RU2836467C2 - Il-4/il-13 pathway inhibitors for higher efficacy in treating malignant neoplasms - Google Patents

Abstract

FIELD: biotechnology.

SUBSTANCE: disclosed are methods of treating or inhibiting tumour growth, including type 2 immune-dependent malignant growth, wherein the methods comprise selecting and administering to an individual in need thereof a therapeutically effective amount of an antibody specifically binding the IL-4 receptor (IL-4R), and a therapeutically effective amount of an antibody specifically binding programmed death protein 1 (PD-1).

EFFECT: invention promotes tumour regression, inhibits tumour growth, reduces the tumour load and/or prevents tumour recurrence in the patient.

43 cl, 12 dwg, 1 tbl, 7 ex

Description

FIELD OF INVENTION

[001] The present invention relates to methods of treating malignancies comprising administering to an individual in need thereof a therapeutically effective amount of an IL-4/IL-13 pathway inhibitor in combination with a therapeutically effective amount of a programmed death protein 1 (PD-1) inhibitor. In some embodiments, the IL-4/IL-13 pathway inhibitor enhances the antitumor efficacy of PD-1 blockade.

LEVEL OF TECHNOLOGY

[002] Type 2 immunity promotes tumor growth and metastasis in various types of malignancies. Several type 2-associated cytokines, chemokines, or receptors are involved in the development of malignancies and are overexpressed in various human tumors. For example, IL-4, IL-13, IL-4Rα, and IL-33 are overexpressed in a number of human malignancies and are associated with poor prognosis in bladder, breast, and ovarian cancers (Joshi et al., Cancer Med. , 3(6):1615–28, 2014; Formentini et al., Int J Colorectal Dis ., 27(10):1369–76, 2012); Tong et al., Mol Oncol . 10(1):113–25, 2015). Type 2 immunity is characterized by the production of a number of type 2 cytokines and chemokines (e.g., IL-4, IL-13, thymic stromal lymphopoietin (TSLP), IL-33) and is typically observed in tissues during allergic responses or parasitic infections. Type 2 immune responses also contribute to tissue repair responses and fibrosis (Wynn, Nat Rev Immunol ., 15(5):271–82, 2015; Gieseck et al., Nat Rev Immunol ., 18(1):62–76, 2017).

[003] Pancreatic ductal adenocarcinoma (PDAC) is the third most common cause of cancer-related death in the United States, with a 5-year survival rate of 8%. This extremely poor prognosis is primarily due to the fact that PDAC is refractory to most therapies. In particular, immune checkpoint blockade (ICB) immunotherapy (eg, anti-PD-1 or anti-cytotoxic T-lymphocyte antigen 4 (CTLA-4)) does not provide clinical response in patients with PDAC. A key component of the robust drug resistance in PDAC is its unique desmoplastic stroma, a dense fibrous and stromal reaction environment composed of tumor-associated fibroblasts, immune cells, and large amounts of extracellular matrix components that acts as a physical barrier to drug perfusion and T cell infiltration and creates a highly immunosuppressive environment that inhibits T cell function (Olive et al., Science , 324(5933):1457–61, 2009; Provenzano et al., Cancer Cell , 21(3):418–29, 2012; Feig et al., Proc Natl Acad Sci US A. , 110(50):20212–17, 2013).

[004] Preclinical studies have shown that genetic ablation or blockade of IL-13, IL-4R, or TSLP can reduce tumor growth and metastasis in pancreatic cancer, breast cancer, and colorectal cancer. In addition, serum IL-4 and TSLP have been shown to predict survival in patients with pancreatic cancer (Piro et al., Oncoimmunology 6(9):e1322242, 2017; De Monte et al., J Exp Med , 208(3):469–78, 2011). Type 2 immune-dependent tumorigenesis involves induction of fibrosis, inhibition of antitumor immune surveillance, or increased glucose and glutamine metabolism in malignant cells (DeNardo et al., Cancer Cell, 16(2):91–102, 2009; Liou et al., Cell Rep. , 19(7):1322–33, 2017; Pedroza-Gonzalez et al., J Exp Med , 208(3):479–90, 2011; Venmar et al., Biochim Biophys Acta , 1853(5):1219–28, 2015; Ostrand-Rosenberg et al., J. Immunol ., 165:6015–19, 2000). Taken together, these data suggest that type 2 immunity promotes tumor growth and metastasis in various types of malignancies.

[005] Thus, there is a need for new and effective therapies for the treatment of malignant neoplasms, including types and subtypes of malignant neoplasms characterized by a type 2 immune response and malignant neoplasms resistant to previous therapy.

ESSENCE OF THE INVENTION

[006] In one aspect, the present invention relates to a method of treating or inhibiting the growth of a tumor, comprising: (a) selecting an individual with a tumor; and (b) administering to the individual in need thereof a therapeutically effective amount of an IL-4/IL-13 pathway inhibitor and a therapeutically effective amount of a programmed death protein 1 (PD-1) inhibitor. In one embodiment, the individual has a tumor selected from the group consisting of colorectal cancer, ovarian cancer, prostate cancer, bladder cancer, breast cancer, malignant neoplasm of the brain, cervical cancer, bladder cancer, anal cancer, uterine cancer, colon cancer, liver cancer, pancreatic cancer, lung cancer, endometrial cancer, malignant neoplasm of the bone, testicular cancer, skin cancer, kidney cancer, stomach cancer, esophageal cancer, head and neck cancer, salivary gland cancer, myeloma, Hodgkin's lymphoma, non-Hodgkin's lymphoma, follicular lymphoma, small lymphocytic lymphoma, lymphoplasmacytoid lymphoma, marginal zone cell lymphoma, mantle cell lymphoma, diffuse large B-cell lymphoma, B-cell lymphomas, Hodgkin's disease, Burkitt's lymphoma, acute lymphoblastic leukemia, hairy cell leukemia, and B-cell chronic lymphocytic leukemia. In one embodiment, the tumor comprises a type 2 immune-dependent malignancy. In another embodiment, the type 2 immune-dependent malignancy comprises pancreatic cancer, breast cancer, colorectal cancer, ovarian cancer, brain cancer, skin cancer, prostate cancer, kidney cancer, lung cancer, Hodgkin's lymphoma, or bladder cancer. In another embodiment, the tumor comprises pancreatic cancer. In one embodiment, the tumor is primary, metastatic, or recurrent. In one embodiment, the individual is treated with one or more anti-tumor therapies. In one embodiment, the individual has a tumor that is resistant to or inadequately responsive to prior therapy. In one embodiment, the individual has a tumor that is resistant or unresponsive to treatment with an immune checkpoint inhibitor (e.g., a PD-1 inhibitor, a LAG3 inhibitor, etc.).

[007] In one embodiment, the IL-4/IL-13 pathway inhibitor is selected from the group consisting of an anti-IL-4 antibody, an anti-IL-13 antibody, an anti-IL-4/IL-13 bispecific antibody, an IL-4 receptor (IL-4R) inhibitor, an IL-4 trap, an IL-13 trap, and an anti-IL-4R antibody. In another embodiment, the IL-4/IL-13 pathway inhibitor is an anti-IL-4 antibody (e.g., pascolizumab). In another embodiment, the IL-4/IL-13 pathway inhibitor is an anti-IL-13 antibody (e.g., tralokinumab, lebrikizumab, dextrecumab, GSK679586, or MEDI7836). In another embodiment, the IL-4/IL-13 pathway inhibitor is an IL-4/IL-13 bispecific antibody (e.g., romilkimab). In another embodiment, the IL-4/IL-13 pathway inhibitor is an IL-4R inhibitor (e.g., an IL-4 mutein such as pitrakinra or an IL-4R antibody). In another embodiment, the IL-4/IL-13 pathway inhibitor is an IL-4R antibody. In another embodiment, the IL-4/IL-13 pathway inhibitor is an IL-4 trap or an IL-13 trap.

[008] In another embodiment, an anti-IL-4R antibody comprises a heavy chain variable region (HCVR) comprising the amino acid sequence of SEQ ID NO: 1 and a light chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO: 2. In another embodiment, the HCVR comprises three heavy chain complementarity determining regions (CDRs) (HCDR1, HCDR2, and HCDR3) and the LCVR comprises three light chain CDRs (LCDR1, LCDR2, and LCDR3), wherein: HCDR1 has the amino acid sequence of SEQ ID NO: 3; HCDR2 has the amino acid sequence of SEQ ID NO: 4; HCDR3 has the amino acid sequence of SEQ ID NO: 5; LCDR1 has the amino acid sequence of SEQ ID NO: 6; LCDR2 has the amino acid sequence of SEQ ID NO: 7; and LCDR3 has the amino acid sequence of SEQ ID NO: 8. In another embodiment, the anti-IL-4R antibody comprises a heavy chain and a light chain, wherein the heavy chain has the amino acid sequence of SEQ ID NO: 9. In another embodiment, the anti-IL-4R antibody comprises a heavy chain and a light chain, wherein the light chain has the amino acid sequence of SEQ ID NO: 10. In another embodiment, the anti-IL-4R antibody comprises a heavy chain and a light chain, wherein the heavy chain has the amino acid sequence of SEQ ID NO: 9 and the light chain has the amino acid sequence of SEQ ID NO: 10. In another embodiment, the IL-4/IL-13 pathway inhibitor is dupilumab or a bioequivalent thereof. In another embodiment, the IL-4/IL-13 pathway inhibitor is selected from the group consisting of dupilumab, pascolizumab, AMG317, MEDI2045, MEDI9314, tralokinumab, lebrikizumab, anrukinzumab, dectrecumab, GSK679586, MEDI7836, romilkimab, IL-4 trap, IL-13 trap, AER-003, and pitrakinra.

[009] In one embodiment, the PD-1 inhibitor is selected from an anti-PD-1 antibody, an anti-PD-L1 antibody, and an anti-PD-L2 antibody. In another embodiment, the PD-1 inhibitor is an anti-PD-1 antibody comprising an HCVR comprising the amino acid sequence of SEQ ID NO: 11 and an LCVR comprising the amino acid sequence of SEQ ID NO: 12. In another embodiment, the HCVR comprises three heavy chain CDRs (HCDR1, HCDR2, and HCDR3), and the LCVR comprises three light chain CDRs (LCDR1, LCDR2, and LCDR3), wherein: HCDR1 has the amino acid sequence of SEQ ID NO: 13; HCDR2 has the amino acid sequence of SEQ ID NO: 14; HCDR3 has the amino acid sequence of SEQ ID NO: 15; LCDR1 has the amino acid sequence of SEQ ID NO: 16; LCDR2 has the amino acid sequence of SEQ ID NO: 17; and LCDR3 has the amino acid sequence of SEQ ID NO: 18. In another embodiment, the anti-PD-1 antibody comprises the HCVR/LCVR sequence pair of SEQ ID NO: 11/12. In another embodiment, the anti-PD-1 antibody comprises a heavy chain and a light chain, wherein the heavy chain has the amino acid sequence of SEQ ID NO: 19. In another embodiment, the anti-PD-1 antibody comprises a heavy chain and a light chain, wherein the light chain has the amino acid sequence of SEQ ID NO: 20. In another embodiment, the anti-PD-1 antibody comprises a heavy chain and a light chain, wherein the heavy chain has the amino acid sequence of SEQ ID NO: 19 and the light chain has the amino acid sequence of SEQ ID NO: 20. In another embodiment, the PD-1 inhibitor is cemiplimab or a bioequivalent thereof. In another embodiment, the PD-1 inhibitor is an anti-PD-1 antibody selected from the group consisting of cemiplimab, nivolumab, pembrolizumab, pidilizumab, MEDI0608, BI 754091, PF-06801591, sintilimab, AGEN2034, spartalizumab, camrelizumab, JNJ-63723283, and MCLA-134. In another embodiment, the PD-1 inhibitor is an anti-PD-L1 antibody selected from the group consisting of H1H8314N, avelumab, atezolizumab, durvalumab, MDX-1105, LY3300054, FAZ053, STI-1014, CX-072, KN035, and CK-301.

[010] In another embodiment, one or more doses of an IL-4/IL-13 pathway inhibitor are administered in combination with one or more doses of an anti-PD-1 antibody. In another embodiment, the at least one dose of the IL-4/IL-13 pathway inhibitor comprises about 0.1 to about 50 mg/kg of the subject's body weight. In another embodiment, the at least one dose of the IL-4/IL-13 pathway inhibitor comprises about 0.05 to about 1000 mg of the inhibitor. In another embodiment, the at least one dose of the PD-1 inhibitor comprises about 0.1 mg/kg to about 20 mg/kg of the subject's body weight. In another embodiment, the at least one dose of the PD-1 inhibitor comprises about 0.05 to about 500 mg of the inhibitor. In another embodiment, the IL-4/IL-13 pathway inhibitor is administered prior to the PD-1 inhibitor. In another embodiment, the IL-4/IL-13 pathway inhibitor is administered after the PD-1 inhibitor. In another embodiment, the IL-4/IL-13 pathway inhibitor is administered concurrently with the PD-1 inhibitor. In another embodiment, the method promotes tumor regression, delays tumor growth, reduces the number of tumor cells, reduces tumor burden, and/or prevents tumor recurrence in a patient. In another embodiment, the method promotes at least about 10% greater tumor regression in a treated individual compared to an untreated individual or an individual treated with any inhibitor as monotherapy. In another embodiment, the method results in at least a 30% or greater reduction in tumor cells or tumor size compared to an untreated individual or an individual treated with any inhibitor as monotherapy.

[011] In another embodiment, the method further comprises using at least one additional therapeutic agent or method of therapy. In another embodiment, the additional therapeutic agent or method of therapy comprises chemotherapy, cyclophosphamide, surgery, radiation therapy, a tumor vaccine, a LAG3 inhibitor, a CTLA-4 inhibitor, a GITR agonist, a TIM3 inhibitor, a BTLA inhibitor, a TIGIT inhibitor, a CD47 inhibitor, an IDO inhibitor, a VEGF antagonist, an Ang2 inhibitor, a TGFβ inhibitor, an EGFR inhibitor, a VISTA inhibitor, a CD38 inhibitor, a CD40 agonist, a CSF1R inhibitor, a CCR2 inhibitor, a CXCR4 inhibitor, a CXCR2 inhibitor, a CCR4 inhibitor, a CXCL12 inhibitor, a CD28 activator, a costimulatory receptor agonist, an anti-tumor antigen antibody, an anti-CD3/anti-CD20 bispecific antibody, GM-CSF, a cytotoxin, a chemotherapeutic agent, an oncolytic virus, IL-6R inhibitor, IL-10 inhibitor, cytokine or its derivative (e.g., IL-12 or IL-2), ADC, chimeric antigen receptor T cells, anti-inflammatory drug, NSAID, and/or dietary supplement.

[012] In another aspect, the present invention relates to a pharmaceutical delivery system comprising: (a) a pharmaceutical composition comprising an IL-4/IL-13 pathway inhibitor and a pharmaceutically acceptable carrier; and (b) a pharmaceutical composition comprising a programmed death protein 1 (PD-1) inhibitor and a pharmaceutically acceptable carrier. In one embodiment, pharmaceutical compositions (a) and (b) are separate from one another. In another embodiment, pharmaceutical composition (a) comprises one or more doses of an IL-4/IL-13 pathway inhibitor. In another embodiment, at least one dose comprises about 5-1000 mg of an IL-4/IL-13 pathway inhibitor. In another embodiment, pharmaceutical composition (b) comprises one or more doses of a PD-1 inhibitor. In another embodiment, at least one dose comprises about 5-500 mg of a PD-1 inhibitor. In another embodiment, the IL-4/IL-13 pathway inhibitor is selected from the group consisting of an anti-IL-4 antibody, an anti-IL-13 antibody, an anti-IL-4/IL-13 bispecific antibody, an IL-4 receptor (IL-4R) inhibitor, an IL-4 trap, an IL-13 trap, and an anti-IL-4R antibody. In another embodiment, the IL-4/IL-13 pathway inhibitor is selected from the group consisting of dupilumab, pascolizumab, AMG317, MEDI2045, MEDI9314, tralokinumab, lebrikizumab, anrukinzumab, dectrecumab, GSK679586, MEDI7836, romilkimab, IL-4 trap, IL-13 trap, AER-003, and pitrakinra. In another embodiment, the PD-1 inhibitor is selected from the group consisting of an anti-PD-1 antibody, an anti-PD-L1 antibody, and an anti-PD-L2 antibody. In another embodiment, the PD-1 inhibitor is an anti-PD-1 antibody selected from the group consisting of cemiplimab, nivolumab, pembrolizumab, pidilizumab, MEDI0608, BI 754091, PF-06801591, sintilimab, AGEN2034, spartalizumab, camrelizumab, JNJ-63723283, and MCLA-134. In another embodiment, the PD-1 inhibitor is an anti-PD-L1 antibody selected from the group consisting of H1H8314N, avelumab, atezolizumab, durvalumab, MDX-1105, LY3300054, FAZ053, STI-1014, CX-072, KN035, and CK-301.

[013] In another embodiment, the pharmaceutical delivery system further comprises at least one additional therapeutic agent selected from the group consisting of cyclophosphamide, a tumor vaccine, a LAG3 inhibitor, a CTLA-4 inhibitor, a GITR agonist, a CD28 activator, a TIM3 inhibitor, a BTLA inhibitor, a TIGIT inhibitor, a CD38 inhibitor, a CD47 inhibitor, an IDO inhibitor, a VEGF antagonist, an Ang2 inhibitor, a TGFβ inhibitor, an EGFR inhibitor, a VISTA inhibitor, a CD40 agonist, a CSF1R inhibitor, a CCR2 inhibitor, a CXCR4 inhibitor, a CXCR2 inhibitor, a CCR4 inhibitor, a CXCL12 inhibitor, a costimulatory receptor agonist, an anti-tumor antigen antibody, an anti-CD3/anti-CD3 bispecific antibody, CD20, GM-CSF, cytotoxin, chemotherapeutic agent, IL-6R inhibitor, IL-10 inhibitor, oncolytic virus, cytokine, ADC, chimeric antigen receptor T cells, anti-inflammatory drug, NSAID and dietary supplement. In another embodiment, the pharmaceutical delivery system is for use in treating or inhibiting tumor growth. In another embodiment, the tumor comprises a type 2 immune-dependent malignancy.

[014] In another aspect, the present invention relates to a kit comprising a pharmaceutical delivery system comprising: (a) a pharmaceutical composition comprising an IL-4/IL-13 pathway inhibitor and a pharmaceutically acceptable carrier; (b) a pharmaceutical composition comprising a PD-1 inhibitor and a pharmaceutically acceptable carrier; and written instructions for using the IL-4/IL-13 pathway inhibitor in combination with a PD-1 inhibitor to treat or inhibit tumor growth.

[015] In another aspect, the present invention relates to an IL-4/IL-13 pathway inhibitor for use in a method of treating or inhibiting tumor growth in combination with a PD-1 inhibitor, comprising administering to an individual in need thereof a therapeutically effective amount of each inhibitor. In another aspect, the present invention relates to a PD-1 inhibitor for use in a method of treating or inhibiting tumor growth in combination with an IL-4/IL-13 pathway inhibitor, comprising administering to an individual in need thereof a therapeutically effective amount of each inhibitor.

BRIEF DESCRIPTION OF DRAWINGS

[016] Figure 1A is a line graph showing the reduction in tumor volume from tumor volumes of 30-110 ^mm3 at baseline according to the study described in Example 1. The symbol (*) indicates the degree of statistical significance relative to isotype controls (IgG).

[017] Figure 1B is a bar graph showing the percentage change in tumor volume in individual animals from tumor volumes of 30-110 ^mm3 at baseline to day 38, according to the study described in Example 1.

[018] Figure 2A is a line graph showing the reduction in tumor volume from tumor volumes of 50-110 ^mm3 at baseline according to the replication study described in Example 1. The symbol (*) indicates the degree of statistical significance relative to isotype controls (IgG).

[019] Figure 2B is a bar graph showing the percentage change in tumor volume in individual animals from tumor volumes of 50-110 ^mm3 at baseline to day 38, according to the repeat study described in Example 1.

[020] Figure 3A is a bar graph showing IL-4 and IL-13 mRNA expression relative to normal in Balb/c mice implanted subcutaneously (Sub-Cu) or orthotopically (Ortho) with syngeneic pancreatic tumor organoid cells engineered to express oncogenic KRAS with concomitant loss of function of the tumor suppressor P53 (KP cells), as described in Example 3.

[021] Figure 3B is a bar graph showing IL-4 cytokine production in Balb/c mice implanted subcutaneously (Sub-Cu) or orthotopically (Ortho) with syngeneic pancreatic tumor organoid cells engineered to express oncogenic KRAS with concomitant loss of function of the tumor suppressor P53 (KP cells), as described in Example 3.

[022] Figure 4A is a line graph showing the reduction in tumor volume from tumor volumes of 70-100 ^mm3 at baseline to day 40 in accordance with the study described in Example 4. The symbol (*) indicates the degree of statistical significance relative to isotype controls (IgG).

[023] Figure 4B is a bar graph showing the percentage change in tumor volume in individual animals from tumor volumes of 70-100 ^mm3 at baseline to day 40, according to the study described in Example 4.

[024] Figure 4C is a pair of bar graphs showing the levels of the cytokines interferon gamma (IFNγ) and tumor necrosis factor alpha (TNFα) measured in tumor lysates at the end of the study described in Example 4.

[025] Figure 5A is a bar graph showing the percentage change in tumor volume in individual animals from 50-100 ^mm3 tumor volumes at baseline to the end of the study, consistent with the study described in Example 5.

[026] Figure 5B is a bar graph showing the hyaluronic acid (HA) content of pancreatic tumors as determined by histochemical staining using biotinylated HA-binding protein (HABP) in four study groups: ISO/ISO, IL-4R/ISO, ISO/PD-1, and IL-4R/PD-1, according to the study described in Example 5.

[027] Figure 5C is a bar graph showing CD8 ⁺ T cell infiltration in the tumor bed as determined by histological quantification of the tumor infiltrating lymphocyte (TIL) marker CD8 in four study groups: ISO/ISO, IL-4R/ISO, ISO/PD-1, and IL-4R/PD-1, in accordance with the study described in Example 5.

DETAILED DESCRIPTION

[028] It should be understood that the present invention is not limited to the specific methods and experimental conditions described, since such methods and conditions may vary. It should also be understood that the terminology used in this description is intended to describe only particular embodiments and not to be limiting, and that the scope of the present invention will be limited only by the claims.

[029] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the invention pertains. As used herein, the term "about" when used with respect to a specific recited numerical value means that the value may vary from the recited value by no more than 1%. For example, as used herein, the expression "about 100" includes 99 and 101 and all values in between (e.g., 99.1, 99.2, 99.3, 99.4, etc.).

[030] Although any methods and materials similar or equivalent to those described herein can be used in the practice of the present invention, the preferred methods and materials are described below.

[031] The present invention includes methods of treating a cancer comprising administering to an individual in need thereof a therapeutically effective amount of an IL-4/IL-13 pathway inhibitor in combination with a therapeutically effective amount of a programmed death protein 1 (PD-1) inhibitor. In some embodiments, the disclosed methods target type 2 immune-dependent pro-oncogenic mechanisms of cancer development, thereby providing a novel and highly effective method of treating or inhibiting tumor growth, including tumors exhibiting desmoplastic features. In some embodiments, the disclosed methods treat type 2 immune-dependent malignancies using IL-4R blockade or other immunotherapeutic agents targeting type 2 immune response pathways in combination with antineoplastic agents, including, but not limited to, ICBs (e.g., chemotherapy, bispecific antibodies, antibody-drug conjugates (ADCs), and chimeric antigen receptor (CAR) T cells).

[032] As used herein, the term "Type 2 immune gene signature" refers to the upregulation or overexpression of one or more Type 2 cytokines, Type 2 chemokines, Type 2 receptors, and Type 2 downstream target genes, including, but not limited to: IL-13RA2, IL-25, IL-17RB, SERPINB2, CCL24, CEACAM1, CCL1, MUC5B, CCL26, IL-13RA1, POSTN, IL-6R, CCL18, CCL17, FCER2, CCR3, IL-6, CCL8, CRLF2, IL-33, TSLP, IL-4R, CCR4, PTGDR, FCER1A, IL-1RL1, DPP4, IL-4, IL-5, and IL-13. As shown in Example 3, for example, the expression of IL-4 and IL-13 genes was found to be extensively upregulated in an in vivo model of pancreatic cancer, a type 2 immune-dependent malignancy.

[033] As used herein, the term "type 2 immune-dependent malignancies" refers to malignancies or subtypes of malignancies, such as human malignancies or subtypes of human malignancies, that exhibit a strong type 2 immune-dependent gene signature.

[034] As used herein, the term "type 2 immune pathway blockade" refers to an immunotherapy that targets any gene in the type 2 immune gene signature.

Methods of treating or inhibiting tumor growth

[035] The present invention includes methods for treating, ameliorating, or reducing the severity of at least one symptom or sign, or inhibiting the growth of a cancer in an individual. In this aspect, the disclosed methods comprise selecting an individual with a tumor and administering to the individual in need thereof a therapeutically effective amount of an IL-4/IL-13 pathway inhibitor (e.g., an anti-IL-4 antibody, an anti-IL-13 antibody, an anti-IL-4/IL-13 bispecific antibody, an IL-4 receptor (IL-4R) inhibitor, an anti-IL-4R antibody, or any other "IL-4/IL-13 pathway inhibitor" as provided herein) and a therapeutically effective amount of a programmed death protein 1 (PD-1) inhibitor (e.g., an antibody that specifically binds to PD-1, PD-L1 and/or PD-L2, or any other "PD-1 inhibitor" as provided herein).

[036] In the present description, references to any particular anti-IL-4R antibody and/or any particular anti-PD-1 antibody are provided to illustrate a typical IL-4/IL-13 pathway inhibitor and a typical PD-1 inhibitor, respectively, and are not intended to limit the scope of the invention, as combinations of other IL-4/IL-13 pathway inhibitors and PD-1 inhibitors may also be used.

[037] As used herein, the terms "treating," "treating," or the like, mean alleviating or reducing the severity of at least one symptom or sign, eliminating the causative factor of the symptoms on a temporary or permanent basis, delaying or inhibiting tumor growth, reducing the number of tumor cells or tumor load, promoting tumor regression, causing a tumor to shrink, necrose, and/or disappear, preventing tumor recurrence, preventing or inhibiting metastasis, inhibiting metastatic tumor growth, eliminating the need for surgery, shrinking a tumor and facilitating surgery, increasing the resection rate, and/or increasing the survival time of an individual. In many embodiments, the terms "tumor," "malignancy," and "malignant tumor" are used interchangeably and mean one or more tumors.

[038] In some embodiments, the tumor comprises a type 2 immune-dependent malignancy. Non-limiting examples of type 2 immune-dependent malignancies include pancreatic cancer (e.g., PDAC), breast cancer, colorectal cancer, ovarian cancer, lung cancer, lymphoma, and bladder cancer.

[039] As used herein, the term "individual in need thereof" means a human or non-human mammal exhibiting one or more symptoms or signs of a cancer and/or diagnosed with a cancer, including a solid tumor, and in need of treatment. In many embodiments, the terms "individual" and "patient" are used interchangeably. The methods provided herein may include the step of selecting an individual in need thereof. For example, a human may be diagnosed with a primary or metastatic tumor and/or one or more symptoms or signs, including, but not limited to, enlarged lymph nodes, abdominal bloating, chest pain/pressure, unexplained weight loss, fever, night sweats, persistent fatigue, loss of appetite, enlarged spleen, itching. The term includes individuals with a primary, established, or recurrent tumor. In particular embodiments, the expression includes people having a solid tumor and/or in need of treatment. In other embodiments, the expression includes people having a tumor (e.g., lymphomas or leukemias) and/or in need of treatment. The expression also includes individuals with primary or metastatic tumors (malignancies at an advanced stage). The expression also includes individuals with tumors including immune-dependent type 2 malignancy, including, but not limited to, pancreatic cancer, non-small cell lung cancer, squamous cell lung carcinoma, and Hodgkin's lymphoma.

[040] In some embodiments, the expression includes patients with a tumor that is resistant or refractory or inadequately controlled by prior therapy (e.g., treatment with an antineoplastic agent such as IL-4/IL-13 pathway inhibitor monotherapy, PD-1 inhibitor monotherapy, carboplatin, or docetaxel). In some embodiments, the expression includes patients with a tumor that has been treated with one or more lines of prior therapy (e.g., surgically resected) but has subsequently recurred. In some embodiments, the expression includes individuals with a tumor that are not candidates for curative surgery or curative radiation therapy, or for whom conventional antineoplastic therapy is not recommended, such as due to toxic side effects.

[041] In other embodiments, the term "individual in need thereof" includes patients with a cancer that has been treated but has subsequently recurred or metastasized. For example, patients with a tumor may be treated with one or more anti-tumor agents resulting in tumor regression; however, they then experience a relapse of the cancer that is resistant to the one or more anti-tumor agents (e.g., a chemotherapy-resistant cancer) and can be treated with the methods of the present invention. In some embodiments, the term includes patients that exhibit upregulation of at least one cytokine, such as IL-4 and/or IL-13. In some embodiments, the term includes patients that exhibit increased production of at least one cytokine, such as increased production of IL-4. In some embodiments, the term includes patients that exhibit increased hyaluronic acid (HA) content in the tumor.

[042] In some embodiments, the present invention includes methods of treating, delaying, or inhibiting tumor growth. In some embodiments, the present invention includes methods of promoting tumor regression. In some embodiments, the present invention includes methods of reducing the number of tumor cells or reducing tumor burden. In some embodiments, the present invention includes methods of increasing the survival of a treated individual, increasing the response, or increasing the duration of a response. In some embodiments, the present invention includes methods of treating or inhibiting tumor growth in individuals with pancreatic cancer, breast cancer, colorectal cancer, brain cancer, skin cancer, kidney cancer, ovarian cancer, lung cancer (e.g., non-small cell lung cancer), Hodgkin's lymphoma, or bladder cancer.

[043] In some embodiments, the disclosed methods comprise administering a therapeutically effective amount of an IL-4/IL-13 pathway inhibitor and a therapeutically effective amount of a PD-1 inhibitor in combination with an additional therapeutic agent or method of therapy (e.g., a treatment regimen or method of treatment). In some embodiments, the disclosed methods comprise administering an additional therapeutic agent, such as an antineoplastic drug. As used herein, the term "antineoplastic drug" refers to any agent that can be used to treat a cancer, including, but not limited to, cytotoxins and agents such as antimetabolites, alkylating agents, anthracyclines, antibiotics, antimitotic agents, procarbazine, hydroxyurea, asparaginase, corticosteroids, mitotane (O,P′-(DDD)), biologics (e.g., antibodies and interferons), and radioactive agents. For the purposes of the invention, "cytotoxin or cytotoxic agent" also refers to a chemotherapeutic agent and means any agent that is harmful to cells. Examples include Taxol® (paclitaxel), temozolomide, cytochalasin B, gramicidin D, ethidium bromide, emetine, cisplatin, mitomycin, etoposide, teniposide, vincristine, vinblastine, colchicine, doxorubicin, daunorubicin, dihydroxyanthracindione, mitoxantrone, mithramycin, actinomycin D, 1-dehydrotestosterone, glucocorticoids, procaine, tetracaine, lidocaine, propranolol and puromycin and its analogs or homologues. An additional therapeutic agent or method of therapy can be used to enhance the antitumor efficacy, reduce the toxic effects of one or more methods of therapy and/or reduce the dose of one or more therapeutic agents. In various embodiments, the additional therapeutic agent or method of therapy may include one or more of: chemotherapy, cyclophosphamide, surgery, radiation therapy, a tumor vaccine, a lymphocyte activation gene 3 (LAG3) inhibitor (e.g., anti-LAG3 antibodies), a cytotoxic T lymphocyte antigen 4 (CTLA-4) inhibitor (e.g., ipilimumab), a glucocorticoid-inducible tumor necrosis factor receptor (GITR) agonist (e.g., anti-GITR antibodies), an inhibitor of T-cell immunoglobulin and mucin domain-containing protein 3 (TIM3), an inhibitor of B-lymphocyte and T-lymphocyte attenuator (BTLA), an inhibitor of T-cell immunoreceptor with Ig domains and ITIM (TIGIT), an inhibitor of CD47, an inhibitor of indoleamine 2,3-dioxygenase (IDO), a vascular endothelial growth factor (VEGF) antagonist (e.g., a "VEGF decoy" such as aflibercept or other VEGF inhibitory fusion proteins as described in U.S. Patent No. 7,087,411, or an antibody against VEGF or an antigen-binding fragment thereof (e.g., bevacizumab or ranibizumab), or a small molecule inhibitor of VEGF receptor kinases (e.g., sunitinib, sorafenib, or pazopanib)), an angiopoietin-2 (Ang2) inhibitor (e.g., nesvacumab), a transforming growth factor beta (TGFβ) inhibitor, an epidermal growth factor receptor (EGFR) inhibitor (e.g., erlotinib, cetuximab), a costimulatory receptor agonist (e.g., a CD28 agonist), a VISTA inhibitor, a CD38 inhibitor, a CD40 agonist, a CSF1R inhibitor, a CCR2 inhibitor, a CXCR4 inhibitor, a CXCR2 inhibitor, a CCR4 inhibitor, a CXCL12 inhibitor, an antibody against a tumor-associated antigen [e.g., CA9, CA125, melanoma-associated antigen 3 (MAGE3), carcinoembryonic antigen (CEA), vimentin, tumor-M2-PK, prostate-specific antigen (PSA), mucin-1, MART-1, and CA19-9], an anti-CD3/anti-CD20 bispecific antibody, a vaccine (e.g., bacillus Calmette-Guérin), granulocyte-macrophage colony-stimulating factor (GM-CSF), a cytotoxin, a chemotherapeutic agent, an IL-6R inhibitor (e.g., sarilumab), an IL-10 inhibitor, a cytokine such as IL-2, IL-7, IL-12, IL-21 and IL-15, antibody-drug conjugate (ADC) (e.g. anti-CD19-DM4 ADC and anti-DS6-DM4 ADC), oncolytic virus, chimeric antigen receptor T cells (e.g. CD19-targeted T cells), anti-inflammatory drug such as corticosteroid, non-steroidal anti-inflammatory drug (NSAID), and dietary supplement such as antioxidant.

[044] In some embodiments, the IL-4/IL-13 pathway inhibitor and the PD-1 inhibitor may be administered in combination with a therapy comprising a chemotherapeutic agent (e.g., dacarbazine, temozolomide, cyclophosphamide, docetaxel, doxorubicin, daunorubicin, cisplatin, carboplatin, gemcitabine, methotrexate, mitoxantrone, oxaliplatin, paclitaxel, and vincristine) and/or surgery. In some embodiments, the IL-4/IL-13 pathway inhibitor and the PD-1 inhibitor may be administered in combination with an anti-cancer therapy, including, but not limited to, conventional anti-cancer therapies such as chemotherapy, radiation therapy, surgery, or as described elsewhere herein.

[045] In some embodiments, the disclosed method results in increased inhibition of tumor growth, such as increased tumor regression, in a treated individual. For example, the disclosed methods of administering a therapeutically effective amount of an IL-4/IL-13 pathway inhibitor and a therapeutically effective amount of a PD-1 inhibitor results in at least about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, or about 80% greater tumor regression in a treated individual compared to an untreated individual or an individual treated with the inhibitor as monotherapy.

[046] In some embodiments, the disclosed methods of administering a therapeutically effective amount of an IL-4/IL-13 pathway inhibitor and a therapeutically effective amount of a PD-1 inhibitor result in a delay in tumor growth and progression, such as tumor growth can be delayed by about 3 days, greater than 3 days, about 7 days, greater than 7 days, greater than 15 days, greater than 1 month, greater than 3 months, greater than 6 months, greater than 1 year, greater than 2 years, or greater than 3 years in a treated individual compared to an untreated individual or an individual treated with either inhibitor as monotherapy.

[047] In some embodiments, the disclosed methods of administering a therapeutically effective amount of an IL-4/IL-13 pathway inhibitor and a therapeutically effective amount of a PD-1 inhibitor result in a complete disappearance of all evidence of tumor cells ("complete response"), result in at least a 30% or greater reduction in tumor cells or tumor size ("partial response"), or result in a complete or partial disappearance of tumor cells, including new measurable tumors. Tumor reduction can be measured by any means known in the art, such as by radiography, positron emission tomography (PET), computed tomography (CT), magnetic resonance imaging (MRI), cytology, histology, or molecular genetic analysis.

[048] In some embodiments, the disclosed methods of administering a therapeutically effective amount of an IL-4/IL-13 pathway inhibitor and a therapeutically effective amount of a PD-1 inhibitor result in an improvement in overall survival (OS) or progression-free survival (PFS) of an individual compared to an individual treated with the "standard of care" (SOC) (e.g., chemotherapy, surgery, or radiation therapy). In some embodiments, PFS is improved by at least one month, at least 2 months, at least 3 months, at least 4 months, at least 5 months, at least 6 months, at least 7 months, at least 8 months, at least 9 months, at least 10 months, at least 11 months, at least 1 year, at least 2 years, or at least 3 years compared to an individual who is subjected to any one or more SOC therapies. In some embodiments, OS is increased by at least one month, at least 2 months, at least 3 months, at least 4 months, at least 5 months, at least 6 months, at least 7 months, at least 8 months, at least 9 months, at least 10 months, at least 11 months, at least 1 year, at least 2 years, or at least 3 years compared to an individual who is treated with any one or more SOC therapies.

[049] In some embodiments, the disclosed methods of administering a therapeutically effective amount of an IL-4/IL-13 pathway inhibitor and a therapeutically effective amount of a PD-1 inhibitor result in an increased response and duration of response in a treated individual, such as by greater than 2%, greater than 3%, greater than 4%, greater than 5%, greater than 6%, greater than 7%, greater than 8%, greater than 9%, greater than 10%, greater than 20%, greater than 30%, greater than 40%, or greater than 50% compared to an untreated individual or an individual treated with either inhibitor as monotherapy.

[050] In some embodiments, the combination of administered inhibitors is safe and well tolerated by the individual such that there is no increase in undesirable side effects compared to the individual treated with either inhibitor as monotherapy.

[051] In some embodiments, the disclosed methods comprise administering to an individual a therapeutically effective amount of an IL-4/IL-13 pathway inhibitor and a therapeutically effective amount of a PD-1 inhibitor, wherein the individual exhibits upregulation of at least one cytokine, such as IL-4 and/or IL-13. In some embodiments, the individual exhibits higher upregulation of at least one cytokine (e.g., IL-4 and/or IL-13) compared to an individual not in need of anti-tumor treatment.

[052] In some embodiments, the disclosed methods comprise administering to an individual a therapeutically effective amount of an IL-4/IL-13 pathway inhibitor and a therapeutically effective amount of a PD-1 inhibitor, wherein the individual exhibits increased production of at least one cytokine, such as increased production of IL-4. In some embodiments, the individual exhibits increased production of at least one cytokine (e.g., IL-4) compared to an individual not in need of anti-tumor treatment.

[053] In some embodiments, the disclosed methods of administering to an individual a therapeutically effective amount of an IL-4/IL-13 pathway inhibitor and a therapeutically effective amount of a PD-1 inhibitor result in significantly increased interferon gamma (IFNγ) production compared to an individual treated with either inhibitor as monotherapy. In some such embodiments, these disclosed methods further result in increased tumor regression and anti-tumor activity.

[054] In some embodiments, the disclosed methods comprise administering to an individual a therapeutically effective amount of an IL-4/IL-13 pathway inhibitor and a therapeutically effective amount of a PD-1 inhibitor, wherein the individual exhibits increased hyaluronic acid (HA) levels in a tumor. In some such embodiments, the individual exhibits higher HA levels than an individual who does not have a type 2 immune-dependent malignancy.

[055] In some embodiments, the disclosed methods of administering to an individual a therapeutically effective amount of an IL-4/IL-13 pathway inhibitor and a therapeutically effective amount of a PD-1 inhibitor result in a decrease in hyaluronic acid (HA) content in a tumor.

[056] In some embodiments, the disclosed methods of administering to an individual a therapeutically effective amount of an IL-4/IL-13 pathway inhibitor and a therapeutically effective amount of a PD-1 inhibitor result in a significantly increased tumor infiltrating lymphocyte (TIL) density and, thus, an increase in the cytotoxic anti-tumor efficacy of the combination treatment. In some such embodiments, the disclosed methods further result in increased tumor regression and anti-tumor activity.

IL-4/IL-13 pathway inhibitors

[057] The methods provided herein comprise administering to an individual in need thereof a therapeutically effective amount of an IL-4/IL-13 pathway inhibitor. As used herein, an "IL-4/IL-13 pathway inhibitor" (also referred to herein as an "IL-4/IL-13 pathway antagonist," "IL-4/IL-13 pathway blocker," etc.) is any agent that inhibits or reduces at least one of: (i) IL-4 and/or IL-13 binding to their respective receptors; (ii) IL-4 and/or IL-13 signaling and/or activity; and/or (iii) downstream signaling/activity resulting from IL-4 and/or IL-13 binding to their respective receptors. Non-limiting examples of IL-4/IL-13 pathway inhibitors include anti-IL-4 antibodies (e.g., those described in U.S. Patent No. 7,740,843 and U.S. Patent Application Publication Nos. 20100297110, 20160207995), anti-IL-13 antibodies (e.g., those described in U.S. Patent Nos. 7,501,121, 7,674,459, 7,807,788, 7,910,708, 7,915,388, 7,935,343, 8,088,618, 8,691,233, 9,605,065, U.S. Patent Application Publication Nos. 2006,007,3148, 2008,004,420, and EP2627673B1), bispecific antibodies that bind to IL-4 and IL-13 (e.g., the antibodies described in U.S. Patent No. 8,388,965, U.S. Patent Application Publication Nos. 20110008345, 20130251718, 20160207995), and IL-4 receptor (IL-4R) inhibitors (described below). Accordingly, the portions of the publications cited herein that define IL-4/IL-13 pathway inhibitors are hereby incorporated by reference.

[058] In some embodiments, the IL-4/IL-13 pathway inhibitor may be an antibody, a small molecule, a nucleic acid, a polypeptide, or a functional fragment or variant thereof. Non-limiting examples of suitable IL-4/IL-13 pathway inhibitor antibodies include anti-IL-4 antibodies, anti-IL-13 antibodies and bispecific anti-IL-4/IL-13 antibodies, anti-IL-4R antibodies, and antigen-binding fragments of any of the above antibodies. Other non-limiting examples of suitable IL-4/IL-13 pathway inhibitors include: RNAi molecules such as IL-4 RNAi and IL-13 RNAi molecules, antisense molecules such as IL-4 antisense RNA and IL-13 antisense RNA, and dominant negative proteins such as IL-4 dominant negative protein, IL-13 dominant negative protein.

[059] As used herein, an "IL-4R inhibitor" (also referred to herein as an "IL-4/IL-13 pathway inhibitor," "IL-4Rα antagonist," "IL-4R blocker," "IL-4Rα blocker," etc.) is any agent that binds to or interacts with IL-4Rα or an IL-4R ligand and inhibits or reduces the normal biological signaling function of the IL-4 receptor type 1 and/or type 2. The IL-4 receptor type 1 is a dimeric receptor comprising an IL-4Rα chain and a γc chain. The IL-4 receptor type 2 is a dimeric receptor comprising an IL-4Rα chain and an IL-13Rα1 chain. IL-4 type 1 receptors interact with and are stimulated by IL-4, while IL-4 type 2 receptors interact with and are stimulated by IL-4 and IL-13. Thus, IL-4R inhibitors that may be used in the methods of the present invention may function by blocking IL-4-mediated signaling, IL-13-mediated signaling, or IL-4- and IL-13-mediated signaling. Thus, IL-4R inhibitors of the present invention may prevent the interaction of IL-4 and/or IL-13 with a type 1 or type 2 receptor.

[060] Non-limiting examples of categories of IL-4R inhibitors include IL-4 muteins (e.g., pitrakinra), small molecule IL-4R inhibitors, IL-4R aptamers, peptide inhibitors of IL-4R (e.g., "peptibody" molecules), "receptor antibodies" (e.g., engineered molecules comprising the ligand binding domain of an IL-4R component), and antibodies or antigen-binding fragments of antibodies that specifically bind to human IL-4Rα. Within the scope of the invention, IL-4R inhibitors also include antigen-binding proteins that specifically bind to IL-4 and/or IL-13.

[061] Other non-limiting examples of suitable IL-4/IL-13 pathway inhibitors that can be used in the context of the present invention include, for example, pitrakinra (AER-001; BAY-16-9996), aeroderm (AER-003), and antibodies known in the art as dupilumab, pascolizumab, AMG-317, MILR1444A, CAT-354, QAX576, anrukinzumab (IMA-638), ISIS-369645 (AIR-645), IMA-026, APG-201, CNTO-607, MK-6105, MEDI9314, MEDI2045, tralokinumab, lebrikizumab, romilkimab, and DOM-0910.

Anti-IL-4Rα antibodies and their antigen-binding fragments

[062] In some exemplary embodiments of the present invention, the IL-4/IL-13 pathway inhibitor is an anti-IL-4Rα antibody or antigen-binding fragment thereof. The term "antibody" as used herein includes immunoglobulin molecules comprising four polypeptide chains, two heavy (H) chains and two light (L) chains, linked together by disulfide bonds, as well as multimers thereof (e.g., IgM). In an exemplary antibody, each heavy chain comprises a heavy chain variable region (abbreviated herein as HCVR or V _H ) and a heavy chain constant region. The heavy chain constant region comprises three domains, _CH 1, _CH 2, and _CH 3. Each light chain comprises a light chain variable region (abbreviated herein as LCVR or V _L ) and a light chain constant region. The light chain constant region comprises one domain ( _CL 1). The V _H and V _L regions can be further divided into regions of hypervariability, referred to as complementarity determining regions (CDRs), interspersed with regions that are more conserved, referred to as framework regions (FRs). Each V _H and V _L consists of three CDRs and four FRs, arranged from amino terminus to carboxy terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. In other embodiments of the present invention, the FRs of an antibody (or antigen-binding portion thereof) can be identical to human germline sequences or can be naturally or artificially modified. A consensus amino acid sequence can be determined based on parallel analysis of two or more CDRs.

[063] The term "antibody" throughout the present disclosure includes antigen-binding fragments thereof, i.e., antigen-binding fragments of complete antibody molecules. The terms "antigen-binding portion" of an antibody, "antigen-binding fragment" of an antibody, and the like throughout the present disclosure include any naturally occurring, enzymatically produced, synthetic, or genetically engineered polypeptide or glycoprotein that specifically binds to an antigen to form a complex. Antigen-binding fragments of an antibody can be obtained, for example, from full-length antibody molecules by any suitable standard methods, such as proteolytic cleavage or recombinant genetic engineering techniques involving manipulation and expression of DNA encoding the variable and, optionally, constant domains of the antibody. Such DNA is known and/or readily available, such as from commercial sources, DNA libraries (including, for example, antibody phage libraries), or can be synthesized. DNA can be sequenced and manipulated chemically or by molecular biology techniques, such as to arrange one or more variable and/or constant domains into a suitable configuration, or to insert codons, create cysteine residues, modify, add or delete amino acids, etc.

[064] Non-limiting examples of antigen-binding fragments include: (i) Fab fragments; (ii) F(ab') ₂ fragments; (iii) Fd fragments; (iv) Fv fragments; (v) single-chain Fv molecules (scFv); (vi) dAb fragments; and (vii) minimal recognition units consisting of amino acid residues that mimic a hypervariable region of an antibody (e.g., an isolated complementarity determining region (CDR), such as a CDR3 peptide), or a constrained FR3-CDR3-FR4 peptide. Other engineered molecules such as domain-specific antibodies, single domain antibodies, domain-deleted antibodies, chimeric antibodies, CDR-grafted antibodies, diabodies, triabodies, tetrabodies, minibodies, nanobodies (e.g., monovalent nanobodies, bivalent nanobodies, etc.), small modular immunopharmaceuticals (SMIPs), and shark IgNAR variable domains are also included within the term "antigen-binding moiety" within the scope of the invention.

[065] An antigen-binding fragment of an antibody will typically comprise at least one variable domain. The variable domain may be of any size or amino acid composition, and will typically comprise at least one CDR adjacent to or in frame with one or more framework sequences. In antigen-binding fragments comprising a V _H domain linked to a V _L domain, the V _H and V _L domains may be arranged relative to one another in any suitable order. For example, the variable region may be dimeric and include V _H -V _H , V _H -V _L , or V _L -V _L dimers. Alternatively, an antigen-binding fragment of an antibody may comprise a monomeric V _H or V _L domain.

[066] In some embodiments, an antigen-binding fragment of an antibody may comprise at least one variable domain covalently linked to at least one constant domain. Non-limiting examples of variable and constant domain configurations that may be found in an antigen-binding fragment of an antibody of the present invention include: (i) V _{H -CH} 1; (ii) V _H -CH ₂ ; (iii) V _{H -CH 3} ; (iv) V _H -CH 1- _CH 2; (v) V _H -CH 1- _CH 2- _CH 3; (vi) _{V H} -CH 2- _CH 3; ₍ vii) V _{H -CH} ; (viii) V _{L -CH} 1; (ix) V _L -CH ₂ ; (x) V _{L -CH} 3; (xi) V _{L -CH} 1- _CH 2; (xii) V _{L -CH1} - _CH2 - _CH3 ; (xiii) V _{L -CH2} - _CH3 ; and (xiv) V _{L -CH} . In any configuration of the variable and constant domains, including any of the above exemplary configurations, the variable and constant domains can be linked directly to each other or via a full or partial hinge or linker region. The hinge region can be composed of at least 2 (e.g., 5, 10, 15, 20, 40, 60 or more) amino acids, resulting in flexible or semi-flexible linkage between adjacent variable and/or constant domains in a single polypeptide molecule. In addition, the antigen-binding fragment of an antibody of the present invention may comprise a homodimer or heterodimer (or other multimer) of any of the above configurations of variable and constant domains in non-covalent association with each other and/or with one or more monomeric V _H or V _L domains (e.g., via disulfide bonds).

[067] The term "antibody" throughout the present disclosure also includes multispecific (e.g., bispecific) antibodies. A multispecific antibody or antigen-binding fragment of an antibody will typically comprise at least two different variable domains, wherein each variable domain can specifically bind to a separate antigen or a different epitope on the same antigen. Any multispecific antibody format can be adapted for use in the context of an antibody or antigen-binding fragment of an antibody of the present invention using conventional methods available in the art. For example, the present invention includes methods comprising the use of bispecific antibodies, wherein one arm of the immunoglobulin is specific for IL-4Rα or a fragment thereof, and the other arm of the immunoglobulin is specific for a second therapeutic target or is conjugated to a therapeutic moiety. Non-limiting examples of bispecific formats that can be used in the context of the present invention include, for example, scFv or diabody-based bispecific formats, IgG-scFv fusion proteins, dual variable domain Ig (DVD-Ig), quadroma, knob-into-hole, common light chain (e.g., common light chain with knobs-into-hole, etc.), CrossMab, CrossFab, (SEED)-antibody, leucine zipper, duobody, IgG1/IgG2, dual-acting Fab (DAF)-IgG, and Mab ² (for a review of the above formats, see, for example, Klein et al . 2012, mAbs 4(6):653-663 and references cited therein). Bispecific antibodies can also be constructed using peptide/nucleic acid conjugation, for example, where unnatural amino acids with orthogonal chemical reactivity are used to generate site-specific antibody-oligonucleotide conjugates that then self-assemble into multimeric complexes with defined composition, valence, and geometry (see, for example, Kazane et al ., J. Am. Chem. Soc .,2013, 135(1):340–46).

[068] The antibodies used in the methods of the present invention can be human antibodies. The term "human antibody" throughout the present disclosure is intended to include antibodies having variable and constant regions derived from human germline immunoglobulin sequences. However, the human antibodies of the present invention can include amino acid residues not coding for human germline immunoglobulin sequences (e.g., mutations introduced by random or site-directed mutagenesis in vitro or by somatic mutation in vivo ), such as in the CDRs and in particular CDR3. However, the term "human antibody" throughout the present disclosure is not intended to include antibodies in which CDR sequences derived from the germline of another mammalian species, such as a mouse, are grafted onto human framework sequences.

[069] The antibodies used in the methods of the present invention may be recombinant human antibodies. The term "recombinant human antibody" throughout the present disclosure is intended to include all human antibodies produced, expressed, generated, or isolated by recombinant methods, such as antibodies expressed using a recombinant expression vector that is transfected into a host cell (described below), antibodies isolated from a recombinant, combinatorial human antibody library (described below), antibodies isolated from an animal (e.g., a mouse) transgenic for human immunoglobulin genes (see, for example, Taylor et al. (1992)Nucleic Acid Res., 20:6287-6295), or antibodies produced, expressed, generated, or isolated by any other means involving splicing of human immunoglobulin gene sequences with other DNA sequences. Such recombinant human antibodies comprise variable and constant regions derived from human germline immunoglobulin sequences. However, in some embodiments, such recombinant human antibodies are subjected to mutagenesisin vitro(or, if an animal transgenic for human Ig sequences is used, somatic mutagenesisin vivo), and thus the amino acid sequences of regions V_Hand V_Lrecombinant antibodies are sequences that, although derived from V sequences_Hand V_Lhuman germline antibodies and are related to them may not naturally exist in the human germline antibody repertoirein vivo.

[070] In some embodiments, antibodies used in the methods of the present invention specifically bind to IL-4Rα. The term "specifically binds" or the like means that the antibody or antigen-binding fragment thereof forms a complex with the antigen that is relatively stable under physiological conditions. Methods for determining whether an antibody specifically binds to an antigen are well known in the art and include, for example, equilibrium dialysis, surface plasmon resonance, and the like. For example, an antibody that "specifically binds" to IL-4Rα within the scope of the invention includes antibodies that bind to IL-4Rα or a portion thereof with a K _D of less than about 500 nM, less than about 300 nM, less than about 200 nM, less than about 100 nM, less than about 90 nM, less than about 80 nM, less than about 70 nM, less than about 60 nM, less than about 50 nM, less than about 40 nM, less than about 30 nM, less than about 20 nM, less than about 10 nM, less than about 5 nM, less than about 4 nM, less than about 3 nM, less than about 2 nM, less than about 1 nM, or less than about 0.5 nM, as measured by surface plasmon resonance analysis. However, an isolated antibody that specifically binds to human IL-4Rα may have cross-reactivity with other antigens, such as IL-4Rα molecules from other biological species (non-human).

[071] In some exemplary embodiments, the IL-4/IL-13 pathway inhibitor is an anti-IL-4Rα antibody or antigen-binding fragment thereof that comprises a heavy chain variable region (HCVR), a light chain variable region (LCVR), and/or complementarity determining regions (CDRs) comprising any of the amino acid sequences of the anti-IL-4R antibodies as described in U.S. Patent No. 7,608,693. In some exemplary embodiments, an anti-IL-4Rα antibody or antigen-binding fragment thereof that may be used in the context of the methods of the invention comprises heavy chain complementarity determining regions (HCDRs) of a heavy chain variable region (HCVR) comprising the amino acid sequence of SEQ ID NO: 1 and light chain complementarity determining regions (LCDRs) of a light chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO: 2. In some embodiments, the anti-IL-4Rα antibody or antigen-binding fragment thereof comprises three HCDRs (HCDR1, HCDR2, and HCDR3) and three LCDRs (LCDR1, LCDR2, and LCDR3), wherein HCDR1 comprises the amino acid sequence of SEQ ID NO: 3; HCDR2 comprises the amino acid sequence of SEQ ID NO: 4; HCDR3 comprises the amino acid sequence of SEQ ID NO: 5; LCDR1 comprises the amino acid sequence of SEQ ID NO: 6; LCDR2 comprises the amino acid sequence of SEQ ID NO: 7; and LCDR3 comprises the amino acid sequence of SEQ ID NO: 8. In other embodiments, the anti-IL-4R antibody or antigen-binding fragment thereof comprises an HCVR comprising SEQ ID NO: 1 and an LCVR comprising SEQ ID NO: 2. In some embodiments, the methods of the present invention comprise using an anti-IL-4R antibody, wherein the antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 9. In some embodiments, the anti-IL-4R antibody comprises a light chain comprising the amino acid sequence of SEQ ID NO: 10. An example of an antibody comprising an HCVR comprising the amino acid sequence of SEQ ID NO: 1 and an LCVR comprising the amino acid sequence of SEQ ID NO: 2 is the fully human anti-IL-4R antibody known as dupilumab (DUPIXENT®). In some exemplary embodiments, the methods of the present invention comprise using dupilumab or a bioequivalent thereof. The term "bioequivalent" with respect to dupilumab refers to anti-IL-4R antibodies or IL-4R binding proteins or fragments thereof that are pharmaceutical equivalents or pharmaceutical alternatives that have a rate and/or extent of absorption that does not show significant differences relative to dupilumab when administered at the same molar dose under similar experimental conditions in a single dose or multiple doses. In the context of the present invention, the term refers to antigen-binding proteins that bind to IL-4R that do not have clinically significant differences relative to dupilumab in their safety, purity and/or potency.

[072] In some embodiments of the present invention, an anti-human IL-4R antibody or antigen-binding fragment thereof comprises an HCVR having 90%, 95%, 98%, or 99% sequence identity to SEQ ID NO: 1.

[073] In some embodiments of the present invention, an anti-human IL-4R antibody or antigen-binding fragment thereof comprises an LCVR having 90%, 95%, 98%, or 99% sequence identity to SEQ ID NO: 2.

[074] In some embodiments, the anti-human IL-4R antibody or antigen-binding fragment thereof comprises an HCVR comprising the amino acid sequence of SEQ ID NO: 1, having no more than 5 amino acid substitutions. In some embodiments, the anti-human IL-4R antibody or antigen-binding fragment thereof comprises an LCVR comprising the amino acid sequence of SEQ ID NO: 2, having no more than 2 amino acid substitutions.

[075] Sequence identity can be measured by methods known in the art (e.g., GAP, BESTFIT, and BLAST).

[076] The present invention also includes the use of anti-IL-4R antibodies in methods of treating cancer, wherein the anti-IL-4R antibodies comprise variants of any of the HCVR, LCVR, and/or CDR amino acid sequences provided herein having one or more conservative amino acid substitutions. For example, the present invention includes the use of anti-IL-4R antibodies having HCVR, LCVR, and/or CDR amino acid sequences with, e.g., 10 or less, 8 or less, 6 or less, 4 or less, etc. conservative amino acid substitutions relative to any of the HCVR, LCVR, and/or CDR amino acid sequences provided herein.

[077] Other anti-IL-4Rα antibodies that can be used in the context of the methods of the present invention include, for example, the antibody known in the art as AMG317 (Corren et al ., 2010, Am J Respir Crit Care Med., 181 (8):788-796), or MEDI 9314, or any of the anti-IL-4Rα antibodies described in U.S. Patent No. 7,186,809, U.S. Patent No. 7,605,237, U.S. Patent No. 7,638,606, U.S. Patent No. 8,092,804, U.S. Patent No. 8,679,487, or U.S. Patent No. 8,877,189. Portions of the publications cited herein that define anti-IL-4Rα antibodies are hereby incorporated by reference herein. links.

[078] Anti-IL-4Rα antibodies used in the context of the methods of the present invention may have pH-dependent binding characteristics. For example, an anti-IL-4Rα antibody for use in the methods of the present invention may exhibit reduced binding to IL-4Rα at acidic pH compared to neutral pH. Alternatively, an anti-IL-4Rα antibody of the present invention may exhibit increased binding to its antigen at acidic pH compared to neutral pH. The term "acidic pH" includes pH values of less than about 6.2, such as about 6.0, 5.95, 5.9, 5.85, 5.8, 5.75, 5.7, 5.65, 5.6, 5.55, 5.5, 5.45, 5.4, 5.35, 5.3, 5.25, 5.2, 5.15, 5.1, 5.05, 5.0 or less. As used herein, the term "neutral pH" means a pH of from about 7.0 to about 7.4. The term "neutral pH" includes pH values of about 7.0, 7.05, 7.1, 7.15, 7.2, 7.25, 7.3, 7.35 and 7.4.

[079] In some instances, "reduced binding to IL-4Rα at acidic pH compared to neutral pH" is expressed in terms of the ratio of the _KD value of an antibody binding to IL-4Rα at acidic pH to the _KD value of an antibody binding to IL-4Rα at neutral pH (or vice versa). For example, an antibody or antigen-binding fragment thereof may be considered to exhibit "reduced binding to IL-4Rα at acidic pH compared to neutral pH" for purposes of the present invention if the antibody or antigen-binding fragment thereof exhibits a _KD ratio at acidic/neutral pH of about 3.0 or greater. In some exemplary embodiments, the K _D ratio at acidic/neutral pH of an antibody or antigen-binding fragment of the present invention can be about 3.0, 3.5, 4.0, 4.5, 5.0, 5.5, 6.0, 6.5, 7.0, 7.5, 8.0, 8.5, 9.0, 9.5, 10.0, 10.5, 11.0, 11.5, 12.0, 12.5, 13.0, 13.5, 14.0, 14.5, 15.0, 20.0, 25.0, 30.0, 40.0, 50.0, 60.0, 70.0, 100.0 or more.

[080] Antibodies with pH-dependent binding characteristics can be obtained, for example, by screening a population of antibodies for decreased (or increased) binding to a particular antigen at an acidic pH compared to a neutral pH. In addition, modifications of the antigen-binding domain at the amino acid level can result in antibodies with pH-dependent characteristics. For example, by replacing one or more amino acids of the antigen-binding domain (e.g., in the CDRs) with a histidine residue, one can obtain an antibody with decreased antigen binding at an acidic pH relative to a neutral pH. As used herein, the term "acidic pH" means a pH of 6.0 or less.

PD-1 inhibitors

[081] The methods provided herein comprise administering to an individual in need thereof a therapeutically effective amount of a PD-1 inhibitor. As used herein, the term "PD-1 inhibitor" refers to any molecule capable of inhibiting, blocking, eliminating, or interfering with the activity or expression of PD-1. In some embodiments, the PD-1 inhibitor may be an antibody, a small molecule, a nucleic acid, a polypeptide, or a functional fragment or variant thereof. Non-limiting examples of suitable PD-1 inhibitor antibodies include anti-PD-1 antibodies and antigen-binding fragments thereof, anti-PD-L1 antibodies and antigen-binding fragments thereof, and anti-PD-L2 antibodies and antigen-binding fragments thereof. Other non-limiting examples of suitable PD-1 inhibitors include RNAi molecules such as anti-PD-1 RNAi, anti-PD-L1 RNAi, and anti-PD-L2 RNAi molecules, antisense molecules such as anti-PD-1 antisense RNA, anti-PD-L1 antisense RNA, and anti-PD-L2 antisense RNA, and dominant negative proteins such as dominant negative PD-1 protein, dominant negative PD-L1 protein, and dominant negative PD-L2 protein. Some examples of the above PD-1 inhibitors are described, for example, in U.S. Patent Nos. 9,308,236, 10,011,656, and U.S. Patent Application No. 20170290808, the portions of which defining PD-1 inhibitors are hereby incorporated by reference.

Antibodies against PD-1 and its antigen-binding fragments

[082] In some embodiments, the PD-1 inhibitors used in the methods provided herein are antibodies or antigen-binding fragments thereof that specifically bind to PD-1.

[083] The terms "antibody,""antigen-bindingfragment,""humanantibody,""recombinantantibody," and other related terms are defined below. In the context of anti-PD-1 antibodies and antigen-binding fragments thereof, the present invention includes the use of bispecific antibodies, wherein one arm of the immunoglobulin is specific for PD-1 or a fragment thereof, and the other arm of the immunoglobulin is specific for a second therapeutic target or is conjugated to a therapeutic moiety. Non-limiting examples of bispecific formats that can be used in the context of the present invention include, for example, scFv or diabody-based bispecific formats, IgG-scFv fusion proteins, dual variable domain Ig ((DVD)-Ig), quadroma, knob-into-hole, common light chain (e.g., knob-into-hole common light chain, etc.), CrossMab, CrossFab, (SEED)-body, leucine zipper, duobody, IgG1/IgG2, dual-acting Fab (DAF)-IgG, and Mab ² bispecific formats (for a review of the above formats, see, for example, Klein et al . 2012, mAbs 4(6):653-663 and references cited therein). Bispecific antibodies can also be constructed using peptide/nucleic acid conjugation, for example, where unnatural amino acids with orthogonal chemical reactivity are used to generate site-specific antibody-oligonucleotide conjugates that then self-assemble into multimeric complexes with defined composition, valence, and geometry (see, for example, Kazane et al ., J. Am. Chem. Soc .,2013, 135(1):340–46).

[084] The term "specifically binds" or the like means that the antibody or antigen-binding fragment thereof forms a complex with the antigen that is relatively stable under physiological conditions. Methods for determining whether an antibody specifically binds to an antigen are well known in the art and include, for example, equilibrium dialysis, surface plasmon resonance, and the like. For example, an antibody that "specifically binds" to PD-1, as used herein, includes antibodies that bind to PD-1 or a portion thereof with a K _D of less than about 500 nM, less than about 300 nM, less than about 200 nM, less than about 100 nM, less than about 90 nM, less than about 80 nM, less than about 70 nM, less than about 60 nM, less than about 50 nM, less than about 40 nM, less than about 30 nM, less than about 20 nM, less than about 10 nM, less than about 5 nM, less than about 4 nM, less than about 3 nM, less than about 2 nM, less than about 1 nM, or less than about 0.5 nM, as measured by surface plasmon resonance analysis. However, an isolated antibody that specifically binds to human PD-1 may have cross-reactivity with other antigens, such as PD-1 molecules from other biological species (non-human).

[085] In some exemplary embodiments, the anti-PD-1 antibody or antigen-binding fragment thereof comprises a heavy chain variable region (HCVR), a light chain variable region (LCVR), and/or complementarity determining regions (CDRs) comprising the amino acid sequences of any of the anti-PD-1 antibodies described in U.S. Patent No. 9,987,500, hereby incorporated by reference in its entirety. In some exemplary embodiments, an anti-PD-1 antibody or antigen-binding fragment thereof that may be used in the context of the present invention comprises a heavy chain complementarity determining region (HCDR) of a heavy chain variable region (HCVR) comprising the amino acid sequence of SEQ ID NO: 11 and a light chain complementarity determining region (LCDR) of a light chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO: 12. In some embodiments, an anti-PD-1 antibody or antigen-binding fragment thereof comprises three HCDRs (HCDR1, HCDR2, and HCDR3) and three LCDRs (LCDR1, LCDR2, and LCDR3), wherein HCDR1 comprises the amino acid sequence of SEQ ID NO: 13; HCDR2 comprises the amino acid sequence of SEQ ID NO: 14; HCDR3 comprises the amino acid sequence of SEQ ID NO: 15; LCDR1 comprises the amino acid sequence of SEQ ID NO: 16; LCDR2 comprises the amino acid sequence SEQ ID NO: 17; and LCDR3 comprises the amino acid sequence of SEQ ID NO: 18. In other embodiments, the anti-PD-1 antibody or antigen-binding fragment thereof comprises an HCVR comprising SEQ ID NO: 11 and an LCVR comprising SEQ ID NO: 12. In some embodiments, the methods of the present invention comprise using an anti-PD-1 antibody, wherein the antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 19. In some embodiments, the anti-PD-1 antibody comprises a light chain comprising the amino acid sequence of SEQ ID NO: 20. An example of an antibody comprising an HCVR comprising the amino acid sequence of SEQ ID NO: 11 and an LCVR comprising the amino acid sequence of SEQ ID NO: 12 is the fully human anti-PD-1 antibody known as cemiplimab (also known as REGN2810) (LIBTAYO®).

[086] In some exemplary embodiments, the methods of the present invention comprise using REGN2810 or a bioequivalent thereof. As used herein, the term "bioequivalent" with respect to anti-PD-1 antibodies refers to anti-PD-1 antibodies or PD-1 binding proteins or fragments thereof that are pharmaceutical equivalents or pharmaceutical alternatives that have a rate and/or extent of absorption that does not exhibit significant differences relative to a reference antibody (e.g., REGN2810) when administered at the same molar dose under similar experimental conditions in a single dose or multiple doses; the term "bioequivalent" also includes antigen-binding proteins that bind to PD-1 and that do not exhibit clinically significant differences relative to REGN2810 in terms of safety, purity, and/or potency.

[087] In some embodiments of the present invention, an anti-human PD-1 antibody or antigen-binding fragment thereof comprises an HCVR having 90%, 95%, 98%, or 99% sequence identity to SEQ ID NO: 11.

[088] In some embodiments of the present invention, an anti-human PD-1 antibody or antigen-binding fragment thereof comprises an LCVR having 90%, 95%, 98%, or 99% sequence identity to SEQ ID NO: 12.

[089] In some embodiments, the anti-human PD-1 antibody or antigen-binding fragment thereof comprises an HCVR comprising the amino acid sequence of SEQ ID NO: 11 having no more than 5 amino acid substitutions. In some embodiments, the anti-human PD-1 antibody or antigen-binding fragment thereof comprises an LCVR comprising the amino acid sequence of SEQ ID NO: 12 having no more than 2 amino acid substitutions.

[090] Sequence identity can be measured by methods known in the art (e.g., GAP, BESTFIT, and BLAST).

[091] The present invention also includes the use of anti-PD-1 antibodies in methods of treating cancer, wherein the anti-PD-1 antibodies comprise variants of any of the HCVR, LCVR, and/or CDR amino acid sequences provided herein comprising one or more conservative amino acid substitutions. For example, the present invention includes the use of anti-PD-1 antibodies comprising HCVR, LCVR, and/or CDR amino acid sequences with, for example, 10 or less, 8 or less, 6 or less, 4 or less, etc. conservative amino acid substitutions relative to any of the HCVR, LCVR, and/or CDR amino acid sequences provided herein.

[092] Other anti-PD-1 antibodies that can be used in the context of the methods of the present invention include, for example, those known in the art as nivolumab, pembrolizumab, MEDI0608, pidilizumab, BI 754091, spartalizumab (also known as PDR001), camrelizumab (also known as SHR-1210), sintilimab, AGEN2034, JNJ-63723283, MCLA-134, or any of the anti-PD-1 antibodies described in U.S. Patent Nos. 6,808,710, 7,488,802, 8,008,449, 8,168,757, 8,354,509, 8,609,089, 8,686,119, 8,779,105, 8,900,587, and 9987500 and in Patent Publication Nos. WO2006/121168, WO2009/114335. Portions of the publications cited herein that define anti-PD-1 antibodies are hereby incorporated by reference.

[093] Anti-PD-1 antibodies used in the context of the methods of the present invention may have pH-dependent binding characteristics. For example, an anti-PD-1 antibody for use in the methods of the present invention may exhibit reduced binding to PD-1 at acidic pH compared to neutral pH. Alternatively, an anti-PD-1 antibody of the present invention may exhibit increased binding to its antigen at acidic pH compared to neutral pH. The term "acidic pH" includes pH values of less than about 6.2, such as about 6.0, 5.95, 5.9, 5.85, 5.8, 5.75, 5.7, 5.65, 5.6, 5.55, 5.5, 5.45, 5.4, 5.35, 5.3, 5.25, 5.2, 5.15, 5.1, 5.05, 5.0 or less. As used herein, the term "neutral pH" means a pH of from about 7.0 to about 7.4. The term "neutral pH" includes pH values of about 7.0, 7.05, 7.1, 7.15, 7.2, 7.25, 7.3, 7.35 and 7.4.

[094] In some instances, "reduced binding to PD-1 at acidic pH compared to neutral pH" is expressed in terms of the ratio of the K _D value of the PD-1 binding antibody at acidic pH to the K _D value of the PD-1 binding antibody at neutral pH (or vice versa). For example, an antibody or antigen-binding fragment thereof may be considered to exhibit "reduced binding to PD-1 at acidic pH compared to neutral pH" for purposes of the present invention if the antibody or antigen-binding fragment thereof exhibits an acidic/neutral pH K _D ratio of about 3.0 or greater. In some exemplary embodiments, the K _D ratio at acidic/neutral pH for an antibody or antigen-binding fragment of the present invention can be about 3.0, 3.5, 4.0, 4.5, 5.0, 5.5, 6.0, 6.5, 7.0, 7.5, 8.0, 8.5, 9.0, 9.5, 10.0, 10.5, 11.0, 11.5, 12.0, 12.5, 13.0, 13.5, 14.0, 14.5, 15.0, 20.0, 25.0, 30.0, 40.0, 50.0, 60.0, 70.0, 100.0 or more.

[095] Antibodies with pH-dependent binding characteristics can be obtained, for example, by screening a population of antibodies for decreased (or increased) binding to a particular antigen at an acidic pH compared to a neutral pH. In addition, modifications of the antigen-binding domain at the amino acid level can result in antibodies with pH-dependent characteristics. For example, by replacing one or more amino acids of the antigen-binding domain (e.g., in the CDRs) with a histidine residue, one can obtain an antibody with decreased antigen binding at an acidic pH relative to a neutral pH. As used herein, the term "acidic pH" means a pH of 6.0 or less.

Antibodies against PD-L1 and its antigen-binding fragments

[096] In some embodiments, the PD-1 inhibitors used in the methods provided herein are antibodies or antigen-binding fragments thereof that specifically bind to PD-L1. For example, an antibody that "specifically binds" to PD-L1, as used herein, includes antibodies that bind to PD-L1 or a portion thereof with a _KD of about 1× ^10-8 M or less (e.g., a lower _KD means stronger binding). The term "high-affinity" anti-PD-L1 antibody refers to a mAb having a binding affinity for PD-L1, expressed as a _KD of at least ^10-8 M, preferably ^10-9 M, more preferably ^10-10 M, even more preferably ^10-11 M, even more preferably ^10-12 M, as measured by surface plasmon resonance, such as BIACORE™ or by affinity determination in solution by ELISA. However, an isolated antibody that specifically binds to human PD-L1 may have cross-reactivity with other antigens, such as PD-L1 molecules from another biological species (non-human).

[097] In some exemplary embodiments, the anti-PD-L1 antibody, or antigen-binding fragment thereof, comprises a heavy chain variable region (HCVR), a light chain variable region (LCVR), and/or complementarity determining regions (CDRs) comprising the amino acid sequences of any of the anti-PD-L1 antibodies described in U.S. Patent No. 9,938,345, hereby incorporated by reference in its entirety. In some exemplary embodiments, an anti-PD-L1 antibody or antigen-binding fragment thereof that may be used in the context of the present invention comprises heavy chain complementarity determining regions (HCDRs) of a heavy chain variable region (HCVR) and light chain complementarity determining regions (LCDRs) of a light chain variable region (LCVR), wherein the HCVRs and LCVRs comprise the amino acid sequences of the anti-PD-L1 antibody designated as H1H8314N in U.S. Patent No. 9,938,345. In some embodiments, an anti-PD-L1 antibody or antigen-binding fragment thereof comprises three HCDRs (HCDR1, HCDR2, and HCDR3) and three LCDRs (LCDR1, LCDR2, and LCDR3), wherein HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 comprise the amino acid sequences of the anti-PD-L1 antibody designated as H1H8314N in U.S. Patent No. 9,938,345. In other embodiments, the anti-PD-L1 antibody or antigen-binding fragment thereof comprises HCVRs and LCVRs comprising the amino acid sequences of the anti-PD-L1 antibody designated as H1H8314N in U.S. Patent No. 9,938,345.

[098] In some embodiments of the present invention, an anti-human PD-L1 antibody or antigen-binding fragment thereof comprises an LCVR that has 90%, 95%, 98%, or 99% sequence identity to the amino acid sequence of the LCVR of the anti-PD-L1 antibody designated H1H8314N in U.S. Patent No. 9,938,345.

[099] In some embodiments, an anti-human PD-L1 antibody or antigen-binding fragment thereof comprises an HCVR comprising an amino acid sequence of an anti-PD-L1 antibody designated as H1H8314N in U.S. Patent No. 9,938,345, having no more than 5 amino acid substitutions. In some embodiments, an anti-human PD-L1 antibody or antigen-binding fragment thereof comprises an LCVR comprising an amino acid sequence of an anti-PD-L1 antibody designated as H1H8314N in U.S. Patent No. 9,938,345, having no more than 2 amino acid substitutions.

[0100] Sequence identity can be measured by methods known in the art (e.g., GAP, BESTFIT, and BLAST).

[0101] The present invention also includes the use of anti-PD-L1 antibodies in methods of treating cancer, wherein the anti-PD-L1 antibodies comprise variants of any of the HCVR, LCVR, and/or CDR amino acid sequences provided herein comprising one or more conservative amino acid substitutions. For example, the present invention includes the use of anti-PD-L1 antibodies comprising HCVR, LCVR, and/or CDR amino acid sequences with, for example, 10 or less, 8 or less, 6 or less, 4 or less, etc. conservative amino acid substitutions relative to any of the HCVR, LCVR, and/or CDR amino acid sequences provided herein.

[0102] Other anti-PD-L1 antibodies that can be used in the context of the methods of the present invention include, for example, those known in the art as MDX-1105, atezolizumab (TECENTRIQ™), durvalumab (IMFINZI™), avelumab (BAVENCIO™), LY3300054, FAZ053, STI-1014, CX-072, KN035 (Zhang et al., Cell Discovery , 3, 170004 (March 2017)), CK-301 (Gorelik et al., American Association for Cancer Research Annual Meeting (AACR), 2016-04-04 Abstract 4606), or any of the other anti-PD-L1 antibodies described in Patent Publication Nos. US7943743, US8217149, US9402899, US9624298, US9938345, WO2007/005874, WO2010/077634, WO2013/181452, WO2013/181634, WO2016/149201, WO2017/034916 or EP3177649. Portions of the publications cited herein that define anti-PD-L1 antibodies are hereby incorporated by reference.

Pharmaceutical compositions and administration

[0103] The disclosed methods include administering to an individual in need thereof an IL-4/IL-13 pathway inhibitor in combination with a PD-1 inhibitor, wherein the inhibitors are contained in separate pharmaceutical compositions or a combined (single) pharmaceutical composition. The pharmaceutical compositions of the present invention can be formulated with pharmaceutically acceptable carriers, excipients, and other means to provide suitable transport, delivery, tolerability, and the like. A variety of suitable formulations can be found in a reference book known to all pharmaceutical chemists: Remington's Pharmaceutical Sciences, Mack Publishing Company, Easton, Pa. These formulations include, for example, powders, pastes, ointments, jellies, waxes, oils, lipids, lipid-containing (cationic or anionic) vesicles (such as LIPOFECTIN™), DNA conjugates, anhydrous absorption pastes, oil-in-water and water-in-oil emulsions, carbowax emulsions (polyethylene glycols of different molecular weights), semi-solid gels, and semi-solid mixtures containing carbowax. See also Powell et al.,1998,J Pharm Sci Technol, 52:238-311.

[0104] In some embodiments, the pharmaceutical compositions of the present invention comprise a therapeutically effective amount of an IL-4/IL-13 pathway inhibitor (such as an anti-IL-4R antibody) and/or a therapeutically effective amount of a PD-1 inhibitor (such as an anti-PD-1 antibody) and a pharmaceutically acceptable carrier. In some embodiments, the pharmaceutical compositions of the present invention are formulated for administration by injection, such as intravenous injection.

[0105] Various delivery systems are known and can be used to administer the pharmaceutical compositions of the present invention, such as encapsulation in liposomes, microparticles, microcapsules, recombinant cells capable of expressing mutant viruses, receptor-mediated endocytosis (see, for example, Wu et al., 1987,J. Biol. Chem. 262: 4429-4432). Routes of administration include, but are not limited to, intradermal, intramuscular, intraperitoneal, intravenous, subcutaneous, intranasal, epidural, and oral routes. The compositions may be administered by any conventional route, such as by infusion or bolus injection, by absorption through the epithelial or mucocutaneous lining (e.g., oral mucosa, rectal and intestinal mucosa, etc.), and may be administered together with other biologically active agents.

[0106] The pharmaceutical composition of the present invention can be administered subcutaneously or intravenously using a standard needle and syringe. Furthermore, in the context of subcutaneous administration, a pen device can be used to deliver the pharmaceutical composition of the present invention. Such a pen device can be reusable or disposable. A reusable pen device typically uses a replaceable cartridge containing the pharmaceutical composition. After all of the pharmaceutical composition in the cartridge has been administered and the cartridge is empty, the empty cartridge can be easily discarded and replaced with a new cartridge containing the pharmaceutical composition. The pen device can then be reused. A disposable pen device does not have a replaceable cartridge. In contrast, a disposable pen device is supplied pre-filled with the pharmaceutical composition contained in a reservoir in the device. After the reservoir with the pharmaceutical composition is emptied, the entire device is discarded.

[0107] In some cases, one or both of the pharmaceutical compositions can be administered using a controlled release system. In one embodiment, a pump can be used. In another embodiment, polymeric materials can be used. See Medical Applications of Controlled Release, Langer and Wise (eds.), 1974, CRC Pres., Boca Raton, Fla. In another embodiment, the controlled release system can be placed near the target of the composition, and thus only a fraction of the systemic dose is required (see, (e.g., Goodson, 1984, Medical Applications of Controlled Release, vol. 2, pp. 115-138). Other controlled release systems are described in Langer, 1990,Science249:1527-1533.

[0108] Suitable injectable preparations may include dosage forms for intravenous, subcutaneous, intradermal and intramuscular injections, drip infusion, etc. These injectable preparations can be prepared by known methods. For example, the injectable preparations can be prepared, for example, by dissolving, suspending or emulsifying the above-described antibody or salt thereof in a sterile aqueous medium or oil medium conventionally used for injections. As the aqueous injection medium, there can be used, for example, physiological saline, isotonic solution containing glucose and other auxiliary agents, etc., which can be used in combination with a suitable solubilizer such as alcohol (e.g. ethanol), polyol (e.g. propylene glycol, polyethylene glycol), nonionic surfactant [e.g. polysorbate 80, HCO-50 (adduct of polyoxyethylene (50 mol) and hydrogenated castor oil)], etc. As the oil medium, there can be used, for example, sesame oil, soybean oil, etc., which can be used in combination with a solubilizer such as benzyl benzoate, benzyl alcohol, etc. The thus obtained injection preparation is preferably filled into a suitable ampoule.

[0109] Preferably, the pharmaceutical compositions for oral or parenteral use described above are prepared in unit dose dosage forms suitable for the dose of the active ingredients. Such unit dose dosage forms include, for example, tablets, pills, capsules, injections (ampoules), suppositories, etc.

[0110] Injectable formulations of the pharmaceutical compositions can be prepared by known methods. For example, an injectable formulation can be prepared by, for example, dissolving, suspending or emulsifying an inhibitor (e.g., an anti-PD-1 antibody or an anti-IL-4R antibody) or a salt thereof in a sterile aqueous medium or an oil medium conventionally used for injection. As an aqueous medium for injection, for example, a physiological saline solution, an isotonic solution containing glucose and other auxiliary agents, etc. can be used, which can be used in combination with a suitable solubilizer such as an alcohol (e.g., ethanol), a polyol (e.g., propylene glycol, polyethylene glycol), a nonionic surfactant [e.g., polysorbate 80, HCO-50 (adduct of polyoxyethylene (50 mol) and hydrogenated castor oil)], etc. As an oil medium, for example, sesame oil, soybean oil, etc. can be used, which can be used in combination with a solubilizer such as benzyl benzoate, benzyl alcohol, etc. The injectable formulation thus obtained is preferably filled into a suitable injection ampoule. In some embodiments, the injectable formulation may include a concentration of an inhibitor, such as an anti-PD-1 antibody or an anti-IL-4R antibody, and one or more pharmaceutically acceptable solvents (e.g., distilled water, saline, etc.).

[0111] Examples of pharmaceutical compositions comprising an anti-IL-4R antibody that may be used in the context of the present invention are described, for example, in U.S. Patent No. 8,945,559, the portions of which defining pharmaceutical compositions comprising an anti-IL-4R antibody are hereby incorporated by reference herein. Examples of pharmaceutical compositions comprising an anti-PD-1 antibody that may be used in the context of the present invention are described, for example, in U.S. Patent Application No. 2019/0040137, the portions of which defining pharmaceutical compositions comprising an anti-PD-1 antibody are hereby incorporated by reference herein.

Pharmaceutical delivery system

[0112] In some embodiments, pharmaceutical compositions for use in the disclosed methods can be provided in a pharmaceutical delivery system comprising: (i) a pharmaceutical composition comprising a therapeutically effective amount of an IL-4/IL-13 pathway inhibitor; and (ii) a pharmaceutical composition comprising a therapeutically effective amount of a PD-1 inhibitor. In this aspect of the invention, the pharmaceutical compositions are separate from one another, but can be provided in a single pharmaceutical delivery system or as a kit, i.e., a pharmaceutical delivery system with accompanying written instructions for using an IL-4/IL-13 pathway inhibitor in combination with a PD-1 inhibitor to treat or inhibit tumor growth, as described herein. In some embodiments, the pharmaceutical delivery system comprises one or more doses of an IL-4/IL-13 pathway inhibitor. In some embodiments, the pharmaceutical delivery system comprises one or more doses of a PD-1 inhibitor.

[0113] In some embodiments, the pharmaceutical delivery system further comprises one or more additional therapeutic agents, such as any of the additional therapeutic agents noted above. For example, a pharmaceutical delivery system comprising a pharmaceutical composition comprising an IL-4/IL-13 pathway inhibitor and a pharmaceutical composition comprising a PD-1 inhibitor may further comprise one or more additional therapeutic agents selected from cyclophosphamide, a tumor vaccine, a LAG3 inhibitor (e.g., an anti-LAG3 antibody), a CTLA-4 inhibitor (e.g., ipilimumab), a GITR agonist (e.g., an anti-GITR antibody), a TIM3 inhibitor, a CD28 activator, a BTLA inhibitor, a TIGIT inhibitor, a CD38 inhibitor, a CD47 inhibitor, an IDO inhibitor, a VEGF antagonist (e.g., a "VEGF decoy" such as aflibercept or other VEGF inhibitory fusion proteins described in U.S. Patent No. S 7,087,411), or an anti-VEGF antibody or antigen-binding fragment thereof. (eg, bevacizumab or ranibizumab) or a small molecule VEGF receptor kinase inhibitor (eg, sunitinib, sorafenib, or pazopanib)), Ang2 inhibitor (eg, nesvacumab), TGFβ inhibitor, EGFR inhibitor (eg, erlotinib, cetuximab), costimulatory receptor agonist (eg, CD28 agonist), VISTA inhibitor, CD40 agonist, CSF1R inhibitor, CCR2 inhibitor, CXCR4 inhibitor, CXCR2 inhibitor, CCR4 inhibitor, CXCL12 inhibitor, tumor-associated antigen antibody [eg, CA9, CA125, MAGE3, CEA, vimentin, tumor-M2-PK, PSA, mucin-1, MART-1, and CA19-9], bispecific anti-CD3/anti-CD20 antibodies, an oncolytic virus, a vaccine (e.g., bacillus Calmette-Guerin), GM-CSF, a cytotoxin, a chemotherapeutic agent, an IL-6R inhibitor (e.g., sarilumab), an IL-10 inhibitor, a cytokine such as IL-2, IL-7, IL-12, IL-21, and IL-15, an ADC (e.g., an anti-CD19-DM4 ADC and an anti-DS6-DM4 ADC), a chimeric antigen receptor T cell (e.g., a CD19-targeted T cell), an anti-inflammatory drug such as a corticosteroid, an NSAID, and a nutritional supplement such as an antioxidant. In one embodiment, the pharmaceutical delivery system is used in combination with surgery in a subject in need thereof.

Administration schemes

[0114] In some embodiments, the disclosed methods comprise sequentially administering a therapeutically effective amount of an IL-4/IL-13 pathway inhibitor in combination with a therapeutically effective amount of a PD-1 inhibitor to an individual in need thereof, wherein each inhibitor is administered to the individual at one or more doses, such as as part of a specific therapeutic administration regimen. In some embodiments, the methods of the present invention comprise administering the inhibitors for additive or synergistic activity for the treatment of a cancer, preferably a type 2 immune-dependent cancer such as pancreatic cancer, breast cancer, colorectal cancer, ovarian cancer, brain cancer, skin cancer, prostate cancer, kidney cancer, lung cancer, Hodgkin's lymphoma, or bladder cancer.

[0115] As used herein, the term "sequential administration" means that each dose of the inhibitor is administered to the individual at a different time point, such as on different days separated by a predetermined interval (e.g., hours, days, weeks, or months). In some embodiments, the disclosed methods comprise sequentially administering to the individual one initial dose of an IL-4/IL-13 pathway inhibitor, followed by one or more secondary doses of an IL-4/IL-13 pathway inhibitor, and optionally one or more tertiary doses of an IL-4/IL-13 pathway inhibitor. In some embodiments, the methods further comprise sequentially administering to the individual one initial dose of a PD-1 inhibitor, followed by one or more secondary doses of a PD-1 inhibitor, and optionally one or more tertiary doses of a PD-1 inhibitor.

[0116] In some embodiments, the therapeutic administration regimen comprises administering one or more doses of an IL-4/IL-13 pathway inhibitor in combination with one or more doses of a PD-1 inhibitor. In some embodiments, the one or more doses of the IL-4/IL-13 pathway inhibitor and/or the one or more doses of the PD-1 inhibitor are administered to the individual at a frequency of about once per day, once every two days, once every three days, once every four days, once every five days, once every six days, once per week, once every two weeks, once every three weeks, once every four weeks, once per month, once every two months, once every three months, once every four months, or less frequently.

[0117] As used herein, the term "in combination with" means that the IL-4/IL-13 pathway inhibitor is administered before, after, or simultaneously with the PD-1 inhibitor. The term "in combination with" also includes sequential or simultaneous administration of the IL-4/IL-13 pathway inhibitor and the PD-1 inhibitor.

[0118] For example, if the IL-4/IL-13 pathway inhibitor is administered "prior" to the PD-1 inhibitor, the IL-4/IL-13 pathway inhibitor can be administered greater than 150 hours, about 150 hours, about 100 hours, about 72 hours, about 60 hours, about 48 hours, about 36 hours, about 24 hours, about 12 hours, about 10 hours, about 8 hours, about 6 hours, about 4 hours, about 2 hours, about 1 hour, about 30 minutes, or about 15 minutes before administration of the PD-1 inhibitor. If the IL-4/IL-13 pathway inhibitor is administered "after" the PD-1 inhibitor, the IL-4/IL-13 pathway inhibitor may be administered about 15 minutes, about 30 minutes, about 1 hour, about 2 hours, about 4 hours, about 6 hours, about 8 hours, about 10 hours, about 12 hours, about 24 hours, about 36 hours, about 48 hours, about 60 hours, about 72 hours, or more than 72 hours after administration of the PD-1 inhibitor. Administering an IL-4/IL-13 pathway inhibitor "concurrently" with a PD-1 inhibitor means that the IL-4/IL-13 pathway inhibitor is administered to the individual in a separate dosage form within 15 minutes (before, after, or simultaneously) of the administration of the PD-1 inhibitor, or is administered to the individual as a single combination dosage form containing both the IL-4/IL-13 pathway inhibitor and the PD-1 inhibitor.

[0119] In some embodiments, the described methods include the use of an additional (e.g., third) therapeutic agent or method of therapy, such as any of the agents or methods of therapy described herein.

[0120] As used herein, the terms "initial," "secondary," "tertiary," etc., refer to the temporal order of administration. Thus, an "initial dose" is a dose administered at the beginning of a treatment regimen (also referred to as a "base dose"); a "secondary dose" is a dose administered after the initial dose; and a "tertiary dose" is a dose administered after the secondary dose. The initial, secondary, and tertiary doses may contain the same amount of an IL-4/IL-13 pathway inhibitor or PD-1 inhibitor. However, in some embodiments, the amount contained in the initial, secondary, and/or tertiary doses varies (e.g., is adjusted upward or downward, as needed) during treatment. In some embodiments, one or more (e.g., 1, 2, 3, 4, or 5) doses are administered at the beginning of the treatment regimen as "loading doses" followed by doses that are administered less frequently (e.g., "maintenance doses"). For example, an IL-4/IL-13 pathway inhibitor or a PD-1 inhibitor can be administered to a patient with a cancer at a loading dose of about 1 mg/kg to about 20 mg/kg followed by one or more maintenance doses of about 0.1 mg/kg to about 10 mg/kg of the patient's body weight.

[0121] In one exemplary embodiment of the present invention, each secondary and/or tertiary dose is administered ½-14 weeks or more (e.g., ½, 1, 1½, 2, 2½, 3, 3½, 4, 5, 5½, 6, 6½, 7, 7½, 8, 8½, 9, 9½, 10, 10½ or more weeks) after the immediately preceding dose. As used herein, the phrase "immediately preceding dose" in a sequence of multiple administrations means a dose of an anti-PD-1 antibody administered to an individual before the next dose in the sequence is administered, with no intermediate doses.

[0122] Similarly, an "initial treatment cycle" is a treatment cycle administered at the beginning of a treatment regimen; a "secondary treatment cycle" is a treatment cycle administered after the initial treatment cycle; and a "tertiary treatment cycle" is a treatment cycle administered after the secondary treatment cycle. In the context of the present invention, treatment cycles may be the same or different from each other.

Dose

[0123] In some embodiments, at least one dose of an IL-4/IL-13 pathway inhibitor comprises about 0.1-50 mg/kg, such as about 0.1-10 mg/kg of the subject's body weight. For example, the at least one dose can comprise about 0.1, 1, 0.3, 3, 4, 5, 6, 7, 8, 9, or 10 mg/kg of the subject's body weight. In some embodiments, at least one dose of the IL-4/IL-13 pathway inhibitor comprises about 0.05-1000 mg of the IL-4/IL-13 pathway inhibitor, such as about 5, 10, 15, 20, 25, 40, 45, 50, 60, 70, 80, 90, 100, 200, 300, 400, 500, 600, 700, 800, 900 mg or more of the IL-4/IL-13 pathway inhibitor. In one embodiment, the IL-4/IL-13 pathway inhibitor is REGN668 (dupilumab).

[0124] In some embodiments, the at least one dose of the PD-1 inhibitor comprises about 0.1-20 mg/kg of the subject's body weight, such as about 0.1, 1, 0.3, 3, 4, 5, 6, 7, 8, 9, or 10 mg/kg of the subject's body weight. In some embodiments, the at least one dose of the PD-1 inhibitor comprises about 0.05-500 mg of the PD-1 inhibitor, such as about 5, 10, 15, 20, 25, 40, 45, 50, 60, 70, 80, 90, 100 mg or more of the PD-1 inhibitor. In one embodiment, the PD-1 inhibitor is REGN2810 (cemiplimab).

[0125] The amounts of an IL-4/IL-13 pathway inhibitor and a PD-1 inhibitor administered to an individual in the methods provided herein comprise a therapeutically effective amount. As used herein, the term "therapeutically effective amount" means an amount of each inhibitor that results in one or more of: (a) reducing the severity or duration of a symptom or sign of a cancer, such as a tumor lesion; (b) inhibiting tumor growth or increasing tumor necrosis, shrinking a tumor, and/or causing a tumor to disappear; (c) delaying tumor growth and progression; (d) inhibiting tumor metastasis; (e) preventing recurrence of tumor growth; (f) increasing survival of an individual with a cancer; and/or (g) a reduction in the use of or need for conventional anticancer therapy (e.g., eliminating the need for surgery or reducing or eliminating the use of chemotherapeutic or cytotoxic agents) compared to an untreated individual or an individual treated with any inhibitor as monotherapy.

[0126] In the case of an IL-4/IL-13 pathway inhibitor (e.g., an anti-IL-4R antibody), the therapeutically effective amount can be from about 0.05 mg to about 1000 mg, such as about 0.05 mg, about 0.1 mg, about 1.0 mg, about 1.5 mg, about 2.0 mg, about 10 mg, about 20 mg, about 30 mg, about 40 mg, about 50 mg, about 60 mg, about 70 mg, about 80 mg, about 90 mg, about 100 mg, about 110 mg, about 120 mg, about 130 mg, about 140 mg, about 150 mg, about 160 mg, about 170 mg, about 180 mg, about 190 mg, about 200 mg, about 210 mg, about 220 mg, about 230 mg, about 240 mg, about 250 mg, about 260 mg, about 270 mg, about 280 mg, about 290 mg, about 300 mg, about 310 mg, about 320 mg, about 330 mg, about 340 mg, about 350 mg, about 360 mg, about 370 mg, about 380 mg, about 390 mg, about 400 mg, about 410 mg, about 420 mg, about 430 mg, about 440 mg, about 450 mg, about 460 mg, about 470 mg, about 480 mg, about 490 mg, about 500 mg, about 510 mg, about 520 mg, about 530 mg, about 540 mg, about 550 mg, about 560 mg, about 570 mg, about 580 mg, about 590 mg, about 600 mg, about 610 mg, about 620 mg, about 630 mg, about 640 mg, about 650 mg, about 660 mg, about 670 mg, about 680 mg, about 690 mg, about 700 mg, about 710 mg, about 720 mg, about 730 mg, about 740 mg, about 750 mg, about 760 mg, about 770 mg, about 780 mg, about 790 mg, about 800 mg, about 810 mg, about 820 mg, about 830 mg, about 840 mg, about 850 mg, about 860 mg, about 870 mg, about 880 mg, about 890 mg, about 900 mg, about 910 mg, about 920 mg, about 930 mg, about 940 mg, about 950 mg, about 960 mg, about 970 mg, about 980 mg, about 990 mg, or about 1000 mg of an IL-4/IL-13 pathway inhibitor. In some embodiments, the individual is administered 10 mg, 25 mg, 50 mg, 75 mg, 150 mg, 300 mg, 600 mg, or 900 mg of an IL-4/IL-13 pathway inhibitor.

[0127] In the case of a PD-1 inhibitor (e.g., an anti-PD-1 antibody), the therapeutically effective amount can be from about 0.05 mg to about 500 mg, from about 1 mg to about 500 mg, from about 10 mg to about 450 mg, from about 50 mg to about 400 mg, from about 75 mg to about 350 mg, or from about 100 mg to about 300 mg of the antibody. For example, in various embodiments, the amount of the PD-1 inhibitor is about 0.05 mg, about 0.1 mg, about 1.0 mg, about 1.5 mg, about 2.0 mg, about 5 mg, about 10 mg, about 15 mg, about 20 mg, about 30 mg, about 40 mg, about 50 mg, about 60 mg, about 70 mg, about 80 mg, about 90 mg, about 100 mg, about 110 mg, about 120 mg, about 130 mg, about 140 mg, about 150 mg, about 160 mg, about 170 mg, about 180 mg, about 190 mg, about 200 mg, about 210 mg, about 220 mg, about 230 mg, about 240 mg, about 250 mg, about 260 mg, about 270 mg, about 280 mg, about 290 mg, about 300 mg, about 310 mg, about 320 mg, about 330 mg, about 340 mg, about 350 mg, about 360 mg, about 370 mg, about 380 mg, about 390 mg, about 400 mg, about 410 mg, about 420 mg, about 430 mg, about 440 mg, about 450 mg, about 460 mg, about 470 mg, about 480 mg, about 490 mg, or about 500 mg of a PD-1 inhibitor.

[0128] In some embodiments, the amount of an IL-4/IL-13 pathway inhibitor (e.g., an anti-IL-4R antibody) and/or a PD-1 inhibitor (e.g., an anti-PD-1 antibody) in a single dose administered to an individual may be less than a therapeutically effective amount, i.e., a subtherapeutic dose. For example, if a therapeutically effective amount of an inhibitor comprises 3 mg/kg, a subtherapeutic dose comprises an amount less than 3 mg/kg, such as 2 mg/kg, 1.5 mg/kg, 1 mg/kg, 0.5 mg/kg, or 0.3 mg/kg. As defined herein, the term "subtherapeutic dose" refers to an amount of an inhibitor that does not, by itself, result in a therapeutic effect. However, in some embodiments, multiple subtherapeutic doses of an inhibitor may be administered to collectively achieve a therapeutic effect in an individual.

EXAMPLES

[0129] The present invention will now be described by way of the following examples. The use of these and other examples anywhere in this description is illustrative only and does not in any way limit the scope and spirit of the present invention or any illustrative form. Likewise, the invention is not limited to any specific preferred embodiments presented herein. In fact, modifications and variations of the invention may be apparent to those skilled in the art upon reading the description, and they can be practiced without departing from the spirit and scope thereof. Accordingly, the present invention is limited only by the claims, together with the full scope of equivalents to which the claims pertain. Furthermore, while attempts have been made to ensure accuracy with respect to the numbers (e.g., amounts, temperatures, etc.) used, some experimental errors and deviations must be taken into account. Unless otherwise indicated, parts are parts by weight, molecular weight is average molecular weight, temperature is in degrees Celsius, and pressure is at or near atmospheric pressure.

Example 1: Increased efficiency in vivo Anti-IL-4R antibodies in combination with anti-PD-1 antibody in a pancreatic cancer model

[0130] This example relates to a study that demonstrated enhanced antitumor efficacy of IL-4R blockade alone and in combination with anti-PD-1 therapy in vivo using a subcutaneous implantation model of KP organoid cells, a mouse model of human pancreatic cancer, which is a type 2 immune-dependent malignancy. In this study, 8-10 mice per treatment group were implanted with KP tumor organoid cells, and treatment of tumor-bearing mice was initiated when initial tumor volumes ranged from 30 to 110 mm ³ .

[0131] The IL-4/IL-13 pathway inhibitor used in this example was a mouse anti-IL-4R antibody designated REGN1103, which is a mouse surrogate antibody of a human monoclonal antibody designated REGN668 (also known as dupilumab) that targets human IL-4R. REGN1103 comprises the HCVR/LCVR amino acid sequence pair of SEQ ID NO: 21/22 and has an affinity for mouse IL-4R in a range similar to the affinity of dupilumab for human IL-4R. Additionally, REGN1103 inhibits IL-4- and IL-13-dependent cell line proliferation with an IC ₅₀ of 1.9 nM and 11 pM, respectively.

[0132] The PD-1 inhibitor used in this example was a mouse anti-PD-1 antibody clone designated RPMI-14 (Bio X Cell), which is a surrogate anti-PD-1 antibody commonly used for syngeneic mouse tumor models.

Study design

[0133] To assess whether the type 2 immune response is activated in in vivo pancreatic cancer models, IL-4 and IL-13 expression levels and IL-4 cytokine production were assessed in implantation models of pancreatic cancer. Syngeneic tumor organoid cells engineered to express oncogenic KRAS with concomitant loss of function of the tumor suppressor P53 (KP) were implanted subcutaneously or orthotopically into Balb/c mice, and tumors were harvested 4 weeks after implantation.

[0134] The subcutaneous implantation model of KP organoid cells was also used to evaluate the antitumor efficacy of IL-4R blockade as monotherapy and/or in combination with anti-PD-1 ICB therapy in vivo .

Materials and methods

[0135]Animals.Mice Balb/c was purchased from Jackson Laboratories and experiments were performed according to IACUC-approved guidelines.

[0136] Preparation of KP organoids. Normal pancreatic epithelial organoids were isolated from wild-type Balb/c mouse pancreatic tissue. Pancreatic tissue was minced and digested in collagenase P buffer (Fisher Sci, 50-100-3398) at 37°C for 15 min. The cell suspension was passed through a 500 μm filter and then again through a 100 μm filter. The cells were then seeded on Matrigel (VWR, 354234) overlaid with complete organoid medium; DME/F12 supplemented with 1× P/S/L-glutamine, 1× B27 (Invitrogen), 1 mM N-acetylcysteine (Sigma-Aldrich), 10 nM h-gastrin (Sigma-Aldrich), 50 ng/ml mEGF (Life Technologies), 1 μg/ml mRSPO1 (Peprotech), 25 ng/ml h-noggin (PeproTech), 100 ng/ml hFGF10 (PeproTech), and 10 mM nicotinamide (Sigma-Aldrich) (Boj et al., Mol Cell Oncol , 3(1):e1014757, 2015).

[0137] The resulting organoids were then maintained in growth factor-reduced Matrigel containing complete organoid medium supplemented with 10 nM Rock inhibitor Y-27632 (Sigma-Aldrich, Y0503). For passage, organoids were washed out of Matrigel using cell isolation solution (Corning) for 2 hours on ice and then dissociated into cell suspension using Accutase. Passage was performed at a 1:4 ratio once per week.

[0138] For lentiviral transduction of pancreatic organoids, they were dissociated into single cell suspensions and transduced using lentiviruses to express KRas ^V12 and to lose p53 expression or to express p53 ^R175H using the spin infection method in the presence of polybrene (8 μg/ml).

[0139] Transduced KP organoids were then orthotopically implanted into the pancreatic head of syngeneic Balb/c mice (0.1-0.5E6 cells in 50:50 PBS/Matrigel solution per implantation), resulting in tumor growth for months after implantation.

[0140] The resulting KP tumors were harvested and pancreatic cancer cells were isolated using collagenase XI/dispase solution overnight at 37°C. The cells were then washed in PBS and resuspended in growth factor-reduced Matrigel containing complete organoid medium.

Treatment

[0141] In each study group (listed in Table 1 below), 8-10 mice were implanted with KP tumor organoid cells and treatment was initiated when tumors had matured (tumor volumes 30-110 ^mm3 at baseline). Mice were injected intraperitoneally with 25 mg/kg REGN1103 (anti-mIL-4R antibody) or an isotype control mIgG1 Ab in combination with 10 mg/kg anti-mPD1 antibody (RPMI-14 clone) or an isotype control rat IgG2a, beginning on day 13 after KP cell injection (0.1E6 to 0.5E6 cells), every 3-4 days for 7 doses.

Table 1

Group Treatment 1 ISO/ISO Ab mIgG1, 25 mg/kg + Rat IgG2a, 10 mg/kg 2 IL-4R/ISO Anti-IL-4R antibody (REGN1103), 25 mg/kg+rat IgG2a, 10 mg/kg 3 ISO/PD-1 Ab mIgG1, 25 mg/kg + antibody against PD-1 (RPMI-14, BioXCell), 10 mg/kg 4 IL-4R/PD-1 Anti-IL-4R antibody (REGN1103), 25 mg/kg + anti-PD-1 antibody (RPMI-14, BioXCell), 10 mg/kg

[0142] Balb/c mice were injected subcutaneously with 1E5-2.5E5 syngeneic KP organoid cells in 50:50 PBS/Matrigel solution. Cells were tested using the IMPACT assay prior to in vivo implantation. Tumors were measured twice weekly using calipers and volume was calculated using the formula (L*W ² )/2, where L is the longest diameter and W is the perpendicular diameter.

Results

[0143] It was found that IL-4 and IL-13 gene expression was extensively upregulated in orthotopically and subcutaneously implanted tumors relative to normal pancreas, which lacks IL-4 and IL-13 gene expression. Similarly, IL-4 cytokine production, as measured by ELISA, was extensively upregulated in the orthotopic and subcutaneous tumor models, whereas IL-4 cytokine production was not observed in normal pancreas. These data suggest that the KP organoid cell implantation models of pancreatic cancer express high levels of IL-4 and IL-13, which are two key cytokines associated with type 2 immunity, thus suggesting that type 2 immune response is activated in these in vivo models of pancreatic cancer.

[0144] Additionally, anti-IL-4R treatment enhanced the anti-tumor efficacy of PD-1 blockade in vivo . Results from the treatment study in tumor volumes of 30-110 ^mm3 at baseline are shown in Figures 1A and 1B. Results in a subset of treated mice bearing tumors of 50-110 ^mm3 at baseline are shown in Figures 2A and 2B. Anti-mPD-1 monotherapy demonstrated partial anti-tumor efficacy, with 4 of 8 mice experiencing tumor regression, including 2 complete responses, while anti-mIL-4R monotherapy did not demonstrate anti-tumor activity. However, combination anti-IL-4R and anti-PD-1 treatment demonstrated significantly enhanced anti-tumor efficacy, with 7 of 10 mice experiencing tumor regression, including 6 complete responses. This was an unexpectedly excellent result.

[0145] Thus, IL-4/IL-13 type 2 immune responses in type 2 immune-dependent malignancies can be effectively targeted by blockade of the IL-4/IL-13 pathway. This study shows that anti-IL-4R therapy significantly enhances the antitumor efficacy of anti-PD-1 therapy to an unexpected extent in a subcutaneous animal model of pancreatic cancer, typical of type 2 immune-dependent malignancies.

Example 2: Human pancreatic cancer and subtypes of other human malignancies exhibit a strong type 2 immune signature

[0146] To assess the degree of activation of the type 2 immune response in different types of human malignancies, a type 2 immunity gene signature was constructed, including type 2 immunity-associated cytokines, chemokines and their receptors, as well as downstream type 2 immunity target genes (IL-13RA2, IL-25, IL-17RB, SERPINB2, CCL24, CEACAM1, CCL1, MUC5B, CCL26, IL-13RA1, POSTN, IL-6R, CCL18, CCL17, FCER2, CCR3, IL-6, CCL8, CRLF2, IL-33, TSLP, IL-4R, CCR4, PTGDR, FCER1A, IL-1RL1, DPP4, IL-4, IL-5 and IL-13). This type 2 immunity signature was applied to the expression profiles of multiple human cancer types available in The Cancer Genome Atlas (TCGA) database, namely BRCA (breast cancer), COAD (colon adenocarcinoma), GBM (glioblastoma multiforme), KIRC (renal cell carcinoma), LGG (low grade glioma), LUAD (lung adenocarcinoma), LUSC (lung squamous cell carcinoma), OV (ovarian cancer), PAAD (pancreatic adenocarcinoma), PRAD (prostate adenocarcinoma), and SKCM (skin melanoma). The upper quantile of normalized expression values in FPKM (fragments per kb of transcript per million mapped reads) was used as the initial input. Z-scores were obtained for the expression of each gene in all tumor samples represented in the heatmap. To calculate the Th2 signature score, the maximum Z-score (highest Z-score) was set as +4; and the minimum Z-score (lowest Z-score) was set as -4. The sum of the Z-scores for each gene in the Th2 signature gene list was then calculated for each sample. The combined expression (Z-score) represents the Th2 signature level of the sample.

[0147] Analysis of this type 2 immune signature heat map revealed that pancreatic cancer, non-small cell lung cancer, and lung squamous cell carcinoma exhibited the highest overall expression of this type 2 immune signature with minimal heterogeneity. Subtypes of other cancer types also exhibited a strong type 2 immune signature, including breast cancer and colon cancer subtypes. However, other cancer types (e.g., glioblastoma multiforme (GBM), glioma, melanoma, prostate cancer) exhibited overall low expression of the signature. Overall, these data suggest strong type 2 immune activity in human pancreatic cancer and lung cancer, as well as in subtypes of other human cancer types, including breast cancer and colon cancer.

Example 3: Cytokines IL-4 and IL-13 are extensively upregulated and produced in a pancreatic cancer model in vivo

[0148] This example relates to the study of whether the type 2 immune response is activated in in vivo models of pancreatic cancer by assessing the expression levels of IL-4 and IL-13 and the production of the cytokine IL-4 in implantation models of pancreatic cancer. The materials and methods described in Example 1 regarding the animals and the production of KP organoids are the same in Example 3.

[0149] Balb/c mice were implanted subcutaneously (Sub-Cu) or orthotopically (Ortho) with syngeneic pancreatic tumor organoid cells engineered to express oncogenic KRAS with concomitant loss of function of the tumor suppressor P53 (KP cells), and tumors were harvested 4 weeks after implantation (n=4-5 mice per condition).

[0150] Cytokine measurements . Tumor samples were resuspended in Tissue Protein Extraction Reagent (T-PER; Thermo Fisher, Waltham, Mass) supplemented with protease inhibitor cocktail and mechanically homogenized with a TissueLyser II (Qiagen, Hilden, Germany). Total protein content of tumor protein extracts was measured using the Bradford assay (Pierce™ Protein Assay at 660 nm (Thermo Fisher, Waltham, Mass)). Cytokine concentrations were determined using custom MSD kits and a QuickPlex SQ120 plate reader (Meso Scale Discovery, Rockville, MD).

Results

[0151] IL-4 and IL-13 gene expression were found to be extensively upregulated in orthotopically and subcutaneously implanted tumors relative to normal pancreas, which lacks IL-4 and IL-13 gene expression (Figure 3A). Similarly, IL-4 cytokine production, as measured by ELISA, was extensively upregulated in orthotopically and subcutaneously implanted tumor models, whereas IL-4 cytokine production was not observed in normal pancreas (Figure 3B). These data indicate that KP pancreatic cancer tumor organoid cell implantation models express high levels of IL-4 and IL-13, two key cytokines associated with type 2 immunity, thus demonstrating that type 2 immune response is activated in these in vivo pancreatic cancer models. Thus, IL-4/IL-13-induced type 2 immune responses can be effectively targeted by IL-4R blockade (Harb et al., Clin Exp Allergy , 50(1):5–14, 2019; Le Floc'h et al., Allergy , Dec 2019).

Example 4: Combined anti-IL-4R and anti-PD-1 blockade consistently results in enhanced antitumor efficacy in vivo in larger tumors

[0152] This example is a replication of the study from Example 1 and evaluates the efficacy of combination treatment against IL-4R/PD-1 against larger tumors. Specifically, in this study, treatment was initiated when the tumors had developed volumes in the range of 70 to 100 mm ³ .

Results

[0153] Anti-IL-4R monotherapy demonstrated a trend toward tumor growth control, while anti-mPD-1 monotherapy did not demonstrate overall anti-tumor activity. In contrast, anti-IL-4R/PD-1 combination therapy demonstrated significantly increased anti-tumor efficacy relative to monotherapy controls, controlling tumor growth in 4 of 7 mice. The results of this study are shown in Figures 4A and 4B.

[0154] In addition, systemic levels of the cytokines interferon gamma (IFNγ) and tumor necrosis factor alpha (TNFα) were measured in tumor lysates at the end-of-study time point. IFNγ and TNFα production were significantly increased in the combination treatment group compared to the monotherapy or isotype control groups (Figure 4C). Specifically, the highest intratumoral IFNγ levels (>50 pg/mL) corresponded to the most responsive tumors in each study group. Overall, the above results suggest that combination treatment against IL-4R/PD-1 induced enhanced IFNγ-dependent antitumor activity.

Example 5: Combined IL-4R and PD-1 blockade demonstrates immunomodulatory activity against tumor stroma and tumor

[0155] This example is a repeat of the study from Example 1, but examined the stroma and immunomodulatory activity of combination treatment against IL-4R/PD-1 in mice bearing PDAC tumors. Histological analysis of the stromal contents was performed in this short-term treatment endpoint study in which 8-10 mice per treatment group were implanted with KP tumor organoid cells. Treatment was initiated when tumor volumes reached 50-100 mm ³ . Tumors were harvested after 2 treatment doses to avoid excessive tumor regression, allowing for unbiased histological analysis. The percentage change in tumor volume across treatment groups is shown in Figure 5A.

[0156] Hyaluronic acid (HA) is a significant component of the pancreatic tumor microenvironment, shown to inhibit drug perfusion into the tumor bed (Provenzano et al., Cancer Cell , 21(3):418–29, 2012, Jacobetz et al., Gut , 62(1):112–20, 2013). Enzymatic depletion of HA has been shown to enhance cytotoxic antitumor efficacy in preclinical models of PDAC and has been investigated in patients with pancreatic cancer in combination with chemotherapy or immune checkpoint blockade. HA content in the pancreatic tumor microenvironment was determined by histochemical staining using biotinylated HA-binding protein (HABP) and digital pathology analysis (Figure 5B).

[0157] Immunostaining. Specimens were fixed in 10% neutral formalin for 16-24 h and transferred to 70% ethanol for paraffin embedding. All staining was performed on 5 μm paraffin sections of mouse tissue.

[0158] For immunostaining, sections were deparaffinized, rehydrated, and antigen retrieval was performed with RTU AR Citra solution (Biogenex, San Ramon, CA). H ₂ O ₂ was used to block endogenous peroxidases. Nonspecific protein binding was blocked with 10% goat serum (Sigma, St Louis, MA). Sections were incubated with anti-CD8 primary antibody (Cell Signaling Technologies, Danvers, MA) overnight at 4°C and horseradish peroxidase-conjugated secondary antibody (Cell Signaling Technologies). The remaining steps were performed using the appropriate Vectastain Elite ABC kits (Vector Labs) and DAB peroxidase substrate (Vector Labs) and counterstained with hematoxylin. Digital quantification of HABP deposition and CD8 ⁺ T cell density was performed using the HALO image analysis platform from Indica Labs.

[0159] HA histochemistry . Biotinylated HA binding protein (bHABP, Millipore Danvers MA USA) was used. Sections were deparaffinized, rehydrated, and blocked with 3% bovine serum albumin (BSA) before incubation with bHABP (1:200 in 1% BSA) overnight at 4°C. The remaining steps were performed using the Vectastain Elite ABC kit (Vector Labs. Burlingame CA) and DAB peroxidase substrate (Vector Labs) counterstained with hematoxylin.

Results

[0160] Figure 5A shows the percent change in tumor volume (relative to baseline) at the end of the study. HA content was significantly reduced in the anti-IL-4R monotherapy group and the anti-IL-4R/PD-1 combination group compared to the anti-PD-1 monotherapy and isotype control groups (Figure 5B). This reduction in HA content was associated with increased CD8 ⁺ T cell infiltration in the tumor bed, particularly in the anti-IL-4R/PD-1 combination group, as demonstrated by histological quantification of the tumor infiltrating lymphocyte (TIL) marker CD8 (Figure 5C). Furthermore, increased T cell infiltration was associated with significant synergistic antitumor efficacy in the anti-IL-4R/PD-1 combination group.

[0161] The above data suggest that in the context of immune-polarized type 2 pancreatic fibroids, IL-4R blockade has antitumor activity through a dual mechanism of action: (i) potent stromal modulation and regulation of HA deposition; and (ii) significant enhancement of TIL infiltration when combined with anti-PD-1 therapeutics.

Example 6: Efficacy of anti-human PD-1 antibody in combination with anti-IL-4R antibody against implanted tumors

[0162] For the experiments in this example, humanized mice expressing the extracellular portion of human PD-1 and the transmembrane and intracellular portions of the mouse version of the protein were generated using VelociGene® technology (Valenzuela et al 2003, Nat. Biotechnol . 21:652-659; also see U.S. Patent No. 9,987,500).

[0163] An example of an anti-PD-1 antibody used for this study was a fully human antibody (cemiplimab) that specifically binds to human PD-1 and comprises HCDR1-HCDR2-HCDR3-LCDR1-LCDR2-LCDR3 SEQ ID NOs: 13-14-15-16-17-18 and HCVR/LCVR SEQ ID NOs: 11/12. An example of an IL-4/IL-13 pathway inhibitor used in this example is described in Example 1.

[0164] Pancreatic epithelial organoids were isolated from a PD-1-humanized mouse and processed as described in Example 1 to generate KP tumor organoid cells (pancreatic cancer cells).

[0165] PD-1-humanized mice were implanted with 1.5 x 10 ⁶ KP tumor organoid cells on day 0 and treatment was started when tumors developed. Mice were injected intraperitoneally with 25 mg/kg REGN1103 (anti-mIL-4R antibody) or isotype control Ab in combination with 10 mg/kg cemiplimab or isotype control starting on day 13 after KP cell injection (0.1E6 to 0.5E6 cells) every 3-4 days for 7 doses. Tumor volumes were monitored by caliper measurements twice weekly throughout the experiment.

[0166] Anti-IL-4R treatment enhances the antitumor efficacy of cemiplimab. Tumor regression and complete responses were observed in mice treated with the combination treatment compared to cemiplimab monotherapy.

Example 7: Clinical trial

[0167] In a clinical trial, patients with pancreatic cancer, lung cancer, multiple myeloma, breast cancer, and colon cancer were given cemiplimab in combination with dupilumab. Patients given cemiplimab and dupilumab had greater tumor inhibition than patients given either drug alone.

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LIST OF SEQUENCES

<110> Regeneron Pharmaceuticals, Inc.

<120> IL-4/IL-13 PATHWAY INHIBITORS FOR ENHANCED EFFECTIVENESS

IN THE TREATMENT OF MALIGNANT NEOPLASMS

<130> 10529P1-US

<160> 22

<170> FastSEQ for Windows version 4.0

<210> 1

<211> 124

<212> PROTEIN

<213> Artificial sequence

<220>

<223>HCVR R668

<400> 1

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Glu Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Gly Ser Gly Phe Thr Phe Arg Asp Tyr

20 25 30

Ala Met Thr Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ser Ile Ser Gly Ser Gly Gly Asn Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Lys Asp Arg Leu Ser Ile Thr Ile Arg Pro Arg Tyr Tyr Gly Leu

100 105 110

Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser

115 120

<210> 2

<211> 112

<212> PROTEIN

<213> Artificial sequence

<220>

<223>LCVR R668

<400> 2

Asp Ile Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Pro Gly

1 5 10 15

Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Leu Tyr Ser

20 25 30

Ile Gly Tyr Asn Tyr Leu Asp Trp Tyr Leu Gln Lys Ser Gly Gln Ser

35 40 45

Pro Gln Leu Leu Ile Tyr Leu Gly Ser Asn Arg Ala Ser Gly Val Pro

50 55 60

Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile

65 70 75 80

Ser Arg Val Glu Ala Glu Asp Val Gly Phe Tyr Tyr Cys Met Gln Ala

85 90 95

Leu Gln Thr Pro Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105 110

<210> 3

<211> 8

<212> PROTEIN

<213> Artificial sequence

<220>

<223>HCDR1 R668

<400> 3

Gly Phe Thr Phe Arg Asp Tyr Ala

1 5

<210> 4

<211> 8

<212> PROTEIN

<213> Artificial sequence

<220>

<223>HCDR2 R668

<400> 4

Ile Ser Gly Ser Gly Gly Asn Thr

1 5

<210> 5

<211> 18

<212> PROTEIN

<213> Artificial sequence

<220>

<223>HCDR3 R668

<400> 5

Ala Lys Asp Arg Leu Ser Ile Thr Ile Arg Pro Arg Tyr Tyr Gly Leu

1 5 10 15

Asp Val

<210> 6

<211> 11

<212> PROTEIN

<213> Artificial sequence

<220>

<223> LCDR1 R668

<400> 6

Gln Ser Leu Leu Tyr Ser Ile Gly Tyr Asn Tyr

1 5 10

<210> 7

<211> 3

<212> PROTEIN

<213> Artificial sequence

<220>

<223> LCDR2 R668

<400> 7

Leu Gly Ser

1

<210> 8

<211> 9

<212> PROTEIN

<213> Artificial sequence

<220>

<223> LCDR3 R668

<400> 8

Met Gln Ala Leu Gln Thr Pro Tyr Thr

1 5

<210> 9

<211> 451

<212> PROTEIN

<213> Artificial sequence

<220>

<223>HC R668

<400> 9

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Glu Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Gly Ser Gly Phe Thr Phe Arg Asp Tyr

20 25 30

Ala Met Thr Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ser Ile Ser Gly Ser Gly Gly Asn Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Lys Asp Arg Leu Ser Ile Thr Ile Arg Pro Arg Tyr Tyr Gly Leu

100 105 110

Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr

115 120 125

Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser

130 135 140

Glu Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu

145 150 155 160

Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His

165 170 175

Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser

180 185 190

Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys

195 200 205

Asn Val Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu

210 215 220

Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Leu

225 230 235 240

Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu

245 250 255

Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser

260 265 270

Gln Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu

275 280 285

Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr

290 295 300

Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn

305 310 315 320

Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser

325 330 335

Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln

340 345 350

Val Tyr Thr Leu Pro Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val

355 360 365

Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val

370 375 380

Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro

385 390 395 400

Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr

405 410 415

Val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val

420 425 430

Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu

435 440 445

Ser Leu Gly

450

<210> 10

<211> 219

<212> PROTEIN

<213> Artificial sequence

<220>

<223> LC R668

<400> 10

Asp Ile Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Pro Gly

1 5 10 15

Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Leu Tyr Ser

20 25 30

Ile Gly Tyr Asn Tyr Leu Asp Trp Tyr Leu Gln Lys Ser Gly Gln Ser

35 40 45

Pro Gln Leu Leu Ile Tyr Leu Gly Ser Asn Arg Ala Ser Gly Val Pro

50 55 60

Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile

65 70 75 80

Ser Arg Val Glu Ala Glu Asp Val Gly Phe Tyr Tyr Cys Met Gln Ala

85 90 95

Leu Gln Thr Pro Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105 110

Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu

115 120 125

Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe

130 135 140

Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln

145 150 155 160

Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser

165 170 175

Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu

180 185 190

Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser

195 200 205

Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys

210 215

<210> 11

<211> 117

<212> PROTEIN

<213> Artificial sequence

<220>

<223>HCVR R2810

<400> 11

Glu Val Gln Leu Leu Glu Ser Gly Gly Val Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asn Phe

20 25 30

Gly Met Thr Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Gly Ile Ser Gly Gly Gly Arg Asp Thr Tyr Phe Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Lys Gly Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Val Lys Trp Gly Asn Ile Tyr Phe Asp Tyr Trp Gly Gln Gly Thr Leu

100 105 110

Val Thr Val Ser Ser

115

<210> 12

<211> 107

<212> PROTEIN

<213> Artificial sequence

<220>

<223>LCVR R2810

<400> 12

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Ser Ile Thr Ile Thr Cys Arg Ala Ser Leu Ser Ile Asn Thr Phe

20 25 30

Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Asn Leu Leu Ile

35 40 45

Tyr Ala Ala Ser Ser Leu His Gly Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Arg Thr Leu Gln Pro

65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Ser Asn Thr Pro Phe

85 90 95

Thr Phe Gly Pro Gly Thr Val Val Asp Phe Arg

100 105

<210> 13

<211> 8

<212> PROTEIN

<213> Artificial sequence

<220>

<223>HCDR1R2810

<400> 13

Gly Phe Thr Phe Ser Asn Phe Gly

1 5

<210> 14

<211> 8

<212> PROTEIN

<213> Artificial sequence

<220>

<223>HCDR2R2810

<400> 14

Ile Ser Gly Gly Gly Arg Asp Thr

1 5

<210> 15

<211> 10

<212> PROTEIN

<213> Artificial sequence

<220>

<223>HCDR3R2810

<400> 15

Val Lys Trp Gly Asn Ile Tyr Phe Asp Tyr

1 5 10

<210> 16

<211> 6

<212> PROTEIN

<213> Artificial sequence

<220>

<223> LCDR1 R2810

<400> 16

Leu Ser Ile Asn Thr Phe

1 5

<210> 17

<211> 3

<212> PROTEIN

<213> Artificial sequence

<220>

<223> LCDR2 R2810

<400> 17

Ala Ala Ser

1

<210> 18

<211> 9

<212> PROTEIN

<213> Artificial sequence

<220>

<223> LCDR3 R2810

<400> 18

Gln Gln Ser Ser Asn Thr Pro Phe Thr

1 5

<210> 19

<211> 444

<212> PROTEIN

<213> Artificial sequence

<220>

<223>HC R2810

<400> 19

Glu Val Gln Leu Leu Glu Ser Gly Gly Val Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asn Phe

20 25 30

Gly Met Thr Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Gly Ile Ser Gly Gly Gly Arg Asp Thr Tyr Phe Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Lys Gly Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Val Lys Trp Gly Asn Ile Tyr Phe Asp Tyr Trp Gly Gln Gly Thr Leu

100 105 110

Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu

115 120 125

Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly Cys

130 135 140

Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser

145 150 155 160

Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser

165 170 175

Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser

180 185 190

Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn

195 200 205

Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro Pro Cys Pro

210 215 220

Pro Cys Pro Ala Pro Glu Phe Leu Gly Gly Pro Ser Val Phe Leu Phe

225 230 235 240

Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val

245 250 255

Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu Val Gln Phe

260 265 270

Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro

275 280 285

Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr

290 295 300

Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val

305 310 315 320

Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala

325 330 335

Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Gln

340 345 350

Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly

355 360 365

Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro

370 375 380

Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser

385 390 395 400

Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg Trp Gln Glu

405 410 415

Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His

420 425 430

Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys

435 440

<210> 20

<211> 214

<212> PROTEIN

<213> Artificial sequence

<220>

<223>LC R2810

<400> 20

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Ser Ile Thr Ile Thr Cys Arg Ala Ser Leu Ser Ile Asn Thr Phe

20 25 30

Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Asn Leu Leu Ile

35 40 45

Tyr Ala Ala Ser Ser Leu His Gly Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Arg Thr Leu Gln Pro

65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Ser Asn Thr Pro Phe

85 90 95

Thr Phe Gly Pro Gly Thr Val Val Asp Phe Arg Arg Thr Val Ala Ala

100 105 110

Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly

115 120 125

Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala

130 135 140

Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln

145 150 155 160

Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser

165 170 175

Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr

180 185 190

Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser

195 200 205

Phe Asn Arg Gly Glu Cys

210

<210> 21

<211> 117

<212> PROTEIN

<213> Artificial sequence

<220>

<223>HCVR R1103

<400> 21

Glu Val Gln Leu Gln Gln Ser Gly Pro Glu Leu Val Lys Pro Gly Ala

1 5 10 15

Ser Val Arg Met Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr

20 25 30

Asn Ile His Trp Val Lys Gln Ser His Gly Lys Ser Leu Glu Trp Ile

35 40 45

Gly Tyr Ile Tyr Pro Asn Asn Gly Asp Asn Gly Tyr Asn Gln Lys Phe

50 55 60

Arg Gly Lys Ala Thr Leu Thr Val Asp Lys Ser Ser Ser Thr Ala Tyr

65 70 75 80

Met Glu Leu Arg Ser Leu Thr Ser Asp Asp Ser Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Gly Arg Leu Arg Tyr Phe Asp Val Trp Gly Thr Gly Thr Thr

100 105 110

Val Thr Val Ser Ser

115

<210> 22

<211> 111

<212> PROTEIN

<213> Artificial sequence

<220>

<223>LCVR R1103

<400> 22

Asn Ile Val Leu Thr Gln Ser Pro Ala Ser Leu Ala Val Ser Leu Gly

1 5 10 15

Gln Arg Ala Thr Ile Ser Cys Arg Ala Ser Glu Ser Val Asp Asn Tyr

20 25 30

Gly His Ser Phe Met His Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro

35 40 45

Lys Leu Leu Ile Tyr Leu Ala Ser Asn Leu Glu Ser Gly Val Pro Ala

50 55 60

Arg Phe Ser Gly Ser Gly Ser Arg Thr Asp Phe Thr Leu Thr Leu Asp

65 70 75 80

Pro Val Glu Ala Asp Asp Ala Ala Thr Tyr Tyr Cys Gln Gln Tyr Asn

85 90 95

Glu Asp Pro Pro Thr Phe Gly Ser Gly Thr Lys Leu Glu Ile Lys

100 105 110

<---

Claims (46)

1. A method for treating or inhibiting the growth of a type 2 immune-dependent malignancy, comprising: (a) selection of an individual with type 2 immune-dependent malignancy; and (b) administering to an individual in need thereof a therapeutically effective amount of an antibody that specifically binds the IL-4 receptor (IL-4R) and a therapeutically effective amount of an antibody that specifically binds programmed death protein 1 (PD-1), wherein the anti-PD-1 antibody comprises three heavy chain complementarity determining regions (CDRs) (HCDR1, HCDR2 and HCDR3) of a heavy chain variable region (HCVR) and three light chain CDRs (LCDR1, LCDR2 and LCDR3) of a light chain variable region (LCVR), wherein HCDR1 comprises the amino acid sequence of SEQ ID NO: 13, HCDR2 comprises the amino acid sequence of SEQ ID NO: 14, HCDR3 comprises the amino acid sequence of SEQ ID NO: 15, LCDR1 comprises the amino acid sequence of SEQ ID NO: 16, LCDR2 comprises the amino acid sequence of SEQ ID NO: 17 and LCDR3 comprises the amino acid sequence of SEQ ID NO: 18. 2. The method of claim 1, wherein the type 2 immune-dependent malignancy comprises pancreatic cancer, breast cancer, colorectal cancer, ovarian cancer, brain cancer, skin cancer, prostate cancer, kidney cancer, lung cancer, Hodgkin's lymphoma, or bladder cancer. 3. The method according to claim 1 or 2, wherein the type 2 immune-dependent malignancy comprises pancreatic cancer. 4. The method according to claim 1 or 2, wherein the type 2 immune-dependent malignancy comprises non-small cell lung cancer. 5. The method according to claim 1 or 2, wherein the type 2 immune-dependent malignancy comprises squamous cell carcinoma of the lung. 6. The method according to any one of paragraphs. 1-5, wherein the malignant neoplasm is primary, metastatic or recurrent. 7. The method of any one of claims 1-6, wherein the individual has been treated with a previous anti-tumor therapeutic agent or therapy. 8. The method according to any one of claims 1-7, wherein the individual is treated with a PD-1 inhibitor. 9. The method according to any one of claims 1-8, wherein the tumor is resistant or unresponsive to previous treatment with a therapeutic agent or therapy. 10. The method according to any one of claims 1-9, wherein the individual exhibits upregulation of at least one cytokine. 11. The method of claim 10, wherein the at least one cytokine comprises at least one of IL-4 and IL-13. 12. The method according to any one of claims 1-11, wherein the individual exhibits increased production of at least one cytokine. 13. The method of claim 12, wherein at least one cytokine comprises IL-4. 14. The method according to any one of claims 1-13, wherein the individual has an increased content of hyaluronic acid (HA) in a malignant tumor. 15. The method according to any one of claims 1-14, wherein the anti-IL-4R antibody comprises an HCVR comprising three complementarity determining regions (CDRs) of the heavy chain (HCDR1, HCDR2 and HCDR3) and an LCVR comprising three CDRs of the light chain (LCDR1, LCDR2 and LCDR3), wherein HCDR1 comprises the amino acid sequence of SEQ ID NO: 3; HCDR2 comprises the amino acid sequence of SEQ ID NO: 4; HCDR3 comprises the amino acid sequence of SEQ ID NO: 5; LCDR1 comprises the amino acid sequence of SEQ ID NO: 6; LCDR2 comprises the amino acid sequence of SEQ ID NO: 7 and LCDR3 comprises the amino acid sequence of SEQ ID NO: 8. 16. The method according to any one of claims 1-15, wherein the HCVR of the anti-IL-4R antibody comprises the amino acid sequence of SEQ ID NO: 1 and the LCVR of the anti-IL-4R antibody comprises the amino acid sequence of SEQ ID NO: 2. 17. The method according to any one of claims 1-16, wherein the anti-IL-4R antibody comprises a heavy chain and a light chain, wherein the heavy chain comprises the amino acid sequence of SEQ ID NO: 9. 18. The method according to any one of claims 1-16, wherein the anti-IL-4R antibody comprises a heavy chain and a light chain, wherein the light chain comprises the amino acid sequence of SEQ ID NO: 10. 19. The method according to any one of claims 1-16, wherein the anti-IL-4R antibody comprises a heavy chain and a light chain, wherein the heavy chain comprises the amino acid sequence of SEQ ID NO: 9 and the light chain comprises the amino acid sequence of SEQ ID NO: 10. 20. The method according to any one of claims 1-19, wherein the anti-IL-4R antibody is dupilumab or a bioequivalent thereof. 21. The method according to any one of claims 1-20, wherein the HCVR of the anti-PD-1 antibody comprises the amino acid sequence of SEQ ID NO: 11 and the LCVR of the anti-PD-1 antibody comprises the amino acid sequence of SEQ ID NO: 12. 22. The method according to any one of claims 1-21, wherein the anti-PD-1 antibody comprises a heavy chain and a light chain, wherein the heavy chain comprises the amino acid sequence of SEQ ID NO: 19. 23. The method according to any one of claims 1-21, wherein the anti-PD-1 antibody comprises a heavy chain and a light chain, wherein the light chain comprises the amino acid sequence of SEQ ID NO: 20. 24. The method according to any one of claims 1-21, wherein the anti-PD-1 antibody comprises a heavy chain and a light chain, wherein the heavy chain comprises the amino acid sequence of SEQ ID NO: 19 and the light chain comprises the amino acid sequence of SEQ ID NO: 20. 25. The method according to any one of claims 1-24, wherein the anti-PD-1 antibody is cemiplimab or a bioequivalent. 26. The method according to any one of claims 1-25, wherein one or more doses of an anti-IL-4R antibody are administered in combination with one or more doses of an anti-PD-1 antibody. 27. The method of claim 26, wherein at least one dose of the IL-4R antibody comprises from about 0.1 to about 50 mg/kg of the individual's body weight. 28. The method of claim 26, wherein at least one dose of the anti-IL-4R antibody comprises from about 0.05 to about 1000 mg. 29. The method of any one of claims 26-28, wherein each dose of the anti-IL-4R antibody is administered 0.5-12 weeks after the immediately preceding dose. 30. The method of any one of claims 26-29, wherein at least one dose of the anti-PD-1 antibody comprises from about 0.1 to about 20 mg/kg of the subject's body weight. 31. The method of any one of paragraphs 26-29, wherein at least one dose of the anti-PD-1 antibody comprises from about 0.05 to about 500 mg. 32. The method of claim 30 or 31, wherein each dose of the anti-PD-1 antibody is administered 0.5 to 12 weeks after the immediately preceding dose. 33. The method according to any one of claims 1-32, wherein the anti-IL-4R antibody is administered simultaneously with the anti-PD-1 antibody. 34. The method according to any one of claims 1-32, wherein the anti-IL-4R antibody is administered before the anti-PD-1 antibody. 35. The method according to any one of claims 1-32, wherein the anti-IL-4R antibody is administered after the anti-PD-1 antibody. 36. The method according to any one of claims 1-35, wherein the method promotes tumor regression, delays tumor growth, reduces the number of tumor cells, reduces tumor load, and/or prevents tumor recurrence in a patient. 37. The method of any one of claims 1-36, wherein the method promotes at least about 10% greater tumor regression in the treated individual compared to an untreated individual or an individual treated with any antibody as monotherapy. 38. The method of any one of claims 1-36, wherein the method results in at least a 30% or greater reduction in tumor cells or tumor size compared to an untreated individual or an individual treated with any antibody as monotherapy. 39. The method according to any one of claims 1-38, further comprising using at least one additional therapeutic agent or method of therapy. 40. The method of claim 39, wherein the additional therapeutic agent or method of therapy comprises chemotherapy, cyclophosphamide, surgery, radiation therapy, a tumor vaccine, a LAG3 inhibitor, a CTLA-4 inhibitor, a GITR agonist, a TIM3 inhibitor, a BTLA inhibitor, a TIGIT inhibitor, a CD38 inhibitor, a CD47 inhibitor, an IDO inhibitor, a VEGF antagonist, an Ang2 inhibitor, a TGFβ inhibitor, an EGFR inhibitor, a VISTA inhibitor, a CD40 agonist, a CSF1R inhibitor, a CCR2 inhibitor, a CXCR4 inhibitor, a CXCR2 inhibitor, a CCR4 inhibitor, a CXCL12 inhibitor, a CD28 activator, a costimulatory receptor agonist, an anti-tumor antigen antibody, an anti-CD3/anti-CD20 bispecific antibody, GM-CSF, a cytotoxin, chemotherapeutic agent, oncolytic virus, IL-6R inhibitor, IL-10 inhibitor, cytokine, ADC, chimeric antigen receptor T cell, anti-inflammatory drug, NSAID, and/or dietary supplement. 41. A method of treating or inhibiting the growth of a type 2 immune-dependent malignancy, comprising: (a) selecting an individual with a type 2 immune-dependent malignancy; and (b) administering to the individual in need thereof a therapeutically effective amount of an antibody that specifically binds the IL-4 receptor (IL-4R), wherein the IL-4R antibody comprises an HCVR comprising three heavy chain complementarity determining regions (CDRs) (HCDR1, HCDR2, and HCDR3) and an LCVR comprising three light chain CDRs (LCDR1, LCDR2, and LCDR3), wherein HCDR1 comprises the amino acid sequence of SEQ ID NO: 3; HCDR2 comprises the amino acid sequence of SEQ ID NO: 4; HCDR3 comprises the amino acid sequence of SEQ ID NO: 5; LCDR1 comprises the amino acid sequence of SEQ ID NO: 6; LCDR2 comprises the amino acid sequence of SEQ ID NO: 7 and LCDR3 comprises the amino acid sequence of SEQ ID NO: 8. 42. The method according to claim 41, wherein the HCVR of the anti-IL-4R antibody comprises the amino acid sequence of SEQ ID NO: 1 and the LCVR of the anti-IL-4R antibody comprises the amino acid sequence of SEQ ID NO: 2. 43. The method according to claim 41 or 42, wherein the anti-IL-4R antibody comprises a heavy chain and a light chain, wherein the heavy chain comprises the amino acid sequence of SEQ ID NO: 9 and the light chain comprises the amino acid sequence of SEQ ID NO: 10.