1 antibody is PD1-2. Even more preferably, this dosing schedule is utilized for the treatment of gastrointestinal cancer, melanoma, bladder cancer or lung cancer (including gastrointestinal cancer, melanoma, bladder cancer and lung cancer that are refractory or resistant to checkpoint inhibitor therapy) or any solid tumor that is refractory or resistant to checkpoint inhibitor therapy. This dosing schedule is particularly preferably utilized when the LRP5 / LRP6 antagonist is LRP5 / LRP6#1 and the anti-PD-1 antibody is PD1-3. Even more preferably, this dosing schedule is utilized for the treatment of gastrointestinal cancer, melanoma, bladder cancer or lung cancer (including gastrointestinal cancer, melanoma, bladder cancer and lung cancer that are refractory or resistant to checkpoint inhibitor therapy) or any solid tumor that is refractory or resistant to checkpoint inhibitor therapy. This dosing schedule is particularly preferably utilized when the LRP5 / LRP6 antagonist is LRP5 / LRP6#5 and the anti-PD-1 antibody is PD1-3. Even more preferably, this dosing schedule is utilized for the treatment of gastrointestinal cancer, melanoma, bladder cancer or lung cancer (including gastrointestinal cancer, melanoma, bladder cancer and lung cancer that are refractory or resistant to checkpoint inhibitor therapy) or any solid tumor that is refractory or resistant to checkpoint inhibitor therapy. This dosing schedule is particularly preferably utilized when the LRP5 / LRP6 antagonist is LRP5 / LRP6#6 and the anti-PD-1 antibody is PD1-3.Even more preferably, this dosing schedule is utilized for the treatment of gastrointestinal cancer, melanoma, bladder cancer or lung cancer (including gastrointestinal cancer, melanoma, bladder cancer and lung cancer that are refractory or resistant to checkpoint inhibitor therapy) or any solid tumor that is refractory or resistant to checkpoint inhibitor therapy.

[0188] Preferably, both the LRP5 / LRP6 antagonist described herein (e.g., about 0.5 to 10 mg / kg patient body weight) and the anti-PD1 antibody described herein (e.g., any one dose of 2, 3, 4, or 5 mg / kg patient body weight) are administered (by intravenous infusion or subcutaneously simultaneously or contemporaneously) every 3 or 4 weeks during a first period (e.g., corresponding to the first or second administration), and then the anti-PD1 antibody is administered, for example, every 3 or 4 weeks (e.g., by intravenous infusion or subcutaneously) during a second period. This dosing schedule is particularly preferably utilized when the LRP5 / LRP6 antagonist is LRP5 / LRP6#1 and the anti-PD-1 antibody is PD1-1. Even more preferably, this dosing schedule is utilized for the treatment of gastrointestinal cancer, melanoma, bladder cancer, or lung cancer (including gastrointestinal cancer, melanoma, bladder cancer, and lung cancer refractory or resistant to checkpoint inhibitor therapy) or any solid tumor refractory or resistant to checkpoint inhibitor therapy. This dosing schedule is particularly preferably utilized when the LRP5 / LRP6 antagonist is LRP5 / LRP6#5 and the anti-PD-1 antibody is PD1-1. Even more preferably, this dosing schedule is utilized for the treatment of gastrointestinal cancer, melanoma, bladder cancer, or lung cancer (including gastrointestinal cancer, melanoma, bladder cancer, and lung cancer refractory or resistant to checkpoint inhibitor therapy) or any solid tumor refractory or resistant to checkpoint inhibitor therapy. This dosing schedule is particularly preferably utilized when the LRP5 / LRP6 antagonist is LRP5 / LRP6#6 and the anti-PD-1 antibody is PD1-1. Even more preferably, this dosing schedule is utilized for the treatment of gastrointestinal cancer, melanoma, bladder cancer, or lung cancer (including gastrointestinal cancer, melanoma, bladder cancer, and lung cancer refractory or resistant to checkpoint inhibitor therapy) or any solid tumor refractory or resistant to checkpoint inhibitor therapy. This dosing schedule is particularly preferably utilized when the LRP5 / LRP6 antagonist is LRP5 / LRP6#1 and the anti-PD-1 antibody is PD1-2.Even more preferably, this administration schedule is used for the treatment of gastrointestinal cancer, melanoma, bladder cancer or lung cancer (including gastrointestinal cancer, melanoma, bladder cancer and lung cancer that are refractory or resistant to checkpoint inhibitor therapy) or any solid tumor that is refractory or resistant to checkpoint inhibitor therapy. This administration schedule is particularly preferably used when the LRP5 / LRP6 antagonist is LRP5 / LRP6#5 and the anti-PD-1 antibody is PD1-2. Even more preferably, this administration schedule is used for the treatment of gastrointestinal cancer, melanoma, bladder cancer or lung cancer (including gastrointestinal cancer, melanoma, bladder cancer and lung cancer that are refractory or resistant to checkpoint inhibitor therapy) or any solid tumor that is refractory or resistant to checkpoint inhibitor therapy. This administration schedule is particularly preferably used when the LRP5 / LRP6 antagonist is LRP5 / LRP6#6 and the anti-PD-1 antibody is PD1-2. Even more preferably, this administration schedule is used for the treatment of gastrointestinal cancer, melanoma, bladder cancer or lung cancer (including gastrointestinal cancer, melanoma, bladder cancer and lung cancer that are refractory or resistant to checkpoint inhibitor therapy) or any solid tumor that is refractory or resistant to checkpoint inhibitor therapy. This administration schedule is particularly preferably used when the LRP5 / LRP6 antagonist is LRP5 / LRP6#1 and the anti-PD-1 antibody is PD1-3. Even more preferably, this administration schedule is used for the treatment of gastrointestinal cancer, melanoma, bladder cancer or lung cancer (including gastrointestinal cancer, melanoma, bladder cancer and lung cancer that are refractory or resistant to checkpoint inhibitor therapy) or any solid tumor that is refractory or resistant to checkpoint inhibitor therapy. This administration schedule is particularly preferably used when the LRP5 / LRP6 antagonist is LRP5 / LRP6#5 and the anti-PD-1 antibody is PD1-3.Even more preferably, this dosing schedule is utilized for the treatment of gastrointestinal cancer, melanoma, bladder cancer or lung cancer (including gastrointestinal cancer, melanoma, bladder cancer and lung cancer refractory or resistant to checkpoint inhibitor therapy) or any solid tumor refractory or resistant to checkpoint inhibitor therapy. This dosing schedule is particularly preferably utilized when the LRP5 / LRP6 antagonist is LRP5 / LRP6#6 and the anti-PD-1 antibody is PD1-3. Even more preferably, this dosing schedule is utilized for the treatment of gastrointestinal cancer, melanoma, bladder cancer or lung cancer (including gastrointestinal cancer, melanoma, bladder cancer and lung cancer refractory or resistant to checkpoint inhibitor therapy) or any solid tumor refractory or resistant to checkpoint inhibitor therapy.

[0189] In some embodiments of the invention, both the LRP5 / LRP6 antagonist and the anti-PD1 antibody described herein are administered every 3 or 4 weeks (by intravenous infusion or subcutaneously simultaneously or contemporaneously) during a first period (e.g., corresponding to the first or second administration), and then the anti-PD1 antibody is administered weekly, bi-weekly, tri-weekly or monthly (e.g., by intravenous infusion or subcutaneously) during a second period.

[0190] Depending on the disease being treated, the combination therapy defined herein can be used by itself or, in particular, in further combination with one or more additional therapeutic agents selected from chemotherapeutic agents or therapeutically active compounds that inhibit angiogenesis, signal transduction pathways or mitotic checkpoints in cancer cells.

[0191] The additional therapeutic agent(s) can be administered simultaneously with the administration of the LRP5 / LRP6 antagonist and / or the PD1 antibody, optionally as a component of the same pharmaceutical preparation or before or after the same administration.

[0192] This / These additional therapeutic agent(s) can be selected (but not limited to) from the following.

[0193] · Immunotherapeutic agents, for example, the following checkpoint inhibitors: TIM3, PD-L1, PD-L2, CTLA-4, VISTA, BTLA, TIGIT, CD160, LAIR1, 2B4, CEACAM modulators of · Cancer vaccines, · DNA damaging agents, · Inhibitors of angiogenesis, · Inhibitors of signal transduction pathways, · Inhibitors of the mitotic checkpoint and · Hormones, hormone analogs, and antihormonal agents (e.g., tamoxifen, toremifene, raloxifene, fulvestrant, megestrol acetate, flutamide, nilutamide, bicalutamide, aminoglutethimide, cyproterone acetate, finasteride, buserelin acetate, fludrocortisone, fluoxymesterone, medroxyprogesterone, octreotide), aromatase inhibitors (e.g., anastrozole, letrozole, exemestane, atamestane), LHRH agonists and antagonists (e.g., goserelin acetate, leuprolide), inhibitors of growth factors (growth factors, e.g., "platelet-derived growth factor (PDGF)", "fibroblast growth factor (FGF)", "vascular endothelial growth factor (VEGF)", "epidermal growth factor (EGF)", "insulin-like growth factor (IGF)", "human epidermal growth factor (HER, e.g., HER2, HER3, HER4)", and "hepatocyte growth factor (HGF)", etc.), e.g., "growth factor" antibodies, "growth factor receptor" antibodies, and tyrosine kinase inhibitors, e.g., cetuximab, gefitinib, imatinib, lapatinib, bosutinib, and trastuzumab, etc., antimetabolites (e.g., folic acid antagonists, e.g., methotrexate, raltitrexed, pyrimidine analogs, e.g., 5-fluorouracil (5-FU), capecitabine, and gemcitabine, purine and adenosine analogs, e.g., mercaptopurine, thioguanine, cladribine, and pentostatin, cytarabine (ara C), fludarabine); antitumor antibiotics (e.g., anthracyclines, e.g., doxorubicin, doxil (pegylated liposomal doxorubicin hydrochloride), myocet (non-pegylated liposomal doxorubicin), daunorubicin, epirubicin, and idarubicin, mitomycin-C, bleomycin, dactinomycin, plicamycin, streptozocin), platinum derivatives (e.g., cisplatin, oxaliplatin, carboplatin), alkylating agents (e.g., estramustine, mechlorethamine, melphalan, chlorambucil, busulfan, dacarbazine, cyclophosphamide, ifosfamide, temozolomide, nitrosoureas, e.g., carmustine and lomustine, etc., thiotepa), antimitotic agents (e.g., vinca alkaloids, e.g., vinblastine,Vincindine, vinorelbine, vincristine, etc., and taxanes, such as paclitaxel, docetaxel), angiogenesis inhibitors (e.g., tasquinimod), tubulin inhibitors, DNA synthesis inhibitors (e.g., sapacitabine), PARP inhibitors, topoisomerase inhibitors (e.g., epipodophyllotoxins, such as etoposide and etopophos, teniposide, amsacrine, topotecan, irinotecan, mitoxantrone, etc.), serine / threonine kinase inhibitors (e.g., PDK1 inhibitors, Raf inhibitors, A-Raf inhibitors, B-Raf inhibitors, C-Raf inhibitors, mTOR inhibitors, mTORC1 / 2 inhibitors, PI3K inhibitors, PI3Kα inhibitors, dual mTOR / PI3K inhibitors, STK33 inhibitors, AKT inhibitors, PLK1 inhibitors, CDK inhibitors, aurora kinase inhibitors), tyrosine kinase inhibitors (e.g., PTK2 / FAK inhibitors), protein-protein interaction inhibitors (e.g., IAP activators, Mcl-1, MDM2 / MDMX), MEK inhibitors (e.g., pimasertib), ERK inhibitors, FLT3 inhibitors (e.g., quizartinib), BRD4 inhibitors, IGF-1R inhibitors, TRAILR2 agonists, Bcl-xL inhibitors, Bcl-2 inhibitors (e.g., venetoclax), Bcl-2 / Bcl-xL inhibitors, ErbB receptor inhibitors, BCR-ABL inhibitors, ABL inhibitors, Src inhibitors, rapamycin analogs (e.g., everolimus, temsirolimus, ridaforolimus, sirolimus), androgen synthesis inhibitors (e.g., abiraterone, TAK-700), androgen receptor inhibitors (e.g., enzalutamide, ARN-509), immunotherapy (e.g., Sipuleucel-T), DNMT inhibitors (e.g., SGI110, temozolomide, bosutinib), HDAC inhibitors (e.g., vorinostat, entinostat, panobinostat, panobinostat), ANG1 / 2 inhibitors (e.g., trebananib), CYP17 inhibitors (e.g., galeterone), radiopharmaceuticals (e.g., radium-223, alpharadin), immunotherapeutic agents (e.g., poxvirus-based vaccines, ipilimumab, immune checkpoint inhibitors), and various chemotherapeutic agents, such as amifostine, anagrelide, cladribine, filgrastim, interferon, interferon alpha, leucovorin,Rituximab, procarbazine, levamisole, mesna, mitotane, pamidronate and porfimer, 2-chlorodeoxyadenosine, 2-fluorodeoxycytidine, 2-methoxyestradiol, 2C4, 3-arotinoid, 131-I-TM-601, 3CPA, 7-ethyl-10-hydroxycamptothecin, 16-aza-epothilone B, ABT-199, ABT-263 / navitoclax, ABT-737, A105972, A204197, aldesleukin, alisertib / MLN8237, alitretinoin, arovastatin-7, altretamine, albosidinib, amonafide, anthrapyrazole, AG-2037, AP-5280, apazicon, apomine, alanose, algrabin, arzoxifene, atamestane, atrasentan, auristatin PE, AVLB, AZ10992, ABX-EGF, AMG-479 (ganitumab), AMG-232, AMG-511, AMG2520765, AMG2112819, ARRY162, ARRY438162, ARRY-300, ARRY-142886 / AZD-6244 (selumetinib), ARRY-704 / AZD-8330, ATSP-7041, AR-12, AR-42, AS-703988, AXL-1717, AZD-1480, AZD-4547, AZD-8055, AZD-5363, AZD-6244, AZD-7762, ARQ-736, ARQ680, AS-703026 (primasertib), abastin, AZD-2014, azacitidine (5-aza), aza-epothilone B, azonafide, barasertib / AZD1152, BAY-43-9006, BAY80-6946, BBR-3464, BBR-3576, bevacizumab, BEZ-235 / dactolisib, bilicodar dicitrate, bilinapant, BCX-1777, BKM-120 / buparlisib, bleomycin, BLP-25, BMS-184476, BMS-247550, BMS-188797, BMS-275291, BMS-663513, BMS-754807, BNP-1350, BNP-7787, BIBW2992 / afatinib, BIBF1120 / nintedanib, BI836845, BI2536, BI6727 / volasertib, BI836845, BI847325, BI853520, BIIB-022,Bleomycinic acid, Bleomycin A, Bleomycin B, Brivanib, Bryostatin-1, Bortezomib, Brostallicin, Busulfan, BYL-719 / Alpelisib, CA-4 prodrug, CA-4, Cabazitaxel, Cabozantinib, CapCell, Calcitriol, Canertinib, Cyclophosphamide, Capecitabine, Carboxyphthalato platinum, CCI-779, CC-115, CC-223, CEP-701, CEP-751, CBT-1, Cefixime, Ceflatonin, Ceftriaxone, Celecoxib, Selumetinib, Semadotin, CGM-097, CH4987655 / RO-4987655, Chlorotrianisene, Siramesine, Cyclosporine, CD20 antibody, CDA-II, CDC-394, CKD-602, CKI-27, Clofarabine, Colchicine, Combretastatin A4, COT inhibitor, CHS-828, CH-5132799, CLL-zeta, CMT-3, Cryptophycin 52, CPI-613, CTP-37, CTLA-4 monoclonal antibody (e.g., Ipilimumab), CP-461, Crizotinib, CV-247, Cyanomorpholinodoxorubicin, Cytarabine, D24851, Dasatinib, Decitabine, Doxorubicin, Doxorubicin, Deoxycholformycin, Depsipeptide, Deoxyepothilone B, Dexamethasone, Dexrazoxane, Diethylstilbestrol, Difluoromotecan, Doxorubicin, DMDC, Drastatin 10, Dranidazole, DS-7423, DS-3032, E7010, E-6201, Edatrexate, Edotreotide, Efaproxiral, Eflornithine, EGFR inhibitor, EKB-569, EKB-509, Enzastaurin, Elescromol, Elsamitrucin, Epothilone B, Epratuzumab, EPZ-004777, ER-86526, Erlotinib, ET-18-OCH3, Ethynylcytidine, Ethinylestradiol, Exatecan, Exatecan mesylate, Exemestane, Exisulind, Fenretinide, Figitumumab, Floxuridine, Folic acid, FOLFOX, FOLFOX4, FOLFIRI, Formestane, Hostamatinib, Hotemustin, Galarubicin, Gallium maltolate, Ganetespib, Gefitinib, Gemtuzumab, Gemtuzumab ozogamicin, Gimatecan,Glufosfamide, GCS-IOO, GDC-0623, GDC-0941 (Pictrelisib), GDC-0980, GDC-0032, GDC-0068, GDC-0349, GDC-0879, G17DT Immunogen, GMK, GMX-1778, GPX-100, gp100-Peptide Vaccine, GSK-5126766, GSK-690693, GSK-1120212 (Trametinib), GSK-1995010, GSK-2118436 (Dabrafenib), GSK-2126458, GSK-2132231A, GSK-2334470, GSK-2110183, GSK-2141795, GSK-2636771, GSK-525762A / I-BET-762, GW2016, Granisetron, Herceptin, Hexamethylmelamine, Histamine, Homoharringtonine, Hyaluronic Acid, Hydroxyurea, Hydroxyprogesterone Caproate, HDM-201, Ibandronate, Ibritumomab, Ibrutinib / PCI-32765, Idasanutlin, Idatrexate, Idelalisib / CAL-101, Idenestrol, IDN-5109, IGF-1R Inhibitor, IMC-1C11, IMC-A12 (Cixutumumab), Immunol, Indisulam, Interferon Alpha-2a, Interferon Alpha-2b, Pegylated Interferon Alpha-2b, Interleukin-2, INK-1117, INK-128, INSM-18, Ionaflunib, Iproplatin, Irofulven, Isohomohalicondrin-B, Isoflavone, Isotretinoin, Ixabepilone, JRX-2, JSF-154, JQ-1, J-107088, Conjugated Estrogen, Kahalide F, Ketoconazole, KW-2170, KW-2450, KU-55933, LCL-161, Lobaplatin, Leflunomide, Lenalidomide, Lenograstim, Leuprolide, Luprelin, Lexidronam, LGD-1550, Linezolid, Lovastatin, Lutetium Texaphyrin, Lomustine, Lonidamine, Losoxantrone, LU223651, Lurbinectedin, Lurtotecan, LY-S6AKT1, LY-2780301, LY-2109761 / Galnac-Serum, Melphalan, Marimastat, Masoprocol, Mechlorethamine, MEK Inhibitor, MEK-162,Methyltestosterone, Methylprednisolone, MEDI-573, MEN-10755, MDX-H210, MDX-447, MDX-1379, MGV, Midostaurin, Minodronic acid, Mitomycin, Mibobulin, MK-2206, MK-0646 (Darotuzumab), MLN518, MLN-0, 128, MLN-2480, Motexafin gadolinium, MS-209, MS-275, MX6, Nerydro-nate, Neratinib, Nexavar, Neovastat, Nilotinib, Nimesulide, Nitroglycerin, Noratopexed, Norelin, N-acetylcysteine, NU-7441 06-Benzylguanine, Oblimersen, Omeprazole, Olaparib, Oncolophage, oncoVEX GM-CSF, ormaplatin, ortataxel, OX44 antibody, OSI-027, OSI-906 (linitinib), 4-1BB antibody, oxantrazole, estrogen, onapristone, palbociclib / PD-0332991, panitumumab, panobinostat, patupilone, pazopanib, pegfilgrastim, PCK-3145, pegfilgrastim, PBI-1402, PBI-05204, PD0325901, PD-1 and PD-L1 antibodies (e.g., pembrolizumab, nivolumab, pidilizumab, MEDI-4736 / dulvalumab, RG-7446 / atezolizumab), PD-616, PEG-paclitaxel, albumin-stabilized paclitaxel, PEP-005, PF-05197281, PF-05212384, PF-04691502, PF-3758309, PHA-665752, PHT-427, P-04, PKC412, P54, PI-88, pelitinib, pemetrexed, pentrix, perifosine, perillyl alcohol, pertuzumab, pebonexostat, PI3K inhibitor, PI3K / mTOR inhibitor, PG-TXL, PG2, PLX-4032 / RO-5185426 (vemurafenib), PLX-3603 / RO-5212054, PT-100, PWT-33597, PX-866, picoplatin, pivaloyloxymethyl butyrate, pixantrone, phenoxodiol O, PKI166, premetrexed, plicamycin, polyprenic acid, ponatinib, porfiromycin, posaconazole, prednisone, prednisolone, PRT-062607, quinamed, quinupristin, quizartinib / AC220, R115777, RAF-265, lamotrigine, lamprenase, RDEA-119 / BAY869766, RDEA-436, levomycetin analog, receptor tyrosine kinase (RTK) inhibitor, lenalidomide, RG-7167, RG-7112, RG-7304, RG-7421, RG-7321, RG-7356, RG7440, RG-7775, lysokin, rhu-MAb, rigosertib, lymphabort, risedronate, rituximab, robatumumab, rofecoxib, romidepsin, RO-4929097, RO-31-7453, RO-5126766, RO-5068760, RPR109881A, rubidazone, rubitecan,R-flurbiprofen, RX-0201, LY2835219 (Luminespib), S-9788, sabatolimab, SAHA (Vorinostat), sapacitabine, SAR-405838, sargramostim, satraplatin, SB-408075, SB-431542, Se-015 / Ve-015, SU5416, SU6668, SDX-101, selinexor, semustine, theocaltitol, SM-11355, SN-38, SN-4071, SR-27897, SR-31747, SR-13668, SRL-172, sorafenib, spiroplatin, squalamine, STF-31, suberanilohydroxamic acid, stent, T900607, T138067, TAE-684, TAK-733, TAS-103, tasidotin, talaporfin, tanespimycin, tarceva, tariquidar, tacrolimus, taxotere, taxoprexin, tazarotene, tegafur, temozolomide, tesmilifene, testosterone, testosterone propionate, tesmilifene, tetraplatin, tetrodotoxin, tiazofurin, tipifarnib, tirapazamine, tocladesine, tomudex, tremofene, tosedostat, trabectedin, TransMID-107, tretinoin, trastuzumab, tremelimumab, tretinoin, triacetyluridine, triapine, triciribine, trimethoprim, TLK-286TXD258, ticilimumab / tirilimumab, uridine, valproic acid, valrubicin, vandetanib, batranib, vincristine, vinflunine, virulizin, besimodegib, boscalid, WX-UK1, WX-554, bexarotene, XAV-939, Xeloda, XELOX, XL-147, XL-228, XL-281, XL-518 / R-7420 / GDC-0973, XL-765, YM-511, YM-598, ZD-4190, ZD-6474, ZD-4054, ZD-0473, ZD-6126, ZD-9331, ZDI839, ZSTK-474, zoledronic acid and zosuquidar.,

[0194] In some embodiments, the described combination therapy comprises an LRP5 / LRP6 antagonist and an anti-PD-1 antibody described herein without any additional chemotherapeutic agent.

[0195] Proliferative disorder / cancer The combinations, compositions, kits, uses, methods and compounds for use of the invention (including all embodiments) are useful for the treatment and / or prevention of proliferative disorders, particularly cancer.

[0196] In certain embodiments, the combinations, compositions, kits, uses, methods and compounds for use of the invention (including all embodiments) are useful for the treatment of proliferative disorders, particularly cancer.

[0197] As used herein, "proliferative disorder" refers to a condition in which cell proliferation is increased beyond normal levels. For example, proliferative disorders or diseases include malignant diseases (e.g., esophageal cancer, colon cancer, biliary tract cancer) and non-malignant diseases (e.g., atherosclerosis, benign hyperplasia, benign prostatic hyperplasia).

[0198] In preferred embodiments, the proliferative disorder is cancer. In preferred embodiments, the cancer is characterized by having a mutated / inactivated RNF43 or an activated R-spondin fusion transcript.

[0199] Cancers are classified in two ways: by the type of tissue in which the cancer develops (tissue type) and by the primary site or location in the body where the cancer first develops. The most common sites where cancers develop include the skin, lung, breast, prostate, colon and rectum, cervix and uterus, and the hematological compartment.

[0200] The combinations, compositions, kits, uses, methods, and compounds for use of the invention (including all embodiments) are useful for a variety of proliferative disorders, particularly, for example, the following: · Gastrointestinal cancers, such as esophageal cancer (e.g., gastroesophageal junction cancer), gastric (stomach) cancer, hepatocellular carcinoma, biliary tract cancer (e.g., cholangiocarcinoma), gallbladder cancer, pancreatic cancer or colorectal cancer (CRC), · melanoma, · bladder cancer and · lung cancer (e.g., NSCLC) It can be useful for the treatment of cancers including, but not limited to, these.

[0201] In some embodiments of the present invention, the combinations, compositions, kits, uses, methods, and compounds for use of the present invention (including all embodiments) are used to treat gastrointestinal cancers, preferably esophageal cancer (e.g., gastroesophageal junction cancer), gastric (stomach) cancer, hepatocellular carcinoma, biliary tract cancer (e.g., cholangiocarcinoma), gallbladder cancer, pancreatic cancer, or colorectal cancer (CRC). These cancers are particularly preferably treated with LRP5 / LRP6#1 as an LRP5 / LRP6 antagonist and PD1-1 as an anti-PD-1 antibody. These cancers are particularly preferably treated with LRP5 / LRP6#5 as an LRP5 / LRP6 antagonist and PD1-1 as an anti-PD-1 antibody. These cancers are particularly preferably treated with LRP5 / LRP6#6 as an LRP5 / LRP6 antagonist and PD1-1 as an anti-PD-1 antibody. These cancers are particularly preferably treated with LRP5 / LRP6#1 as an LRP5 / LRP6 antagonist and PD1-2 as an anti-PD-1 antibody. These cancers are particularly preferably treated with LRP5 / LRP6#5 as an LRP5 / LRP6 antagonist and PD1-2 as an anti-PD-1 antibody. These cancers are particularly preferably treated with LRP5 / LRP6#6 as an LRP5 / LRP6 antagonist and PD1-2 as an anti-PD-1 antibody. These cancers are particularly preferably treated with LRP5 / LRP6#1 as an LRP5 / LRP6 antagonist and PD1-3 as an anti-PD-1 antibody. These cancers are particularly preferably treated with LRP5 / LRP6#5 as an LRP5 / LRP6 antagonist and PD1-3 as an anti-PD-1 antibody. These cancers are particularly preferably treated with LRP5 / LRP6#6 as an LRP5 / LRP6 antagonist and PD1-3 as an anti-PD-1 antibody.

[0202] In some embodiments of the present invention, the combinations, compositions, kits, uses, methods, and compounds for use of the present invention (including all embodiments) are used for the treatment of melanoma. These cancers are particularly preferably treated with LRP5 / LRP6#1 as an LRP5 / LRP6 antagonist and PD1-1 as an anti-PD-1 antibody. These cancers are particularly preferably treated with LRP5 / LRP6#5 as an LRP5 / LRP6 antagonist and PD1-1 as an anti-PD-1 antibody. These cancers are particularly preferably treated with LRP5 / LRP6#6 as an LRP5 / LRP6 antagonist and PD1-1 as an anti-PD-1 antibody. These cancers are particularly preferably treated with LRP5 / LRP6#1 as an LRP5 / LRP6 antagonist and PD1-2 as an anti-PD-1 antibody. These cancers are particularly preferably treated with LRP5 / LRP6#5 as an LRP5 / LRP6 antagonist and PD1-2 as an anti-PD-1 antibody. These cancers are particularly preferably treated with LRP5 / LRP6#6 as an LRP5 / LRP6 antagonist and PD1-2 as an anti-PD-1 antibody. These cancers are particularly preferably treated with LRP5 / LRP6#1 as an LRP5 / LRP6 antagonist and PD1-3 as an anti-PD-1 antibody. These cancers are particularly preferably treated with LRP5 / LRP6#5 as an LRP5 / LRP6 antagonist and PD1-3 as an anti-PD-1 antibody. These cancers are particularly preferably treated with LRP5 / LRP6#6 as an LRP5 / LRP6 antagonist and PD1-3 as an anti-PD-1 antibody.

[0203] In some embodiments of the present invention, the combinations, compositions, kits, uses, methods and compounds for use of the present invention (including all embodiments) are used in the treatment of bladder cancer. These cancers are particularly preferably treated with LRP5 / LRP6#1 as an LRP5 / LRP6 antagonist and PD1-1 as an anti-PD-1 antibody. These cancers are particularly preferably treated with LRP5 / LRP6#5 as an LRP5 / LRP6 antagonist and PD1-1 as an anti-PD-1 antibody. These cancers are particularly preferably treated with LRP5 / LRP6#6 as an LRP5 / LRP6 antagonist and PD1-1 as an anti-PD-1 antibody. These cancers are particularly preferably treated with LRP5 / LRP6#1 as an LRP5 / LRP6 antagonist and PD1-2 as an anti-PD-1 antibody. These cancers are particularly preferably treated with LRP5 / LRP6#5 as an LRP5 / LRP6 antagonist and PD1-2 as an anti-PD-1 antibody. These cancers are particularly preferably treated with LRP5 / LRP6#6 as an LRP5 / LRP6 antagonist and PD1-2 as an anti-PD-1 antibody. These cancers are particularly preferably treated with LRP5 / LRP6#1 as an LRP5 / LRP6 antagonist and PD1-3 as an anti-PD-1 antibody. These cancers are particularly preferably treated with LRP5 / LRP6#5 as an LRP5 / LRP6 antagonist and PD1-3 as an anti-PD-1 antibody. These cancers are particularly preferably treated with LRP5 / LRP6#6 as an LRP5 / LRP6 antagonist and PD1-3 as an anti-PD-1 antibody.

[0204] In some embodiments of the present invention, the combinations, compositions, kits, uses, methods, and compounds for use of the present invention (including all embodiments) are used for the treatment of lung cancer (e.g., non-small cell lung cancer NSCLC). These cancers are particularly preferably treated with LRP5 / LRP6#1 as an LRP5 / LRP6 antagonist and PD1-1 as an anti-PD-1 antibody. These cancers are particularly preferably treated with LRP5 / LRP6#5 as an LRP5 / LRP6 antagonist and PD1-1 as an anti-PD-1 antibody. These cancers are particularly preferably treated with LRP5 / LRP6#6 as an LRP5 / LRP6 antagonist and PD1-1 as an anti-PD-1 antibody. These cancers are particularly preferably treated with LRP5 / LRP6#1 as an LRP5 / LRP6 antagonist and PD1-2 as an anti-PD-1 antibody. These cancers are particularly preferably treated with LRP5 / LRP6#5 as an LRP5 / LRP6 antagonist and PD1-2 as an anti-PD-1 antibody. These cancers are particularly preferably treated with LRP5 / LRP6#6 as an LRP5 / LRP6 antagonist and PD1-2 as an anti-PD-1 antibody. These cancers are particularly preferably treated with LRP5 / LRP6#1 as an LRP5 / LRP6 antagonist and PD1-3 as an anti-PD-1 antibody. These cancers are particularly preferably treated with LRP5 / LRP6#5 as an LRP5 / LRP6 antagonist and PD1-3 as an anti-PD-1 antibody. These cancers are particularly preferably treated with LRP5 / LRP6#6 as an LRP5 / LRP6 antagonist and PD1-3 as an anti-PD-1 antibody.

[0205] In a further embodiment of the present invention, the combinations, compositions, kits, uses, methods and compounds for use of the present invention (including all embodiments) are for the treatment of cancer patients who are treatment-naïve with respect to treatment with checkpoint inhibitors or immunomodulators, i.e., patients who are treatment-naïve with respect to treatment with, for example, an anti-PD-1 antibody (e.g., (i) a gastrointestinal cancer, such as esophageal cancer, gastric cancer, hepatocellular carcinoma, biliary tract cancer, gallbladder cancer, pancreatic cancer or colorectal cancer, (ii) melanoma, (iii) bladder cancer or (iv) lung cancer patients). In one embodiment, the cancer is characterized by having a mutated / inactivated RNF43 or an activated R-spondin fusion transcript. These cancers are particularly preferably treated with LRP5 / LRP6#1 as an LRP5 / LRP6 antagonist and PD1-1 as an anti-PD-1 antibody. These cancers are particularly preferably treated with LRP5 / LRP6#5 as an LRP5 / LRP6 antagonist and PD1-1 as an anti-PD-1 antibody. These cancers are particularly preferably treated with LRP5 / LRP6#6 as an LRP5 / LRP6 antagonist and PD1-1 as an anti-PD-1 antibody. These cancers are particularly preferably treated with LRP5 / LRP6#1 as an LRP5 / LRP6 antagonist and PD1-2 as an anti-PD-1 antibody. These cancers are particularly preferably treated with LRP5 / LRP6#5 as an LRP5 / LRP6 antagonist and PD1-2 as an anti-PD-1 antibody. These cancers are particularly preferably treated with LRP5 / LRP6#6 as an LRP5 / LRP6 antagonist and PD1-2 as an anti-PD-1 antibody. These cancers are particularly preferably treated with LRP5 / LRP6#1 as an LRP5 / LRP6 antagonist and PD1-3 as an anti-PD-1 antibody. These cancers are particularly preferably treated with LRP5 / LRP6#5 as an LRP5 / LRP6 antagonist and PD1-3 as an anti-PD-1 antibody. These cancers are particularly preferably treated with LRP5 / LRP6#6 as an LRP5 / LRP6 antagonist and PD1-3 as an anti-PD-1 antibody.

[0206] In a further embodiment of the present invention, the combinations, compositions, kits, uses, methods and compounds for use of the present invention (including all embodiments) are for the treatment of cancer patients who relapse during, subsequent to or after treatment with a checkpoint inhibitor or immunomodulator, i.e., for example, patients who relapse during, subsequent to or after treatment with a PD-1 antagonist, e.g., an anti-PD-1 antibody (e.g., (i) patients suffering from gastrointestinal cancer, e.g., esophageal cancer, gastric cancer, hepatocellular carcinoma, biliary tract cancer, gallbladder cancer, pancreatic cancer or colorectal cancer, (ii) melanoma, (iii) bladder cancer or (iv) lung cancer). In one embodiment, the cancer is characterized by having a mutated / inactivated RNF43 or an activated R-spondin fusion transcript. These cancers are particularly preferably treated with LRP5 / LRP6#1 as an LRP5 / LRP6 antagonist and PD1-1 as an anti-PD-1 antibody. These cancers are particularly preferably treated with LRP5 / LRP6#5 as an LRP5 / LRP6 antagonist and PD1-1 as an anti-PD-1 antibody. These cancers are particularly preferably treated with LRP5 / LRP6#6 as an LRP5 / LRP6 antagonist and PD1-1 as an anti-PD-1 antibody. These cancers are particularly preferably treated with LRP5 / LRP6#1 as an LRP5 / LRP6 antagonist and PD1-2 as an anti-PD-1 antibody. These cancers are particularly preferably treated with LRP5 / LRP6#5 as an LRP5 / LRP6 antagonist and PD1-2 as an anti-PD-1 antibody. These cancers are particularly preferably treated with LRP5 / LRP6#6 as an LRP5 / LRP6 antagonist and PD1-2 as an anti-PD-1 antibody. These cancers are particularly preferably treated with LRP5 / LRP6#1 as an LRP5 / LRP6 antagonist and PD1-3 as an anti-PD-1 antibody. These cancers are particularly preferably treated with LRP5 / LRP6#5 as an LRP5 / LRP6 antagonist and PD1-3 as an anti-PD-1 antibody. These cancers are particularly preferably treated with LRP5 / LRP6#6 as an LRP5 / LRP6 antagonist and PD1-3 as an anti-PD-1 antibody.

[0207] The therapeutic applicability of the combination therapy of the present invention can include the first, second, third or further stages of treatment of patients (e.g., patients suffering from (i) gastrointestinal cancer, such as esophageal cancer, gastric cancer, hepatocellular carcinoma, biliary tract cancer, gallbladder cancer, pancreatic cancer or colorectal cancer, (ii) melanoma, (iii) bladder cancer or (iv) lung cancer). The cancer may be metastatic, recurrent, relapsing, resistant or refractory to one or more anti-cancer treatments. For this reason, the patient may be treatment-naive or may have received one or more previous anti-cancer therapies in which the disease was not completely cured.

[0208] Also, patients having recurrence and / or resistance to one or more anti-cancer agents (e.g., single components of a combination or standard chemotherapeutic agents) can receive the combination treatment of the present invention, e.g., as an additional combination or as a replacement treatment, e.g., in the second or third stage treatment cycles (optionally, further in combination with one or more other anti-cancer agents).

[0209] Accordingly, part of the disclosed combination therapy of the present invention is effective for treating subjects (e.g., patients suffering from (i) gastrointestinal cancer, such as esophageal cancer, gastric cancer, hepatocellular carcinoma, biliary tract cancer, gallbladder cancer, pancreatic cancer or colorectal cancer, (ii) melanoma, (iii) bladder cancer or (iv) lung cancer) in whom the cancer has recurred or the cancer has become drug-resistant or multi-drug resistant or the cancer has failed one or two or more stages of monotherapy or combination therapy with one or more anti-cancer agents (e.g., single components of a combination or standard chemotherapeutic agents).

[0210] Cancer that initially responded to an anti-cancer agent may recur and become resistant to the anti-cancer agent, e.g., when the anti-cancer agent is no longer effective in treating the subject having the cancer despite increased dosing of the anti-cancer agent. Cancer that has developed resistance to two or more anti-cancer agents is said to be multi-drug resistant.

[0211] In a preferred embodiment, the combinations, compositions, kits, uses, methods and compounds for use of the present invention (including all embodiments) are used in the treatment of cancer patients (e.g., (i) gastrointestinal cancers such as esophageal cancer, gastric cancer, hepatocellular carcinoma, biliary tract cancer, gallbladder cancer, pancreatic cancer or colorectal cancer, (ii) melanoma, (iii) bladder cancer or (iv) lung cancer) previously treated with one or more immune checkpoint inhibitors and / or immunomodulators, e.g., one or more PD-1 antagonists such as anti-PD1 antibodies. In one embodiment, the cancer is characterized by having a mutated / inactivated RNF43 or an activated R-spondin fusion transcript. These cancers are particularly preferably treated with LRP5 / LRP6#1 as an LRP5 / LRP6 antagonist and PD1-1 as an anti-PD-1 antibody. These cancers are particularly preferably treated with LRP5 / LRP6#5 as an LRP5 / LRP6 antagonist and PD1-1 as an anti-PD-1 antibody. These cancers are particularly preferably treated with LRP5 / LRP6#6 as an LRP5 / LRP6 antagonist and PD1-1 as an anti-PD-1 antibody. These cancers are particularly preferably treated with LRP5 / LRP6#1 as an LRP5 / LRP6 antagonist and PD1-2 as an anti-PD-1 antibody. These cancers are particularly preferably treated with LRP5 / LRP6#5 as an LRP5 / LRP6 antagonist and PD1-2 as an anti-PD-1 antibody. These cancers are particularly preferably treated with LRP5 / LRP6#6 as an LRP5 / LRP6 antagonist and PD1-2 as an anti-PD-1 antibody. These cancers are particularly preferably treated with LRP5 / LRP6#1 as an LRP5 / LRP6 antagonist and PD1-3 as an anti-PD-1 antibody. These cancers are particularly preferably treated with LRP5 / LRP6#5 as an LRP5 / LRP6 antagonist and PD1-3 as an anti-PD-1 antibody. These cancers are particularly preferably treated with LRP5 / LRP6#6 as an LRP5 / LRP6 antagonist and PD1-3 as an anti-PD-1 antibody.

[0212] In a further preferred embodiment, the combinations, compositions, kits, uses, methods and compounds for use of the invention (including all embodiments) are used for the treatment of cancer patients refractory or resistant to checkpoint inhibitor therapy (e.g., treatment with one or more immune checkpoint inhibitors and / or immunomodulators, e.g., one or more PD-1 antagonists, e.g., anti-PD1 antibodies), such as patients suffering from (i) gastrointestinal cancer, such as esophageal cancer, gastric cancer, hepatocellular carcinoma, biliary tract cancer, gallbladder cancer, pancreatic cancer or colorectal cancer, (ii) melanoma, (iii) bladder cancer or (iv) lung cancer. In one embodiment, the cancer is characterized by having a mutated / inactivated RNF43 or an activated R-spondin fusion transcript. These cancers are particularly preferably treated with LRP5 / LRP6#1 as an LRP5 / LRP6 antagonist and PD1-1 as an anti-PD-1 antibody. These cancers are particularly preferably treated with LRP5 / LRP6#5 as an LRP5 / LRP6 antagonist and PD1-1 as an anti-PD-1 antibody. These cancers are particularly preferably treated with LRP5 / LRP6#6 as an LRP5 / LRP6 antagonist and PD1-1 as an anti-PD-1 antibody. These cancers are particularly preferably treated with LRP5 / LRP6#1 as an LRP5 / LRP6 antagonist and PD1-2 as an anti-PD-1 antibody. These cancers are particularly preferably treated with LRP5 / LRP6#5 as an LRP5 / LRP6 antagonist and PD1-2 as an anti-PD-1 antibody. These cancers are particularly preferably treated with LRP5 / LRP6#6 as an LRP5 / LRP6 antagonist and PD1-2 as an anti-PD-1 antibody. These cancers are particularly preferably treated with LRP5 / LRP6#1 as an LRP5 / LRP6 antagonist and PD1-3 as an anti-PD-1 antibody. These cancers are particularly preferably treated with LRP5 / LRP6#5 as an LRP5 / LRP6 antagonist and PD1-3 as an anti-PD-1 antibody. These cancers are particularly preferably treated with LRP5 / LRP6#6 as an LRP5 / LRP6 antagonist and PD1-3 as an anti-PD-1 antibody.

[0213] In an alternative preferred embodiment, the combinations, compositions, kits, uses, methods and compounds for use of the invention (including all embodiments) are used for the treatment of cancer patients suffering from any solid tumor that is refractory or resistant to checkpoint inhibitor therapy (e.g., treatment with one or more immune checkpoint inhibitors and / or immunomodulators, e.g., one or more PD-1 antagonists, e.g., an anti-PD1 antibody). In one embodiment, the cancer is characterized by having a mutated / inactivated RNF43 or an activated R-spondin fusion transcript. These cancers are particularly preferably treated with LRP5 / LRP6#1 as an LRP5 / LRP6 antagonist and PD1-1 as an anti-PD-1 antibody. These cancers are particularly preferably treated with LRP5 / LRP6#5 as an LRP5 / LRP6 antagonist and PD1-1 as an anti-PD-1 antibody. These cancers are particularly preferably treated with LRP5 / LRP6#6 as an LRP5 / LRP6 antagonist and PD1-1 as an anti-PD-1 antibody. These cancers are particularly preferably treated with LRP5 / LRP6#1 as an LRP5 / LRP6 antagonist and PD1-2 as an anti-PD-1 antibody. These cancers are particularly preferably treated with LRP5 / LRP6#5 as an LRP5 / LRP6 antagonist and PD1-2 as an anti-PD-1 antibody. These cancers are particularly preferably treated with LRP5 / LRP6#6 as an LRP5 / LRP6 antagonist and PD1-2 as an anti-PD-1 antibody. These cancers are particularly preferably treated with LRP5 / LRP6#1 as an LRP5 / LRP6 antagonist and PD1-3 as an anti-PD-1 antibody. These cancers are particularly preferably treated with LRP5 / LRP6#5 as an LRP5 / LRP6 antagonist and PD1-3 as an anti-PD-1 antibody. These cancers are particularly preferably treated with LRP5 / LRP6#6 as an LRP5 / LRP6 antagonist and PD1-3 as an anti-PD-1 antibody. Examples of solid tumors are well known in the art. Similarly, the terms refractory or resistant are also known to those skilled in the art and are used herein according to the definitions utilized in the art.

[0214] Tumors that are refractory or resistant to immune checkpoint inhibitor therapy are herein also referred to as "immune therapy-resistant tumors" or "immune therapy-resistant non-T cell inflammatory tumors". In recent years, it has been found that in the microenvironment of many tumors, high expression of specific immune cells can be found. This is referred to in the art as the "T cell inflammatory phenotype", and it has been observed that tumors with this phenotype are correlated with tumors that are sensitive to treatment with multiple immunotherapies including therapeutic vaccines and checkpoint blockade antibodies, such as anti-PD-1 antibodies. On the other hand, certain tumors lack this expression of immune cells in their microenvironment. These tumors are referred to in the art as "non-T cell inflammatory tumors", and it has been found that these tumors lack clinical benefit, particularly from immunotherapy with anti-PD-1 antibodies. According to the present invention, the latter type of tumors having active Wnt signaling are preferred targets for the claimed combination therapies. These cancers are particularly preferably treated with LRP5 / LRP6#1 as an LRP5 / LRP6 antagonist and PD1-1 as an anti-PD-1 antibody. These cancers are particularly preferably treated with LRP5 / LRP6#5 as an LRP5 / LRP6 antagonist and PD1-1 as an anti-PD-1 antibody. These cancers are particularly preferably treated with LRP5 / LRP6#6 as an LRP5 / LRP6 antagonist and PD1-1 as an anti-PD-1 antibody. These cancers are particularly preferably treated with LRP5 / LRP6#1 as an LRP5 / LRP6 antagonist and PD1-2 as an anti-PD-1 antibody. These cancers are particularly preferably treated with LRP5 / LRP6#5 as an LRP5 / LRP6 antagonist and PD1-2 as an anti-PD-1 antibody. These cancers are particularly preferably treated with LRP5 / LRP6#6 as an LRP5 / LRP6 antagonist and PD1-2 as an anti-PD-1 antibody. These cancers are particularly preferably treated with LRP5 / LRP6#1 as an LRP5 / LRP6 antagonist and PD1-3 as an anti-PD-1 antibody. These cancers are particularly preferably treated with LRP5 / LRP6#5 as an LRP5 / LRP6 antagonist and PD1-3 as an anti-PD-1 antibody.These cancers are particularly preferably treated with LRP5 / LRP6#6 as an LRP5 / LRP6 antagonist and PD1-3 as an anti-PD-1 antibody.

[0215] The present invention is not limited to the scope according to the specific embodiments described herein. In addition to what is described herein, various modifications of the present invention will be apparent to those skilled in the art from this disclosure. Such modifications are intended to be within the scope of the appended claims.

[0216] All patent applications cited herein are hereby incorporated by reference in their entirety.

[0217] Example 1 Antitumor activity of exemplary LRP5 / LRP6 in combination with a mouse antibody against PD-1 in a subcutaneous syngeneic mouse model obtained from the mammary cancer cell line EMT6 of Balb / c mice The efficacy of an exemplary LRP5 / 6 antagonist was tested as a single agent and in combination with a mouse antibody against PD-1 in a syngeneic model derived from an s.c. cell line of mouse mammary cancer (EMT6).

[0218] BALB / cJBomTac mice were used in this study. 1 × 10 6 individual EMT6 mammary cancer cells were injected per mouse to establish tumors. Tumor volume was measured at least three times per week using calipers. Treatment was initiated when the median tumor volume reached approximately 200 mm 3 and ended 30 days later.

[0219] Animals with 10 tumors were treated twice weekly intravenously (i.v.) with an exemplary LRP5 / LRP6, twice weekly i.p. with an exemplary mouse PD-1 antibody, or with a combination of both compounds. 10 animals were used in a vehicle / isotype control treatment group. Animals were euthanized at the end of the study for ethical reasons based on tumor mass (tumor ≧ 1.5 cm 3 ).

[0220] Cell EMT6 cells were obtained from ATCC (catalog number ATCC® CRL2755™). Master cell bank (MCB) and working cell bank (WCB) were established. The cells were cultured in T175 tissue culture flasks at 37 °C and 5% CO2. The medium used was Waymouth's MB752 / 1 supplemented with 15% fetal bovine serum (HyClone® fetal bovine serum, characterized; catalog number SH30071.03; Thermo Scientific) and 2 mM L-glutamine (200 mM L-glutamine (100×); Ref25030-024; Gibco by Life Technologies). The cultures were split every 2 - 3 days at a ratio of 1:10 / 1:15.

[0221] Mouse The mice were 7 - 8-week-old BALB / cJBomTac purchased from Taconic, Denmark. After arrival at the animal facility, the mice were acclimatized to the ambient conditions for at least 5 days before being used in the experiments. They were housed in 10 groups of Macrolon® type III cages under standardized conditions of 21.5 ± 1.5 °C and 55 ± 10% humidity. Standard irradiated food (PROVIMI KLIBA) and autoclaved tap water were provided ad libitum. Each mouse was identified using a microchip implanted subcutaneously under isoflurane anesthesia. Cage cards indicating the study number, animal number, compound and dose level, route of administration and schedule were left with the animals throughout the study.

[0222] Administration of Test Compounds The LRP5 / LRP6 antagonist was suspended in histidine buffer pH 6.5 and administered i.v. at a dose of 10 mg / kg twice a week for the first 2 weeks at an application volume of 10 mL / kg per mouse. The PD-1 antibody was diluted in PBS and injected intraperitoneally at a dose of 10 mg / kg twice a week at a volume of 10 mL / kg per mouse until the end of the study.

[0223] Monitoring of Tumor Growth and Disease Progression The tumor diameter was measured three times a week (Monday, Wednesday, and Friday) using calipers. The volume of each tumor [mm 3 was calculated according to the formula "tumor volume = length × diameter2 × π / 6". To monitor the side effects of the treatment, the mice were examined daily for abnormalities and their body weights were measured daily. The animals were sacrificed at the end of the study. Animals with necrotic tumors or tumor sizes exceeding 1500 mm 3 were sacrificed at the beginning of the study for ethical reasons.

[0224] Results Treatment of ETM6 tumors with a mouse antibody against PD-1 resulted in moderate tumor intestinal obstruction. The combination of an LRP5 / LRP6 antagonist and a PD-1 antibody significantly improved the efficacy compared to single-agent treatment. Compared to single-agent treatment in which tumor shrinkage was observed in only 1 out of 10 mice, tumor shrinkage was brought about in 4 out of 9 mice. The results demonstrating the synergistic effect of combination administration compared to single-agent treatment are shown in Figure 1. The combination of an LRP5 / LRP6 antagonist and a PD-1 antibody increased the survival rate reported in Table 5 as the period (number of days) until the tumor volume reached at least 500 mm 3 compared to single-agent treatment.

[0225] Table 5 shows the antitumor activity of an exemplary LRP5 / LRP6 antagonist as a single agent and in combination with a mouse antibody against PD-1. The median value refers to the period (number of days) until the tumor volume reached at least 500 mm 3 from the start of treatment.

[0226]

Table 10

[0227] Furthermore, histological analysis of samples from mice showing tumor shrinkage (i.e., the tumor volume at the end of the test was smaller compared to that at the start of treatment) was performed. In particular, tumors were collected from all groups and fixed in 10% NBF (neutral buffered formalin solution, 10%) for FFPE (formalin-fixed paraffin-embedded). For morphological evaluation, histological analysis was performed on FFPE tumor tissues by hematoxylin-eosin (HE) staining. Evidence of tumors at the end of the test for tissues from the sites where tumors were previously observed was not reported only in the combination group (3 out of 9 mice). From this, it was shown that a pathological complete response can be achieved only by the combined treatment of an LRP5 / 6 antagonist and a PD-1 antibody compared to monotherapy (Table 6).

[0228] Table 6 shows the anti-tumor activity of exemplary LRP5 / 6 antagonists as monotherapy and in combination with a mouse antibody against PD-1. A complete response at the end of the study refers to the absence of evidence of remaining cancer even upon histological examination of tissues from the sites where tumors were previously observed, compared to a partial response where tumor cells are detected.

[0229]

Table 11

[0230] Example 2 Increased tumor T cell infiltration in a subcutaneous syngeneic mouse model obtained from the mammary cancer cell line EMT6 of Balb / c mice in combination with a mouse antibody against PD-1 of an exemplary LRP5 / LRP6 antagonist The ability of an exemplary LRP5 / LRP6 antagonist to attract T cell infiltration in tumors was tested as monotherapy and in combination with a mouse antibody against PD-1 in a syngeneic model (EMT6) derived from an s.c. cell line of mouse mammary cancer (EMT6).

[0231] CD8-positive T cells were analyzed in tumors on day 16 from mice treated with the agent alone and in combination with a murine antibody against PD-1 as reported in Example 1. Tumors were collected from all groups, fixed in 10% NBF for FFPE tissues, and immunohistochemistry (IHC) was performed using a rat monoclonal antibody against CD8a (53-6.7, eBioscience™, working dilution 1:200) with a standard protocol to detect CD8-positive T cells. Quantitative evaluation was performed using HALO™ image analysis software, and the significance level was determined using Graph Pad Prism software. A statistical significant difference between groups was considered when the adjusted p-value was less than 0.05. The results are shown in Figure 2.

[0232] Example 3 Combined effect of LRP5 / LRP6 antagonist and anti-human PD-1 antibody in 3D spheroids To further evaluate the combined effect of an anti-LRP5 / LRP6 antagonist (LRP5 / LRP6#5 defined above, also shown as SEQ ID NO: 65) and the anti-human PD-1 antibody of the present invention (PD1-3 defined in Table 3 above) in Wnt-driven immune suppression, in vitro co-culture of tumor cells, activated human PBMCs and Wnt ligand (Wnt3a) was used, and tumor cell viability was measured as a readout.

[0233] For this purpose, tumor cells (NCI-H1437) stably transfected to express the red fluorescent protein (mKate2) and cultured in 3D as spheroids with activated human PBMCs and Wnt3a ligand (0.5 μg / ml) ligand were treated with 1000 nM LRP5 / LRP6 antagonist and 200 nM anti-PD-1 antibody, and cell viability was measured at the indicated time points after compound addition.

[0234] 3.1 Study design To establish an in vitro co - culture assay of tumor cells (NCI - H1437 non - small cell lung cancer cell line) and human PBMCs, NCI - H1437 cells were stably transfected to express the red fluorescent protein (mKate2) and cultured in 3D as spheroids. To perform the co - culture assay, NCI - H1437 mKate2 cells were seeded into a 96 - well Spheroid Microplate (5000 cells / well). The NCI - H1437 mKate2 cells were seeded at a volume of 200 μl of RPMI - 1640 + Glutamax medium (containing 10% FCShi) per well. Four days later, spheroids were formed, 100 μl of the medium was removed from each well, and 100 μl of RPMI1640 medium + Glutamax (+10% FCShi) with or without 3×10 5 PBMCs (activated with anti - CD3 antibody and anti - CD28 antibody (1 μg / ml) for 72 hours) was added to the appropriate wells.

[0235] Spheroids with or without PBMCs were exposed to either an anti - LRP5 / LRP6 antagonist, Wnt3a, an anti - human PD - 1 antibody, or an isotype of the anti - human PD - 1 antibody (as a control) as monotherapy or in combination. These compounds were added only once on day 0 (4 days after seeding the tumor cells into the microplate).

[0236] Twelve hours after adding the compounds, the first measurement of mKate2 fluorescence was taken and used to determine the cell viability of the tumor spheroids. This time point was used as the baseline (100%) and compared with the following measurements (taken at time intervals between 12 and 48 hours). The fluorescence of mKate2 (excitation: 590 nm; emission 635 nm) was measured using an EnVision 2100 MULTILABEL READER (PerkinElmer). In the experiment, biological replicates were performed 6 times until day 2, 5 times on day 3 and day 4, and 4 times on day 7 and day 8, in the presence or absence of PBMCs and treatment of spheroids.

[0237] Reagents and tissue culture materials ·PBS (Gibco; 14190 - 094) ·Trypsin - EDTA (Gibco; 043 - 90317FU) ·Ultra - LEAF™ Purified Anti - Human CD3 Antibody (Biolegend; 300332) ·Ultra - LEAF™ Purified Anti - Human CD28 Antibody (Biolegend; 302934) ·RPMI1640 + Glutamax (Gibco; 61870 - 010) ·RPMI1640 (Gibco; A10491 - 01) ·FCS (HyClone; SH30084.03) ·WNT3a (R&D 5036 - WN / CF; Lot SVH181610A) ·Stem cell donor: B001000527; Lot: 1812180182

[0238] 3.2 Culturing of NCI - H1437MKATE2 NCI - H1437mKate2 cells were cultured using RPMI1640 (Gibco; A10491 - 01) + 10% FCS. The cells were split once a week (1:10), and the medium was changed over time. For sub - culturing, the cells were detached from the cell culture flask using trypsin - EDTA (Gibco; 043 - 90317FU) in PBS. The medium was removed, and 5 ml of trypsin was added at 37°C for about 5 minutes. Visual checks were performed every minute to confirm if the cells were already detached. After detachment, the cell / trypsin solution was mixed with 45 ml of culture medium containing 10% FCS and centrifuged at 400×g for 5 minutes at room temperature. The cell pellet was resuspended in an appropriate amount of medium and either counted for co - culture assay or split 1:10 for culturing. The cells were cultured at 37°C and 5% CO2.

[0239] 3.3 Thawing of PBMC and PBMC Activation One vial containing PBMC (stem cell donor: B001000527; lot: 1812180182) was thawed at RT until a very small amount of ice chips remained, and then poured into a 50 ml Falcon containing 20 ml of cold (2 - 8°C) RPMI - 1640 + Glutamax. After vortexing, the Falcon tube was centrifuged at 400×g for 5 minutes. Then, the supernatant was discarded and the PBMC pellet was resuspended in 1 - 2 ml of assay medium (RPMI1640 + glutamax + 10% FCShi).

[0240] The cells were counted and activated with anti - CD3 antibody and anti - CD28 antibody (1 μg / ml) for 72 hours (5×10^6 cells / ml). After 72 hours, the activated PBMC were centrifuged at 400×g for 5 minutes. The cell pellet was resuspended in 1 - 2 ml of RPMI - 1640 + Glutamax medium (containing 10% FCShi). Finally, the cells were counted and diluted to 3×10^6 cells / ml for the co - culture assay.

[0241] 3.4 Sphere viability change: Measurement and analysis The EnVision 2100 MULTILABEL READER (PerkinElmer) was used to determine the change in cell viability of NCI - H1437mKate2 spheres. The fluorescence of mKate2 was measured at an excitation of 590 nm, an emission of 635 nm, and a measurement height of 4.1 mm. For analysis, the mean of the background (medium only) was subtracted from the measured values, the % change for each well was calculated, and the new measured value of the well (after subtracting the background) was compared to the baseline measured value (12 hours after adding the compound and PBMC). The standard deviation shown is the % standard deviation of the % change at the corresponding treatment and time point. The % change in the obtained viability values was transferred to Graph Pad software and analyzed by applying two - way ANOVA in combination with Bonferroni's multiple comparison test to determine statistical significance.

[0242] 3.5 Statistical analysis The significance level was determined using Graph Pad Prism software. The (adjusted) p - value is* less than 0.05 for ** less than 0.01 for *** less than 0.001 for and **** less than 0.0001 for, a statistically significant difference between groups was considered to be observed.

[0243] 3.6 Results Figure 3A shows the effects of treatment with Wnt3a ligand, LRP5 / LRP6 antagonist, or anti-human PD-1 antibody on the survival rate of tumor spheroids co-cultured with activated PMBC. Treatment with Wnt3a resulted in a significant improvement in the survival rate of tumor spheroids detected at any time point between 4 and 8 days (inhibition of PBMC-mediated tumor cell killing). Treatment with LRP5 / LRP6 antagonist or anti-human PD-1 antibody had no significant effect on the survival rate of tumor spheroids compared to isotype treatment (control).

[0244] Figure 3B shows the effects of treatment with LRP5 / LRP6 antagonist as monotherapy or in combination with anti-human PD-1 antibody in the presence of Wnt3a ligand. Treatment with LRP5 / LRP6 antagonist as monotherapy suppressed the Wnt3a-mediated increase in tumor spheroid survival rate (a significant effect was reported 4 - 8 days after treatment initiation, Tum / PBMC 1:3+LRP5 / 6+WNT3a+iso vs. Tum / PBMC 1:3+iso). Therefore, treatment with LRP5 / LRP6 antagonist in the presence of Wnt3a ligand restored the PBMC-mediated inhibition of tumor spheroid survival rate.

[0245] Combined treatment with an LRP5 / LRP6 antagonist and an anti-human PD-1 antibody results in a significant decrease in tumor spheroid viability compared to treatment with the LRP5 / LRP6 antagonist alone (a significant effect is reported 7 - 8 days after treatment initiation, Tum / PBMC 1:3+LRP5 / 6+WNT3a+PD1 vs Tum / PBMC 1:3+LRP5 / 6+WNT3a+iso). Thus, combined treatment with an LRP5 / LRP6 antagonist and an anti-human PD-1 antibody results in an improvement in PBMC-mediated tumor cell killing compared to treatment with the LRP5 / LRP6 antagonist alone.

[0246] 3.7 Discussion From these results, it is shown that blockade of LRP5 and LRP6 in combination with a PD-1 antagonist results in PBMC-mediated killing of tumor spheroids. These data, together with the data shown in Examples 1 and 2, indicate that the combination therapy of the present invention has potent anti-tumor activity.

Claims

1. A polypeptide capable of specifically binding to LRP5 and LRP6 for use in a method for treating and / or preventing a hyperproliferative disease, preferably cancer, comprising: The method includes administering to a patient in need thereof a polypeptide capable of specifically binding to LRP5 and LRP6 in combination with a PD-1 antibody; Here, the polypeptide capable of specifically binding to LRP5 and LRP6 is (i) the following CDR sequences: CDR1: TYTVG (=SEQ ID NO: 40) CDR2: AIRRGSSTYYADSVKG (=SEQ ID NO: 41) CDR3: DTRTVALLQYRYDY (=SEQ ID NO:42) (a) a first immunoglobulin single variable domain (ISVD) comprising The following CDR sequences: CDR1: SYAMG (=SEQ ID NO:49) CDR2: AISWSGGSTYYADSVKG (=SEQ ID NO:50) CDR3: SPIPYGSLLRRRNNYDY (=SEQ ID NO:51) a polypeptide comprising a second ISVD (b) comprising: (ii) the following CDR sequences: CDR1: SYAMG (=SEQ ID NO:43) CDR2: AIRSGRRTYYADSVKG (=SEQ ID NO:44) CDR3: ARVRSSTRYNTGTWWWEY (=SEQ ID NO: 45) A first ISVD (a) including The following CDR sequences: CDR1: SYAMG (=SEQ ID NO:49) CDR2: AISWSGGSTYYADSVKG (=SEQ ID NO:50) CDR3: SPIPYGSLLRRRNNYDY (=SEQ ID NO:51) a polypeptide comprising a second ISVD (b) comprising: (iii) the following CDR sequences: CDR1: RYTMG (=SEQ ID NO: 46) CDR2: AIVRSGGSTYYADSVKG (=SEQ ID NO: 47) CDR3: DRRGRGENYILLYSSGRYEY (=SEQ ID NO:48) A first ISVD (a) including The following CDR sequences: CDR1: SYAMG (=SEQ ID NO:49) CDR2: AISWSGGSTYYADSVKG (=SEQ ID NO:50) CDR3: SPIPYGSLLRRRNNYDY (=SEQ ID NO:51) a polypeptide comprising a second ISVD (b) comprising: (iv) the following CDR sequences: CDR1: TYTVG (=SEQ ID NO: 40) CDR2: AIRRGSSTYYADSVKG (=SEQ ID NO: 41) CDR3: DTRTVALLQYRYDY (=SEQ ID NO:42) A first ISVD (a) including The following CDR sequences: CDR1: SYAMG (=SEQ ID NO:52) CDR2: AISWRSGSTYYADSVKG (=SEQ ID NO:53) CDR3: DPRGYGVAYVSAYYEY (=SEQ ID NO:54) a polypeptide comprising a second ISVD (b) comprising: (v) the following CDR sequences: CDR1: SYAMG (=SEQ ID NO:43) CDR2: AIRSGRRTYYADSVKG (=SEQ ID NO:44) CDR3: ARVRSSTRYNTGTWWWEY (=SEQ ID NO: 45) A first ISVD (a) including The following CDR sequences: CDR1: SYAMG (=SEQ ID NO:52) CDR2: AISWRSGSTYYADSVKG (=SEQ ID NO:53) CDR3: DPRGYGVAYVSAYYEY (=SEQ ID NO:54) and a polypeptide comprising a second ISVD (b) comprising (vi) the following CDR sequences: CDR1: RYTMG (=SEQ ID NO: 46) CDR2: AIVRSGGSTYYADSVKG (=SEQ ID NO: 47) CDR3: DRRGRGENYILLYSSGRYEY (=SEQ ID NO:48) A first ISVD (a) including The following CDR sequences: CDR1: SYAMG (=SEQ ID NO:52) CDR2: AISWRSGSTYYADSVKG (=SEQ ID NO:53) CDR3: DPRGYGVAYVSAYYEY (=SEQ ID NO:54) A polypeptide comprising a second ISVD (b) comprising is selected from the group consisting of Here, the PD-1 antibody is (i) an anti-PD1 antibody comprising a heavy chain CDR comprising the amino acid sequences of SEQ ID NO: 1 (HCDR1), SEQ ID NO: 2 (HCDR2), and SEQ ID NO: 3 (HCDR3), and a light chain CDR comprising the amino acid sequences of SEQ ID NO: 4 (LCDR1), SEQ ID NO: 5 (LCDR2), and SEQ ID NO: 6 (LCDR3); (ii) an anti-PD1 antibody comprising a heavy chain CDR comprising the amino acid sequences of SEQ ID NO: 7 (HCDR1), SEQ ID NO: 8 (HCDR2), and SEQ ID NO: 9 (HCDR3), and a light chain CDR comprising the amino acid sequences of SEQ ID NO: 10 (LCDR1), SEQ ID NO: 11 (LCDR2), and SEQ ID NO: 12 (LCDR3); and (iii) an anti-PD1 antibody comprising a heavy chain CDR comprising the amino acid sequences of SEQ ID NO: 13 (HCDR1), SEQ ID NO: 14 (HCDR2), and SEQ ID NO: 15 (HCDR3), and a light chain CDR comprising the amino acid sequences of SEQ ID NO: 16 (LCDR1), SEQ ID NO: 17 (LCDR2), and SEQ ID NO: 18 (LCDR3); Selected from the group consisting of: Polypeptides.

2. A method for treating and / or preventing a hyperproliferative disease, preferably cancer, comprising the steps of: The method comprises administering to a patient in need thereof a therapeutically effective amount of a polypeptide capable of specifically binding to LRP5 and LRP6 and a therapeutically effective amount of a PD-1 antibody; Here, the polypeptide capable of specifically binding to LRP5 and LRP6 is (i) the following CDR sequences: CDR1: TYTVG (=SEQ ID NO: 40) CDR2: AIRRGSSTYYADSVKG (=SEQ ID NO: 41) CDR3: DTRTVALLQYRYDY (=SEQ ID NO:42) (a) a first immunoglobulin single variable domain (ISVD) comprising The following CDR sequences: CDR1: SYAMG (=SEQ ID NO:49) CDR2: AISWSGGSTYYADSVKG (=SEQ ID NO:50) CDR3: SPIPYGSLLRRRNNYDY (=SEQ ID NO:51) a polypeptide comprising a second ISVD (b) comprising: (ii) the following CDR sequences: CDR1: SYAMG (=SEQ ID NO:43) CDR2: AIRSGRRTYYADSVKG (=SEQ ID NO:44) CDR3: ARVRSSTRYNTGTWWWEY (=SEQ ID NO: 45) A first ISVD (a) including The following CDR sequences: CDR1: SYAMG (=SEQ ID NO:49) CDR2: AISWSGGSTYYADSVKG (=SEQ ID NO:50) CDR3: SPIPYGSLLRRRNNYDY (=SEQ ID NO:51) a polypeptide comprising a second ISVD (b) comprising: (iii) the following CDR sequences: CDR1: RYTMG (=SEQ ID NO: 46) CDR2: AIVRSGGSTYYADSVKG (=SEQ ID NO: 47) CDR3: DRRGRGENYILLYSSGRYEY (=SEQ ID NO:48) A first ISVD (a) including The following CDR sequences: CDR1: SYAMG (=SEQ ID NO:49) CDR2: AISWSGGSTYYADSVKG (=SEQ ID NO:50) CDR3: SPIPYGSLLRRRNNYDY (=SEQ ID NO:51) a polypeptide comprising a second ISVD (b) comprising: (iv) the following CDR sequences: CDR1: TYTVG (=SEQ ID NO: 40) CDR2: AIRRGSSTYYADSVKG (=SEQ ID NO: 41) CDR3: DTRTVALLQYRYDY (=SEQ ID NO:42) A first ISVD (a) including The following CDR sequences: CDR1: SYAMG (=SEQ ID NO:52) CDR2: AISWRSGSTYYADSVKG (=SEQ ID NO:53) CDR3: DPRGYGVAYVSAYYEY (=SEQ ID NO:54) a polypeptide comprising a second ISVD (b) comprising: (v) the following CDR sequences: CDR1: SYAMG (=SEQ ID NO:43) CDR2: AIRSGRRTYYADSVKG (=SEQ ID NO:44) CDR3: ARVRSSTRYNTGTWWWEY (=SEQ ID NO: 45) A first ISVD (a) including The following CDR sequences: CDR1: SYAMG (=SEQ ID NO:52) CDR2: AISWRSGSTYYADSVKG (=SEQ ID NO:53) CDR3: DPRGYGVAYVSAYYEY (=SEQ ID NO:54) and a polypeptide comprising a second ISVD (b) comprising (vi) the following CDR sequences: CDR1: RYTMG (=SEQ ID NO: 46) CDR2: AIVRSGGSTYYADSVKG (=SEQ ID NO: 47) CDR3: DRRGRGENYILLYSSGRYEY (=SEQ ID NO:48) A first ISVD (a) including The following CDR sequences: CDR1: SYAMG (=SEQ ID NO:52) CDR2: AISWRSGSTYYADSVKG (=SEQ ID NO:53) CDR3: DPRGYGVAYVSAYYEY (=SEQ ID NO:54) A polypeptide comprising a second ISVD (b) comprising is selected from the group consisting of Here, the PD-1 antibody is (i) an anti-PD1 antibody comprising a heavy chain CDR comprising the amino acid sequences of SEQ ID NO: 1 (HCDR1), SEQ ID NO: 2 (HCDR2), and SEQ ID NO: 3 (HCDR3), and a light chain CDR comprising the amino acid sequences of SEQ ID NO: 4 (LCDR1), SEQ ID NO: 5 (LCDR2), and SEQ ID NO: 6 (LCDR3); (ii) an anti-PD1 antibody comprising a heavy chain CDR comprising the amino acid sequences of SEQ ID NO: 7 (HCDR1), SEQ ID NO: 8 (HCDR2), and SEQ ID NO: 9 (HCDR3), and a light chain CDR comprising the amino acid sequences of SEQ ID NO: 10 (LCDR1), SEQ ID NO: 11 (LCDR2), and SEQ ID NO: 12 (LCDR3); and (iii) an anti-PD1 antibody comprising a heavy chain CDR comprising the amino acid sequences of SEQ ID NO: 13 (HCDR1), SEQ ID NO: 14 (HCDR2), and SEQ ID NO: 15 (HCDR3), and a light chain CDR comprising the amino acid sequences of SEQ ID NO: 16 (LCDR1), SEQ ID NO: 17 (LCDR2), and SEQ ID NO: 18 (LCDR3); Selected from the group consisting of: method.

3. 1. A PD-1 antibody for use in a method for treating and / or preventing a hyperproliferative disease, preferably cancer, comprising: The method includes administering to a patient in need thereof a PD-1 antibody in combination with a polypeptide capable of specifically binding to LRP5 and LRP6; Here, the polypeptide capable of specifically binding to LRP5 and LRP6 is (i) the following CDR sequences: CDR1: TYTVG (=SEQ ID NO: 40) CDR2: AIRRGSSTYYADSVKG (=SEQ ID NO: 41) CDR3: DTRTVALLQYRYDY (=SEQ ID NO:42) (a) a first immunoglobulin single variable domain (ISVD) comprising The following CDR sequences: CDR1: SYAMG (=SEQ ID NO:49) CDR2: AISWSGGSTYYADSVKG (=SEQ ID NO:50) CDR3: SPIPYGSLLRRRNNYDY (=SEQ ID NO:51) a polypeptide comprising a second ISVD (b) comprising: (ii) the following CDR sequences: CDR1: SYAMG (=SEQ ID NO:43) CDR2: AIRSGRRTYYADSVKG (=SEQ ID NO:44) CDR3: ARVRSSTRYNTGTWWWEY (=SEQ ID NO: 45) A first ISVD (a) including The following CDR sequences: CDR1: SYAMG (=SEQ ID NO:49) CDR2: AISWSGGSTYYADSVKG (=SEQ ID NO:50) CDR3: SPIPYGSLLRRRNNYDY (=SEQ ID NO:51) a polypeptide comprising a second ISVD (b) comprising: (iii) the following CDR sequences: CDR1: RYTMG (=SEQ ID NO: 46) CDR2: AIVRSGGSTYYADSVKG (=SEQ ID NO: 47) CDR3: DRRGRGENYILLYSSGRYEY (=SEQ ID NO:48) A first ISVD (a) including The following CDR sequences: CDR1: SYAMG (=SEQ ID NO:49) CDR2: AISWSGGSTYYADSVKG (=SEQ ID NO:50) CDR3: SPIPYGSLLRRRNNYDY (=SEQ ID NO:51) a polypeptide comprising a second ISVD (b) comprising: (iv) the following CDR sequences: CDR1: TYTVG (=SEQ ID NO: 40) CDR2: AIRRGSSTYYADSVKG (=SEQ ID NO: 41) CDR3: DTRTVALLQYRYDY (=SEQ ID NO:42) A first ISVD (a) including The following CDR sequences: CDR1: SYAMG (=SEQ ID NO:52) CDR2: AISWRSGSTYYADSVKG (=SEQ ID NO:53) CDR3: DPRGYGVAYVSAYYEY (=SEQ ID NO:54) a polypeptide comprising a second ISVD (b) comprising: (v) the following CDR sequences: CDR1: SYAMG (=SEQ ID NO:43) CDR2: AIRSGRRTYYADSVKG (=SEQ ID NO:44) CDR3: ARVRSSTRYNTGTWWWEY (=SEQ ID NO: 45) A first ISVD (a) including The following CDR sequences: CDR1: SYAMG (=SEQ ID NO:52) CDR2: AISWRSGSTYYADSVKG (=SEQ ID NO:53) CDR3: DPRGYGVAYVSAYYEY (=SEQ ID NO:54) and a polypeptide comprising a second ISVD (b) comprising (vi) the following CDR sequences: CDR1: RYTMG (=SEQ ID NO: 46) CDR2: AIVRSGGSTYYADSVKG (=SEQ ID NO: 47) CDR3: DRRGRGENYILLYSSGRYEY (=SEQ ID NO:48) A first ISVD (a) including The following CDR sequences: CDR1: SYAMG (=SEQ ID NO:52) CDR2: AISWRSGSTYYADSVKG (=SEQ ID NO:53) CDR3: DPRGYGVAYVSAYYEY (=SEQ ID NO:54) A polypeptide comprising a second ISVD (b) comprising is selected from the group consisting of Here, the PD-1 antibody is (i) an anti-PD1 antibody comprising a heavy chain CDR comprising the amino acid sequences of SEQ ID NO: 1 (HCDR1), SEQ ID NO: 2 (HCDR2), and SEQ ID NO: 3 (HCDR3), and a light chain CDR comprising the amino acid sequences of SEQ ID NO: 4 (LCDR1), SEQ ID NO: 5 (LCDR2), and SEQ ID NO: 6 (LCDR3); (ii) an anti-PD1 antibody comprising a heavy chain CDR comprising the amino acid sequences of SEQ ID NO: 7 (HCDR1), SEQ ID NO: 8 (HCDR2), and SEQ ID NO: 9 (HCDR3), and a light chain CDR comprising the amino acid sequences of SEQ ID NO: 10 (LCDR1), SEQ ID NO: 11 (LCDR2), and SEQ ID NO: 12 (LCDR3); and (iii) an anti-PD1 antibody comprising a heavy chain CDR comprising the amino acid sequences of SEQ ID NO: 13 (HCDR1), SEQ ID NO: 14 (HCDR2), and SEQ ID NO: 15 (HCDR3), and a light chain CDR comprising the amino acid sequences of SEQ ID NO: 16 (LCDR1), SEQ ID NO: 17 (LCDR2), and SEQ ID NO: 18 (LCDR3); Selected from the group consisting of: PD-1 antibody.

4. Use of a polypeptide capable of specifically binding to LRP5 and LRP6 for the manufacture of a pharmaceutical composition for use in a method for treating and / or preventing a hyperproliferative disease, preferably cancer, comprising: A polypeptide capable of specifically binding to LRP5 and LRP6 is used in combination with a PD-1 antibody; Here, the polypeptide capable of specifically binding to LRP5 and LRP6 is (i) the following CDR sequences: CDR1: TYTVG (=SEQ ID NO: 40) CDR2: AIRRGSSTYYADSVKG (=SEQ ID NO: 41) CDR3: DTRTVALLQYRYDY (=SEQ ID NO:42) (a) a first immunoglobulin single variable domain (ISVD) comprising The following CDR sequences: CDR1: SYAMG (=SEQ ID NO:49) CDR2: AISWSGGSTYYADSVKG (=SEQ ID NO:50) CDR3: SPIPYGSLLRRRNNYDY (=SEQ ID NO:51) a polypeptide comprising a second ISVD (b) comprising: (ii) the following CDR sequences: CDR1: SYAMG (=SEQ ID NO:43) CDR2: AIRSGRRTYYADSVKG (=SEQ ID NO:44) CDR3: ARVRSSTRYNTGTWWWEY (=SEQ ID NO: 45) A first ISVD (a) including The following CDR sequences: CDR1: SYAMG (=SEQ ID NO:49) CDR2: AISWSGGSTYYADSVKG (=SEQ ID NO:50) CDR3: SPIPYGSLLRRRNNYDY (=SEQ ID NO:51) a polypeptide comprising a second ISVD (b) comprising: (iii) the following CDR sequences: CDR1: RYTMG (=SEQ ID NO: 46) CDR2: AIVRSGGSTYYADSVKG (=SEQ ID NO: 47) CDR3: DRRGRGENYILLYSSGRYEY (=SEQ ID NO:48) A first ISVD (a) including The following CDR sequences: CDR1: SYAMG (=SEQ ID NO:49) CDR2: AISWSGGSTYYADSVKG (=SEQ ID NO:50) CDR3: SPIPYGSLLRRRNNYDY (=SEQ ID NO:51) a polypeptide comprising a second ISVD (b) comprising: (iv) the following CDR sequences: CDR1: TYTVG (=SEQ ID NO: 40) CDR2: AIRRGSSTYYADSVKG (=SEQ ID NO: 41) CDR3: DTRTVALLQYRYDY (=SEQ ID NO:42) A first ISVD (a) including The following CDR sequences: CDR1: SYAMG (=SEQ ID NO:52) CDR2: AISWRSGSTYYADSVKG (=SEQ ID NO:53) CDR3: DPRGYGVAYVSAYYEY (=SEQ ID NO:54) a polypeptide comprising a second ISVD (b) comprising: (v) the following CDR sequences: CDR1: SYAMG (=SEQ ID NO:43) CDR2: AIRSGRRTYYADSVKG (=SEQ ID NO:44) CDR3: ARVRSSTRYNTGTWWWEY (=SEQ ID NO: 45) A first ISVD (a) including The following CDR sequences: CDR1: SYAMG (=SEQ ID NO:52) CDR2: AISWRSGSTYYADSVKG (=SEQ ID NO:53) CDR3: DPRGYGVAYVSAYYEY (=SEQ ID NO:54) and a polypeptide comprising a second ISVD (b) comprising (vi) the following CDR sequences: CDR1: RYTMG (=SEQ ID NO: 46) CDR2: AIVRSGGSTYYADSVKG (=SEQ ID NO: 47) CDR3: DRRGRGENYILLYSSGRYEY (=SEQ ID NO:48) A first ISVD (a) including The following CDR sequences: CDR1: SYAMG (=SEQ ID NO:52) CDR2: AISWRSGSTYYADSVKG (=SEQ ID NO:53) CDR3: DPRGYGVAYVSAYYEY (=SEQ ID NO:54) A polypeptide comprising a second ISVD (b) comprising is selected from the group consisting of Here, the PD-1 antibody is (i) an anti-PD1 antibody comprising a heavy chain CDR comprising the amino acid sequences of SEQ ID NO: 1 (HCDR1), SEQ ID NO: 2 (HCDR2), and SEQ ID NO: 3 (HCDR3), and a light chain CDR comprising the amino acid sequences of SEQ ID NO: 4 (LCDR1), SEQ ID NO: 5 (LCDR2), and SEQ ID NO: 6 (LCDR3); (ii) an anti-PD1 antibody comprising a heavy chain CDR comprising the amino acid sequences of SEQ ID NO: 7 (HCDR1), SEQ ID NO: 8 (HCDR2), and SEQ ID NO: 9 (HCDR3), and a light chain CDR comprising the amino acid sequences of SEQ ID NO: 10 (LCDR1), SEQ ID NO: 11 (LCDR2), and SEQ ID NO: 12 (LCDR3); and (iii) an anti-PD1 antibody comprising a heavy chain CDR comprising the amino acid sequences of SEQ ID NO: 13 (HCDR1), SEQ ID NO: 14 (HCDR2), and SEQ ID NO: 15 (HCDR3), and a light chain CDR comprising the amino acid sequences of SEQ ID NO: 16 (LCDR1), SEQ ID NO: 17 (LCDR2), and SEQ ID NO: 18 (LCDR3); Selected from the group consisting of: use.

5. Use of an antibody against PD-1 for the manufacture of a pharmaceutical composition for use in a method for treating and / or preventing a hyperproliferative disease, preferably cancer, comprising: The PD-1 antibody is used in combination with a polypeptide capable of specifically binding to LRP5 and LRP6, Here, the polypeptide capable of specifically binding to LRP5 and LRP6 is (i) the following CDR sequences: CDR1: TYTVG (=SEQ ID NO: 40) CDR2: AIRRGSSTYYADSVKG (=SEQ ID NO: 41) CDR3: DTRTVALLQYRYDY (=SEQ ID NO:42) (a) a first immunoglobulin single variable domain (ISVD) comprising The following CDR sequences: CDR1: SYAMG (=SEQ ID NO:49) CDR2: AISWSGGSTYYADSVKG (=SEQ ID NO:50) CDR3: SPIPYGSLLRRRNNYDY (=SEQ ID NO:51) a polypeptide comprising a second ISVD (b) comprising: (ii) the following CDR sequences: CDR1: SYAMG (=SEQ ID NO:43) CDR2: AIRSGRRTYYADSVKG (=SEQ ID NO:44) CDR3: ARVRSSTRYNTGTWWWEY (=SEQ ID NO: 45) A first ISVD (a) including The following CDR sequences: CDR1: SYAMG (=SEQ ID NO:49) CDR2: AISWSGGSTYYADSVKG (=SEQ ID NO:50) CDR3: SPIPYGSLLRRRNNYDY (=SEQ ID NO:51) a polypeptide comprising a second ISVD (b) comprising: (iii) the following CDR sequences: CDR1: RYTMG (=SEQ ID NO: 46) CDR2: AIVRSGGSTYYADSVKG (=SEQ ID NO: 47) CDR3: DRRGRGENYILLYSSGRYEY (=SEQ ID NO:48) A first ISVD (a) including The following CDR sequences: CDR1: SYAMG (=SEQ ID NO:49) CDR2: AISWSGGSTYYADSVKG (=SEQ ID NO:50) CDR3: SPIPYGSLLRRRNNYDY (=SEQ ID NO:51) a polypeptide comprising a second ISVD (b) comprising: (iv) the following CDR sequences: CDR1: TYTVG (=SEQ ID NO: 40) CDR2: AIRRGSSTYYADSVKG (=SEQ ID NO: 41) CDR3: DTRTVALLQYRYDY (=SEQ ID NO:42) A first ISVD (a) including The following CDR sequences: CDR1: SYAMG (=SEQ ID NO:52) CDR2: AISWRSGSTYYADSVKG (=SEQ ID NO:53) CDR3: DPRGYGVAYVSAYYEY (=SEQ ID NO:54) a polypeptide comprising a second ISVD (b) comprising: (v) the following CDR sequences: CDR1: SYAMG (=SEQ ID NO:43) CDR2: AIRSGRRTYYADSVKG (=SEQ ID NO:44) CDR3: ARVRSSTRYNTGTWWWEY (=SEQ ID NO: 45) A first ISVD (a) including The following CDR sequences: CDR1: SYAMG (=SEQ ID NO:52) CDR2: AISWRSGSTYYADSVKG (=SEQ ID NO:53) CDR3: DPRGYGVAYVSAYYEY (=SEQ ID NO:54) and a polypeptide comprising a second ISVD (b) comprising (vi) the following CDR sequences: CDR1: RYTMG (=SEQ ID NO: 46) CDR2: AIVRSGGSTYYADSVKG (=SEQ ID NO: 47) CDR3: DRRGRGENYILLYSSGRYEY (=SEQ ID NO:48) A first ISVD (a) including The following CDR sequences: CDR1: SYAMG (=SEQ ID NO:52) CDR2: AISWRSGSTYYADSVKG (=SEQ ID NO:53) CDR3: DPRGYGVAYVSAYYEY (=SEQ ID NO:54) A polypeptide comprising a second ISVD (b) comprising is selected from the group consisting of Here, the PD-1 antibody is (i) an anti-PD1 antibody comprising a heavy chain CDR comprising the amino acid sequences of SEQ ID NO: 1 (HCDR1), SEQ ID NO: 2 (HCDR2), and SEQ ID NO: 3 (HCDR3), and a light chain CDR comprising the amino acid sequences of SEQ ID NO: 4 (LCDR1), SEQ ID NO: 5 (LCDR2), and SEQ ID NO: 6 (LCDR3); (ii) an anti-PD1 antibody comprising a heavy chain CDR comprising the amino acid sequences of SEQ ID NO: 7 (HCDR1), SEQ ID NO: 8 (HCDR2), and SEQ ID NO: 9 (HCDR3), and a light chain CDR comprising the amino acid sequences of SEQ ID NO: 10 (LCDR1), SEQ ID NO: 11 (LCDR2), and SEQ ID NO: 12 (LCDR3); and (iii) an anti-PD1 antibody comprising a heavy chain CDR comprising the amino acid sequences of SEQ ID NO: 13 (HCDR1), SEQ ID NO: 14 (HCDR2), and SEQ ID NO: 15 (HCDR3), and a light chain CDR comprising the amino acid sequences of SEQ ID NO: 16 (LCDR1), SEQ ID NO: 17 (LCDR2), and SEQ ID NO: 18 (LCDR3); Selected from the group consisting of: use.

6. A polypeptide capable of specifically binding to LRP5 and LRP6, a PD-1 antibody, and optionally one or more pharma- ceutically acceptable carriers, excipients and / or vehicles; Here, the polypeptide capable of specifically binding to LRP5 and LRP6 is (i) the following CDR sequences: CDR1: TYTVG (=SEQ ID NO: 40) CDR2: AIRRGSSTYYADSVKG (=SEQ ID NO: 41) CDR3: DTRTVALLQYRYDY (=SEQ ID NO:42) (a) a first immunoglobulin single variable domain (ISVD) comprising The following CDR sequences: CDR1: SYAMG (=SEQ ID NO:49) CDR2: AISWSGGSTYYADSVKG (=SEQ ID NO:50) CDR3: SPIPYGSLLRRRNNYDY (=SEQ ID NO:51) a polypeptide comprising a second ISVD (b) comprising: (ii) the following CDR sequences: CDR1: SYAMG (=SEQ ID NO:43) CDR2: AIRSGRRTYYADSVKG (=SEQ ID NO:44) CDR3: ARVRSSTRYNTGTWWWEY (=SEQ ID NO: 45) A first ISVD (a) including The following CDR sequences: CDR1: SYAMG (=SEQ ID NO:49) CDR2: AISWSGGSTYYADSVKG (=SEQ ID NO:50) CDR3: SPIPYGSLLRRRNNYDY (=SEQ ID NO:51) a polypeptide comprising a second ISVD (b) comprising: (iii) the following CDR sequences: CDR1: RYTMG (=SEQ ID NO: 46) CDR2: AIVRSGGSTYYADSVKG (=SEQ ID NO: 47) CDR3: DRRGRGENYILLYSSGRYEY (=SEQ ID NO:48) A first ISVD (a) including The following CDR sequences: CDR1: SYAMG (=SEQ ID NO:49) CDR2: AISWSGGSTYYADSVKG (=SEQ ID NO:50) CDR3: SPIPYGSLLRRRNNYDY (=SEQ ID NO:51) a polypeptide comprising a second ISVD (b) comprising: (iv) the following CDR sequences: CDR1: TYTVG (=SEQ ID NO: 40) CDR2: AIRRGSSTYYADSVKG (=SEQ ID NO: 41) CDR3: DTRTVALLQYRYDY (=SEQ ID NO:42) A first ISVD (a) including The following CDR sequences: CDR1: SYAMG (=SEQ ID NO:52) CDR2: AISWRSGSTYYADSVKG (=SEQ ID NO:53) CDR3: DPRGYGVAYVSAYYEY (=SEQ ID NO:54) a polypeptide comprising a second ISVD (b) comprising: (v) the following CDR sequences: CDR1: SYAMG (=SEQ ID NO:43) CDR2: AIRSGRRTYYADSVKG (=SEQ ID NO:44) CDR3: ARVRSSTRYNTGTWWWEY (=SEQ ID NO: 45) A first ISVD (a) including The following CDR sequences: CDR1: SYAMG (=SEQ ID NO:52) CDR2: AISWRSGSTYYADSVKG (=SEQ ID NO:53) CDR3: DPRGYGVAYVSAYYEY (=SEQ ID NO:54) and a polypeptide comprising a second ISVD (b) comprising (vi) the following CDR sequences: CDR1: RYTMG (=SEQ ID NO: 46) CDR2: AIVRSGGSTYYADSVKG (=SEQ ID NO: 47) CDR3: DRRGRGENYILLYSSGRYEY (=SEQ ID NO:48) A first ISVD (a) including The following CDR sequences: CDR1: SYAMG (=SEQ ID NO:52) CDR2: AISWRSGSTYYADSVKG (=SEQ ID NO:53) CDR3: DPRGYGVAYVSAYYEY (=SEQ ID NO:54) A polypeptide comprising a second ISVD (b) comprising is selected from the group consisting of Here, the PD-1 antibody is (i) an anti-PD1 antibody comprising a heavy chain CDR comprising the amino acid sequences of SEQ ID NO: 1 (HCDR1), SEQ ID NO: 2 (HCDR2), and SEQ ID NO: 3 (HCDR3), and a light chain CDR comprising the amino acid sequences of SEQ ID NO: 4 (LCDR1), SEQ ID NO: 5 (LCDR2), and SEQ ID NO: 6 (LCDR3); (ii) an anti-PD1 antibody comprising a heavy chain CDR comprising the amino acid sequences of SEQ ID NO: 7 (HCDR1), SEQ ID NO: 8 (HCDR2), and SEQ ID NO: 9 (HCDR3), and a light chain CDR comprising the amino acid sequences of SEQ ID NO: 10 (LCDR1), SEQ ID NO: 11 (LCDR2), and SEQ ID NO: 12 (LCDR3); and (iii) an anti-PD1 antibody comprising a heavy chain CDR comprising the amino acid sequences of SEQ ID NO: 13 (HCDR1), SEQ ID NO: 14 (HCDR2), and SEQ ID NO: 15 (HCDR3), and a light chain CDR comprising the amino acid sequences of SEQ ID NO: 16 (LCDR1), SEQ ID NO: 17 (LCDR2), and SEQ ID NO: 18 (LCDR3); Selected from the group consisting of: Pharmaceutical compositions.

7. 7. A pharmaceutical composition according to claim 6 for use in a method for treating and / or preventing a hyperproliferative disease, preferably cancer.

8. In one or more containers, - a first pharmaceutical composition or dosage form comprising a polypeptide capable of specifically binding to LRP5 and LRP6, and optionally one or more pharma- ceutically acceptable carriers, excipients and / or vehicles; a second pharmaceutical composition or dosage form comprising a PD-1 antibody and, optionally, one or more pharma- ceutically acceptable carriers, excipients and / or vehicles; and - possibly including a package insert containing printed instructions, Here, the polypeptide capable of specifically binding to LRP5 and LRP6 is (i) the following CDR sequences: CDR1: TYTVG (=SEQ ID NO: 40) CDR2: AIRRGSSTYYADSVKG (=SEQ ID NO: 41) CDR3: DTRTVALLQYRYDY (=SEQ ID NO:42) (a) a first immunoglobulin single variable domain (ISVD) comprising The following CDR sequences: CDR1: SYAMG (=SEQ ID NO:49) CDR2: AISWSGGSTYYADSVKG (=SEQ ID NO:50) CDR3: SPIPYGSLLRRRNNYDY (=SEQ ID NO:51) a polypeptide comprising a second ISVD (b) comprising: (ii) the following CDR sequences: CDR1: SYAMG (=SEQ ID NO:43) CDR2: AIRSGRRTYYADSVKG (=SEQ ID NO:44) CDR3: ARVRSSTRYNTGTWWWEY (=SEQ ID NO: 45) A first ISVD (a) including The following CDR sequences: CDR1: SYAMG (=SEQ ID NO:49) CDR2: AISWSGGSTYYADSVKG (=SEQ ID NO:50) CDR3: SPIPYGSLLRRRNNYDY (=SEQ ID NO:51) a polypeptide comprising a second ISVD (b) comprising: (iii) the following CDR sequences: CDR1: RYTMG (=SEQ ID NO: 46) CDR2: AIVRSGGSTYYADSVKG (=SEQ ID NO: 47) CDR3: DRRGRGENYILLYSSGRYEY (=SEQ ID NO:48) A first ISVD (a) including The following CDR sequences: CDR1: SYAMG (=SEQ ID NO:49) CDR2: AISWSGGSTYYADSVKG (=SEQ ID NO:50) CDR3: SPIPYGSLLRRRNNYDY (=SEQ ID NO:51) a polypeptide comprising a second ISVD (b) comprising: (iv) the following CDR sequences: CDR1: TYTVG (=SEQ ID NO: 40) CDR2: AIRRGSSTYYADSVKG (=SEQ ID NO: 41) CDR3: DTRTVALLQYRYDY (=SEQ ID NO:42) A first ISVD (a) including The following CDR sequences: CDR1: SYAMG (=SEQ ID NO:52) CDR2: AISWRSGSTYYADSVKG (=SEQ ID NO:53) CDR3: DPRGYGVAYVSAYYEY (=SEQ ID NO:54) a polypeptide comprising a second ISVD (b) comprising: (v) the following CDR sequences: CDR1: SYAMG (=SEQ ID NO:43) CDR2: AIRSGRRTYYADSVKG (=SEQ ID NO:44) CDR3: ARVRSSTRYNTGTWWWEY (=SEQ ID NO: 45) A first ISVD (a) including The following CDR sequences: CDR1: SYAMG (=SEQ ID NO:52) CDR2: AISWRSGSTYYADSVKG (=SEQ ID NO:53) CDR3: DPRGYGVAYVSAYYEY (=SEQ ID NO:54) and a polypeptide comprising a second ISVD (b) comprising (vi) the following CDR sequences: CDR1: RYTMG (=SEQ ID NO: 46) CDR2: AIVRSGGSTYYADSVKG (=SEQ ID NO: 47) CDR3: DRRGRGENYILLYSSGRYEY (=SEQ ID NO:48) A first ISVD (a) including The following CDR sequences: CDR1: SYAMG (=SEQ ID NO:52) CDR2: AISWRSGSTYYADSVKG (=SEQ ID NO:53) CDR3: DPRGYGVAYVSAYYEY (=SEQ ID NO:54) A polypeptide comprising a second ISVD (b) comprising is selected from the group consisting of Here, the PD-1 antibody is (i) an anti-PD1 antibody comprising a heavy chain CDR comprising the amino acid sequences of SEQ ID NO: 1 (HCDR1), SEQ ID NO: 2 (HCDR2), and SEQ ID NO: 3 (HCDR3), and a light chain CDR comprising the amino acid sequences of SEQ ID NO: 4 (LCDR1), SEQ ID NO: 5 (LCDR2), and SEQ ID NO: 6 (LCDR3); (ii) an anti-PD1 antibody comprising a heavy chain CDR comprising the amino acid sequences of SEQ ID NO: 7 (HCDR1), SEQ ID NO: 8 (HCDR2), and SEQ ID NO: 9 (HCDR3), and a light chain CDR comprising the amino acid sequences of SEQ ID NO: 10 (LCDR1), SEQ ID NO: 11 (LCDR2), and SEQ ID NO: 12 (LCDR3); and (iii) an anti-PD1 antibody comprising a heavy chain CDR comprising the amino acid sequences of SEQ ID NO: 13 (HCDR1), SEQ ID NO: 14 (HCDR2), and SEQ ID NO: 15 (HCDR3), and a light chain CDR comprising the amino acid sequences of SEQ ID NO: 16 (LCDR1), SEQ ID NO: 17 (LCDR2), and SEQ ID NO: 18 (LCDR3); Selected from the group consisting of: kit.

9. A kit according to claim 8 for use in a method for treating and / or preventing a hyperproliferative disease, preferably cancer.

10. A polypeptide capable of specifically binding to LRP5 and LRP6, (i) a polypeptide comprising a first ISVD comprising the amino acid sequence of SEQ ID NO: 58 and a second ISVD comprising the sequence of SEQ ID NO: 61; (ii) a polypeptide comprising a first ISVD comprising the amino acid sequence of SEQ ID NO: 59 and a second ISVD comprising the sequence of SEQ ID NO: 61; (iii) a polypeptide comprising a first ISVD comprising the sequence of SEQ ID NO: 60 and a second ISVD comprising the sequence of SEQ ID NO: 61; (iv) a polypeptide comprising a first ISVD comprising the amino acid sequence of SEQ ID NO: 58 and a second ISVD comprising the sequence of SEQ ID NO: 62; (v) a polypeptide comprising a first ISVD comprising the amino acid sequence of SEQ ID NO: 59 and a second ISVD comprising the sequence of SEQ ID NO: 62; and (vi) a polypeptide comprising a first ISVD comprising the amino acid sequence of SEQ ID NO: 60 and a second ISVD comprising the sequence of SEQ ID NO:

62. is selected from the group consisting of Preferably, the polypeptide capable of specifically binding to LRP5 and LRP6 for use according to claim 1 further comprises an Alb11 domain comprising the amino acid sequence of SEQ ID NO: 63, the method of treatment according to claim 2, the PD-1 antibody for use according to claim 3, the use according to claim 4 or 5, the pharmaceutical composition according to claim 6 or 7, or the kit according to claim 8 or 9.

11. A polypeptide capable of specifically binding to LRP5 and LRP6 for use according to claim 1, comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 64, SEQ ID NO: 65 and SEQ ID NO:

66. A polypeptide capable of specifically binding to LRP5 and LRP6 for use according to claim 1, a method of treatment according to claim 2, a PD-1 antibody for use according to claim 3, a use according to claim 4 or 5, a pharmaceutical composition according to claim 6 or 7, or a kit according to claim 8 or 9.

12. The PD-1 antibody (i) an antibody having a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 19 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 20; (ii) an antibody having a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 21 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 22; (iii) an antibody having a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 23 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 24; (iv) an antibody having a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 25 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 26; and (v) an antibody having a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 27 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 28; A polypeptide capable of specifically binding to LRP5 and LRP6 for use according to claim 1, 10 or 11, a method of treatment according to claim 2, 10 or 11, a PD-1 antibody for use according to claim 3, 10 or 11, a use according to claim 4, 5, 10 or 11, a pharmaceutical composition according to claim 6, 7, 10 or 11, or a kit according to claim 8, 9, 10 or 11.

13. The PD-1 antibody (i) an antibody having a heavy chain comprising the amino acid sequence of SEQ ID NO: 29 and a light chain comprising the amino acid sequence of SEQ ID NO: 30; (ii) an antibody having a heavy chain comprising the amino acid sequence of SEQ ID NO: 31 and a light chain comprising the amino acid sequence of SEQ ID NO: 32; (iii) an antibody having a heavy chain comprising the amino acid sequence of SEQ ID NO: 33 and a light chain comprising the amino acid sequence of SEQ ID NO: 34; (iv) an antibody having a heavy chain comprising the amino acid sequence of SEQ ID NO: 35 and a light chain comprising the amino acid sequence of SEQ ID NO: 36; and (v) an antibody having a heavy chain comprising the amino acid sequence of SEQ ID NO: 37 and a light chain comprising the amino acid sequence of SEQ ID NO: 38 A polypeptide capable of specifically binding to LRP5 and LRP6 for use according to claim 1, 10 or 11, a method of treatment according to claim 2, 10 or 11, a PD-1 antibody for use according to claim 3, 10 or 11, a use according to claim 4, 5, 10 or 11, a pharmaceutical composition according to claim 6, 7, 10 or 11, or a kit according to claim 8, 9, 10 or 11.

14. A polypeptide capable of specifically binding to LRP5 and LRP6 for use according to claim 1, 10, 11, 12 or 13, wherein the PD-1 antibody is administered simultaneously, contemporaneously, sequentially, consecutively, alternatingly or separately with a polypeptide capable of specifically binding to LRP5 and LRP6. A method of treatment according to claim 2, 10, 11, 12 or 13, a PD-1 antibody for use according to claim 3, 10, 11, 12 or 13, a use according to claim 4, 5, 10, 11, 12 or 13, or a kit for use according to claim 9, 10, 11, 12 or 13.

15. The polypeptide capable of specifically binding to LRP5 and LRP6 and the PD-1 antibody are administered in the following treatment regimen: (i) a first treatment period in which a polypeptide capable of specifically binding to LRP5 and LRP6 and a PD-1 antibody are administered simultaneously or contemporaneously, preferably every 3 or 4 weeks; (ii) a second treatment period in which only the PD-1 antibody is administered, and no polypeptide capable of specifically binding to LRP5 and LRP6 is administered, preferably in which the PD-1 antibody is administered every 3 or 4 weeks; A polypeptide capable of specifically binding to LRP5 and LRP6 for use according to claim 1, 10, 11, 12 or 13, a method of treatment according to claim 2, 10, 11, 12 or 13, a PD-1 antibody for use according to claim 3, 10, 11, 12 or 13, a use according to claim 4, 5, 10, 11, 12 or 13, or a kit for use according to claim 9, 10, 11, 12 or 13.

16. When a polypeptide capable of specifically binding to LRP5 and LRP6 and a PD-1 antibody are administered every 3 weeks, the first treatment period is 3 weeks or 6 weeks, or When the polypeptide capable of specifically binding to LRP5 and LRP6 and the PD-1 antibody are administered every 4 weeks, the first treatment period is 4 weeks or 8 weeks.

16. A polypeptide capable of specifically binding to LRP5 and LRP6, a method of treatment, a PD-1 antibody for use, a kit for use, or a kit for use.

17. The polypeptide capable of specifically binding to LRP5 and LRP6 for use according to claim 14, 15 or 16, wherein the administration is intravenous administration, the method of treatment, the PD-1 antibody for use, the use, the pharmaceutical composition for use or the kit for use.

18. The polypeptide capable of specifically binding to LRP5 and LRP6 for use according to any one of claims 1, 10 to 17, the method of treatment according to any one of claims 2, 10 to 17, the PD-1 antibody for use according to any one of claims 3, 10 to 17, the use according to any one of claims 4, 5, 10 to 17, the pharmaceutical composition for use according to any one of claims 7, 10 to 17, or the kit for use according to any one of claims 9 to 17, wherein the hyperproliferative disease to be treated is a cancer selected from the group consisting of gastrointestinal cancer, melanoma tumor, bladder cancer and lung cancer (e.g., NSCLC).

19. The polypeptide capable of specifically binding to LRP5 and LRP6 for use according to claim 18, wherein the gastrointestinal cancer is esophageal cancer (e.g., gastroesophageal junction cancer), gastric (gastric) cancer, hepatocellular carcinoma, biliary tract cancer (e.g., bile duct cancer), gallbladder cancer, pancreatic cancer or colorectal cancer (CRC). Method of treatment, PD-1 antibody for use, use, pharmaceutical composition for use, or kit for use.

20. The polypeptide capable of specifically binding to LRP5 and LRP6 for use according to claim 18 or 19, wherein the cancer is an immunotherapy-resistant tumor, the method of treatment, the PD-1 antibody for use, the use, the pharmaceutical composition for use, or the kit for use.

21. The polypeptide capable of specifically binding to LRP5 and LRP6 for use according to any one of claims 1, 10 to 17, the method of treatment according to any one of claims 2, 10 to 17, the PD-1 antibody for use according to any one of claims 3, 10 to 17, the use according to any one of claims 4, 5, 10 to 17, the pharmaceutical composition for use according to any one of claims 7, 10 to 17, or the kit for use according to any one of claims 9 to 17, wherein the hyperproliferative disease to be treated is an immunotherapy-resistant solid tumor.

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