Combination therapy comprising Anti-CTLA4 antibodies and chemotherapy for cancer treatment

Abstract

1 BNT ref [P1819WO01] / / C&F ref. 240673WO A b s t r a c t The present invention relates to an anti-CTLA4 antibody or fragment thereof and / or a chemotherapy agent for use in a method of treating cancer in a subject in need thereof, wherein the subject is administered a combination comprising the anti-CTLA4 antibody or fragment thereof and the chemotherapy agent. The invention further provides a composition 5 and a kit of parts comprising the anti-CTLA4 antibody or fragment thereof and the chemotherapy agent. The invention further concerns a method of treating cancer in a subject in need thereof, wherein the method comprises administering to the subject the anti-CTLA4 antibody or fragment thereof and the chemotherapy agent. The invention further provides a combination of the anti-CTLA4 antibody or fragment thereof and the chemotherapy agent. 10

Description

[0001] COMBINATION THERAPY COMPRISING ANTI-CTLA4 ANTIBODIES AND CHEMOTHERAPY FOR CANCER TREATMENT

[0002] FIELD OF THE INVENTION

[0003] The present invention relates to methods for treating cancer in a subject using a combination of an anti-CTLA4 antibody and a chemotherapy agent. The invention further provides a composition, and a combination of the anti-CTLA4 antibody and the chemotherapy agent.

[0004] BACKGROUND OF THE INVENTION

[0005] The immune system of humans and other mammals provides protection against infection and disease through mechanisms of innate and adaptive immunity. The evolutionary ancient innate immune system provides a rapid, i.e., within minutes, but non-specific immune response which relies on invariant receptors that recognize common molecular patterns associated with pathogens (antigens). In contrast, the immune response of the adaptive immune system is considerably slower, i.e., taking days to weeks, but involves highly specific antigen receptors on B cells (B lymphocytes) and T cells (T lymphocytes) for pathogen recognition. Like the innate immune system, the adaptive immune system also comprises a humoral immune and a cell-mediated immune response. The humoral response is primarily driven by antibodies produced by B cells, which are able to recognize and neutralize foreign target antigens. In contrast, the cell-mediated immune response involves the activation of macrophages, neutrophils, natural killer cells (NK), and antigen-specific cytotoxic T cells, and the release of various cytokines in response to the recognition of an antigen.

[0006] Besides the protection against pathogens, the immune system plays a pivotal role in cancer prevention, development, and defense (Gonzalez et al., Genes Dev 2018, 32(19-20): 1267-

[0007] 1

[0008] BNT ref [P1819WO01] / / C&F ref. 240673WO 1284). Therefore, the field of cancer immunotherapy has attracted great attention from both the scientific and clinical communities over the past two decades. The overarching concept of cancer immunotherapy is to activate, induce and / or enhance a specific immune response in patients to control and / or eliminate the cancer disease. Cancer immunotherapies can generally be categorized into active and passive immunization strategies, depending on their ability to (re)-activate the immune system against cancer cells. Passive forms of cancer immunotherapy include tumor-targeting monoclonal antibodies (mAbs) and adoptively transferred T cells, while anti-cancer vaccines and immune checkpoint inhibitors are considered active forms of cancer immunotherapy (Galluzi et al., Oncotarget 2014, 5(24): 12472-12508).

[0009] Immune checkpoints play a pivotal role in the regulation of the immune response in tumor microenvironments. Immune checkpoints act as gatekeepers of the immune system modulating the nature, magnitude, and duration of the immune response and maintaining self-tolerance. One of the key inhibitory immune checkpoints is Cytotoxic T lymphocyte antigen-4 (CTLA4), also known as CD 152 (cluster of differentiation 152). The interaction between the B7.1 (CD80) (Freeman et al., J Immunol 1989, 143(8): 2714-22) and B7.2 (CD86) (Freeman et al., Science 1993, 262(5135): 909-11; Hathcock et al., Science 1993, 262(5135): 905-7; Wu et al., J Exp Med 1993, 178(5): 1789-93) ligands of antigen presenting cells and the CD28 and CTLA4 receptors (Leach et al., Science 1996, 271(5256): 1734-6; Linsley et al, J Exp Med 1991, 174(3): 561-9; Linsley et al., Proc Natl Acad Sci U S A 1990, 87(13): 5031-5) of T cells governs the activation or downregulation of T cells. CTLA4 is recognized as a key regulator of the adaptive immune response, having a central role in the maintenance of peripheral tolerance and in shaping the repertoire of emerging T cell responses and, is therefore a therapeutic target for the treatment of cancer and inflammation.

[0010] Treatment with anti-CTLA4 antibodies has been shown to be a powerful tool for enhancing anti-tumor immunity in preclinical models (Leach et al., Science 1996, 271(5256): 1734-6). Monotherapy with an antibody against CTLA4 promoted rejection of transplantable tumors of various origins.

[0011] BNT ref [P1819WO01] / / C&F ref. [240673WO] Based on promising preclinical tumor model studies, the clinical potential of antibodies against CTLA4 has been explored in different human malignancies. Unlike other checkpoint inhibitors such as anti-PD-l / PD-Ll antibodies, the anti-CTLA4 antibody, ipilimumab (YERVOY®), has gained market approval for one indication (melanoma) as a monotherapy. The toxicity profile significantly limits its dose and exposure that are required for achieving higher efficacy benefit. The less optimal dose may explain the consistently lower response rate than an anti-PD-1 antibody in head-to-head comparison studies in melanoma and its failure as a monotherapy in multiple Phase III clinical trials in other cancer indications. Despite its approval for multiple cancer indication as a combination therapy with nivolumab (Opdivo®, an anti-PD-1 antibody), the incidence of grade 3 / 4 immunotherapy-related adverse effects (irAEs) (e.g., up to 73-90% of patients with melanoma receiving ipilimumab / nivolumab as a neo-adjuvant therapy) remains high. CTLA4 remains a valid and attractive immunotherapy target, however, the less favorable safety profile significantly limits its clinical usage.

[0012] The molecular basis underlying irAEs and cancer immunotherapeutic effects (CITE) of anti-CTLA4 antibodies is traditionally viewed as antagonizing the endogenous function of CTLA4. In both mice and humans, genetic inactivation of CTLA4 caused severe autoimmune diseases. Therefore, an effective antagonist of CTLA4 is expected to likely induce autoimmune diseases. If inactivation of CTLA4 is necessary, then irAEs are expected to be a necessary price for cancer immunity.

[0013] Therefore, there is a desire to improve the therapeutic potential of anti-CTLA4 antibodies by increasing CITE efficacy while reducing the associated irAEs. In the past, different therapeutic immune check inhibitors were combined to enhance anti-tumor activity, particularly against poorly immunogenic tumors. However, this approach is associated with the risk of further increasing the autoimmune side effects further highlighting the need to selectively modulate cancer immunity without enhancing autoimmunity. Immune checkpoint inhibitors were also combined with other anti-cancer therapies, such as chemotherapies, radiation therapies, or anti-angiogenic agents (Vafaei et al., Cancer Cell Int, 2022, 22:2). Also, in these combination treatment approaches, anti-CTLA4 antibodies

[0014] BNT ref [P1819WO01] / / C&F ref. [240673WO] with improved therapeutic efficacy and reduced irAEs are desired, as the combined treatment-associated side-effects often lead to unacceptable toxicity.

[0015] Besides autoimmunity -related toxicity of immune checkpoint cancer immunotherapy, primary or acquired resistance of patients to treatment with immune checkpoint inhibitors is another central challenge in the field of cancer immunotherapy. For patients with primary or acquired resistance to treatment with immune checkpoint inhibitors, further cancer therapy options are limited with most being palliative. A clinical trial involving non-small cell lung cancer (NSCLC) patients with acquired resistance to anti-PD-l / anti-PD-Ll antibody treatment demonstrated that further treatment with a combination of an anti- CTLA4 antibody (tremelimumab, Imjudo®) and an anti-PD-Ll antibody (durvalumab (Imfinzi®) showed only minimal treatment success, i.e., an overall response rate of 4%.

[0016] Furthermore, the applicability of cancer immunotherapy is limited for patients who lack oncogenic driver mutations that can be targeted by cancer immunotherapeutic approaches.

[0017] Therefore, there is an unmet medical need for improved cancer immunotherapeutic methods and new and improved cancer therapies addressing the challenges posed by, e.g., autoimmune side effects, resistance to treatment with immune checkpoint inhibitors, and lack of targetable oncogenic driver mutations.

[0018] SUMMARY OF THE INVENTION

[0019] Against the aforementioned background, it is therefore an object of the present invention to provide safe and effective immunotherapeutic treatment options for subjects afflicted with cancer. It is a further object of the present invention to provide effective therapeutic means for the treatment of cancer with reduced autoimmune side effects. It is also an object of the present invention to provide improved immunotherapeutic treatment options for subjects afflicted with cancer and resistance to treatment with immune checkpoint inhibitors. It is a further object of the present invention to provide improved cancer treatment methods for subjects afflicted with cancer who lack targetable oncogenic driver mutations. It is a further object of the present invention to provides safe and effective combination treatment options

[0020] BNT ref [P1819WO01] / / C&F ref. [240673WO] for the treatment of cancer in patients with resistance to treatment with immune checkpoint inhibitors and without targetable oncogenic driver mutations.

[0021] These objects are achieved by the invention set forth in the claims and embodiments explained in more detail below.

[0022] The invention concerns an anti-CTLA4 antibody or fragment thereof for use in a method of treating cancer in a subject in need thereof, wherein the method comprises administering to the subject: a) the anti-CTLA4 antibody or fragment thereof; and b) a chemotherapy agent.

[0023] The invention further concerns a chemotherapy agent for use in a method of treating cancer in a subject in need thereof, wherein the method comprises administering to the subject: a) the chemotherapy agent or fragment thereof; and b) an anti-CTLA4 antibody.

[0024] Challenges of cancer immunotherapy with immune checkpoint inhibitors are the undesired side effects associated with the treatment, such as immune-related adverse events and the resistance of patients to the treatment. In the research underlying the invention, the applicants surprisingly found that the combined use of the anti-CTLA4 antibody and the chemotherapy agent provided an unexpected beneficial effect in the treatment of cancer, while in parallel providing a safe treatment.

[0025] This surprising effect results in longer overall survival, longer progression-free survival, longer freedom from disease progression (stable disease state), reduction of tumor-related symptoms, and / or reduction of need for pain medications during and / or following the anticancer therapy (anti-CTLA4 antibody and chemotherapy agent combination therapy). The effect can be reflected in an improved quality of life, such as mobility, strength of appetite, and / or psychological status.

[0026] BNT ref [P1819WO01] / / C&F ref. [240673WO] The invention further provides an anti-CTLA4 antibody or fragment thereof and a chemotherapy agent for use in a method of treating cancer in a subject in need thereof, wherein the method comprises administering to the subject: a) the anti-CTLA4 antibody or fragment thereof; and b) the chemotherapy agent.

[0027] The invention also concerns a composition comprising: a) an anti-CTLA4 antibody or fragment thereof; and b) a chemotherapy agent.

[0028] The invention further provides a method of treating cancer in a subject in need thereof, wherein the method comprises administering to the subject: a) an anti-CTLA4 antibody or fragment thereof; and b) a chemotherapy agent.

[0029] The invention also provides a combination of an anti-CTLA4 antibody or fragment thereof and a chemotherapy agent.

[0030] Advantageous embodiments of the invention are indicated in the dependent claims and in the following.

[0031] DETAILED DESCRIPTION

[0032] Although certain embodiments of the present invention are described in detail below, it is to be understood that this invention is not limited to the particular embodiments, methodologies, protocols and reagents described herein as these may vary within the scope set by the claims. It is also to be understood that terminology used herein is for the purpose of describing particular embodiments only, and is not intended to limit the scope of the present invention which is defined by the appended claims. Unless defined otherwise, all technical and scientific terms used herein have the same meanings as commonly understood by one of ordinary skill in the art.

[0033] BNT ref [P1819WO01] / / C&F ref. [240673WO] In the following description, certain elements of the present invention will be described. These elements may be discussed with specific embodiments, however, it should be understood that they may be combined in any manner and in any number to create additional embodiments. The variously described examples, features and particular embodiments should not be construed to limit the present invention to only the explicitly described embodiments or to the explicitly described combination of features. This description should be understood to disclose and encompass embodiments which combine the explicitly described embodiments with any number of the disclosed and / or preferred elements. Furthermore, any permutations and combinations of all described elements in this application should be considered disclosed by this description unless the context indicates otherwise.

[0034] The above objects are achieved by the following embodiments in accordance with the invention:

[0035] 1. An anti-CTLA4 antibody or fragment thereof for use in a method of treating cancer in a subject in need thereof, wherein the method comprises administering to the subject: a) the anti-CTLA4 antibody; and b) a chemotherapy agent.

[0036] 2. A chemotherapy agent for use in a method of treating cancer in a subject in need thereof, wherein the method comprises administering to the subject: a) the chemotherapy agent; and b) an anti-CTLA4 antibody or fragment thereof.

[0037] 3. The anti-CTLA4 antibody or fragment thereof for use according to embodiment 1 or the chemotherapy agent for use according to embodiment 2, wherein the anti-

[0038] BNT ref [P1819WO01] / / C&F ref. [240673WO] CTLA4 antibody or fragment thereof and the chemotherapy agent are administered separately. The anti-CTLA4 antibody or fragment thereof for use according to embodiment 1 or 3 or the chemotherapy agent for use according to embodiment 2 or 3, wherein the anti-CTLA4 antibody or fragment thereof and the chemotherapy agent are administered concurrently or consecutively. The anti-CTLA4 antibody or fragment thereof for use according to embodiments 1, 3 or 4 or the chemotherapy agent for use according to embodiments 2-4, wherein the anti-CTLA4 antibody or fragment thereof and the chemotherapy agent are administered intravenously. The anti-CTLA4 antibody or fragment thereof for use according to embodiments 1 or 3-5 or the chemotherapy agent for use according to embodiments 2-5, wherein the anti-CTLA4 antibody or fragment thereof and the chemotherapy agent are administered via an intravenous injection or an intravenous infusion, preferably an intravenous infusion. The anti-CTLA4 antibody or fragment thereof for use according to embodiments 1 or 3-6 or the chemotherapy agent for use according to embodiments 2-6, wherein the anti-CTLA4 antibody or fragment thereof dosage administered ranges from about 0.1 mg / kg to about 45 mg / kg body weight, preferably from about 1 mg / kg to about 30 mg / kg body weight, more preferably from about 2 mg / kg to about 20 mg / kg body weight. The anti-CTLA4 antibody or fragment thereof for use according to embodiments 1 or 3-7 or the chemotherapy agent for use according to embodiments 3-7, wherein the anti-CTLA4 antibody or fragment thereof dosage administered is independently selected from about 0.1 mg / kg body weight, about 0.3 mg / kg body weight, about 1.0 mg / kg body weight, about 3.0 mg / kg body weight, about 6 mg / kg body weight,

[0039] BNT ref [P1819WO01] / / C&F ref. [240673WO] about 10 mg / kg body weight, about 12 mg / kg body weight, about 15 mg / kg body weight, and about 20 mg / kg body weight.

[0040] 9. The anti-CTLA4 antibody or fragment thereof for use according to embodiments 1 or 3-8 or the chemotherapy agent for use according to embodiments 2-8, wherein the chemotherapy agent dosage administered ranges from about 0.1 to about 300 mg / m2body surface area, preferably from about 1 to about 200 mg / m2body surface area, more preferably from about 10 to about 100 mg / m2body surface area.

[0041] 10. The anti-CTLA4 antibody or fragment thereof for use according to embodiments 1 or 3-9 or the chemotherapy agent for use according to embodiments 2-9, wherein the chemotherapy agent dosage administered is independently selected from about 0.1 mg / m2body surface area, about 0.3 mg / m2body surface area, about 1 mg / m2body surface area, about 10 mg / m2body surface area, about 22.5 mg / m2body surface area, about 25 mg / m2body surface area, about 27.5 mg / m2body surface area, about 30 mg / m2body surface area, about 32.5 mg / m2body surface area, about 35 mg / m2body surface area, about 37.5 mg / m2body surface area, about 40 mg / m2body surface area about 50 mg / m2body surface area, about 55 mg / m2body surface area, about 60 mg / m2body surface area, about 65 mg / m2body surface area, about 70 mg / m2body surface area, about 75 mg / m2body surface area, about 100 mg / m2body surface area, and about 125 mg / m2body surface area.

[0042] 11. The anti-CTLA4 antibody or fragment thereof for use according to embodiments 1 or 3-10 or the chemotherapy agent for use according to embodiments 2-10, wherein a treatment cycle is repeated at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, or 17 times.

[0043] 12. The anti-CTLA4 antibody or fragment thereof for use according to embodiments 1 or 3-11 or the chemotherapy agent for use according to embodiments 2-11, wherein each treatment cycle has up to about 28 days, preferably up to about 21 days, more preferably about 21 days.

[0044] BNT ref [P1819WO01] / / C&F ref. [240673WO] 13. The anti-CTLA4 antibody or fragment thereof for use according to embodiments 1 or 3-12 or the chemotherapy agent for use according to embodiments 2-12, wherein the anti-CTLA4 antibody or fragment thereof is administered about every 6 weeks, preferably about every 4 weeks, more preferably about every 3 weeks.

[0045] 14. The anti-CTLA4 antibody or fragment thereof for use according to embodiments 1 or 3-13 or the chemotherapy agent for use according to embodiments 2-13, wherein the chemotherapy agent is administered about every 6 weeks, preferably about every 4 weeks, more preferably about every 3 weeks.

[0046] 15. The anti-CTLA4 antibody or fragment thereof for use according to embodiments 1 or 3-14 or the chemotherapy agent for use according to embodiments 2-14, wherein the chemotherapy agent is administered up to the 8thtreatment cycle, preferably up to the 7thtreatment cycle, more preferably up to the 6thtreatment cycle, even more preferably up to the 5thtreatment cycle, and most preferably up to the 4thtreatment cycle.

[0047] 16. The anti-CTLA4 antibody or fragment thereof for use according to embodiments 1 or 3-15 or the chemotherapy agent for use according to embodiments 2-15, wherein the anti-CTLA4 antibody or fragment thereof and the chemotherapy agent are administered up to the 8thtreatment cycle, preferably up to the 7thtreatment cycle, more preferably up to the 6thtreatment cycle, even more preferably up to the 5thtreatment cycle, and most preferably up to the 4thtreatment cycle, followed by consecutive administration of the anti-CTLA4 antibody or fragment thereof.

[0048] 17. The anti-CTLA4 antibody or fragment thereof for use according to embodiments 1 or 3-16 or the chemotherapy agent for use according to embodiments 2-16, wherein the anti-CTLA4 antibody or fragment thereof is administered for 17 treatment cycles or up to 1 year.

[0049] BNT ref [P1819WO01] / / C&F ref. [240673WO] 18. The anti-CTLA4 antibody or fragment thereof for use according to embodiments 1 or 3-17 or the chemotherapy agent for use according to embodiments 2-17, wherein the anti-CTLA4 antibody or fragment thereof dosage administered at least during the 1stand 2ndtreatment cycle ranges from about 2 mg / kg to about 20 mg / kg body weight, preferably from about 4 mg / kg to about 15 mg / kg body weight, more preferably from about 6 mg / kg to about 12 mg / kg body weight.

[0050] 19. The anti-CTLA4 antibody or fragment thereof for use according to embodiments 1 or 3-18 or the chemotherapy agent for use according to embodiments 2-18, wherein the anti-CTLA4 antibody or fragment thereof dosage administered at least during the 3rdand 4thcycle ranges from about 2 mg / kg to about 20 mg / kg, preferably from about 2 mg / kg to about 10 mg / kg, more preferably from about 3 mg / kg to about 8 mg / kg, most preferably from about 4 mg / kg to about 7 mg / kg.

[0051] 20. The anti-CTLA4 antibody or fragment thereof for use according to embodiments 1 or 3-19 or the chemotherapy agent for use according to embodiments 2-19, wherein the chemotherapy agent dosage administered during the 1st, the 2nd, the 3rd, and the 4thcycle ranges from about 1 to about 200 mg / m2body surface area, preferably from about 10 to about 150 mg / m2body surface area, more preferably from about 25 to about 100 mg / m2body surface area, most preferably from about 30 to about 80 mg / m2body surface area.

[0052] 21. The anti-CTLA4 antibody or fragment thereof for use according to embodiments 1 or 3-20 or the chemotherapy agent for use according to embodiments 2-20, wherein the anti-CTLA4 antibody or fragment thereof and / or the chemotherapy agent are formulated with one or more pharmaceutically acceptable carriers, diluents and / or excipients.

[0053] 22. The anti-CTLA4 antibody or fragment thereof for use according to embodiments 1 or 3-21 or the chemotherapy agent for use according to embodiments 2-21, wherein the subject has not previously been treated for cancer.

[0054] BNT ref [P1819WO01] / / C&F ref. [240673WO] 23. The anti-CTLA4 antibody or fragment thereof for use according to embodiments 1 or 3-21 or the chemotherapy agent for use according to embodiments 2-21, wherein the subject has been previously treated for cancer, preferably wherein the subject had at least one previous chemotherapy treatment.

[0055] 24. The anti-CTLA4 antibody or fragment thereof for use according to embodiment 23 or the chemotherapy agent for use according to embodiment 23, wherein the at least one previous chemotherapy treatment comprises or is a platinum-containing chemotherapy.

[0056] 25. The anti-CTLA4 antibody or fragment thereof for use according to embodiments 1 or 3-24 or the chemotherapy agent for use according to embodiments 2-24, wherein overall survival is increased in said subject compared to the chemotherapy agent or the anti-CTLA4 antibody or fragment thereof treatment alone.

[0057] 26. The anti-CTLA4 antibody or fragment thereof for use according to embodiments 1 or 3-25 or the chemotherapy agent for use according to embodiments 2-25, wherein median progression-free survival is increased in said subject compared to the chemotherapy agent or the anti-CTLA4 antibody or fragment thereof treatment alone.

[0058] 27. The anti-CTLA4 antibody or fragment thereof for use according to embodiments 1 or 3-26 or the chemotherapy agent for use according to embodiments 2-26, wherein the cancer comprises one or more solid tumors.

[0059] 28. The anti-CTLA4 antibody or fragment thereof for use according to embodiments 1 or 3-27 or the chemotherapy agent for use according to embodiments 2-27, wherein the cancer is selected from melanoma, lung cancer, human papillomavirus (HPV)- induced cancer, breast cancer, hepatocellular carcinoma, ovarian cancer such as ovarian carcinoma, prostate cancer such as prostate carcinoma, Hodgkin's or nonHodgkin's lymphoma, acute myelogenic Leukemia, chronic myelogenic Leukemia,

[0060] BNT ref [P1819WO01] / / C&F ref. [240673WO] acute lymphocytic Leukemia, chronic lymphocytic Leukemia, or renal cell carcinoma. The anti-CTLA4 antibody or fragment thereof for use according to embodiments 1 or 3-28 or the chemotherapy agent for use according to embodiments 2-28, wherein the cancer is non-small cell lung cancer (NSCLC). The anti-CTLA4 antibody or fragment thereof for use according to embodiment 29 or the chemotherapy agent for use according to embodiment 29, wherein the NSCLC has a squamous histology. The anti-CTLA4 antibody or fragment thereof for use according to embodiment 29 or the chemotherapy agent for use according to embodiment 29, wherein the NSCLC has a non-squamous histology. The anti-CTLA4 antibody or fragment thereof for use according to embodiments 29-31 or the chemotherapy agent for use according to embodiments 29-31, wherein the NSCLC has locally advanced or metastasized. The anti-CTLA4 antibody or fragment thereof for use according to embodiments 29-32 or the chemotherapy agent for use according to embodiments 29-32, wherein the NSCLC is negative for targetable NSCLC driver mutations. The anti-CTLA4 antibody or fragment thereof for use according to embodiment 33 or the chemotherapy agent for use according to embodiment 33, wherein the NSCLC is negative for EGFR and / or ALK mutations. The anti-CTLA4 antibody or fragment thereof for use according to embodiments 29-32 or the chemotherapy agent for use according to embodiments 29-32, wherein the NSCLC is positive for KRAS mutations.

[0061] BNT ref [P1819WO01] / / C&F ref. [240673WO] The anti-CTLA4 antibody or fragment thereof for use according to embodiments 1 or 3-35 or the chemotherapy agent for use according to embodiments 2-35, wherein the chemotherapy agent comprises or is selected from cisplatin, oxaliplatin, carboplatin, lurbinectedin, topotecan, paclitaxel, nanoparticle albumin-bound paclitaxel (nab -paclitaxel), pemetrexed, 5-fluoruracil, irinotecan, etoposide, gemcitabine, docetaxel or combinations thereof. The anti-CTLA4 antibody or fragment thereof for use according to embodiment 36 or the chemotherapy agent for use according to embodiment 36, wherein the chemotherapy agent comprises or is docetaxel. The anti-CTLA4 antibody or fragment thereof for use according to embodiments 1 or 3-37 or the chemotherapy agent for use according to embodiments 2-37, wherein the subject has previously not been treated with docetaxel. The anti-CTLA4 antibody or fragment thereof for use according to embodiments 1 or 3-38 or the chemotherapy agent for use according to embodiments 2-38, wherein the anti-CTLA4 antibody or fragment thereof comprises: a) a heavy chain variable region comprising (1) a complementarity-determining region 1 (HCDR1) comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 1, (2) a complementarity-determining region 2 (HCDR2) comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 3, 4 or 5, and (3) a complementarity-determining region 3 (HCDR3) comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 9; and b) a light chain variable region comprising: (1) a complementarity-determining region 1 (LCDR1) comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 2, (2) a complementarity-determining region 2 (LCDR2) comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 6, 7, or 8, and (3) a complementarity-determining region 3 (LCDR3) comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 10.

[0062] BNT ref [P1819WO01] / / C&F ref. [240673WO] The anti-CTLA4 antibody or fragment thereof for use according to embodiment 39 or the chemotherapy agent for use according to embodiment 39, wherein the anti- CTLA4 antibody or fragment thereof comprises: a) a heavy chain variable region comprising (1) a HCDR1 comprising or consisting of an amino acid sequence is set forth in SEQ ID NO: 1, (2) a HCDR2 comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 3, and (3) a HCDR3 comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 9; and b) a light chain variable region comprising: (1) a LCDR1 comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 2, (2) a LCDR2 comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 7, and (3) a LCDR3 comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 10. The anti-CTLA4 antibody or fragment thereof for use according to embodiment 39 or the chemotherapy agent for use according to embodiment 39, wherein the anti- CTLA4 antibody or fragment thereof comprises: a) a heavy chain variable region comprising (1) a HCDR1 comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 1, (2) a HCDR2 comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 5, and (3) a HCDR3 comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 9; and b) a light chain variable region comprising: (1) a LCDR1 comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 2, (2) a LCDR2 comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 7, and (3) a LCDR3 comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 10.

[0063] BNT ref [P1819WO01] / / C&F ref. [240673WO] The anti-CTLA4 antibody or fragment thereof for use according to embodiment 39 or the chemotherapy agent for use according to embodiment 39, wherein the anti- CTLA4 antibody or fragment thereof comprises: a) a heavy chain variable region comprising (1) a HCDR1 comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 1, (2) a HCDR2 comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 5, and (3) a HCDR3 comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 9; and b) a light chain variable region comprising: (1) a LCDR1 comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 2, (2) a LCDR2 comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 8, and (3) a LCDR3 comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 10. The anti-CTLA4 antibody or fragment thereof for use according to embodiments 1 or 3-42 or the chemotherapy agent for use according to embodiments 2-42, wherein the anti-CTLA4 antibody or fragment thereof comprises: a) a heavy chain variable domain comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 11, 12, or 13, or an amino acid sequence comprising at least 80%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 11, 12, or 13; and b) a light chain variable domain comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 14, 15, or 16, or an amino acid sequence comprising at least 80%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 14, 15, or 16.

[0064] BNT ref [P1819WO01] / / C&F ref. [240673WO] The anti-CTLA4 antibody or fragment thereof for use according to embodiment 43 or the chemotherapy agent for use according to embodiment 43, wherein the anti- CTLA4 antibody or fragment thereof comprises: a) a heavy chain variable domain comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 11 or an amino acid sequence comprising at least 80%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 11; and b) a light chain variable domain comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 15 or an amino acid sequence comprising at least 80%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 15. The anti-CTLA4 antibody or fragment thereof for use according to embodiment 43 or the chemotherapy agent for use according to embodiment 43, wherein the anti- CTLA4 antibody or fragment thereof comprises: a) a heavy chain variable domain comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 13 or an amino acid sequence comprising at least 80%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 13; and b) a light chain variable domain comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 15 or an amino acid sequence comprising at least 80%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 15. The anti-CTLA4 antibody or fragment thereof for use according to embodiment 43 or the chemotherapy agent for use according to embodiment 43, wherein the anti- CTLA4 antibody or fragment thereof comprises: a) a heavy chain variable domain comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 13 or an amino acid sequence comprising at least 80%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 13; and

[0065] BNT ref [P1819WO01] / / C&F ref. [240673WO] b) a light chain variable domain comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 16 or an amino acid sequence comprising at least 80%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 16. The anti-CTLA4 antibody or fragment thereof for use according to embodiments 1, 3-46 or the chemotherapy agent for use according to embodiments 2-46, wherein the anti-CTLA4 antibody comprises a constant region preferably derived from a human antibody, preferably the constant region is selected from the constant region of human IgGl, IgG2, IgG3 or IgG4. The anti-CTLA4 antibody or fragment thereof for use according to embodiment 47 or the chemotherapy agent for use according to embodiment 47, wherein the anti- CTLA4 antibody comprises an IgGl Fc domain, preferably comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 17 or 18 or an amino acid sequence comprising at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 17 or 18. The anti-CTLA4 antibody or fragment thereof for use according to embodiments 1, 3-48 or the chemotherapy agent for use according to embodiments 2-48, wherein the anti-CTLA4 antibody comprises: a) a heavy chain comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 19, 21, or 23 or an amino acid sequence comprising at least 80%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 19, 21, or 23; and b) a light chain comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 25, 27, or 29 or an amino acid sequence comprising at least 80%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 25, 27, or 29.

[0066] BNT ref [P1819WO01] / / C&F ref. [240673WO] The anti-CTLA4 antibody or fragment thereof for use according to embodiment 49 or the chemotherapy agent for use according to embodiment 49, wherein the anti- CTLA4 antibody comprises: a) a heavy chain comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 19 or an amino acid sequence comprising at least 80%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 19; and b) a light chain comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 27 or an amino acid sequence comprising at least 80%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 27. The anti-CTLA4 antibody or fragment thereof for use according to embodiment 49 or the chemotherapy agent for use according to embodiment 49, wherein the anti- CTLA4 antibody comprises: a) a heavy chain comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 23 or an amino acid sequence comprising at least 80%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 23; and b) a light chain comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 27 or an amino acid sequence comprising at least 80%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 27. The anti-CTLA4 antibody or fragment thereof for use according to embodiment 49 or the chemotherapy agent for use according to embodiment 49, wherein the anti- CTLA4 antibody comprises: a) a heavy chain comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 23 or an amino acid sequence comprising at least 80%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 23; and

[0067] BNT ref [P1819WO01] / / C&F ref. [240673WO] b) a light chain comprising or consisting of amino acid sequence set forth in SEQ ID NO: 29 or an amino acid sequence comprising at least 80%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 29. The anti-CTLA4 antibody or fragment thereof for use according to embodiments 1, 3-52 or the chemotherapy agent for use according to embodiments 2-52, wherein the anti-CTLA4 antibody or fragment thereof is capable of binding to human CTLA4. The anti-CTLA4 antibody or fragment thereof for use according to embodiments 1, 3-53 or the chemotherapy agent for use according to embodiments 2-53, wherein the anti CTLA4 antibody or fragment thereof is a humanized anti-CTLA4 antibody or fragment thereof. The anti-CTLA4 antibody or fragment thereof for use according to embodiments 1, 3-46, 53, 54, or the chemotherapy agent for use according to embodiments 2-46, 53, 54, wherein the anti-CTLA4 antibody fragment is an antigen-binding fragment or a variant thereof. The anti-CTLA4 antibody or fragment thereof for use according to embodiment 55 or the chemotherapy agent for use according to embodiment 55, wherein the antigen binding fragment or variant thereof is selected from the group consisting of a Fab, a Fab’, a F(ab’)2, a scFV, a diabody, a triabody, a minibody, and a single-domain antibody (sdAB), and variants thereof. An anti-CTLA4 antibody or fragment thereof and a chemotherapy agent for use in a method of treating cancer in a subj ect in need thereof, wherein the method comprises administering to the subject: a) the anti-CTLA4 antibody or fragment thereof; and

[0068] BNT ref [P1819WO01] / / C&F ref. [240673WO] b) the chemotherapy agent. A composition comprising: a) an anti-CTLA4 antibody or fragment thereof; and b) a chemotherapy agent. The composition according to embodiment 58, comprising the anti-CTLA4 antibody or fragment thereof in a range from about 0.1 mg / kg to about 45 mg / kg body weight, preferably from about 1 mg / kg to about 30 mg / kg body weight, more preferably from about 2 mg / kg to about 20 mg / kg body weight, even more preferably from about 4 mg / kg to about 15 mg / kg body weight, and most preferably from about 5 mg / kg to about 12 mg / kg body weight. The composition according to embodiment 58 or 59, comprising the chemotherapy agent in a range from about 0.1 to about 300 mg / m2body surface area, preferably from about 1 to about 200 mg / m2body surface area, more preferably from about 10 to about 150 mg / m2body surface area, even more preferably from about 25 to about 100 mg / m2body surface area, and most preferably from about 30 to about 80 mg / m2body surface area. The composition according to embodiment 60, comprising about 75 mg / m2body surface area, 55 mg / m2body surface area, or 37.5 mg / m2body surface area of the chemotherapy agent. The composition according to embodiments 58-61, comprising one or more pharmaceutically acceptable carriers, diluents and / or excipients. The composition according to embodiments 58-62, wherein the composition is a pharmaceutical composition.

[0069] BNT ref [P1819WO01] / / C&F ref. [240673WO] 64. A method of treating cancer in a subject in need thereof, wherein the method comprises administering to the subject: a) an anti-CTLA4 antibody or fragment thereof; and b) a chemotherapy agent.

[0070] 65. A combination of an anti-CTLA4 antibody or fragment thereof and a chemotherapy agent.

[0071] Definitions

[0072] The terms indicated for explanation of the invention have the following meaning, unless otherwise indicated in the description or the embodiments. Additional definitions are set forth throughout the detailed description.

[0073] Terms “a” and “an” and “the” and similar reference used in the context of describing the invention (especially in the context of the embodiments) are to be construed to cover both the singular and the plural, unless otherwise indicated herein or clearly contradicted by context.

[0074] The terms “about” or “approximately” as used herein denotes a range of ±10% of a reference value. For examples, “about 10” defines a range of 9 to 11. In general, those skilled in the art, familiar with the context, will appreciate the relevant degree of variance encompassed by “about” or “approximately” in that context.

[0075] The term “plurality” refers to the state of being plural.

[0076] Unless expressly specified otherwise, the term “comprising” is used in the context of the present disclosure to indicate that further members may optionally be present in addition to the members of the list introduced by “comprising”. It is, however, contemplated as specific embodiments of the present invention that each time the term “comprising” is used,

[0077] BNT ref [P1819WO01] / / C&F ref. [240673WO] this shall also encompass the possibility of no further members being present, i.e., for the purpose of this embodiment “comprising” can be understood as having the meaning of “consisting of’.

[0078] Preferably, the terms used herein are defined as described in “A multilingual glossary of biotechnological terms: (IUPAC Recommendations)”, H.G.W. Leuenberger, B. Nagel, and H. K51bl, Eds., (1995) Helvetica Chimica Acta, CH-4010 Basel, Switzerland.

[0079] The practice of the present disclosure will employ, unless otherwise indicated, conventional methods of biochemistry, cell biology, immunology, and recombinant DNA techniques which are explained in the literature in the field (cf., e.g., Molecular Cloning: A Laboratory Manual, 4th Edition, M.R. Green, J. Sambrook et al. eds., Cold Spring Harbor Laboratory Press, Cold Spring Harbor 2012).

[0080] Recitation of ranges of values herein is merely intended to serve as a shorthand method of referring individually to each separate value falling within the range. Unless otherwise indicated herein, each individual value is incorporated into the specification as if it was individually recited herein. All methods described herein can be performed in any suitable order unless otherwise indicated herein or otherwise clearly contradicted by context. The use of any and all examples, or exemplary language (e.g., “such as”), provided herein is intended merely to better illustrate the invention and does not pose a limitation on the scope of the invention otherwise claimed. No language in the specification should be construed as indicating any non-claimed element essential to the practice of the invention.

[0081] Several documents are cited throughout the text of this specification. Each of the documents cited herein (including all patents, patent applications, scientific publications, manufacturer's specifications, instructions, etc.), whether supra or infra, are hereby incorporated by reference in their entirety. Nothing herein is to be construed as an admission that the invention is not entitled to antedate such disclosure by virtue of prior invention.

[0082] The terms “polynucleotide” and “nucleic acid” can be used interchangeably herein to refer to polymers of nucleotides. The term “polynucleotide” comprises deoxyribonucleic acid

[0083] BNT ref [P1819WO01] / / C&F ref. [240673WO] (DNA), ribonucleic acid (RNA), combinations thereof, and modified forms thereof. The term comprises genomic DNA, cDNA, mRNA, recombinantly produced and chemically synthesized molecules. In some embodiments, a polynucleotide is DNA. In some embodiments, a polynucleotide is RNA. A polynucleotide may be present as a mixture of DNA and RNA. A polynucleotide may be further present as a single-stranded or doublestranded and linear or covalently circularly closed molecule. A polynucleotide can be isolated. The term “isolated polynucleotide “ means, according to the present invention, that the polynucleotide (i) was amplified in vitro, for example via polymerase chain reaction (PCR) for DNA or in vitro transcription (using, e.g., an RNA polymerase) for RNA, (ii) was produced recombinantly by cloning, (iii) was purified, for example, by cleavage and separation by gel electrophoresis, or (iv) was synthesized, for example, by chemical synthesis.

[0084] The term “RNA” relates to a nucleic acid molecule which includes ribonucleotide residues. In preferred embodiments, the RNA contains all or a majority of ribonucleotide residues. As used herein, “ribonucleotide” refers to a nucleotide with a hydroxyl group at the 2'- position of a P-D-ribofuranosyl group. RNA encompasses without limitation, double stranded RNA, single stranded RNA, isolated RNA such as partially purified RNA, essentially pure RNA, synthetic RNA, recombinantly produced RNA, as well as modified RNA that differs from naturally occurring RNA by the addition, deletion, substitution and / or alteration of one or more nucleotides. Such alterations may refer to addition of nonnucleotide material to internal RNA nucleotides or to the end(s) of RNA. It is also contemplated herein that nucleotides in RNA may be non-standard nucleotides, such as chemically synthesized nucleotides or deoxynucleotides. For the present invention, these altered / modified nucleotides can be referred to as analogs of naturally occurring nucleotides, and the corresponding RNAs containing such altered / modified nucleotides (i.e., altered / modified RNAs) can be referred to as analogs of naturally occurring RNAs. A molecule contains “a majority of ribonucleotide residues” if the content of ribonucleotide residues in the molecule is more than 50% (such as at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%), based on the total number of nucleotide residues in the molecule. The total number of nucleotide residues in a molecule is the sum

[0085] BNT ref [P1819WO01] / / C&F ref. [240673WO] of all nucleotide residues (irrespective of whether the nucleotide residues are standard (i.e., naturally occurring) nucleotide residues or analogs thereof). The term “RNA” further includes mRNA, tRNA, ribosomal RNA (rRNA), small nuclear RNA (snRNA), selfamplifying RNA (saRNA), trans-amplifying RNA (taRNA), single-stranded RNA (ssRNA), dsRNA, inhibitory RNA (such as antisense ssRNA, small interfering RNA (siRNA), or microRNA (miRNA)), activating RNA (such as small activating RNA) and immunostimulatory RNA (isRNA). In some embodiments, “RNA” refers to mRNA. RNA as described herein may comprise in vitro transcribed RNA (IVT-RNA) and may be obtained by in vitro transcription of an appropriate DNA template. The promoter for controlling transcription can be any promoter for any RNA polymerase. A DNA template for in vitro transcription may be obtained by cloning of a nucleic acid, in particular cDNA, and introducing it into an appropriate vector for in vitro transcription. The cDNA may be obtained by reverse transcription of RNA.

[0086] The term “mRNA” refers to messenger RNA that comprises a coding sequence encoding a polypeptide. An mRNA may further comprise non-coding sequences such as a 5 ’cap, a 5’UTR, a 3’UTR, a Kozak sequence, an FI element or a poly(A) tail or any combination thereof. According to the present disclosure, the term “mRNA” means “messenger-RNA” and includes a “transcript” which may be generated by using a DNA template. Generally, mRNA encodes a peptide or polypeptide. mRNA is single-stranded but may contain self- complementary sequences that allow parts of the mRNA to fold and pair with itself to form double helices. According to the present disclosure, “dsRNA” means double-stranded RNA and is RNA with two partially or completely complementary strands.

[0087] Herein, the term “DNA” relates to a nucleic acid molecule which is entirely or at least substantially composed of deoxyribonucleotide residues. In preferred embodiments, the DNA contains all or a majority of deoxyribonucleotide residues. As used herein, “deoxyribonucleotide” refers to a nucleotide which lacks a hydroxyl group at the 2’- position of a P-D-ribofuranosyl group. DNA encompasses without limitation, double stranded DNA, single stranded DNA, isolated DNA such as partially purified DNA, essentially pure DNA, synthetic DNA, recombinantly produced DNA, as well as modified DNA that differs from naturally occurring DNA by the addition, deletion, substitution

[0088] BNT ref [P1819WO01] / / C&F ref. [240673WO] and / or alteration of one or more nucleotides. Such alterations may refer to addition of nonnucleotide material to internal DNA nucleotides or to the end(s) of DNA. It is also contemplated herein that nucleotides in DNA may be non-standard nucleotides, such as chemically synthesized nucleotides or ribonucleotides.

[0089] The term “nucleoside” relates to compounds which can be thought of as nucleotides without a phosphate group. While a nucleoside is a nucleobase linked to a sugar (e.g., ribose or deoxyribose), a nucleotide is composed of a nucleoside and one or more phosphate groups. Examples of nucleosides include cytidine, uridine, pseudouridine, adenosine, and guanosine. The five standard nucleosides which usually make up naturally occurring nucleic acids are uridine, adenosine, thymidine, cytidine and guanosine. The five nucleosides are commonly abbreviated to their one letter codes U, A, T, C and G, respectively. However, thymidine is more commonly written as “dT” (“d” represents “deoxy”) as it contains a 2'-deoxyribofuranose moiety rather than the ribofuranose ring found in uridine. This is because thymidine is found in deoxyribonucleic acid (DNA) and not ribonucleic acid (RNA). Conversely, uridine is found in RNA and not DNA. The remaining three nucleosides may be found in both RNA and DNA. In RNA, they would be represented as A, C and G, whereas in DNA they would be represented as dA, dC and dG. A modified purine (A or G) or pyrimidine (C, T, or U) base moiety is, in some embodiments, modified by one or more alkyl groups, e.g., one or more C1-4 alkyl groups, e.g., one or more methyl groups. Particular examples of modified purine or pyrimidine base moieties include N7-alkyl-guanine, N6-alkyl-adenine, 5-alkyl-cytosine, 5-alkyl-uracil, and N(l)-alkyl-uracil, such as N7-CI-4 alkyl-guanine, N6-CI-4 alkyl-adenine, 5-C1-4 alkylcytosine, 5-C1-4 alkyl-uracil, and N(1)-CI-4 alkyl-uracil, preferably N7-methyl-guanine, N6- methyl-adenine, 5-methyl-cytosine, 5-methyl-uracil, and N(l)-methyl-uracil.

[0090] The term “in vitro transcription” or “IVT” means that the transcription (i.e., the generation of RNA) is conducted in a cell-free manner. I.e., IVT does not use living / cultured cells but rather the transcription machinery extracted from cells (e.g., cell lysates or the isolated components thereof, including an RNA polymerase (preferably T7, T3 or SP6 polymerase)).

[0091] BNT ref [P1819WO01] / / C&F ref. [240673WO] The term “codon-optimized” refers to the alteration of codons in the coding region of a nucleic acid molecule to reflect the typical codon usage of a host organism without preferably altering the amino acid sequence encoded by the nucleic acid molecule. Within the context of the present invention, coding regions are preferably codon-optimized for optimal expression in a subject to be treated using the RNA molecules described herein. Codon-optimization is based on the finding that the translation efficiency is also determined by a different frequency in the occurrence of tRNAs in cells. Thus, the sequence of RNA may be modified such that codons for which frequently occurring tRNAs are available are inserted in place of “rare codons”.

[0092] The term “analogue” refers to a substance that shares one or more particular structural features, elements, components, or moieties with a reference substance. Typically, an “analogue” shows significant structural similarity with the reference substance, for example sharing a core or consensus structure, but also differs in certain discrete ways. In some embodiments, an analogue is a substance that can be generated from the reference substance, e.g., by chemical manipulation of the reference substance. In some embodiments, an analogue is a substance that can be generated through performance of a synthetic process substantially similar to (e.g., sharing a plurality of steps with) one that generates the reference substance. In some embodiments, an analogue is or can be generated through performance of a synthetic process different from that used to generate the reference substance.

[0093] As used herein, the term “encode” or “encoding” refers to sequence information of a first molecule that guides production of a second molecule having a defined sequence of nucleotides (e.g., mRNA) or a defined sequence of amino acids. For example, a DNA molecule can encode an RNA molecule (e.g., by a transcription process that includes a DNA-dependent RNA polymerase enzyme). An RNA molecule can encode a polypeptide (e.g., by a translation process). Thus, a gene, a cDNA, or a single-stranded RNA (e.g., an mRNA) encodes a polypeptide if transcription and translation of mRNA corresponding to that gene produces the polypeptide in a cell or other biological system. In some embodiments, a coding region of a single-stranded RNA encoding a target polypeptide agent refers to a coding strand, the nucleotide sequence of which is identical to the mRNA

[0094] BNT ref [P1819WO01] / / C&F ref. [240673WO] sequence of such a target polypeptide agent. In some embodiments, a coding region of a single-stranded RNA encoding a target polypeptide agent refers to a non-coding strand of such a target polypeptide agent, which may be used as a template for transcription of a gene or cDNA. As is understood in the art, the phrase “nucleic acid encoding a peptide or protein” means that the nucleic acid, if present in the appropriate environment, for example within a cell and / or in a cell-free translation system, can direct the assembly of amino acids to produce the peptide or protein via a process of translation.

[0095] The terms “identity” or “sequence identity” refer to the overall relatedness between polynucleotide molecules (e.g., DNA molecules and / or RNA molecules) and / or polypeptide molecules. “Sequence identity” between two nucleic acid sequences indicates the percentage of nucleotides that are identical between the sequences. The terms “% identical”, “% identity” or similar terms refer to the percentage of nucleotides or amino acids which are identical in an optimal alignment between the sequences to be compared. Said percentage is purely statistical, and the differences between the two sequences may be but are not necessarily randomly distributed over the entire length of the sequences to be compared. Comparisons of two sequences are usually carried out by comparing the sequences, after optimal alignment, with respect to a segment or “window of comparison”, in order to identify local regions of corresponding sequences. The optimal alignment for a comparison may be carried out manually or with the aid of the local homology algorithm by Smith and Waterman, 1981, Ads App. Math. 2, 482, with the aid of the local homology algorithm by Needleman and Wunsch, 1970, J. Mol. Biol. 48, 443, with the aid of the similarity search algorithm by Pearson and Lipman, 1988, Proc. Natl Acad. Sei. USA 88, 2444, or with the aid of computer programs using said algorithms (GAP, BESTFIT, FASTA, BLASTP, BLASTN and TFASTA in Wisconsin Genetics Software Package, Genetics Computer Group, 575 Science Drive, Madison, Wis.). For example, percent identity of two sequences is determined using the BLASTN or BLASTP algorithm, as available on the United States National Center for Biotechnology Information (NCBI) website (e.g., at the website blast.ncbi.nlm.nih.gov / Blast.cgi?PAGE_TYPE=BlastSearch&BLAST _SPEC=blast2seq&LINK_LOC=align2seq). The algorithm parameters used for BLASTN algorithm on the NCBI website may include: (i) Expect Threshold set to 10; (ii) Word Size

[0096] BNT ref [P1819WO01] / / C&F ref. [240673WO] set to 28; (iii) Max matches in a query range set to O; (iv) Match / Mismatch Scores set to 1, -2; (v) Gap Costs set to Linear; and (vi) the filter for low complexity regions being used. The algorithm parameters used for BLASTP algorithm on the NCBI website may include: (i) Expect Threshold set to 10; (ii) Word Size set to 3; (iii) Max matches in a query range set to O; (iv) Matrixset to BLOSUM62; (v) Gap Costs set to Existence: 11 Extension: 1; and (vi) conditional compositional score matrix adjustment. Percentage identity is obtained by determining the number of identical positions at which the sequences to be compared correspond, dividing this number by the number of positions compared (e.g., the number of positions in the reference sequence) and multiplying this result by 100. In some embodiments, the degree of identity is given for a region which is at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90% or about 100% of the entire length of the reference sequence. For example, if the reference nucleic acid or amino acid sequence consists of 200 nucleotides or amino acids, the degree of identity is given for at least about 20, at least about 40, at least about 60, at least about 80, at least about 100, at least about 120, at least about 140, at least about 160, at least about 180, or about 200 nucleotides or amino acids, in some embodiments in continuous nucleotides or amino acids. In some embodiments, the degree of identity is given for the entire length of the reference sequence. Nucleic acid sequences or amino acid sequences having a particular degree of identity to a given nucleic acid sequence or amino acid sequence, respectively, may have at least one functional property of said given sequence, e.g., and in some instances, are functionally equivalent to said given sequence. One important property includes an immunogenic property, in particular when administered to a subject. In some embodiments, a nucleic acid sequence or amino acid sequence having a particular degree of identity to a given nucleic acid sequence or amino acid sequence is functionally equivalent to the given sequence.

[0097] The terms “polypeptide,” “peptide,” and “protein” are used interchangeably herein to refer to polymers of amino acids.

[0098] The term “recombinant” when used in the context of a polynucleotide means a polynucleotide having nucleotide sequences that are not naturally joined together and can

[0099] BNT ref [P1819WO01] / / C&F ref. [240673WO] be made by artificially combining two otherwise separated segments of sequence. This artificial combination is often accomplished by chemical synthesis or, more commonly, by the artificial manipulation of isolated segments of nucleic acids, for example, by genetic engineering techniques. Recombinant polynucleotides include vectors comprising an amplified or assembled polynucleotide, which can be used to transform or transfect a suitable host cell. A host cell that comprises the recombinant polynucleotide is referred to as a “recombinant host cell.” The polynucleotide is then expressed in the recombinant host cell to produce a “recombinant polypeptide.” A recombinant polynucleotide can also comprise a non-coding function.

[0100] The term “full length” with respect to a given polypeptide means the form of the polypeptide naturally translated from the coding DNA sequence, beginning with the ATG start codon, which encodes the first methionine in the amino acid sequence, and ending at the TGA, TAG, or TTA stop codon, or whichever stop codon employed by the organism.

[0101] The term “gene” refers to a DNA sequence in a chromosome that codes for a protein. In some embodiments, a gene includes coding sequence (i.e., sequence that encodes a particular protein); in some embodiments, a gene includes non-coding sequence. In some particular embodiments, a gene may include both coding (e.g., exonic) and non-coding (e.g., intronic) sequences. In some embodiments, a gene may include one or more regulatory elements that, for example, may control or impact one or more aspects of gene expression (e.g., cell-type- specific expression, inducible expression, etc.).

[0102] The term “carrier” refers to a component which may be natural, synthetic, organic, inorganic in which the active ingredients of the invention are combined in order to facilitate, enhance or enable administration of the anti-CTLA4 antibody or fragment thereof and / or the chemotherapy agent. A carrier as used herein may be one or more compatible solid or liquid fillers, diluents or encapsulating substances, which are suitable for administration to the subject. Suitable carriers include, without limitation, sterile water, Ringer, Ringer lactate, sterile sodium chloride solution, isotonic saline, polyalkylene glycols, hydrogenated naphthalenes and, in particular, biocompatible lactide polymers, lactide / glycolide copolymers or polyoxy ethylene / polyoxy-propylene copolymers.

[0103] BNT ref [P1819WO01] / / C&F ref. [240673WO] Pharmaceutically acceptable carriers, excipients or diluents for therapeutic use are well known in the pharmaceutical art, and are described, for example, in Remington's Pharmaceutical Sciences, Mack Publishing Co. (A. R Gennaro edit. 1985). Pharmaceutical carriers, excipients or diluents can be selected with regard to the intended route of administration and standard pharmaceutical practice.

[0104] The term “excipient” is a substance which may be present in a formulation of the anti- CTLA4 antibody or fragment thereof and / or the chemotherapy agent but is not an active ingredient. Examples of excipients include, without limitation, carriers, binders, diluents, lubricants, thickeners, surface active agents, preservatives, stabilizers, emulsifiers, buffers, flavoring agents, or colorants.

[0105] The term “pharmaceutically acceptable carrier” or “pharmaceutically acceptable excipient” means solvents, dispersion media, coatings, antibacterial agents and antifungal agents, isotonic agents, and absorption delaying agents, and the like, that are compatible with pharmaceutical administration. The use of such media and agents for pharmaceutically active substances is well known in the art. In certain embodiments, the pharmaceutically acceptable carrier or excipient is not naturally occurring. The term “pharmaceutically acceptable” refers to the non-toxicity of a material which does not interact with the action of the active component of the pharmaceutical composition.

[0106] The term “diluent” relates to a diluting and / or thinning agent. Moreover, the term “diluent” includes any one or more of fluid, liquid or solid suspension and / or mixing media. Nonlimiting examples of suitable diluents include ethanol, glycerol, and water.

[0107] The term “subject” relates to a human of either gender (a male or a female). The subject may be of any age. In some embodiments, the subject is female. In some embodiments, the subject is male. In some embodiments, the subject is a subject having cancer, in particular a female subject having cancer and / or a male subject having cancer. The subject can be a patient.

[0108] The term “treating” when used in the context of a disease or disease condition means ameliorating, improving or remedying a disease, disorder, or symptom of a disease or

[0109] BNT ref [P1819WO01] / / C&F ref. [240673WO] condition associated with the disease, or can mean completely or partially stopping, on a molecular level, the biochemical basis of the disease. It describes an act that leads to the elimination, reduction, alleviation, reversal, or prevention or delay of onset or recurrence of any symptom of a disease.

[0110] The term “pharmaceutical composition” relates to a composition comprising an active agent or a therapeutically effective agent, preferably together with pharmaceutically acceptable carriers, diluents and / or excipients. Said pharmaceutical composition is useful for treating, preventing, or reducing the severity of a disease by administration of said pharmaceutical composition to a subject. For example, a pharmaceutical composition can comprise the CTLA4-antibody and / or the chemotherapy agent disclosed herein.

[0111] The term “cancer” refers to a neoplasm or tumor resulting from abnormal uncontrolled growth of cells. The term “tumor” as it applies to a subject diagnosed with, or suspected of having, a cancer refers to a malignant or potentially malignant neoplasm or tissue mass of any size and includes primary tumors and secondary neoplasms. A solid tumor is an abnormal growth or mass of tissue that usually does not contain cysts or liquid areas. Different types of solid tumors are named for the type of cells that form them. Examples of solid tumors are sarcomas, carcinomas, and lymphomas. Leukemias (cancers of the blood) generally do not form solid tumors (National Cancer Institute, Dictionary of Cancer Terms).

[0112] The term “oncogenic driver mutation(s)” as used herein refers to mutations that are responsible for both the initiation and maintenance of a cancer. These mutations are often found in genes that encode for signaling proteins that are critical for maintaining normal cellular proliferation and survival.

[0113] The term “NSCLC driver mutation” as used herein refers to oncogenic driver mutations of non-small cell lung cancer (NSCLC). Non-limiting examples of NSCLC driver mutations are listed in Luo et al., Transl Respir Med. 2013, 1 (1): 6, and are e.g., epidermal growth factor receptor (EGFR), MET, anaplastic lymphoma kinase (ALK), c-ROS oncogene 1, v- raf murine sarcoma viral oncogene homolog B, neurotrophic receptor tyrosine kinase,

[0114] BNT ref [P1819WO01] / / C&F ref. [240673WO] human epidermal growth factor 2, neuregulin-1, and rearranged during transfection, in particular EGFR gene mutations and ALK gene mutations.

[0115] The term “CTLA4” or “CTLA-4” relates to cytotoxic T lymphocyte antigen-4. “CTLA4” or “CTLA-4” are used interchangeably herein. CTLA4 is expressed on the surface of cells and, if it is located at the surface of said cells, is accessible to binding by CTLA4 specific antibodies. “Cell surface” is used in accordance with its normal meaning in the art, and thus includes the outside of the cell which is accessible to binding by proteins and other molecules. For example, a transmembrane protein having one or more extracellular portions is considered as being expressed on the cell surface.

[0116] The term “immune cell” means any cell of hematopoietic lineage involved in regulating an immune response against an antigen (e.g., a bacterial or viral infection or an auto-antigen). In some embodiments, an immune cell is a leukocyte, such as a white blood cell. Immune cells include neutrophils, eosinophils, basophils, lymphocytes, and / or monocytes. Lymphocytes include T lymphocytes (T cells) and B lymphocytes (B cells). Immune cells can also be dendritic cells, natural killer (NK) cells, and / or a mast cell.

[0117] The term “antibody” refers to a molecule that possesses an antigen-binding site. The term encompasses functional antibody fragments or antigen-binding fragments, such as a Fab, a Fab’, a F(ab’)2, a scFV, a rlgG, a diabody, a triabody, a minibody and a single-domain antibody (sdAB). The antibody can comprise a “variable region”. The terms “variable region” and “variable domain” are used interchangeably herein. The term “variable region” is intended to distinguish such domain of the immunoglobulin from domains that are broadly shared by antibodies (such as an antibody Fc domain). The variable region comprises a “hypervariable region” whose residues are responsible for antigen binding, and as used herein means the segment of an antibody which contains three CDRs, designated CDR1, CDR2 and CDR3. A “variable region” of an antibody refers to the variable region of the antibody light chain or the variable region of the antibody heavy chain, either alone or in combination. The variable region of the heavy chain may be referred to as “VH.” The variable region of the light chain may be referred to as “VL.” Typically, the variable regions of both the heavy and light chains comprise three hypervariable regions, the CDRs,

[0118] BNT ref [P1819WO01] / / C&F ref. [240673WO] which are located within relatively conserved framework regions (FR). The CDRs are usually aligned by the framework regions, enabling binding to a specific epitope. In general, from N-terminal to C-terminal, both light and heavy chains variable domains comprise FR1, CDR1, FR2, CDR2, FR3, CDR3, and FR4. The hypervariable region comprises amino acid residues from a “complementarity determining region” or “CDR” (i.e., typically at approximately residues 24-34 (LI), 50-56 (L2) and 89-97 (L3) in the light chain variable domain and at approximately residues 27-35 (Hl), 50-65 (H2) and 95-102 (H3) in the heavy chain variable domain) and / or those residues from a “hypervariable loop” (i.e., residues 26-32 (LI), 50-52 (L2) and 91-96 (L3) in the light chain variable domain and 26-32 (Hl), 53-55 (H2) and 96-101 (H3) in the heavy chain variable domain. The variable domains of the heavy and light chains each contain three CDRs, designated CDR1, CDR2 and CDR3. The precise boundaries of these CDRs can be defined according to various numbering systems known in the art, such as the Kabat numbering system (Kabat et al., Sequences of Proteins of Immunological Interest, 5thEd. Public Health Service, National Institutes of Health, Bethesda, Md., 1991; Kabat et al., J Biol Chem 1977, 252: 6609-6616; Kabat, Adv Prot Chem 1978, 32: 1-75), the Chothia numbering system (Chothia & Lesk, J Mol Biol 1987, 196: 901-917; Chothia et al., Nature 1989, 342: 878-883) or the IMGT numbering system (Lefranc et al., Dev Comparat Immunol 2003, 27: 55-77). For a given antibody, those skilled in the art will readily identify the CDRs defined by each numbering system. Also, the correspondence between different numbering systems is well known to those skilled in the art (Lefranc et al., Dev Comparat Immunol 2003, 27: 55-77). “Framework Region” or “FR” residues are those variable domain residues other than the hypervariable region or CRR residues as herein defined. “Antibody” includes monoclonal antibodies, multi-specific antibodies, human antibodies, humanized antibodies, synthetic antibodies, chimeric antibodies, camelized antibodies, single chain antibodies, disulfide- linked Fvs (sdFv), intrabodies, and anti -idiotypic (anti-Id) antibodies (including, e.g., anti- id and anti-anti-Id antibodies to antibodies disclosed herein). In particular, such antibodies include immunoglobulin molecules of any type (e.g., IgG, IgE, IgM, IgD, IgA and IgY), class (e.g., IgGi, IgG?, IgGs, IgG4, IgAi and IgA?) or subclass. The term antibody further encompasses functional antibody fragments, such as a Fab, a Fab’, a F(ab’)2, a scFV, a rlgG, a diabody, a triabody, a minibody and a single-domain antibody (sdAB).

[0119] BNT ref [P1819WO01] / / C&F ref. [?40673WO] The term “antigen-binding fragment” or “antibody-binding portion” of an antibody refers to one or more portions or fragments of an antibody that contain the antibody’s CDRs and optionally the framework residues that comprise the antibody’s variable domain antigen recognition site, and exhibit an ability to immunospecifically bind an antigen. Examples of antibody fragments encompassed within the term “antigen binding fragment” or “antigen binding portion” include (i) Fab fragments, monovalent fragments consisting of the VL, VH, CL and CH domains; (ii) F(ab’)2 fragments, bivalent fragments comprising two Fab fragments linked by a disulfide bridge at the hinge region; (iii) Fd fragments consisting of the VH and CH domains; (iv) Fv fragments consisting of the VL and VH domains of a single arm of an antibody, (v) dAb fragments (Ward et al., 1989, Nature 341 :544-546), which consist of a VH domain; (vi) isolated complementarity determining regions (CDR), and (vii) combinations of two or more isolated CDRs which may optionally be joined by a synthetic linker. Furthermore, although the two domains of the Fv fragment, VL and VH, are encoded by separate genes, they can be joined, using recombinant methods, by a synthetic linker that enables them to be made as a single protein chain in which the VL and VH regions pair to form monovalent molecules (known as single chain Fv (scFv); see e.g., Bird et al., 1988, Science, 242:423-426; and Huston et al., 1988, Proc. Natl. Acad. Sci. USA, 85:5879-5883). Such single chain antibodies are also intended to be encompassed within the term “antigen-binding fragment” or “antibody-binding portion” of an antibody. Also included within the term “antigen-binding” fragment of an antibody are a rlgG fragments consisting of one heavy chain (HC) and one light chain (LC), i.e., consisting of one of the identical halves of an IgG antibody. A further example are binding-domain immunoglobulin fusion proteins comprising (i) a binding domain polypeptide that is fused to an immunoglobulin hinge region polypeptide, (ii) an immunoglobulin heavy chain CH2 constant region fused to the hinge region, and (iii) an immunoglobulin heavy chain CH3 constant region fused to the CH2 constant region. The binding domain polypeptide can be a heavy chain variable region or a light chain variable region. Binding-domain immunoglobulin fusion proteins are further disclosed in U.S. Patent Application Publication Nos. 2003 / 0118592 and 2003 / 0133939. These antibody fragments can be obtained using conventional techniques known to those with skill in the art, and the fragments are screened for utility in the same manner as are intact antibodies. “Antigenbinding fragment” or “antibody -binding portion” may further include fusion proteins

[0120] BNT ref [P1819WO01] / / C&F ref. [240673WO] comprising the antibody’s variable region antigen recognition site and a heterologous protein (e.g., a toxin, an antigen recognition site for a different antigen, an enzyme, a receptor or receptor ligand, etc.). As used within the present invention, the term “fragment” refers to a peptide or polypeptide comprising an amino acid sequence of at least 5 contiguous amino acid residues, at least 10 contiguous amino acid residues, at least 15 contiguous amino acid residues, at least 20 contiguous amino acid residues, at least 25 contiguous amino acid residues, at least 40 contiguous amino acid residues, at least 50 contiguous amino acid residues, at least 60 contiguous amino residues, at least 70 contiguous amino acid residues, at least 80 contiguous amino acid residues, at least 90 contiguous amino acid residues, at least 100 contiguous amino acid residues, at least 125 contiguous amino acid residues, at least 150 contiguous amino acid residues, at least 175 contiguous amino acid residues, at least 200 contiguous amino acid residues, or at least 250 contiguous amino acid residues.

[0121] The term “human antibody” refers to an antibody that comprises human immunoglobulin protein sequences only. A human antibody may contain murine carbohydrate chains if produced in a mouse, in a mouse cell, or in a hybridoma derived from a mouse cell. Similarly, “mouse antibody” or “rat antibody” refer to an antibody that comprises only mouse or rat immunoglobulin sequences, respectively.

[0122] A “humanized antibody” is an immunoglobulin comprising a human framework region and one or more CDR’s from a non-human (usually a mouse or rat) immunoglobulin. The nonhuman immunoglobulin providing the CDR’s is called the “donor” and the human immunoglobulin providing the framework is called the “acceptor”. Constant regions need not be present, but if they are, they preferably can be substantially identical to human immunoglobulin constant regions, i.e., at least about 85-90%, preferably about 95% or more identical. Hence, all parts of a humanized immunoglobulin, except possibly the CDR’s, are substantially identical to corresponding parts of natural human immunoglobulin sequences. A humanized antibody is an antibody comprising a humanized light chain and a humanized heavy chain immunoglobulin. For example, a humanized antibody would not encompass a typical chimeric antibody, because, e.g., the entire variable region of a chimeric antibody is non-human. One says that the donor antibody has been “humanized”,

[0123] BNT ref [P1819WO01] / / C&F ref. [240673WO] by the process of “humanization”, because the resultant humanized antibody is expected to bind to the same antigen as the donor antibody that provides the CDR’s. For the most part, humanized antibodies are human immunoglobulins (recipient antibody) in which hypervariable region residues of the recipient are replaced by hypervariable region residues from a non-human species (donor antibody) such as mouse, rat, rabbit or a non-human primate having the desired specificity, affinity, and capacity. In some instances, framework region (FR) residues of the human immunoglobulin are replaced by corresponding non- human residues. Furthermore, humanized antibodies may comprise residues which are not found in the recipient antibody or in the donor antibody. These modifications are made to further refine antibody performance. In general, the humanized antibody will comprise substantially all of at least one, and typically two, variable domains, in which all or substantially all of the hypervariable regions correspond to those of a non-human immunoglobulin and all or substantially all of the FRs are those of a human immunoglobulin sequence. The humanized antibody optionally also will comprise at least a portion of an immunoglobulin constant region (Fc), typically that of a human immunoglobulin that immunospecifically binds to an Fc.gamma.RIIB polypeptide, that has been altered by the introduction of amino acid residue substitutions, deletions or additions (i.e., mutations).

[0124] An antibody that “specifically binds to” or “immunospecifically binds to” a specified target protein is an antibody that exhibits preferential binding to that target as compared to other proteins, but this specificity does not require absolute binding specificity. An antibody is considered “specific” for its intended target if its binding is determinative of the presence of the target protein in a sample, e.g., without producing undesired results such as false positives. Antibodies, or binding fragments thereof, useful in the present invention will bind to the target protein with an affinity that is at least two fold greater, preferably at least ten times greater, more preferably at least 20-times greater, and most preferably at least 100-times greater than the affinity with non-target proteins. As used herein, an antibody is said to bind specifically to a polypeptide comprising a given amino acid sequence, e.g., the amino acid sequence of a mature human CTLA4 molecule, if it binds to polypeptides comprising that sequence but preferably does not bind to proteins lacking that sequence.

[0125] BNT ref [P1819WO01] / / C&F ref. [240673WO] “Chimeric antibody” refers to an antibody in which a portion of the heavy and / or light chain is identical with or homologous to corresponding sequences in an antibody derived from a particular species (e.g., human) or belonging to a particular antibody class or subclass, while the remainder of the chain(s) is identical with or homologous to corresponding sequences in an antibody derived from another species (e.g., mouse) or belonging to another antibody class or subclass, as well as fragments of such antibodies, so long as they exhibit the desired biological activity.

[0126] The term “epitope” refers to the part of an antigen that as used herein, refers to an agent that elicits an immune response; and / or an agent that binds to a T cell receptor (e.g., when presented by an MHC molecule) or to an antibody. For example, epitopes are the discrete, three-dimensional sites on an antigen, which are recognized by the immune system. Epitopes usually consist of chemically active surface groupings of molecules such as amino acids or sugar side chains and usually have specific three-dimensional structural characteristics, as well as specific charge characteristics. Conformational and non- conformational epitopes are distinguished in that the binding to the former but not the latter is lost in the presence of denaturing solvents.

[0127] The term “antigen” as used herein, refers to an agent that elicits an immune response; and / or an agent that binds to a T cell receptor (e.g., when presented by an MHC molecule) or to an antibody. In some embodiments, an antigen elicits a humoral response (e.g., including production of antigen-specific antibodies); in some embodiments, an antigen elicits a cellular response (e.g., involving T-cells whose receptors specifically interact with the antigen). In some embodiments, an antigen binds to an antibody and may or may not induce a particular physiological response in an organism. In general, an antigen may be or include any chemical entity such as, for example, a small molecule, a nucleic acid, a polypeptide, a carbohydrate, a lipid, a polymer (in some embodiments other than a biologic polymer [e.g., other than a nucleic acid or amino acid polymer]) etc. In some embodiments, an antigen is or comprises a polypeptide. In some embodiments, an antigen is or comprises a glycan. Those of ordinary skill in the art will appreciate that, in general, an antigen may be provided in isolated or pure form, or alternatively may be provided in crude form (e.g., together with other materials, for example in an extract such as a cellular extract or other relatively crude preparation of an antigen-containing source). In some embodiments,

[0128] BNT ref [P1819WO01] / / C&F ref. [240673WO] antigens utilized in accordance with the present invention are provided in a crude form. In some embodiments, an antigen is a recombinant antigen.

[0129] The terms “chemotherapeutic agent” or “chemotherapeutical agent” or “chemotherapy agent” can be used interchangeably herein. A chemotherapeutic agent is a chemical compound useful in the treatment of cancer. Classes of chemotherapeutic agents include, but are not limited to: alkylating agents, antimetabolites, kinase inhibitors, spindle poison plant alkaloids, cytotoxic / antitumour antibiotics, topoisomerase inhibitors, photosensitizers, anti -estrogens and selective estrogen receptor modulators (SERMs), anti-progesterones, estrogen receptor down-regulators (ERDs), estrogen receptor antagonists, leutinizing horm one-releasing hormone agonists, anti-androgens, aromatase inhibitors, EGFR inhibitors, VEGF inhibitors, anti-sense oligonucleotides that that inhibit expression of genes implicated in abnormal cell proliferation or tumour growth. Chemotherapeutic agents useful in the treatment methods of the present invention include cytostatic and / or cytotoxic agents. Chemotherapeutic agents as used herein do not include antibodies.

[0130] The term “combination therapy” as used herein refers to the anti-CTLA4 antibody or fragment thereof and the chemotherapy agent as disclosed herein for use in a method of treating a subject with cancer. Each component of the combination therapy can be administered separately, concurrently, or consecutively.

[0131] The term “administration route” refers to the means of administration of an active agent or a pharmaceutical composition. The terms “active agent” or “therapeutically effective agent” as used herein refer to compounds that exert a pharmaceutical effect in the treatment of a disease. The anti-CTLA4 antibody or fragment thereof and the chemotherapy agent as disclosed herein can be administered by any administration route known to the skilled person. In general, active agents or pharmaceutical compositions can be administered using any suitable enteral route or parenteral route of administration. The term “enteral route” of administration refers to the administration via any part of the gastrointestinal tract. Examples of enteral routes include oral, mucosal, buccal, and rectal route, or intragastric route. “Parenteral route” of administration refers to a route of administration other than enteral route. Examples of parenteral routes of administration include intravenous, intramuscular, intradermal, intraperitoneal, intratumour, intravesical, intraarterial,

[0132] BNT ref [P1819WO01] / / C&F ref. [240673WO] intrathecal, intracapsular, intraorbital, intracardiac, transtracheal, intraarticular, subcapsular, subarachnoid, intraspinal, epidural and intrastemal, subcutaneous, or topical administration. For example, anti-CTLA4 antibody or fragment thereof and the chemotherapy agent of the present disclosure can be administered using different enteral or parenteral administration routes. The anti-CTLA4 antibody or fragment thereof and the chemotherapy agent of the present disclosure can also be administered using the same administration route, e.g., intravenous administration using an intravenous injection or intravenous infusion.

[0133] The term “separate administration” as used herein refers to the administration of at least two active agents or therapeutically effective agents using different pharmaceutical compositions or formulations. For example, the anti-CTLA4 antibody or fragment thereof and the chemotherapy agent of the present disclosure can be administered using two different pharmaceutical compositions, each comprising either the anti-CTLA4 antibody (or fragment thereof) disclosed herein or the chemotherapy agent disclosed herein.

[0134] The terms “concurrent administration” or “co-administration” as used herein refer to the administration of at least two active agents, therapeutically effective agents, or pharmaceutical compositions which are administered simultaneously or essentially at the same time. “Essentially at the same time” as used herein means within about 1 minute, about 5 minutes, about 10 minutes, about 15 minutes, about 30 minutes, about 1 hour, about 2 hours, or about 6 hours period of each other. For example, the anti-CTLA4 antibody or fragment thereof and the chemotherapy agent of the present disclosure can be administered essentially at the same time using two different pharmaceutical compositions, each individually comprising the anti-CTLA4 antibody (or fragment thereof) or the chemotherapy agent disclosed herein.

[0135] The term “consecutive administration” as used herein refers to the sequential administration of active agents, therapeutically effective agents, or pharmaceutical compositions in treatment cycles. The term consecutive administration includes the sequential administration of the same or of different, i.e., at least two, active agents, therapeutically effective agents, or pharmaceutical compositions.

[0136] BNT ref [P1819WO01] / / C&F ref. [240673WO] The term “treatment cycle” as used herein refers to the repetitive treatment of a subject with an active agent, therapeutically effective agent, or pharmaceutical composition, or combinations thereof. A treatment cycle begins with the administration of a first dosage of an active agent, therapeutically effective agent, or pharmaceutical composition, or combinations thereof and ends either with the administrations of a consecutive or subsequent dosage of the same or another active agent, therapeutically effective agent, or pharmaceutical composition, or combinations thereof or with the end of the treatment. In some embodiments a treatment cycle has at least about 1 day, about 2 days, about 3 days, about 4 days, about 5 days, about 6 days, about 7 days, about 8 days, about 9 days, about 10 days, about 11 days, about 12 days, about 13 days, about 14 days, about 15 days, about 16 days, about 17 days, about 18 days, about 19 days, about 20 days, about 21 days, about 22 days, about 23 days, about 24 days, about 25 days, about 26 days, about 27 days, or about

[0137] 28 days. In some preferred embodiments a treatment cycle has up to about 28 days, preferably up to about 21 days, more preferably about 21 days.

[0138] The terms “platinum-containing chemotherapy” or “platinum-based chemotherapy” refer to the use of chemotherapeutic agent(s) (also known as platins) for treating cancer that are coordination complexes of platinum. Platinum-containing chemotherapeutic agents are alkylating agents that crosslink DNA, resulting in ineffective DNA mismatch repair and generally leading to apoptosis. Examples of platins include cisplatin, carboplatin, and oxaliplatin.

[0139] The term “overall survival” (OS) as used herein refers to the time period between the first treatment of a subject suffering from cancer with an active agent, therapeutically effective agent, or pharmaceutical composition and the death of the subject by any cause. OS as used herein is determined over a period of 48 months.

[0140] The term “progression-free survival” (PFS) as used herein refers to the time period between the first treatment of a subject suffering from cancer with an active agent, therapeutically effective agent, or pharmaceutical composition and the first objective tumor progression according to RECIST vl.l or death from any cause, whichever occurs first. PFS as used herein is determined over a period of 48 months.

[0141] BNT ref [P1819WO01] / / C&F ref. [240673WO] The terms “effective amount” or “therapeutically effective amount” refer to an amount of a given substance that is sufficient in quantity to produce a desired effect, including an improvement or remediation of the disease, disorder, or symptoms of the disease or condition.

[0142] Recitation of ranges of values herein is merely intended to serve as a shorthand method of referring individually to each separate value falling within the range. Unless otherwise indicated herein, each individual value is incorporated into the specification as if it was individually recited herein. All methods described herein can be performed in any suitable order unless otherwise indicated herein or otherwise clearly contradicted by context. The use of any and all examples, or exemplary language (e.g., “such as”), provided herein is intended merely to better illustrate the invention and does not pose a limitation on the scope of the invention otherwise claimed. No language in the specification should be construed as indicating any non-claimed element essential to the practice of the invention.

[0143] All patents, patent applications, and other publications cited in this application are incorporated by reference in the entirety for all purposes.

[0144] The anti-CTLA4 antibody and chemotherapy agent combination therapy of the invention

[0145] The present invention provides an anti-CTLA4 antibody or fragment thereof for use in a method of treating cancer in a subject in need thereof, wherein the method comprises administering to the subject: a) the anti-CTLA4 antibody or fragment thereof; and b) a chemotherapy agent.

[0146] The invention further provides a chemotherapy agent for use in a method of treating cancer in a subject in need thereof, wherein the method comprises administering to the subject: a) the chemotherapy agent or fragment thereof; and b) an anti-CTLA4 antibody.

[0147] BNT ref [P1819WO01] / / C&F ref. [240673WO] The invention also provides an anti-CTLA4 antibody or fragment thereof and a chemotherapy agent for use in a method of treating cancer in a subject in need thereof, wherein the method comprises administering to the subject: a) the anti-CTLA4 antibody or fragment thereof; and b) the chemotherapy agent.

[0148] The present invention provides antibodies or fragments thereof that immunospecifically bind to CTLA4, in particular human CTLA4, preferably expressed on the surface of a cell at an endogenous or transfected concentration. Cytotoxic T lymphocyte antigen-4 (CTLA4) is a regulator of adaptive immune responses, having a role in the maintenance of peripheral tolerance and in shaping the repertoire of emergent T cell responses. CTLA4 is expressed on the surface of cells and, if it is located at the surface of said cells, is accessible to binding by CTLA4-specific antibodies.

[0149] In some embodiments, the anti-CTLA4 antibody or fragment thereof is capable of binding to human CTLA4. In some embodiments, CTLA4 is expressed on the surface of a T cell. In some embodiments, the anti-CTLA4 antibody or the anti-CTLA4 antibody fragment is a monoclonal antibody, a human antibody, a chimeric antibody, or a humanized antibody, or a fragment thereof. In some embodiments, the anti CTLA4 antibody or fragment thereof is a humanized anti-CTLA4 antibody or a fragment thereof. In some preferred embodiments, the anti CTLA4 antibody or fragment thereof is a humanized anti-CTLA4 IgGl monoclonal antibody or a fragment of a humanized anti-CTLA4 IgGl monoclonal antibody.

[0150] In some embodiments, the anti-CTLA4 antibody fragment is an antigen-binding fragment or a variant thereof. In some embodiments, the antigen binding fragment or variant thereof is selected from the group consisting of a Fab, a Fab’, a F(ab’)2, a scFV, a rlgG, a diabody, a triabody, a minibody, and a single-domain antibody (sdAB), and variants thereof.

[0151] Without wishing to be bound by theory, CTLA4 is contemplated to be recycled between the cell surface and endosomes, where it is usually prevented from lysosomal degradation and can recycle back to the cell surface by binding to the lipopolysaccharide-responsive and beige-like anchor (LRBA) protein. The anti-CTLA4 antibodies and fragments thereof

[0152] BNT ref [P1819WO01] / / C&F ref. [240673WO] disclosed herein have reduced autoimmune side effects when used to enhance immune responses and are particularly suitable for treating cancer in subjects, particularly lung cancer, preferably NSCLC.

[0153] In some embodiments, the subject in need for cancer treatment is a human.

[0154] The anti-CTLA4 antibody described herein is combined with a chemotherapy agent for treating subjects afflicted with cancer. Without wishing to be bound by theory, it is contemplated herein that chemotherapy in combination with anti-CTLA antibodies is particularly effective for the treatment of cancer patients with primary or acquired resistance to treatment with immune checkpoint inhibitors, e.g., to treatment anti -PD1 / anti - PD-L1 antibodies, and without targetable NSCLC driver mutations.

[0155] In some embodiments, the chemotherapy agent comprises or is selected from cisplatin, oxaliplatin, carboplatin, lurbinectedin, topotecan, paclitaxel, nanoparticle albumin-bound paclitaxel (nab -paclitaxel), pemetrexed, 5-fluoruracil, irinotecan, etoposide, gemcitabine, docetaxel or combinations thereof. In some preferred embodiments, the chemotherapy agent is docetaxel.

[0156] Surprisingly, the combination method of the present invention comprising administration of the anti-CTLA4 antibody and the chemotherapy agents shows an enhanced anti-cancer efficacy compared to administration of either the anti-CTLA4 antibody or the chemotherapy agent alone. Furthermore, the combination of the anti-CTLA4 antibody and the chemotherapy agents shows good safety profiles in cancer subjects, in particular in subjects with primary or acquired resistance against cancer immunotherapy by immune checkpoint inhibitors and without targetable NSCLC driver mutations. Furthermore, immune-related adverse events are reduced by the combination therapy of the invention. Immune-related adverse events such as skin rash, hepatitis, colitis and endocrinopathies, particularly hypopituitarism are characteristic side effects of anti-cancer therapies based on immune checkpoint inhibitors such as anti-CTLA4 antibodies. It was therefore surprising that the combination therapy of the invention improves the efficiency of the anti-cancer treatment while in parallel reducing undesired side effects, e.g., immune-related adverse events.

[0157] BNT ref [P1819WO01] / / C&F ref. [240673WO] The anti-CTLA4 antibody

[0158] In some embodiments, the anti-CTLA4 antibody or fragment thereof is capable of binding human CTLA4. The term “binding” preferably relates to a specific binding. An antibody usually is capable of binding to a predetermined target if it has a significant affinity for said predetermined target and binds to said predetermined target in standard assays. “Affinity” or “binding affinity” is often measured by equilibrium dissociation constant (KD). Preferably, the term “significant affinity” refers to the binding to a predetermined target with a dissociation constant (KD) of 10'5M or lower, 10'6M or lower, 10'7M or lower, 10'8M or lower, or 10'9M or lower. An antibody is not (substantially) capable of binding to a target if it has no significant affinity for said target and does not bind significantly, in particular does not bind detectably, to said target in standard assays. For example, if the KD for binding of an antibody to the target to which the antibody is capable of binding is 10'9M, the KD for binding to a target for which the antibody has no significant affinity would be is at least around IO’8M, IO’7M, IO’6M, 10’5M, IO’4M, IO’3M, IO’2M, or 10’1M.

[0159] Binding of an antibody to a target can be determined experimentally using any suitable method; see, for example, Berzofsky et al., Antibody- Antigen Interactions In Fundamental Immunology, Paul, W. E., Ed., Raven Press New York, N Y (1984), Kuby, Janis Immunology, W. H. Freeman and Company New York, N Y (1992), and methods described herein. Affinities may be readily determined using conventional techniques, such as by equilibrium dialysis; by using the BIAcore 2000 instrument, using general procedures outlined by the manufacturer; by radioimmunoassay using radiolabeled target antigen; or by another method known to the skilled artisan. The affinity data may be analyzed, for example, by the method of Scatchard et al. (Scatchard et al., Ann NY Acad ScL 1949, 51 :660). The measured affinity of a particular antibody-antigen interaction can vary if measured under different conditions, e.g., salt concentration, pH. Thus, measurements of affinity and other antigen-binding parameters, e.g., KD, IC50, are preferably made with standardized solutions of antibody and antigen, and a standardized buffer. For example, the affinity of an antibody can be evaluated by Octet. Multi -concentration kinetic experiments can be performed on the Octet Red96 system (ForteBio). Anti-hlgG Fc biosensors (ForteBio, #18-5064) can be hydrated in sample diluent (0.1% BSA in PBS and 0.02%

[0160] BNT ref [P1819WO01] / / C&F ref. [240673WO] Tween 20) and preconditioned in pH 1.7 glycine. The antigen can be diluted using a 7- point, 2-fold serial dilution starting at 600 nM with sample diluent. The antibody to be tested can be diluted to 10 pg / mL with sample diluent and then immobilized onto anti-hlgG Fc biosensors for 120 seconds. After baselines are established for 60 seconds in sample diluent, the biosensors can be moved to wells containing the antigen at a series of concentrations to measure the association. Association can be observed (e.g., for 120 seconds) and dissociation can be observed (e.g., for 180 seconds) for each protein of interest in the sample diluent. The binding affinities can be characterized by fitting the kinetic sensorgrams to a monovalent binding model (1 : 1 binding).

[0161] The anti-CTLA4 antibody or fragment thereof disclosed herein is specific for CTLA4 if it is capable of binding to CTLA4 but is not (substantially) capable of binding to other targets.

[0162] The anti-CTLA4 antibody or fragment thereof disclosed herein preferably does not inhibit binding of human CTLA4 to the B7.1 (CD80) and B7.2 (CD86) ligands of antigen presenting cells. In some embodiments, the level of B7.1 and B7.2 on immune cells following anti-CTLA4 treatment is used as a biomarker for measuring the biological activity of anti-CTLA4 antibodies in vivo and monitoring responses to anti-CTLA4 treatment by measuring the level B7.1 and / or B7.2 expression on immune cells, and comparing the level of expression before and after treatment. In some embodiments, the level of B7.1 and / or B7.2 expression is monitored over time during a course of therapy. The therapeutic effect of CTLA4 antibodies disclosed herein is preferably achieved through antibody -mediated depletion of Tregs specifically within tumor microenvironment. The anti-CTLA4 antibodies disclosed herein are preferably not capable of blocking B7-CTLA4 interactions under physiological conditions.

[0163] A fundamental question for the generation of safe and effective anti-CTLA4 antibodies is whether cancer immunotherapeutic effects (CITE) and immunotherapy -related adverse effects (irAE) are intrinsically linked. The classical checkpoint blockade hypothesis stipulated that anti-CTLA4 antibodies promote cancer immunity by blocking a negative signal of B7-CTLA4 interactions to promote naive T cell activation in the lymphoid organ.

[0164] BNT ref [P1819WO01] / / C&F ref. [240673WO] According to this model, therapeutic antibodies are antagonists that functionally inactivate CTLA4-B7 interactions. Since genetic inactivation of CTLA4 expression leads to autoimmune diseases in mouse and human, it was assumed that the irAE would be a necessary price for CITE. However, there is no evidence to date that blocking CTLA-4 interaction with B7.1 and B7.2 is either necessary or sufficient for the CITE of anti-CTLA- 4 antibodies. Without wishing to be bound by theory, it is contemplated that selective depletion of Tregs in the tumor microenvironment constitutes the main mechanism of action of anti-CTLA4 antibodies. It is not relevant whether an antibody is capable of blocking B7-CTLA4 interactions under physiological conditions for the induction of CITE. In some embodiments, the anti-CTLA4 antibody or fragment thereof disclosed herein can induce CITE without blocking B7-CTLA4 interactions.

[0165] In some embodiments, the anti-CTLA4 antibody is not ipilimumab (marketed as YERVOY®).

[0166] In particularly preferred embodiments, the anti-CTLA4 antibody is described in International Patent Application Publication No., WO 2017 / 106372, which is incorporated herein in its entirety.

[0167] In some embodiments, the anti-CTLA4 antibody fragment is an antigen-binding fragment or a variant thereof. In some embodiments, the antigen binding fragment or variant thereof is selected from the group consisting of a Fab, a Fab’, a F(ab’)2, a scFV, a rlgG, a diabody, a triabody, a minibody, and a single-domain antibody (sdAB), and variants thereof.

[0168] In some embodiments, the anti-CTLA4 antibody or fragment thereof comprises: a) a heavy chain variable region comprising (1) a complementarity-determining region 1 (HCDR1) comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 1, (2) a complementarity-determining region 2 (HCDR2) comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 3, 4 or 5, and (3) a complementarity-determining region 3 (HCDR3) comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 9; and

[0169] BNT ref [P1819WO01] / / C&F ref. [240673WO] b) a light chain variable region comprising: (1) a complementarity-determining region 1 (LCDR1) comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 2, (2) a complementarity-determining region 2 (LCDR2) comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 6, 7, or 8, and (3) a complementarity-determining region 3 (LCDR3) comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 10.

[0170] The anti-CTLA4 antibody or fragment thereof may comprise the CDRs listed in table 1.

[0171] Table 1: exemplary CDR sequences of the anti-CTLA4 antibody or fragment thereof

[0172] In some embodiments, the anti-CTLA4 antibody or fragment thereof comprises: a) a heavy chain variable region comprising (1) a HCDR1 comprising or consisting of an amino acid sequence is set forth in SEQ ID NO: 1, (2) a HCDR2 comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 3, and (3) a HCDR3 comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 9; and b) a light chain variable region comprising: (1) a LCDR1 comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 2, (2) a LCDR2 comprising or

[0173] BNT ref [P1819WO01] / / C&F ref. [240673WO] consisting of an amino acid sequence set forth in SEQ ID NO: 7, and (3) a LCDR3 comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 10.

[0174] In some preferred embodiments, the anti-CTLA4 antibody or fragment thereof comprises: a) a heavy chain variable region comprising (1) a HCDR1 comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 1, (2) a HCDR2 comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 5, and (3) a HCDR3 comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 9; and b) a light chain variable region comprising: (1) a LCDR1 comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 2, (2) a LCDR2 comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 7, and (3) a LCDR3 comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 10.

[0175] In some embodiments, the anti-CTLA4 antibody or fragment thereof comprises: a) a heavy chain variable region comprising (1) a HCDR1 comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 1, (2) a HCDR2 comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 5, and (3) a HCDR3 comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 9; and b) a light chain variable region comprising: (1) a LCDR1 comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 2, (2) a LCDR2 comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 8, and (3) a LCDR3 comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 10.

[0176] In some embodiments, the anti-CTLA4 antibody or fragment thereof comprises:

[0177] BNT ref [P1819WO01] / / C&F ref. [240673WO] a) a heavy chain variable domain comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 11, 12, or 13, or an amino acid sequence comprising at least 80%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 11, 12, or 13; and b) a light chain variable domain comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 14, 15, or 16, or an amino acid sequence comprising at least 80%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 14, 15, or 16.

[0178] The anti-CTLA4 antibody or fragment thereof may comprise the heavy and light chain variable domains listed in table 2.

[0179] Table 2: exemplary heavy and light chain variable domain sequences of the anti- CTLA4 antibody or fragment thereof

[0180] In some embodiments, the anti-CTLA4 antibody or fragment thereof comprises: a) a heavy chain variable domain comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 11 or an amino acid sequence comprising at least 80%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 11; and b) a light chain variable domain comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 15 or an amino acid sequence comprising at least 80%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 15.

[0181] In some preferred embodiments, the anti-CTLA4 antibody or fragment thereof comprises:

[0182] BNT ref [P1819WO01] / / C&F ref. [240673WO] a) a heavy chain variable domain comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 13 or an amino acid sequence comprising at least 80%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 13; and b) a light chain variable domain comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 15 or an amino acid sequence comprising at least 80%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 15.

[0183] In some embodiments, the anti-CTLA4 antibody or fragment thereof comprises: a) a heavy chain variable domain comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 13 or an amino acid sequence comprising at least 80%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 13; and b) a light chain variable domain comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 16 or an amino acid sequence comprising at least 80%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 16.

[0184] In some embodiments, the antibody or fragment thereof having the ability to bind to CTLA4 can be a polyclonal, monoclonal antibody or a chimeric antibody, or fragment thereof, optionally having an IgG or IgM isotype of any subclass, such as subclass I. Preferably, the anti-CTLA4 antibody is an IgG or IgM antibody, preferably an IgG antibody, more preferably an IgGl, IgG2, IgG3 or IgG4 antibody, even more preferably and IgGl antibody. The term “monoclonal antibody” as used herein refers to a preparation of antibody molecules of single molecular composition. A monoclonal antibody displays a single binding specificity and affinity. In some embodiments, the monoclonal antibodies are produced by a hybridoma which includes a B cell obtained from a non-human animal, e.g., mouse, fused to an immortalized cell.

[0185] In some embodiments, the anti-CTLA4 antibody or fragment thereof comprises a constant region. In some preferred embodiments, the constant region of the anti-CTLA4 antibody is derived from a human antibody. In some preferred embodiments, the constant region or

[0186] BNT ref [P1819WO01] / / C&F ref. [240673WO] fragment of the constant region of the anti-CTLA4 antibody fragment is derived from a human antibody. In some preferred embodiments, the constant region of the anti-CTLA4 antibody is selected from the constant region of human IgGl, IgG2, IgG3 or IgG4. In some preferred embodiments, the constant region is selected from the constant region or a fragment thereof of human IgGl, IgG2, IgG3 or IgG4.

[0187] In some embodiments, the anti-CTLA4 antibody comprises an IgGl Fc region (or Fc domain). In some preferred embodiments, the anti-CTLA4 antibody comprises an IgGl Fc comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 17 or 18 or an amino acid sequence comprising at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 17 or 18. In some embodiments, the anti-CTLA4 antibody can comprise a mutated Fc region. Relative to the sequence of the IgGl backbone in SEQ ID NO: 17, the mutation may be M135Y, S137T, T139E, S181A, E216A, or K217A, or a combination thereof. These mutations are contemplated to lead to increased antibody dependent cellular cytotoxicity (ADCC) and increased half-life of the antibody in vivo. Preferably, the Fc region of the anti-CTLA4 antibody may comprise all six mutations. In some particularly preferred embodiments, the Fc region comprises the amino acid sequence of SEQ ID NO: 18.

[0188] In some embodiments, the anti-CTLA4 antibody comprises: a) a heavy chain comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 19, 21, or 23 or an amino acid sequence comprising at least 80%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 19, 21, or 23; and b) a light chain comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 25, 27, or 29 or an amino acid sequence comprising at least 80%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 25, 27, or 29.

[0189] The anti-CTLA4 antibody may comprise the heavy and light chains listed in table 3.

[0190] BNT ref [P1819WO01] / / C&F ref. [240673WO] Table 3: exemplary heavy and light chains of the anti-CTLA4 antibody

[0191] In some embodiments, the anti-CTLA4 antibody comprises: a) a heavy chain comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 19 or an amino acid sequence comprising at least 80%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 19; and b) a light chain comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 27 or an amino acid sequence comprising at least 80%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 27. In some preferred embodiments, the anti-CTLA4 antibody comprises: a) a heavy chain comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 23 or an amino acid sequence comprising at least 80%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 23; and b) a light chain comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 27 or an amino acid sequence comprising at least 80%, 90%, 95%, 96%, 97%,

[0192] 98%, or 99% identity to SEQ ID NO: 27.

[0193] BNT ref [P1819WO01] / / C&F ref. [240673WO] In some embodiments, the anti-CTLA4 antibody comprises: a) a heavy chain comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 23 or an amino acid sequence comprising at least 80%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 23; and b) a light chain comprising or consisting of amino acid sequence set forth in SEQ ID NO: 29 or an amino acid sequence comprising at least 80%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 29.

[0194] In some embodiments, the anti-CTLA4 antibody comprises: a) a heavy chain whose amino acid sequence is encoded by a nucleotide sequence set forth in SEQ ID NO: 20, 22, or 24 or a nucleotide sequence having at least 80%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 20, 22, or 24; and b) a light chain whose amino acid sequence is encoded by a nucleotide sequence set forth in SEQ ID NO: 26, 28, or 30 or a nucleotide sequence having at least 80%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 26, 28, or 30.

[0195] In some embodiments, the thymidine in the nucleotide sequences set forth in SEQ ID NO: 20, 22, 24, 26, 28, and 30 is uridine. Meaning, all thymidines present in the nucleotide sequences of SEQ ID NO: 20, 22, 24, 26, 28, and 30 can be uridines.

[0196] In some embodiments, the anti-CTLA4 antibody comprises: a) a heavy chain whose amino acid sequence is encoded by the nucleotide sequence set forth in SEQ ID NO: 20 or a nucleotide sequence having at least 80%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 20; and

[0197] BNT ref [P1819WO01] / / C&F ref. [240673WO] b) a light chain whose amino acid sequence is encoded by the nucleotide sequence set forth in SEQ ID NO: 28 or a nucleotide sequence having at least 80%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 28.

[0198] In some preferred embodiments, the anti-CTLA4 antibody comprises: a) a heavy chain whose amino acid sequence is encoded by the nucleotide sequence set forth in SEQ ID NO: 24 or a nucleotide sequence having at least 80%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 24; and b) a light chain whose amino acid sequence is encoded by the nucleotide sequence set forth in SEQ ID NO: 28 or a nucleotide sequence having at least 80%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 28.

[0199] In some embodiments, the anti-CTLA4 antibody comprises: a) a heavy chain whose amino acid sequence is encoded by the nucleotide sequence set forth in SEQ ID NO: 24 or a nucleotide sequence having at least 80%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 24; and b) a light chain whose amino acid sequence is encoded by the nucleotide sequence set forth in SEQ ID NO: 30 or a nucleotide sequence having at least 80%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 30.

[0200] In some embodiments, the anti-CTLA4 antibody or fragment thereof is a pH-sensitive anti- CTLA4 antibody or a pH-sensitive anti-CTLA4 antibody fragment. For example, the pH- sensitive anti-CTLA4 antibody or anti-CTLA4 antibody fragment can dissociate from CTLA4 at pH 6.5 or below, more preferably pH 5.5 or below. In some preferred embodiments, binding to CTLA4 is reduced at an endosomal pH of 5.5 by more than 50% relative to binding at neutral pH (pH 7.0). Such a reduction may reach more than 75% at lysosomal pH 4.5 as compared to pH 7.0. The antibody- antigen complex preformed at pH 7.0 may dissociate under an acidic environment of pH 4.5-6.0. The reduction in binding

[0201] BNT ref [P1819WO01] / / C&F ref. [240673WO] may also be in comparison to a reference antibody which may be considerably less pH sensitive using the same standard. The reference antibody may be an antibody known in the art such as Ipilimumab or Tremelimumab. In the context of an engineered antibody or antibody fragment, the changes may also be in comparison to a wild-type antibody or antibody fragment which may be considerably less pH sensitive using the same standard. Without wishing to be bound by theory, it is contemplated that a pH-sensitive antibody or antibody fragment is not only safer but also more effective in Treg depletion and tumor rejection than a pH-insensitive CTLA4 antibody (e.g., Ipilimumab) or a fragment thereof. Meaning, Ipilimumab can bind to CTLA4 at a pH of 4-7 and no dissociation can be observed at pH 4-7. pH-insensitive antibodies can cause down-regulation of CTLA4 through lysosomal degradation. CTLA4 down-regulation can cause autoimmune diseases, while in the tumor CTLA4 down-regulation can reduce ADCC activity and thus anti-cancer efficacy. The sensitivity to pH can be measured by any method known to the skilled person. For example, human or monkey-CTLA4-Fc or (0.5 pg / ml) can be coated on ELISA plates at 4°C overnight. Biotinylated anti-CTLA4 antibodies anti-CTLA4 antibody fragments can be added at 1 pg / ml in 1% BSA PBS with pH 4.5-7.0. Two hours later, antibodies and antibody fragments binding with or to CTLA4 can be measured by using HRP -labeled streptavidin. A pH-sensitive anti-CTLA4 antibody or fragment thereof can comprise, for example, the following CDR sequences: a) a heavy chain variable region comprising (1) a complementarity-determining region 1 (HCDR1) comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 1, (2) a complementarity-determining region 2 (HCDR2) comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 3, 4 or 5, and (3) a complementarity-determining region 3 (HCDR3) comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 9; and b) a light chain variable region comprising: (1) a complementarity-determining region 1 (LCDR1) comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 2, (2) a complementarity-determining region 2 (LCDR2) comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 6, 7, or 8, and (3) a complementarity-determining region 3 (LCDR3) comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 10.

[0202] BNT ref [P1819WO01] / / C&F ref. [240673WO] Although the following provides considerations regarding the mechanism underlying the therapeutic efficacy of antibodies of the disclosure it is not to be considered as limiting to the invention in any way. In some embodiments, the antibodies and antibody fragments described herein are contemplated to be a highly selective, humanized monoclonal immunoglobulin G1 (IgGl)-kappa isotype antibody against CTLA4 with a robust antitumor activity and lower autoimmune toxicity in comparison to ipilimumab. The disclosed antibodies preferably can dissociate from CTLA4 under low pH in endosomes to allow both CTLA4 and the antibody to escape from lysosomal degradation and recycle to the cell surface. Unlike ipilimumab that down-regulates CTLA4 expression on Treg cells, the antibodies can keep a high-level expression of CTLA4 on Treg cells through this recycling mechanism and makes Treg cells a better target for antibody-dependent cellular cytotoxicity, particularly in the tumor microenvironment (TME). The selective elimination of Treg cells in the tumor microenvironment and maintenance of CTLA4 expression in Treg cells in the peripheral tissues by the anti-CTLA4 antibody is contemplated to form the cellular and molecular basis for more potent tumor rejection and low toxicity. For example, the anti-CTLA4 antibody described herein dissociates from CTLA4 in endosomes, allows normal recycling of both antibodies and CTLA4, which lead to a much -reduced autoimmune toxicity. The preservation of the recycling of both CTLA4 and the anti- CTLA4 antibody facilitates more potent ADCC to eliminate Treg cells in the tumor microenvironment and induces strong CITE. ADCC preferably occurs when antibodies bind to antigens such as CTLA4 on Treg cells and the antibody Fc domains engage Fc receptors (FcR) on the surface of immune effector cells.

[0203] In some embodiments, the anti-CTLA4 antibody is an antibody selected from the group consisting of (i) an antibody which is a chimerized or humanized form of the antibody defined by the sequence identifiers above, (ii) an antibody having the specificity of the antibody defined by the sequence identifiers above, and (iii) an antibody comprising the antigen binding portion or antigen binding site, in particular the variable region, of the antibody defined by the sequence identifiers above or variant thereof and preferably having the specificity of the antibody defined by the sequence identifiers above.

[0204] BNT ref [P1819WO01] / / C&F ref. [240673WO] In some embodiments, the anti-CTLA4 antibody comprises one or more CDRs, a set of CDRs or a combination of sets of CDRs as described herein and comprises said CDRs together with their intervening framework regions. Construction of antibodies made by recombinant DNA techniques may result in the introduction of residues N- or C-terminal to the variable regions encoded by linkers introduced to facilitate cloning or other manipulation steps, including the introduction of linkers to join variable regions to further protein sequences including immunoglobulin heavy chains, other variable domains (for example in the production of diabodies) or protein labels. In one embodiment an antibody comprising one or more CDRs, a set of CDRs or a combination of sets of CDRs as described herein comprises said CDRs in a human antibody framework.

[0205] It will be appreciated by those skilled in the art that in particular the sequences of the CDR, hypervariable and variable regions can be modified without losing the ability to bind CTLA4. For example, CDR regions will be either identical or highly homologous to the regions of antibodies specified herein. By “highly homologous” it is contemplated that from 1 to 5, preferably from 1 to 4, such as 1 to 3 or 1 or 2 substitutions may be made in the CDRs. In addition, the hypervariable and variable regions may be modified so that they show substantial homology with the regions of antibodies specifically disclosed herein.

[0206] It will be appreciated by those skilled in the art that the anti-CTLA4 antibody can comprise variants of the sequence(s) disclosed herein without losing the ability to bind CTLA4. Amino acid insertion variants comprise insertions of single or two or more amino acids in a particular amino acid sequence. In the case of amino acid sequence variants having an insertion, one or more amino acid residues are inserted into a particular site in an amino acid sequence, although random insertion with appropriate screening of the resulting product is also possible.

[0207] Amino acid addition variants comprise amino- and / or carboxy-terminal fusions of one or more amino acids, such as 1, 2, 3, 5, 10, 20, 30, 50, or more amino acids. Amino acid deletion variants are characterized by the removal of one or more amino acids from the sequence, such as by removal of 1, 2, 3, 5, 10, 20, 30, 50, or more amino acids. The deletions may be in any position of the protein. Amino acid substitution variants are

[0208] BNT ref [P1819WO01] / / C&F ref. [240673WO] characterized by at least one residue in the sequence being removed and another residue being inserted in its place. Preference is given to the modifications being in positions in the amino acid sequence which are not conserved between homologous proteins or peptides and / or to replacing amino acids with other ones having similar properties. Preferably, amino acid changes in protein variants are conservative amino acid changes, i.e., substitutions of similarly charged or uncharged amino acids. A conservative amino acid change involves substitution of one of a family of amino acids which are related in their side chains. Naturally occurring amino acids are generally divided into four families: acidic (aspartate, glutamate), basic (lysine, arginine, histidine), non-polar (alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, tryptophan), and uncharged polar (glycine, asparagine, glutamine, cysteine, serine, threonine, tyrosine) amino acids. Phenylalanine, tryptophan, and tyrosine are sometimes classified jointly as aromatic amino acids.

[0209] Antibodies and antibody fragments described herein and useful in the methods described herein can be produced by a variety of techniques, including conventional monoclonal antibody methodology, e.g., the standard somatic cell hybridization technique of Kohler and Milstein, Nature 1975, 256: 495. Although somatic cell hybridization procedures are preferred, in principle, other techniques for producing monoclonal antibodies can be employed, e.g., viral or oncogenic transformation of B -lymphocytes or phage display techniques using libraries of antibody genes.

[0210] In some embodiments, an animal system for preparing hybridomas that secrete monoclonal antibodies may be a murine system. Hybridoma production in the mouse is a very well- established procedure. Immunization protocols and techniques for isolation of immunized splenocytes for fusion are known in the art. Fusion partners (e.g., murine myeloma cells) and fusion procedures are also known.

[0211] Other preferred animal systems for preparing hybridomas that secrete monoclonal antibodies are the rat and the rabbit system (e.g., described in Spieker-Polet et al., Proc Natl Acad Sci U.S.A. 1995, 92: 9348; see also Rossi et al., Am J Clin Pathol 2005, 124: 295).

[0212] Yet another strategy for generating monoclonal antibodies is to directly isolate genes encoding antibodies from lymphocytes producing antibodies of defined specificity. For

[0213] BNT ref [P1819WO01] / / C&F ref. [240673WO] details of recombinant antibody engineering see also Welschof and Kraus, Recombinant antibodies for cancer therapy ISBN-0-89603-918-8 and Bemiy K.C. Lo Antibody Engineering ISBN 1 -58829- 092-1.

[0214] To generate antibodies, mice can be immunized with carrier-conjugated peptides derived from the antigen sequence, i.e., the sequence against which the antibodies are to be directed, an enriched preparation of recombinantly expressed antigen or fragments thereof and / or cells expressing the antigen, as described. Alternatively, mice can be immunized with nucleic acid encoding the antigen or fragments thereof. In the event that immunizations using a purified or enriched preparation of the antigen do not result in antibodies, mice can also be immunized with cells expressing the antigen, e.g., a cell line, to promote immune responses.

[0215] The immune response can be monitored over the course of the immunization protocol with plasma and serum samples being obtained by tail vein or retroorbital bleeds. Mice with sufficient titers of immunoglobulin can be used for fusions. Mice can be boosted intraperitonealy or intravenously with antigen expressing cells 3 days before sacrifice and removal of the spleen to increase the rate of specific antibody secreting hybridomas.

[0216] To generate hybridomas producing monoclonal antibodies, splenocytes and lymph node cells from immunized mice can be isolated and fused to an appropriate immortalized cell line, such as a mouse myeloma cell line. The resulting hybridomas can then be screened for the production of antigen-specific antibodies. Individual wells can then be screened by ELISA for antibody secreting hybridomas. By Immunofluorescence and FACS analysis using antigen expressing cells, antibodies with specificity for the antigen can be identified. The antibody secreting hybridomas can be re-plated, screened again, and if still positive for monoclonal antibodies can be subcloned by limiting dilution. The stable subclones can then be cultured in vitro to generate antibody in tissue culture medium for characterization. Antibodies also can be produced in a host cell transfectoma using, for example, a combination of recombinant DNA techniques and gene transfection methods as are well known in the art (Morrison, Science 1985, 229: 1202).

[0217] BNT ref [P1819WO01] / / C&F ref. [240673WO] For example, in some embodiments, the gene(s) of interest, e.g., antibody genes, can be ligated into an expression vector such as a eukaryotic expression plasmid such as used by the GS gene expression system disclosed in International Patent Application Publication Nos. WO 87 / 04462 and WO 89 / 01036 and EP 338 841 A or other expression systems well known in the art. The purified plasmid with the cloned antibody genes can be introduced in eukaryotic host cells such as CHO cells, NS / 0 cells, HEK293T cells or HEK293 cells or alternatively other eukaryotic cells like plant derived cells, fungal or yeast cells. The method used to introduce these genes can be methods described in the art such as electroporation, lipofectine, lipofectamine or others. After introduction of these antibody genes in the host cells, cells expressing the antibody can be identified and selected. These cells represent the transfectomas which can then be amplified for their expression level and upscaled to produce antibodies. Recombinant antibodies can be isolated and purified from these culture supernatants and / or cells.

[0218] Alternatively, the cloned antibody genes can be expressed in other expression systems, including prokaryotic cells, such as microorganisms, e.g., E. coli. Furthermore, the antibodies can be produced in transgenic non-human animals, such as in milk from sheep and rabbits or in eggs from hens, or in transgenic plants; see e.g., Verma, R., et al., J Immunol Meth 1998,216: 165-181; Pollock, et al., J Immunol Meth 1999, 231 : 147-157; and Fischer, R., et al., Biol Chem 1999, 380: 825-839.

[0219] Antibodies interact with target antigens predominantly through amino acid residues that are located in the six heavy and light chain complementarity determining regions (CDRs). For this reason, the amino acid sequences within CDRs are more diverse between individual antibodies than sequences outside of CDRs. Because CDR sequences are responsible for most antibody-antigen interactions, it is possible to express recombinant antibodies that mimic the properties of specific naturally occurring antibodies by constructing expression vectors that include CDR sequences from the specific naturally occurring antibody grafted onto framework sequences from a different antibody with different properties (see, e.g., Riechmann et al., Nature 1998, 332:323-327; Jones et al., Nature 1986, 321 :522-525; and Queen et al., Proc Natl Acad Sci USA 1989, 86: 10029-10033). Such framework sequences can be obtained from public DNA databases that include germline antibody gene

[0220] BNT ref [P1819WO01] / / C&F ref. [240673WO] sequences. These germline sequences will differ from mature antibody gene sequences because they will not include completely assembled variable genes, which are formed by V (D) J joining during B cell maturation. Germline gene sequences will also differ from the sequences of a high affinity secondary repertoire antibody at individual evenly across the variable region.

[0221] The ability of antibodies to bind an antigen can be determined using standard binding assays (e.g., ELISA, Western Blot, Immunofluorescence and flow cytometric analysis).

[0222] To purify antibodies, selected hybridomas can be grown in two-liter spinner- flasks for monoclonal antibody purification. Alternatively, antibodies can be produced in dialysisbased bioreactors. Supernatants can be filtered and, if necessary, concentrated before affinity chromatography with protein G-sepharose or protein A-sepharose. Eluted IgG can be checked by gel electrophoresis and high performance liquid chromatography to ensure purity. The buffer solution can be exchanged into PBS, and the concentration can be determined by OD280 using 1.43 extinction coefficient. The monoclonal antibodies can be ali quoted and stored at -80°C.

[0223] To determine if selected monoclonal antibodies bind to unique epitopes and / or to characterize one or more binding properties, site-directed or multi-site directed mutagenesis can be used.

[0224] To determine the isotype of antibodies, isotype ELISAs with various commercial kits (e.g., Zymed, Roche Diagnostics) can be performed. Wells of microtiter plates can be coated with anti-mouse Ig. After blocking, the plates are reacted with monoclonal antibodies or purified isotype controls, at ambient temperature for two hours. The wells can then be reacted with either mouse IgGl, IgG2a, IgG2b or IgG3, IgA or mouse IgM-specific peroxidase- conjugated probes. After washing, the plates can be developed with ABTS substrate (1 mg / ml) and analyzed at OD of 405-650. Alternatively, the IsoStrip Mouse Monoclonal Antibody Isotyping Kit (Roche, Cat. No. 1493027) may be used as described by the manufacturer.

[0225] BNT ref [P1819WO01] / / C&F ref. [240673WO] In order to demonstrate presence of antibodies in sera of immunized mice or binding of monoclonal antibodies to living cells expressing antigen, flow cytometry can be used. Cell lines expressing naturally or after transfection antigen and negative controls lacking antigen expression (grown under standard growth conditions) can be mixed with various concentrations of monoclonal antibodies in hybridoma supernatants or in PBS containing 1 % FBS, and can be incubated at 4 °C for 30 min. After washing, the APC- or Alexa647- labeled anti IgG antibody can bind to antigen-bound monoclonal antibody under the same conditions as the primary antibody staining. The samples can be analyzed by flow cytometry with a FACS instrument using light and side scatter properties to gate on single, living cells. In order to distinguish antigen-specific monoclonal antibodies from nonspecific binders in a single measurement, the method of co-transfection can be employed. Cells transiently transfected with plasmids encoding antigen and a fluorescent marker can be stained as described above. Transfected cells can be detected in a different fluorescence channel than antibody-stained cells. As the majority of transfected cells express both transgenes, antigen- specific monoclonal antibodies bind preferentially to fluorescence marker expressing cells, whereas non-specific antibodies bind in a comparable ratio to nontransfected cells. An alternative assay using fluorescence microscopy may be used in addition to or instead of the flow cytometry assay. Cells can be stained exactly as described above and examined by fluorescence microscopy.

[0226] In order to demonstrate presence of antibodies in sera of immunized mice or binding of monoclonal antibodies to living cells expressing antigen, immunofluorescence microscopy analysis can be used. For example, cell lines expressing either spontaneously or after transfection antigen and negative controls lacking antigen expression are grown in chamber slides under standard growth conditions in DMEM / F12 medium, supplemented with 10 % fetal calf serum (FCS), 2 raM L-glutamine, 100 lU / ml penicillin and 100 pg / ml streptomycin. Cells can then be fixed with methanol or paraformaldehyde or left untreated. Cells can then be reacted with monoclonal antibodies against the antigen for 30 min. at 25°C. After washing, cells can be reacted with an Alexa555-labelled anti-mouse IgG secondary antibody (Molecular Probes) under the same conditions. Cells can then be examined by fluorescence microscopy.

[0227] BNT ref [P1819WO01] / / C&F ref. [240673WO] Cell extracts from cells expressing antigen and appropriate negative controls can be prepared and subjected to sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis. After electrophoresis, the separated antigens will be transferred to nitrocellulose membranes, blocked, and probed with the monoclonal antibodies to be tested. IgG binding can be detected using anti-mouse IgG peroxidase and developed with ECL substrate.

[0228] Antibodies can be further tested for reactivity with antigen by Immunohistochemistry in a manner well known to the skilled person, e.g., using paraformaldehyde or acetone fixed cryosections or paraffin embedded tissue sections fixed with paraformaldehyde from noncancer tissue or cancer tissue samples obtained from patients during routine surgical procedures or from mice carrying xenografted tumors inoculated with cell lines expressing spontaneously or after transfection antigen. For immunostaining, antibodies reactive to antigen can be incubated followed by horseradish-peroxidase conjugated goat anti-mouse or goat anti-rabbit antibodies (DAKO) according to the vendors instructions. The testing of monoclonal antibody activity in vitro will provide an initial screening prior to testing in vivo models.

[0229] In some embodiments, the anti-CTLA4 antibody is encoded by one or more nucleic acid molecules.

[0230] The chemotherapy agent

[0231] The methods of treating cancer in a subject disclosed herein comprise administering a chemotherapy agent. Chemotherapy is a type of cancer treatment that uses one or more anti -cancer drugs (chemotherapeutic or chemotherapy agents), usually as part of a standardized chemotherapy regimen. The term chemotherapy has come to connote nonspecific usage of intracellular poisons to inhibit mitosis. The connotation excludes more selective agents that block extracellular signals (signal transduction). The development of therapies with specific molecular or genetic targets, which inhibit growth-promoting signals from classic endocrine hormones (primarily estrogens for breast cancer and androgens for prostate cancer) are now called hormonal therapies. By contrast, other inhibitions of

[0232] BNT ref [P1819WO01] / / C&F ref. [240673WO] growth-signals like those associated with receptor tyrosine kinases are referred to as targeted therapy.

[0233] The use of drugs (whether chemotherapy, hormonal therapy or targeted therapy) constitutes systemic therapy for cancer in that they are introduced into the blood stream and are therefore in principle able to address cancer at any anatomic location in the body. Systemic therapy is often used in conjunction with other modalities that constitute local therapy (i.e., treatments whose efficacy is confined to the anatomic area where they are applied) for cancer such as radiation therapy, surgery or hyperthermia therapy. Traditional chemotherapeutic agents are cytotoxic by means of interfering with cell division (mitosis) but cancer cells vary widely in their susceptibility to these agents. To a large extent, chemotherapy can be thought of as a way to damage or stress cells, which may then lead to cell death if apoptosis is initiated.

[0234] Chemotherapeutic agents include alkylating agents, antimetabolites, anti -microtubule agents, topoisomerase inhibitors, and cytotoxic antibiotics.

[0235] Alkylating agents have the ability to alkylate many molecules, including proteins, RNA and DNA. The subtypes of alkylating agents are the nitrogen mustards, nitrosoureas, tetrazines, aziridines, cisplatins and derivatives, and non-classical alkylating agents. Nitrogen mustards include mechlorethamine, cyclophosphamide, melphalan, chlorambucil, ifosfamide and busulfan. Nitrosoureas include N-Nitroso-N-methylurea (MNU), carmustine (BCNU), lomustine (CCNU) and semustine (MeCCNU), fotemustine and streptozotocin. Tetrazines include dacarbazine, mitozolomide and temozolomide. Aziridines include thiotepa, mytomycin and diaziquone (AZQ). Cisplatin and derivatives include cisplatin, carboplatin and oxaliplatin. They impair cell function by forming covalent bonds with the amino, carboxyl, sulfhydryl, and phosphate groups in biologically important molecules. Non-classical alkylating agents include procarbazine and hexamethylmelamine. In one particularly preferred embodiment, the alkylating agent is cyclophosphamide.

[0236] Anti-metabolites are a group of molecules that impede DNA and RNA synthesis. Many of them have a similar structure to the building blocks of DNA and RNA. Anti -metabolites resemble either nucleobases or nucleosides, but have altered chemical groups. These drugs

[0237] BNT ref [P1819WO01] / / C&F ref. [240673WO] exert their effect by either blocking the enzymes required for DNA synthesis or by becoming incorporated into DNA or RNA. Subtypes of the anti-metabolites are the antifolates, fluoropyrimidines, deoxynucleoside analogues, and thiopurines. The anti-folates include methotrexate and pemetrexed. The fluoropyrimidines include fluorouracil and capecitabine. The deoxynucleoside analogues include cytarabine, gemcitabine, decitabine, azacitidine, fludarabine, nelarabine, cladribine, clofarabine, and pentostatin. The thiopurines include thioguanine and mercaptopurine.

[0238] Anti -microtubule agents block cell division by preventing microtubule function. The vinca alkaloids prevent the formation of the microtubules, whereas the taxanes prevent the microtubule disassembly. Vinca alkaloids include vinorelbine, vindesine, and vinflunine. Taxanes include docetaxel (Taxotere®) and paclitaxel (Taxol®).

[0239] Topoisomerase inhibitors are drugs that affect the activity of two enzymes: topoisomerase I and topoisomerase II and include irinotecan, topotecan, camptothecin, etoposide, doxorubicin, mitoxantrone, teniposide, novobiocin, merbarone, and aclarubicin.

[0240] The cytotoxic antibiotics are a varied group of drugs that have various mechanisms of action. The common theme that they share in their chemotherapy indication is that they interrupt cell division. The most important subgroup is the anthracyclines (e.g., doxorubicin, daunorubicin, epirubicin, idarubicin, pirarubicin, and aclarubicin) and the bleomycins; other prominent examples include mitomycin C, mitoxantrone, and actinomycin.

[0241] In some embodiments, the chemotherapy agent comprises or is selected from alkylating agents, antimetabolites, anti -microtubule agents, topoisomerase inhibitors, and cytotoxic antibiotics.

[0242] In some embodiments, the chemotherapy agent comprises or is selected from platinum- containing chemotherapy agents. The term “platinum-containing chemotherapy” refers to the use of chemotherapeutic agent(s) (also known as platins) used to treat cancer that are coordination complexes of platinum. Platinum-containing chemotherapeutic agents are alkylating agents that crosslink DNA, resulting in ineffective DNA mismatch repair and

[0243] BNT ref [P1819WO01] / / C&F ref. [240673WO] generally leading to apoptosis. Examples of platins include cisplatin, carboplatin, and oxaliplatin.

[0244] In some embodiments, the chemotherapy agent comprises or is selected from cisplatin, oxaliplatin, carboplatin, lurbinectedin, topotecan, paclitaxel, nanoparticle albumin-bound paclitaxel (nab -paclitaxel), pemetrexed, 5-fluoruracil, irinotecan, etoposide, gemcitabine, docetaxel or combinations thereof.

[0245] In some preferred embodiments, the chemotherapy agent comprises or is selected from the group of taxanes. In some embodiments, the chemotherapy agent comprises or is selected from paclitaxel and / or docetaxel. In some embodiments, the chemotherapy agent comprises or is paclitaxel and docetaxel. In some embodiments, the chemotherapy agent is paclitaxel and docetaxel. In some embodiments, the chemotherapy agent comprises or is paclitaxel. In some embodiments, the chemotherapy agent comprises paclitaxel. In some embodiments, the chemotherapy agent is paclitaxel. In some preferred embodiments, the chemotherapy agent comprises or is docetaxel. In some preferred embodiments, the chemotherapy agent comprises docetaxel. In some preferred embodiments, the chemotherapy agent is docetaxel.

[0246] Docetaxel (Taxotere®; Sanofi -Aventis) belongs the class of taxanes which are a class of diterpene compounds that were first derived from natural sources such as plants of the genus Taxus. However, taxanes also some have been synthesized artificially. The principal mechanism of action of the taxane class of drugs is the disruption of microtubule function, thereby inhibiting the process of cell division. Docetaxel promotes microtubule stabilization by promoting the assembly of microtubule dimers into microtubules and preventing depolymerization and thus microtubule disassembly, thereby inhibiting mitosis and proliferation of cancer cells.

[0247] In some preferred embodiments, the term “docetaxel” comprises or consists of the compound (2R,3S)-4-Acetoxy-2a-benzyloxy-13-[3-(N-tert-butoxycarbonyl)amino-2- hydroxy-3-phenyl]propionyl-5p,20-epoxy-l,7p,10P-trihydroxy-9-oxotax-l l-en-13a-ylester.

[0248] BNT ref [P1819WO01] / / C&F ref. [240673WO] The structure of docetaxel may be represented as follows:

[0249] Treatment

[0250] In some embodiments, the anti-CTLA4 antibody or fragment thereof and the chemotherapy agent are administered separately. In some preferred embodiments, a dose of the anti- CTLA4 antibody or fragment thereof and a dose of the chemotherapy agent are administered separately. For example, a dose of the anti-CTLA4 antibody or fragment thereof and a dose of the chemotherapy agent are administered separately using two different pharmaceutical compositions, each individually comprising the anti-CTLA4 antibody (or fragment thereof) disclosed herein, or the chemotherapy agent disclosed herein. In some embodiments, a dose of the anti-CTLA4 antibody or fragment thereof and a dose of the chemotherapy agent are administered using a single pharmaceutical composition.

[0251] In some embodiments, the anti-CTLA4 antibody (or fragment thereof) and the chemotherapy agent are administered concurrently or consecutively. In some embodiments, the anti-CTLA4 antibody (or fragment thereof) and the chemotherapy agent are administered concurrently. This means that a dose of the anti-CTLA4 antibody (or

[0252] BNT ref [P1819WO01] / / C&F ref. [240673WO] fragment thereof) is administered simultaneously or essentially at the same time with a dose of the chemotherapy agent. In some embodiments, the anti-CTLA4 antibody (or fragment thereof) and the chemotherapy agent are administered simultaneously. In some embodiments, the anti-CTLA4 antibody (or fragment thereof) and the chemotherapy agent are administered essentially at the same time. In some embodiments, the anti-CTLA4 antibody or fragment thereof and the chemotherapy agent are administered separately, i.e., using two different pharmaceutical compositions each individually comprising the anti- CTLA4 antibody or fragment thereof or the chemotherapy agent disclosed herein, and concurrently. In some embodiments, the anti-CTLA4 antibody or fragment thereof and the chemotherapy agent are administered separately, i.e., using two different pharmaceutical compositions each individually comprising the anti-CTLA4 antibody or fragment thereof or the chemotherapy agent disclosed herein, and simultaneously. In some embodiments, the anti-CTLA4 antibody or fragment thereof and the chemotherapy agent are administered separately, i.e., using two different pharmaceutical compositions each individually comprising the anti-CTLA4 antibody or fragment thereof or the chemotherapy agent disclosed herein, and essentially at the same time.

[0253] In some embodiments, the anti-CTLA4 antibody or fragment thereof and the chemotherapy agent are administered consecutively. This means that doses of the anti-CTLA4 antibody or fragment thereof and doses of the chemotherapy agent are administered in treatment cycles.

[0254] In some embodiments a treatment cycle has at least about 1 day, about 2 days, about 3 days, about 4 days, about 5 days, about 6 days, about 7 days, about 8 days, about 9 days, about 10 days, about 11 days, about 12 days, about 13 days, about 14 days, about 15 days, about 16 days, about 17 days, about 18 days, about 19 days, about 20 days, about 21 days, about 22 days, about 23 days, about 24 days, about 25 days, about 26 days, about 27 days, or about

[0255] 28 days. In some embodiments a treatment cycle has up to about 28 days. In some embodiments, a treatment cycle has 28 days. In some preferred embodiments, a treatment cycle has up to about 21 days. In some more preferred embodiments, a treatment cycle has about 21 days. In some more preferred embodiments, a treatment cycle has 21 days.

[0256] BNT ref [P1819WO01] / / C&F ref. [240673WO] In some embodiments, a treatment cycle is repeated at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, or 17 times. However, the treatment can be continued until disease progression or the initiation of a new anti-tumor treatment.

[0257] In some embodiments, the anti-CTLA4 antibody (or fragment thereof) is administered about every 6 weeks. In some embodiments, the anti-CTLA4 antibody (or fragment thereof) is administered every 6 weeks. In some preferred embodiments, the anti-CTLA4 antibody (or fragment thereof) is administered about every 4 weeks. In some preferred embodiments, the anti-CTLA4 antibody (or fragment thereof) is administered every 4 weeks. In some more preferred embodiments, the anti-CTLA4 antibody (or fragment thereof) is administered about every 3 weeks. In some more preferred embodiments, the anti-CTLA4 antibody (or fragment thereof) is administered every 3 weeks.

[0258] In some embodiments, the chemotherapy agent is administered about every 6 weeks. In some embodiments, the chemotherapy agent is administered every 6 weeks. In some preferred embodiments, the chemotherapy agent is administered about every 4 weeks. In some preferred embodiments, the chemotherapy agent is administered every 4 weeks. In some more preferred embodiments, the chemotherapy agent is administered about every 3 weeks. In some more preferred embodiments, the chemotherapy agent is administered every 3 weeks.

[0259] In some embodiments, the anti-CTLA4 antibody (or fragment thereof) and the chemotherapy agent are administered about every 6 weeks. In some embodiments, the anti- CTLA4 antibody (or fragment thereof) and the chemotherapy agent are administered every 6 weeks. In some preferred embodiments, the anti-CTLA4 antibody (or fragment thereof) and the chemotherapy agent are administered about every 4 weeks. In some preferred embodiments, the anti-CTLA4 antibody (or fragment thereof) and the chemotherapy agent are administered every 4 weeks. In some more preferred embodiments, the anti-CTLA4 antibody (or fragment thereof) and the chemotherapy agent are administered about every 3 weeks. In some more preferred embodiments, the anti-CTLA4 antibody (or fragment thereof) and the chemotherapy agent are administered every 3 weeks.

[0260] BNT ref [P1819WO01] / / C&F ref. [240673WO] In some embodiments, the chemotherapy agent is administered up to the 8thtreatment cycle. In some preferred embodiments, the chemotherapy agent is administered up to the 7thtreatment cycle. In some more preferred embodiments, the chemotherapy agent is administered more up to the 6thtreatment cycle. In some even more preferred embodiments, the chemotherapy agent is administered up to the 5thtreatment cycle. In some most preferred embodiments, the chemotherapy agent is administered up to the 4thtreatment cycle. In some preferred embodiments, the chemotherapy agent is administered for four treatment cycles.

[0261] The anti-CTLA4 antibody or fragment thereof and the chemotherapy agent can be first administered concurrently, e.g., in up to 4 treatment cycles, followed by consecutive administration of either the anti-CTLA4 antibody (or fragment thereof) or the chemotherapy agent in a subsequent treatment cycle or in subsequent treatment cycles.

[0262] In some embodiments, the anti-CTLA4 antibody (or fragment thereof) and the chemotherapy agent are administered up to the 8thtreatment cycle, followed by consecutive administration of the anti-CTLA4 antibody (or fragment thereof). This means that a dose of the anti-CTLA4 antibody (or fragment thereof) is administered without the chemotherapy agent from the 9thtreatment cycle onwards. In some preferred embodiments, the anti- CTLA4 antibody (or fragment thereof) and the chemotherapy agent are administered up to the 7thtreatment cycle, followed by consecutive administration of the anti-CTLA4 antibody (or fragment thereof). In some more preferred embodiments, the anti-CTLA4 antibody (or fragment thereof) and the chemotherapy agent are administered up to the 6thtreatment cycle, followed by consecutive administration of the anti-CTLA4 antibody (or fragment thereof). In some even more preferred embodiments, the anti-CTLA4 antibody (or fragment thereof) and the chemotherapy agent are administered up to the 5thtreatment cycle, followed by consecutive administration of the anti-CTLA4 antibody (or fragment thereof). In some most preferred embodiments, the anti-CTLA4 antibody (or fragment thereof) and the chemotherapy agent are administered up to the 4thtreatment cycle, followed by consecutive administration of the anti-CTLA4 antibody (or fragment thereof). For example, a dose of the anti-CTLA4 antibody (or fragment thereof) and a dose of the chemotherapy agent are administered concurrently for four treatment cycles, followed by consecutive

[0263] BNT ref [P1819WO01] / / C&F ref. [240673WO] administration of a dose or of doses of the anti-CTLA4 antibody (or fragment thereof) from the 5thtreatment cycle onwards.

[0264] In some embodiments, the anti-CTLA4 antibody (or fragment thereof) is administered for 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, or 17 treatment cycles. In some embodiments, the anti-CTLA4 antibody (or fragment thereof) is administered for up to 17 treatment cycles. In some embodiments, the anti-CTLA4 antibody (or fragment thereof) is administered for 17 treatment cycles or up to 1 year. In some embodiments, the anti- CTLA4 antibody (or fragment thereof) is administered for 17 treatment cycles. In some embodiments, the anti-CTLA4 antibody (or fragment thereof) is administered for up to one year. In some preferred embodiments, the anti-CTLA4 antibody (or fragment thereof) and the chemotherapy agent are administered up to the 4thtreatment cycle, followed by consecutive administration of the anti-CTLA4 antibody (or fragment thereof) for up to 13 treatment cycles or up to one year.

[0265] In some embodiments, the anti-CTLA4 antibody (or fragment thereof) is administered at a dosage ranging from about 0.1 mg / kg to about 45 mg / kg body weight. In some preferred embodiments, the anti-CTLA4 antibody (or fragment thereof) is administered at a dosage ranging from about 1 mg / kg to about 30 mg / kg body weight. In some more preferred embodiments, the anti-CTLA4 antibody (or fragment thereof) is administered at a dosage ranging from about 2 mg / kg to about 20 mg / kg body weight.

[0266] In some embodiments, the anti-CTLA4 antibody (or fragment thereof) is administered at a dosage independently selected from about 0.1 mg / kg body weight, about 0.3 mg / kg body weight, about 1.0 mg / kg body weight, about 3.0 mg / kg body weight, about 6 mg / kg body weight, about 10 mg / kg body weight, about 12 mg / kg body weight, about 15 mg / kg body weight, and about 20 mg / kg body weight.

[0267] In some preferred embodiments, the anti-CTLA4 antibody (or fragment thereof) is administered at a dosage of about 10 mg / kg body weight. In some preferred embodiments, the anti-CTLA4 antibody (or fragment thereof) is administered at a dosage of 10 mg / kg body weight. In some preferred embodiments, the anti-CTLA4 antibody (or fragment thereof) is administered at a dosage of about 6 mg / kg body weight. In some preferred

[0268] BNT ref [P1819WO01] / / C&F ref. [240673WO] embodiments, the anti-CTLA4 antibody (or fragment thereof) is administered at a dosage of 6 mg / kg body weight.

[0269] The subject afflicted with cancer and treated with the anti-CTLA4 antibody disclosed herein can suffer from treatment-related adverse events, e.g., from irAEs. For example, the subject afflicted with cancer and treated with the anti-CTLA4 antibody disclosed herein, e.g., with the anti-CTLA4 antibody denoted as PP4637 herein, may suffer from severe skin reactions or other grade 3 or 4 non-hematological toxicities according to Common Terminology Criteria for Adverse Events (CTCAE) v5.0. In such cases, the dose of the anti-CTLA4 antibody can be reduced. In some embodiments, the anti-CTLA4 antibody (or fragment thereof) is administered at a dosage of about 10 mg / kg body weight, about 6 mg / kg body weight, about 3 mg / kg body weight, or about 1 mg / kg body weight. In some embodiments, the anti-CTLA4 antibody (or fragment thereof) is administered at a dosage of 3 mg / kg body weight. In some embodiments, the anti-CTLA4 antibody (or fragment thereof) is administered at a dosage of 1 mg / kg body weight.

[0270] In some embodiments, the chemotherapy agent is administered at a dosage ranging from about 0.1 to about 300 mg / m2body surface area. In some preferred embodiments, the chemotherapy agent is administered at a dosage ranging from about 1 to about 200 mg / m2body surface area. In some more preferred embodiments, more the chemotherapy agent is administered at a dosage ranging from about 10 to about 100 mg / m2body surface area.

[0271] In some embodiments, the chemotherapy agent is administered at a dosage independently selected from about 0.1 mg / m2body surface area, about 0.3 mg / m2body surface area, about 1 mg / m2body surface area, about 10 mg / m2body surface area, about 22.5 mg / m2body surface area, about 25 mg / m2body surface area, about 27.5 mg / m2body surface area, about 30 mg / m2body surface area, about 32.5 mg / m2body surface area, about 35 mg / m2body surface area, about 37.5 mg / m2body surface area, about 40 mg / m2body surface area about 50 mg / m2body surface area, about 55 mg / m2body surface area, about 60 mg / m2body surface area, about 65 mg / m2body surface area, about 70 mg / m2body surface area, about 75 mg / m2body surface area, about 100 mg / m2body surface area, and about 125 mg / m2body surface area.

[0272] BNT ref [P1819WO01] / / C&F ref. [240673WO] In some preferred embodiments, the chemotherapy agent is administered at a dosage of about 75 mg / m2body surface area. In some preferred embodiments, the chemotherapy agent is administered at a dosage of 75 mg / m2body surface area. The subject afflicted with cancer and treated with the chemotherapy agent disclosed herein may suffer from treatment-related adverse events. For example, the subject afflicted with cancer and treated with docetaxel can suffer from either febrile neutropenia (e.g., neutrophils <500 cells / mm3for more than one week despite support with granulocyte stimulating factor), severe skin reactions or other grade 3 or 4 non-hematological toxicities according to Common Terminology Criteria for Adverse Events (CTCAE) v5.0. In such cases, the dose of the chemotherapy agent, e.g., the dose of docetaxel, can be reduced. In some embodiments, the chemotherapy agent is administered at a dosage of about 37.5 mg / m2body surface area, about 55 mg / m2body surface area, or about 75 mg / m2body surface area. In some embodiments, the chemotherapy agent is administered at a dosage of 37.5 mg / m2body surface area. In some embodiments, the chemotherapy agent is administered at a dosage of 55 mg / m2body surface area.

[0273] In some embodiments, the dosage of the anti-CTLA4 antibody or fragment thereof administered at least during the 1stand 2ndtreatment cycle ranges from about 2 mg / kg to about 20 mg / kg body weight. In some preferred embodiments, the dosage of the anti- CTLA4 antibody or fragment thereof administered at least during the 1stand 2ndtreatment cycle ranges from about 4 mg / kg to about 15 mg / kg body weight. In some more preferred embodiments, the dosage of the anti-CTLA4 antibody or fragment thereof administered at least during the 1stand 2ndtreatment cycle ranges from about 6 mg / kg to about 12 mg / kg body weight.

[0274] In some embodiments, the dosage of the anti-CTLA4 antibody or fragment thereof administered up to the 2ndtreatment cycle ranges from about 2 mg / kg to about 20 mg / kg body weight. In some preferred embodiments, the dosage of the anti-CTLA4 antibody or fragment thereof administered up to the 2ndtreatment cycle ranges from about 4 mg / kg to about 15 mg / kg body weight. In some more preferred embodiments, the dosage of the anti- CTLA4 antibody or fragment thereof administered during the 1stand 2ndtreatment cycle ranges from about 6 mg / kg to about 12 mg / kg body weight.

[0275] BNT ref [P1819WO01] / / C&F ref. [240673WO] In some preferred embodiments, the dosage of the anti-CTLA4 antibody administered during the 1stand 2ndtreatment cycles is 10 mg / kg body weight. In some preferred embodiments, the dosage of the anti-CTLA4 antibody administered during the 1stor 2ndtreatment cycle is 10 mg / kg body weight.

[0276] In some embodiments, the dosage of the anti-CTLA4 antibody or fragment thereof administered at least during the 3rdand 4thcycle ranges from about 2 mg / kg to about 20 mg / kg. In some preferred embodiments, the dosage of the anti-CTLA4 antibody or fragment thereof administered at least during the 3rdand 4thcycle ranges from about 2 mg / kg to about 10 mg / kg. In some more preferred embodiments, the dosage of the anti- CTLA4 antibody or fragment thereof administered at least during the 3rdand 4thcycle ranges from about 3 mg / kg to about 8 mg / kg. In some most preferred embodiments, the dosage of the anti-CTLA4 antibody or fragment thereof administered at least during the 3rdand 4thcycle ranges from about 4 mg / kg to about 7 mg / kg.

[0277] In some preferred embodiments, the dosage of the anti-CTLA4 antibody or fragment thereof administered during the 3rdand 4thcycle is 6 mg / kg body weight. In some preferred embodiments, the dosage of the anti-CTLA4 antibody or fragment thereof administered during the 3rdor 4thcycle is 6 mg / kg body weight.

[0278] In some embodiments, the dosage of the anti-CTLA4 antibody or fragment thereof administered from the 5thcycle onwards ranges from about 2 mg / kg to about 20 mg / kg. In some preferred embodiments, the dosage of the anti-CTLA4 antibody or fragment thereof administered from the 5thcycle onwards ranges from about 2 mg / kg to about 10 mg / kg. In some more preferred embodiments, the dosage of the anti-CTLA4 antibody or fragment thereof administered from the 5thcycle onwards ranges from about 3 mg / kg to about 8 mg / kg. In some most preferred embodiments, the dosage of the anti-CTLA4 antibody or fragment thereof administered from the 5thcycle onwards ranges from about 4 mg / kg to about 7 mg / kg.

[0279] In some preferred embodiments, the dosage of the anti-CTLA4 antibody or fragment thereof administered from the 5thcycle onwards is 6 mg / kg body weight. In some preferred

[0280] BNT ref [P1819WO01] / / C&F ref. [240673WO] embodiments, the dosage of the anti-CTLA4 antibody or fragment thereof administered from the 5thcycle onwards is 6 mg / kg body weight.

[0281] In some embodiments, the dosage of the chemotherapy agent administered during the 1st, the 2nd, the 3rd, and the 4thcycle ranges from about 1 to about 200 mg / m2body surface area. In some preferred embodiments, the dosage of the chemotherapy agent administered during the 1st, the 2nd, the 3rd, and the 4thcycle ranges from about 10 to about 150 mg / m2body surface area. In some more preferred embodiments, the dosage of the chemotherapy agent administered during the 1st, the 2nd, the 3rd, and the 4thcycle ranges from about 25 to about 100 mg / m2body surface area. In some most preferred embodiments, the dosage of the chemotherapy agent administered during the 1st, the 2nd, the 3rd, and the 4thcycle ranges from about 30 to about 80 mg / m2body surface area.

[0282] In some preferred embodiments, the dosage of the chemotherapy agent administered during 1st, the 2nd, the 3rd, and the 4thcycle is 75 mg / m2body surface area. In case in which the subject suffers from treatment-related adverse events as described above, the dose of the chemotherapy agent, e.g., the dose of docetaxel, can be reduced. In some embodiments, the dosage of the chemotherapy agent administered during 1st, the 2nd, the 3rd, and the 4thcycle is about 37.5 mg / m2body surface area, about 55 mg / m2body surface area, or about 75 mg / m2body surface area. In some embodiments, the dosage of the chemotherapy agent administered during 1st, the 2nd, the 3rd, and the 4thcycle is 37.5 mg / m2body surface area. In some embodiments, the dosage of the chemotherapy agent administered during 1st, the 2nd, the 3rd, and the 4thcycle is 55 mg / m2body surface area.

[0283] In some embodiments, the anti-CTLA4 antibody or fragment thereof and the chemotherapy agent are administered as follows:

[0284] • 1stand 2ndtreatment cycle (duration of each treatment cycle: three weeks): administration of the anti-CTLA4 antibody or fragment thereof (dose: 10 mg / kg body weight) and the chemotherapy agent (dose: 75 mg / m2body surface area).

[0285] BNT ref [P1819WO01] / / C&F ref. [240673WO] 3rdand 4thtreatment cycle (duration of each treatment cycle: three weeks): administration of the anti-CTLA4 antibody or fragment thereof (dose: 6 mg / kg body weight) and the chemotherapy agent (dose: 75 mg / m2body surface area).

[0286] • 5thtreatment cycle up to the 17thtreatment cycle or up to one year (duration of each treatment cycle: three weeks): administration of the anti-CTLA4 antibody or fragment thereof (dose: 6 mg / kg body weight).

[0287] In some embodiments, the methods disclosed herein comprise:

[0288] • administering the anti-CTLA4 antibody in a dose of 10 mg / kg, and administering the chemotherapy agent in a dose of 37.5 mg / m2, 55 mg / m2, or 75 mg / m2body surface area, preferably in a 1stand 2ndtreatment cycle;

[0289] • administering the anti-CTLA4 antibody in a dose of 6 mg / kg, and administering the chemotherapy agent in a dose of 37.5 mg / m2, 55 mg / m2, or 75 mg / m2body surface area, preferably in a 3rdand 4thtreatment cycles;

[0290] • administering the anti-CTLA4 antibody in a dose of 6 mg / kg, preferably for up to one year; wherein each treatment cycle preferably is 3 weeks, and wherein the chemotherapy agent preferably is not administered after the 4thtreatment cycle.

[0291] In some embodiments, the methods disclosed herein comprise:

[0292] • administering the anti-CTLA4 antibody in a dose of 10 mg / kg, and administering the chemotherapy agent in a dose of 75 mg / m2body surface area, preferably in a 1stand 2ndtreatment cycle;

[0293] BNT ref [P1819WO01] / / C&F ref. [240673WO] • administering the anti-CTLA4 antibody in a dose of 6 mg / kg, and administering the chemotherapy agent in a dose of 75 mg / m2body surface area, preferably in a 3rdand 4thtreatment cycles;

[0294] • administering the anti-CTLA4 antibody in a dose of 6 mg / kg, preferably for up to one year; wherein each treatment cycle preferably is 3 weeks, and wherein the chemotherapy agent preferably is not administered after the 4thtreatment cycle.

[0295] In some embodiments, the subject has not previously been treated for cancer i.e., it is treatment naive. In some embodiments, the subject has been previously treated for cancer, in particular the subject has had at least one previous chemotherapy treatment. In some embodiments, the at least one previous chemotherapy treatment comprises or is a platinum- containing chemotherapy. In some embodiments, the at least one previous chemotherapy treatment comprises or is a treatment with cisplatin, carboplatin, and / or oxaliplatin.

[0296] In some embodiments, the subject has previously not been treated with docetaxel, i.e., it is treatment naive for docetaxel.

[0297] In some embodiments, overall survival is increased in a subject afflicted with cancer compared to the chemotherapy agent or the anti-CTLA4 antibody or fragment thereof treatment alone. In some embodiments, the median progression-free survival is increased in a subject afflicted with cancer compared to the chemotherapy agent or the anti-CTLA4 antibody or fragment thereof treatment alone.

[0298] In some embodiments, the cancer to be treated by the combination of the present disclosure comprising the anti-CTLA4 antibody as disclosed herein and the chemotherapy agent as disclosed herein comprises one or more solid tumors.

[0299] Examples of cancers include, but are not limited to, carcinoma, lymphoma, blastoma, sarcoma, and leukemia. More particularly, examples of such cancers include bone cancer,

[0300] BNT ref [P1819WO01] / / C&F ref. [240673WO] blood cancer, lung cancer, liver cancer, pancreatic cancer, skin cancer, cancer of the head or neck, cutaneous or intraocular melanoma, uterine cancer, ovarian cancer, rectal cancer, cancer of the anal region, stomach cancer, colon cancer, breast cancer, prostate cancer, uterine cancer, carcinoma of the sexual and reproductive organs, Hodgkin's Disease, cancer of the esophagus, cancer of the small intestine, cancer of the endocrine system, cancer of the thyroid gland, cancer of the parathyroid gland, cancer of the adrenal gland, sarcoma of soft tissue, cancer of the bladder, cancer of the kidney, renal cell carcinoma, carcinoma of the renal pelvis, neoplasms of the central nervous system (CNS), neuroectodermal cancer, spinal axis tumors, glioma, meningioma, and pituitary adenoma.

[0301] In some embodiments, the cancer to be treated by the combination of the present disclosure comprising the anti-CTLA4 antibody as disclosed herein and the chemotherapy agent as disclosed herein is selected from melanoma, lung cancer, human papillomavirus (HPV)- induced cancer, breast cancer, hepatocellular carcinoma, ovarian cancer such as ovarian carcinoma, prostate cancer such as prostate carcinoma, Hodgkin's or non-Hodgkin's lymphoma, acute myelogenic Leukemia, chronic myelogenic Leukemia, acute lymphocytic Leukemia, chronic lymphocytic Leukemia, or renal cell carcinoma. In some preferred embodiments, the cancer is lung cancer. In some preferred embodiments, the cancer is nonsmall cell lung cancer (NSCLC). In some embodiments, the NSCLC has a squamous histology. In some embodiments, the NSCLC has a non-squamous histology. In some embodiments, the NSCLC has locally advanced or metastasized. The skilled person knows how to determine whether the NSCLC has locally advanced or metastasized, e.g., by using X-ray methods, ultrasound methods, positron emission tomography-computed tomography (PET-CT), magnetic resonance imaging (MRI), and / or the parameters defined in Response Evaluation Criteria In Solid Tumors (RECIST) version 1.1.

[0302] In some embodiments, the cancer to be treated by the combination of the present disclosure comprising the anti-CTLA4 antibody as disclosed herein, and the chemotherapy agent as disclosed herein is resistant to treatment with immune checkpoint inhibitors. In some embodiments, the cancer to be treated is resistant to treatment with anti-PDl and / or anti- PD-L1 antibodies. In some embodiments, the cancer to be treated is resistant to treatment with anti-PDl antibodies. In some embodiments, the cancer to be treated is resistant to

[0303] BNT ref [P1819WO01] / / C&F ref. [240673WO] treatment with anti-PD-Ll antibodies. In some embodiments, the subject afflicted with cancer and to be treated with the combination of the present disclosure comprising the anti- CTLA4 antibody as disclosed herein and the chemotherapy agent as disclosed herein has a primary or acquired resistance to treatment with immune checkpoint inhibitors, particularly to treatment with anti-PDl and / or anti-PD-Ll antibodies.

[0304] In some preferred embodiments, the NSCLC is resistant to treatment with anti-PDl and / or anti-PD-Ll antibodies. In some preferred embodiments, the NSCLC is resistant to treatment with anti-PDl antibodies. In some preferred embodiments, the NSCLC is resistant to treatment with anti-PD-Ll antibodies. In some preferred embodiments, the subject afflicted with NSCLC and to be treated with the combination of the present disclosure comprising the anti-CTLA4 antibody as disclosed herein and the chemotherapy agent as disclosed herein has a primary or acquired resistance to treatment with immune checkpoint inhibitors. In some preferred embodiments, the subject afflicted with NSCLC has a primary resistance to treatment with anti-PDl and / or anti-PD-Ll antibodies. In some preferred embodiments, the subject afflicted with NSCLC has a primary resistance to treatment with anti-PDl antibodies. In some preferred embodiments, the subject afflicted with NSCLC has a primary resistance to treatment with anti-PD-Ll antibodies. In some preferred embodiments, the subject afflicted with NSCLC has an acquired resistance to treatment with anti-PDl and / or anti-PD-Ll antibodies. In some preferred embodiments, the subject afflicted with NSCLC has an acquired resistance to treatment with anti-PDl antibodies. In some preferred embodiments, the subject afflicted with NSCLC has an acquired resistance to treatment with anti-PD-Ll antibodies.

[0305] In some embodiments, the NSCLC is negative for targetable NSCLC driver mutations. NSCLC driver mutation are oncogenic driver mutations of NSCLC. Non-limiting examples of NSCLC driver mutations are Epidermal Growth Factor Receptor (EGFR) gene mutations and anaplastic lymphoma kinase (ALK) gene mutations (Luo et al., Transl Respir Med. 2013, 1 (1) :6). In some embodiments, the NSCLC is negative for EGFR and / or ALK mutations. In some embodiments, the NSCLC is negative for EGFR and ALK mutations. In some embodiments, the NSCLC is negative for EGFR mutations. In some embodiments, the NSCLC is negative for ALK mutations. Without wishing to be bound by theory, it is

[0306] BNT ref [P1819WO01] / / C&F ref. [240673WO] contemplated herein that the combination treatment using the anti-CTLA antibodies as disclosed herein and the chemotherapy agent as disclosed herein, is particularly effective for the treatment of cancer patients without targetable NSCLC driver mutations. In some embodiments, the NSCLC can be positive for targetable NSCLC driver mutations. In some embodiments, the NSCLC is positive for Kirsten rat sarcoma (KRAS) mutations.

[0307] In some embodiments, the method does not comprise administering an RNA cancer vaccine comprising at least one RNA. An “RNA cancer vaccine” can be a composition comprising at least one RNA, wherein the composition induces an immune response against cancer cells upon administration to a subject. The RNA may encode tumor-associated or tumorspecific antigens.

[0308] The anti-CTLA4 antibody or fragment thereof, the chemotherapy agent, and the optional additional immunomodulatory agent provided herein can be administered via any suitable route of administration, e.g., parenteral route of administration. “Parenteral route” of administration refers to a route of administration other than enteral route. Examples of parenteral routes of administration include intravenous, intramuscular, intradermal, intraperitoneal, intratumour, intravesical, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, transtracheal, intraarticular, subcapsular, subarachnoid, intraspinal, epidural and intrastemal, subcutaneous, or topical administration.

[0309] The anti-CTLA4 antibody or fragment thereof, the chemotherapy agent, and / or the optional additional immunomodulatory agent of the present disclosure can be administered using any suitable method, such as by oral ingestion, nasogastric tube, gastrostomy tube, injection, infusion, implantable infusion pump, and osmotic pump. The suitable route and method of administration may vary depending on a number of factors such as the specific therapeutic agent being used, the rate of absorption desired, specific formulation or dosage form used, type or severity of the disorder being treated, the specific site of action, and conditions of the subject, and can be readily selected by a person skilled in the art. Administration can be systemic or local.

[0310] In some embodiments, the anti-CTLA4 antibody or fragment thereof, the chemotherapy agent, and / or the optional additional immunomodulatory agent may be administered

[0311] BNT ref [P1819WO01] / / C&F ref. [240673WO] intravenously, intraarterially, subcutaneously, intradermally, dermally, intranodally, or intramuscularly. In some embodiments, the anti-CTLA4 antibody or fragment thereof, the chemotherapy agent, and / or the optional additional immunomodulatory agent may be administered intramuscularly. In some embodiments, the anti-CTLA4 antibody or fragment thereof, the chemotherapy agent, and / or the optional additional immunomodulatory agent are formulated for local administration or systemic administration. Systemic administration may include enteral administration, which involves absorption through the gastrointestinal tract, or parenteral administration. In some embodiments, the anti-CTLA4 antibody or fragment thereof, the chemotherapy agent, and / or the optional additional immunomodulatory agent are formulated for systemic administration. In some embodiments, the systemic administration is by intravenous administration. In some embodiments, the anti-CTLA4 antibody or fragment thereof, the chemotherapy agent, and / or the optional additional immunomodulatory agent are formulated for intramuscular administration. In some embodiments, the anti-CTLA4 antibody or fragment thereof, the chemotherapy agent and / or the optional additional immunomodulatory agent are formulated for intravenous administration.

[0312] In some embodiments, the anti-CTLA4 antibody or fragment thereof, the chemotherapy agent, and / or the optional additional immunomodulatory agent are administered intravenously. In some preferred embodiments, the anti-CTLA4 antibody or fragment thereof, the chemotherapy agent, and / or the optional additional immunomodulatory agent are administered via an intravenous injection or an intravenous infusion. In some more preferred embodiments, the anti-CTLA4 antibody or fragment thereof, the chemotherapy agent, and / or the optional additional immunomodulatory agent are administered via an intravenous infusion. For example, the anti-CTLA4 antibody or fragment thereof, the chemotherapy agent, and / or the optional additional immunomodulatory agent are preferably administered via an intravenous infusion. For example, the anti-CTLA4 antibody or fragment thereof and the chemotherapy agent are preferably administered via an intravenous infusion.

[0313] In some embodiments, the anti-CTLA4 antibody or fragment thereof, the chemotherapy agent, and / or the optional additional immunomodulatory agent are formulated with one or

[0314] BNT ref [P1819WO01] / / C&F ref. [240673WO] more pharmaceutically acceptable carriers, diluents and / or excipients. In some embodiments, the anti-CTLA4 antibody or fragment thereof, the chemotherapy agent, and the optional additional immunomodulatory agent are formulated with one or more pharmaceutically acceptable carriers, diluents and / or excipients. In some embodiments, the anti-CTLA4 antibody or fragment thereof and the chemotherapy agent are formulated with one or more pharmaceutically acceptable carriers, diluents and / or excipients.

[0315] Suitable carriers include, for example, sterile water, Ringer, Ringer lactate, sterile sodium chloride solution, isotonic saline, polyalkylene glycols, hydrogenated naphthalenes and, in particular, biocompatible lactide polymers, lactide / glycolide copolymers or polyoxyethylene / polyoxy-propylene copolymers. Suitable carriers for parenteral administration are preferably isotonic to the blood of the recipient. Non-limiting examples of suitable diluents include ethanol, glycerol, and water. Examples of excipients, include without limitation carriers, binders, diluents, lubricants, thickeners, surface active agents, preservatives, stabilizers, emulsifiers, buffers, flavoring agents, or colorants.

[0316] The composition and kit of parts

[0317] The disclosure further provides a composition comprising: a) an anti-CTLA4 antibody or fragment thereof; and b) a chemotherapy agent.

[0318] The embodiments disclosed herein for the method of treatment comprising the anti-CTLA4 antibody or fragment thereof in combination with the chemotherapy agent can be used in the composition as disclosed herein. Features described herein in more detail in connection with the “anti-CTLA4 antibody or fragment thereof for use in a method of treating, the chemotherapy agent for use in a method of treating, or the anti-CTLA4 antibody or fragment thereof and the chemotherapy agent for use in a method of treating” embodiments equally apply to the corresponding composition embodiments.

[0319] In some embodiments, the composition comprises an additional immunomodulatory agent.

[0320] BNT ref [P1819WO01] / / C&F ref. [240673WO] In some embodiments, the composition comprises one or more pharmaceutically acceptable carriers, diluents and / or excipients.

[0321] In some embodiments, the composition is a pharmaceutical composition. In some preferred embodiments, the anti-CLTA4 antibody or fragment thereof, the chemotherapy agent, and / or the additional immunomodulatory agent are comprised in individual pharmaceutical compositions. For example, the anti-CLTA4 antibody or fragment thereof and the chemotherapy agent of the present disclosure are comprised in two different pharmaceutical compositions, each individually comprising the anti-CLTA4 antibody or fragment thereof or the chemotherapy agent.

[0322] The invention further provides a kit of parts comprising: a) an anti-CTLA4 antibody or fragment thereof; and b) a chemotherapy agent.

[0323] The embodiments disclosed herein for the method of treatment comprising the anti-CTLA4 antibody or fragment thereof in combination with the chemotherapy agent can be used in the kit of parts as disclosed herein. Features described herein in more detail in connection with the “anti-CTLA4 antibody or fragment thereof for use in a method of treating, the chemotherapy agent for use in a method of treating, or the anti-CTLA4 antibody or fragment thereof and the chemotherapy agent for use in a method of treating” embodiments equally apply to the corresponding kit of parts embodiments.

[0324] In some embodiments, the kit of parts comprises and additional immunomodulatory agent.

[0325] In some embodiments, the kit of parts comprises one or more containers comprising the anti-CTLA4 antibody or fragment thereof, the chemotherapy agent, and / or the additional immunomodulatory agent. In some embodiments, the kit of parts comprises one or more containers comprising the anti-CTLA4 antibody or fragment thereof, and / or the chemotherapy agent.

[0326] BNT ref [P1819WO01] / / C&F ref. [240673WO] In some preferred embodiments, the kit of parts comprises individual containers each individually comprising the anti-CTLA4 antibody or fragment thereof, the chemotherapy agent, and / or the additional immunomodulatory agent. In some preferred embodiments, the kit of parts comprises individual containers each individually comprising the anti-CTLA4 antibody or fragment thereof and / or the chemotherapy agent.

[0327] In some embodiments, the kit of parts comprises individual containers comprising combinations of the anti-CTLA4 antibody or fragment thereof, the chemotherapy agent, and / or the additional immunomodulatory agent. In some embodiments, the kit of parts comprises individual containers comprising combinations of the anti-CTLA4 antibody or fragment thereof and / or the chemotherapy agent.

[0328] In some embodiments, the kit of parts comprises instructions for use.

[0329] The invention further provides a combination of an anti-CTLA4 antibody or fragment thereof and a chemotherapy agent.

[0330] The embodiments disclosed herein for the method of treatment comprising the anti-CTLA4 antibody or fragment thereof in combination with the chemotherapy agent can be used in the combination as disclosed herein. Features described herein in more detail in connection with the “anti-CTLA4 antibody or fragment thereof for use in a method of treating, the chemotherapy agent for use in a method of treating, or the anti-CTLA4 antibody or fragment thereof and the chemotherapy agent for use in a method of treating” embodiments equally apply to the corresponding combination embodiments.

[0331] The method of treatment

[0332] The invention also provides method of treating cancer in a subject in need thereof, wherein the method comprises administering to the subject: a) an anti-CTLA4 antibody or fragment thereof; and b) a chemotherapy agent.

[0333] BNT ref [P1819WO01] / / C&F ref. [240673WO] The embodiments disclosed herein for the method of treatment comprising the anti-CTLA4 antibody or fragment thereof in combination with the chemotherapy agent can be used in the method of treating as disclosed herein. Features described herein in more detail in connection with the “anti-CTLA4 antibody or fragment thereof for use in a method of treating, the chemotherapy agent for use in a method of treating, or the anti-CTLA4 antibody or fragment thereof and the chemotherapy agent for use in a method of treating” embodiments equally apply to the corresponding method of treating embodiments.

[0334] SEQUENCE LISTING

[0335] This application contains a Sequence Listing which has been submitted electronically and is hereby incorporated by reference in its entirety. Said Sequence Listing file is named 240673 WO_Sequence Listing.xml and 36.315 bytes in size.

[0336] SEQ ID NOs: 1-10 are exemplary amino acid sequences of heavy and light chain CDR1-3 sequences of the anti-CTLA4 antibody.

[0337] SEQ ID NOs: 11-16 are exemplary amino acid sequences of heavy and light chain variable domains of the anti CTLA4 antibody.

[0338] SEQ ID NO: 17 is an exemplary amino acid sequence of an unmutated Fc region suitable for the anti-CTLA4 antibody.

[0339] SEQ ID NO: 18 is an exemplary amino acid sequence of a mutated Fc region suitable for the anti-CTLA4 antibody.

[0340] SEQ ID NOs: 19, 21, 23, 25, 27, and 29 are exemplary amino acid sequences of the full- length heavy and light chains of the anti-CTLA4 antibody.

[0341] SEQ ID NOs: 20, 22, 24, 26, 28, and 30 are exemplary polynucleotide sequences of the full-length heavy and light chains of the anti-CTLA4 antibody.

[0342] BNT ref [P1819WO01] / / C&F ref. [240673WO] EXAMPLES

[0343] A clinical study is performed to compare the therapeutic effect of using the chemotherapy agent docetaxel as a monotherapy or in combination with the anti-CTLA-4 antibody in the treatment of NSCLC in patients with locally advanced or metastatic NSCLC that is resistant to cancer immunotherapy treatment with anti-PD-l / anti-PD-Ll antibodies and / or is negative for targetable NSCLC diver mutations (except potential KRAS mutations). The patients are subdivided into the following groups:

[0344] • Group Al (anti-CTLA4 antibody monotherapy): anti-CTLA4 antibody is administered in a Q3 week (21 -day) monotherapy schedule for 17 treatment cycles or up to one year.

[0345] • Group A2 (docetaxel monotherapy): docetaxel is administered in a Q3 week (21-day) monotherapy schedule. During these treatment cycles, a single dose is independently selected from 75 mg / m2, 55 mg / m2, or 37.5 mg / m2.

[0346] • Group B (anti-CTLA4 antibody and docetaxel combination therapy): the anti-CTLA4 antibody and docetaxel are administered concurrently in a Q3 week schedule (21 days) for 4 treatment cycles. During treatment cycles 1 to 4, docetaxel is administered in a single dose independently selected from 75 mg / m2, 55 mg / m2, or 37.5 mg / m2. During treatment cycles 1 and 2, the anti-CTLA4 antibody is administered in a single dose of 10 mg / kg, and during treatment cycles 3 and 4, the anti-CTLA4 antibody is administered in a single dose of 6 mg / kg. Subsequently, i.e., from treatment cycle 5 onwards, the anti-CTLA antibody is administered consecutively in a Q3 week monotherapy schedule for up to 13 cycles or up to one year and in a single dose of 6 mg / kg.

[0347] The results surprisingly show an effective and safe treatment and an unexpected beneficial effect compared to the anti-CTLA antibody monotherapy treatment. This demonstrates that the combination therapy comprising the anti-CTLA4 antibody and docetaxel is particularly suitable for the treatment of patients with locally advanced or metastasized NSCLC that is resistant to immune-checkpoint inhibitor cancer immunotherapy and / or negative for targetable NSCLC driver mutations. In particular, the combination unexpectedly provides

[0348] BNT ref [P1819WO01] / / C&F ref. [240673WO] fast progressing patients a good treatment solution while waiting for the beneficial effect of the anti-CTLA4 antibody treatment.

[0349] BNT ref [P1819WO01] / / C&F ref. [240673WO]

Claims

1. C l a i m s1. An anti-CTLA4 antibody or fragment thereof for use in a method of treating cancer in a subject in need thereof, wherein the method comprises administering to the subject: a) the anti-CTLA4 antibody; and b) a chemotherapy agent.

2. A chemotherapy agent for use in a method of treating cancer in a subject in need thereof, wherein the method comprises administering to the subject: a) the chemotherapy agent; and b) an anti-CTLA4 antibody or fragment thereof.

3. The anti-CTLA4 antibody or fragment thereof for use according to claim 1 or the chemotherapy agent for use according to claim 2, wherein the anti-CTLA4 antibody or fragment thereof and the chemotherapy agent are administered separately.

4. The anti-CTLA4 antibody or fragment thereof for use according to claim 1 or 3 or the chemotherapy agent for use according to claim 2 or 3, wherein the anti-CTLA4 antibody or fragment thereof and the chemotherapy agent are administered concurrently or consecutively.

5. The anti-CTLA4 antibody or fragment thereof for use according to claims 1, 3 or 4 or the chemotherapy agent for use according to claims 2-4, wherein the anti-CTLA4 antibody or fragment thereof and the chemotherapy agent are administered intravenously.1BNT ref [P1819WO01] / / C&F ref. 240673WO6. The anti-CTLA4 antibody or fragment thereof for use according to claims 1 or 3-5 or the chemotherapy agent for use according to claims 2-5, wherein the anti-CTLA4 antibody or fragment thereof and the chemotherapy agent are administered via an intravenous injection or an intravenous infusion, preferably an intravenous infusion.

7. The anti-CTLA4 antibody or fragment thereof for use according to claims 1 or 3-6 or the chemotherapy agent for use according to claims 2-6, wherein the anti-CTLA4 antibody or fragment thereof dosage administered ranges from about 0.1 mg / kg to about 45 mg / kg body weight, preferably from about 1 mg / kg to about 30 mg / kg body weight, more preferably from about 2 mg / kg to about 20 mg / kg body weight.

8. The anti-CTLA4 antibody or fragment thereof for use according to claims 1 or 3-7 or the chemotherapy agent for use according to claims 3-7, wherein the anti-CTLA4 antibody or fragment thereof dosage administered is independently selected from about 0.1 mg / kg body weight, about 0.3 mg / kg body weight, about 1.0 mg / kg body weight, about 3.0 mg / kg body weight, about 6 mg / kg body weight, about 10 mg / kg body weight, about 12 mg / kg body weight, about 15 mg / kg body weight, and about 20 mg / kg body weight.

9. The anti-CTLA4 antibody or fragment thereof for use according to claims 1 or 3-8 or the chemotherapy agent for use according to claims 2-8, wherein the chemotherapy agent dosage administered ranges from about 0.1 to about 300 mg / m2body surface area, preferably from about 1 to about 200 mg / m2body surface area, more preferably from about 10 to about 100 mg / m2body surface area.

10. The anti-CTLA4 antibody or fragment thereof for use according to claims 1 or 3-9 or the chemotherapy agent for use according to claims 2-9, wherein the chemotherapy agent dosage administered is independently selected from about 0.1 mg / m2body surface area, about 0.3 mg / m2body surface area, about 1 mg / m2body surface area,BNT ref [P1819WO01] / / C&F ref. [240673WO]about 10 mg / m2body surface area, about 22.5 mg / m2body surface area, about 25 mg / m2body surface area, about 27.5 mg / m2body surface area, about 30 mg / m2body surface area, about 32.5 mg / m2body surface area, about 35 mg / m2body surface area, about 37.5 mg / m2body surface area, about 40 mg / m2body surface area about 50 mg / m2body surface area, about 55 mg / m2body surface area, about 60 mg / m2body surface area, about 65 mg / m2body surface area, about 70 mg / m2body surface area, about 75 mg / m2body surface area, about 100 mg / m2body surface area, and about 125 mg / m2body surface area.

11. The anti-CTLA4 antibody or fragment thereof for use according to claims 1 or 3-10 or the chemotherapy agent for use according to claims 2-10, wherein a treatment cycle is repeated at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, or 17 times.

12. The anti-CTLA4 antibody or fragment thereof for use according to claims 1 or 3-11 or the chemotherapy agent for use according to claims 2-11, wherein each treatment cycle has up to about 28 days, preferably up to about 21 days, more preferably about 21 days.

13. The anti-CTLA4 antibody or fragment thereof for use according to claims 1 or 3-12 or the chemotherapy agent for use according to claims 2-12, wherein the anti-CTLA4 antibody or fragment thereof is administered about every 6 weeks, preferably about every 4 weeks, more preferably about every 3 weeks.

14. The anti-CTLA4 antibody or fragment thereof for use according to claims 1 or 3-13 or the chemotherapy agent for use according to claims 2-13, wherein the chemotherapy agent is administered about every 6 weeks, preferably about every 4 weeks, more preferably about every 3 weeks.BNT ref [P1819WO01] / / C&F ref. [240673WO]15. The anti-CTLA4 antibody or fragment thereof for use according to claims 1 or 3-14 or the chemotherapy agent for use according to claims 2-14, wherein the chemotherapy agent is administered up to the 8thtreatment cycle, preferably up to the 7thtreatment cycle, more preferably up to the 6thtreatment cycle, even more preferably up to the 5thtreatment cycle, and most preferably up to the 4thtreatment cycle.

16. The anti-CTLA4 antibody or fragment thereof for use according to claims 1 or 3-15 or the chemotherapy agent for use according to claims 2-15, wherein the anti-CTLA4 antibody or fragment thereof and the chemotherapy agent are administered up to the 8thtreatment cycle, preferably up to the 7thtreatment cycle, more preferably up to the 6thtreatment cycle, even more preferably up to the 5thtreatment cycle, and most preferably up to the 4thtreatment cycle, followed by consecutive administration of the anti-CTLA4 antibody or fragment thereof.

17. The anti-CTLA4 antibody or fragment thereof for use according to claims 1 or 3-16 or the chemotherapy agent for use according to claims 2-16, wherein the anti-CTLA4 antibody or fragment thereof is administered for 17 treatment cycles or up to 1 year.

18. The anti-CTLA4 antibody or fragment thereof for use according to claims 1 or 3-17 or the chemotherapy agent for use according to claims 2-17, wherein the anti-CTLA4 antibody or fragment thereof dosage administered at least during the 1stand 2ndtreatment cycle ranges from about 2 mg / kg to about 20 mg / kg body weight, preferably from about 4 mg / kg to about 15 mg / kg body weight, more preferably from about 6 mg / kg to about 12 mg / kg body weight.

19. The anti-CTLA4 antibody or fragment thereof for use according to claims 1 or 3-18 or the chemotherapy agent for use according to claims 2-18, wherein the anti-CTLA4 antibody or fragment thereof dosage administered at least during the 3rdand 4thcycle ranges from about 2 mg / kg to about 20 mg / kg, preferably from about 2 mg / kg to aboutBNT ref [P1819WO01] / / C&F ref. [240673WO]10 mg / kg, more preferably from about 3 mg / kg to about 8 mg / kg, most preferably from about 4 mg / kg to about 7 mg / kg.

20. The anti-CTLA4 antibody or fragment thereof for use according to claims 1 or 3-19 or the chemotherapy agent for use according to claims 2-19, wherein the chemotherapy agent dosage administered during the 1st, the 2nd, the 3rd, and the 4thcycle ranges from about 1 to about 200 mg / m2body surface area, preferably from about 10 to about 150 mg / m2body surface area, more preferably from about 25 to about 100 mg / m2body surface area, most preferably from about 30 to about 80 mg / m2body surface area.

21. The anti-CTLA4 antibody or fragment thereof for use according to claims 1 or 3-20 or the chemotherapy agent for use according to claims 2-20, wherein the anti-CTLA4 antibody or fragment thereof and / or the chemotherapy agent are formulated with one or more pharmaceutically acceptable carriers, diluents and / or excipients.

22. The anti-CTLA4 antibody or fragment thereof for use according to claims 1 or 3-21 or the chemotherapy agent for use according to claims 2-21, wherein the subject has not previously been treated for cancer.

23. The anti-CTLA4 antibody or fragment thereof for use according to claims 1 or 3-21 or the chemotherapy agent for use according to claims 2-21, wherein the subject has been previously treated for cancer, preferably wherein the subject had at least one previous chemotherapy treatment.

24. The anti-CTLA4 antibody or fragment thereof for use according to claim 23 or the chemotherapy agent for use according to claim 23, wherein the at least one previous chemotherapy treatment comprises or is a platinum-containing chemotherapy.BNT ref [P1819WO01] / / C&F ref. [240673WO]25. The anti-CTLA4 antibody or fragment thereof for use according to claims 1 or 3-24 or the chemotherapy agent for use according to claims 2-24, wherein overall survival is increased in said subject compared to the chemotherapy agent or the anti-CTLA4 antibody or fragment thereof treatment alone.

26. The anti-CTLA4 antibody or fragment thereof for use according to claims 1 or 3-25 or the chemotherapy agent for use according to claims 2-25, wherein median progression-free survival is increased in said subject compared to the chemotherapy agent or the anti-CTLA4 antibody or fragment thereof treatment alone.

27. The anti-CTLA4 antibody or fragment thereof for use according to claims 1 or 3-26 or the chemotherapy agent for use according to claims 2-26, wherein the cancer comprises one or more solid tumors.

28. The anti-CTLA4 antibody or fragment thereof for use according to claims 1 or 3-27 or the chemotherapy agent for use according to claims 2-27, wherein the cancer is selected from melanoma, lung cancer, human papillomavirus (HPV)-induced cancer, breast cancer, hepatocellular carcinoma, ovarian cancer such as ovarian carcinoma, prostate cancer such as prostate carcinoma, Hodgkin's or non-Hodgkin's lymphoma, acute myelogenic Leukemia, chronic myelogenic Leukemia, acute lymphocytic Leukemia, chronic lymphocytic Leukemia, or renal cell carcinoma.

29. The anti-CTLA4 antibody or fragment thereof for use according to claims 1 or 3-28 or the chemotherapy agent for use according to claims 2-28, wherein the cancer is nonsmall cell lung cancer (NSCLC).

30. The anti-CTLA4 antibody or fragment thereof for use according to claim 29 or the chemotherapy agent for use according to claim 29, wherein the NSCLC has a squamous histology.BNT ref [P1819WO01] / / C&F ref. [240673WO]31. The anti-CTLA4 antibody or fragment thereof for use according to claim 29 or the chemotherapy agent for use according to claim 29, wherein the NSCLC has a non- squamous histology.

32. The anti-CTLA4 antibody or fragment thereof for use according to claims 29-31 or the chemotherapy agent for use according to claims 29-31, wherein the NSCLC has locally advanced or metastasized.

33. The anti-CTLA4 antibody or fragment thereof for use according to claims 29-32 or the chemotherapy agent for use according to claims 29-32, wherein the NSCLC is negative for targetable NSCLC driver mutations.

34. The anti-CTLA4 antibody or fragment thereof for use according to claim 33 or the chemotherapy agent for use according to claim 33, wherein the NSCLC is negative for EGFR and / or ALK mutations.

35. The anti-CTLA4 antibody or fragment thereof for use according to claims 29-32 or the chemotherapy agent for use according to claims 29-32, wherein the NSCLC is positive for KRAS mutations.

36. The anti-CTLA4 antibody or fragment thereof for use according to claims 1 or 3-35 or the chemotherapy agent for use according to claims 2-35, wherein the chemotherapy agent comprises or is selected from cisplatin, oxaliplatin, carboplatin, lurbinectedin, topotecan, paclitaxel, nanoparticle albumin-bound paclitaxel (nab -paclitaxel), pemetrexed, 5-fluoruracil, irinotecan, etoposide, gemcitabine, docetaxel or combinations thereof.BNT ref [P1819WO01] / / C&F ref. [240673WO]37. The anti-CTLA4 antibody or fragment thereof for use according to claim 36 or the chemotherapy agent for use according to claim 36, wherein the chemotherapy agent comprises or is docetaxel.

38. The anti-CTLA4 antibody or fragment thereof for use according to claims 1 or 3-37 or the chemotherapy agent for use according to claims 2-37, wherein the subject has previously not been treated with docetaxel.

39. The anti-CTLA4 antibody or fragment thereof for use according to claims 1 or 3-38 or the chemotherapy agent for use according to claims 2-38, wherein the anti-CTLA4 antibody or fragment thereof comprises: a) a heavy chain variable region comprising (1) a complementarity-determining region 1 (HCDR1) comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 1, (2) a complementarity-determining region 2 (HCDR2) comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 3, 4 or 5, and (3) a complementarity-determining region 3 (HCDR3) comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 9; and b) a light chain variable region comprising: (1) a complementarity-determining region 1 (LCDR1) comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 2, (2) a complementarity-determining region 2 (LCDR2) comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 6, 7, or 8, and (3) a complementarity-determining region 3 (LCDR3) comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 10.

40. The anti-CTLA4 antibody or fragment thereof for use according to claim 39 or the chemotherapy agent for use according to claim 39, wherein the anti-CTLA4 antibody or fragment thereof comprises:BNT ref [P1819WO01] / / C&F ref. [240673WO]a) a heavy chain variable region comprising (1) a HCDR1 comprising or consisting of an amino acid sequence is set forth in SEQ ID NO: 1, (2) a HCDR2 comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 3, and (3) a HCDR3 comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 9; and b) a light chain variable region comprising: (1) a LCDR1 comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 2, (2) a LCDR2 comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 7, and (3) a LCDR3 comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 10.

41. The anti-CTLA4 antibody or fragment thereof for use according to claim 39 or the chemotherapy agent for use according to claim 39, wherein the anti-CTLA4 antibody or fragment thereof comprises: a) a heavy chain variable region comprising (1) a HCDR1 comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 1, (2) a HCDR2 comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 5, and (3) a HCDR3 comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 9; and b) a light chain variable region comprising: (1) a LCDR1 comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 2, (2) a LCDR2 comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 7, and (3) a LCDR3 comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 10.BNT ref [P1819WO01] / / C&F ref. [240673WO]42. The anti-CTLA4 antibody or fragment thereof for use according to claim 39 or the chemotherapy agent for use according to claim 39, wherein the anti-CTLA4 antibody or fragment thereof comprises: a) a heavy chain variable region comprising (1) a HCDR1 comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 1, (2) a HCDR2 comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 5, and (3) a HCDR3 comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 9; and b) a light chain variable region comprising: (1) a LCDR1 comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 2, (2) a LCDR2 comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 8, and (3) a LCDR3 comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 10.

43. The anti-CTLA4 antibody or fragment thereof for use according to claims 1 or 2-42 or the chemotherapy agent for use according to claims 2-42, wherein the anti-CTLA4 antibody or fragment thereof comprises: a) a heavy chain variable domain comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 11, 12, or 13, or an amino acid sequence comprising at least 80%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 11, 12, or 13; and b) a light chain variable domain comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 14, 15, or 16, or an amino acid sequence comprising at least 80%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 14, 15, or 16.BNT ref [P1819WO01] / / C&F ref. [240673WO]44. The anti-CTLA4 antibody or fragment thereof for use according to claim 43 or the chemotherapy agent for use according to claim 43, wherein the anti-CTLA4 antibody or fragment thereof comprises: a) a heavy chain variable domain comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 11 or an amino acid sequence comprising at least 80%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 11; and b) a light chain variable domain comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 15 or an amino acid sequence comprising at least 80%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 15.

45. The anti-CTLA4 antibody or fragment thereof for use according to claim 43 or the chemotherapy agent for use according to claim 43, wherein the anti-CTLA4 antibody or fragment thereof comprises: a) a heavy chain variable domain comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 13 or an amino acid sequence comprising at least 80%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 13; and b) a light chain variable domain comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 15 or an amino acid sequence comprising at least 80%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 15.

46. The anti-CTLA4 antibody or fragment thereof for use according to claim 43 or the chemotherapy agent for use according to claim 43, wherein the anti-CTLA4 antibody or fragment thereof comprises:BNT ref [P1819WO01] / / C&F ref. [240673WO]a) a heavy chain variable domain comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 13 or an amino acid sequence comprising at least 80%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 13; and b) a light chain variable domain comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 16 or an amino acid sequence comprising at least 80%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 16.

47. The anti-CTLA4 antibody or fragment thereof for use according to claims 1, 3-46 or the chemotherapy agent for use according to claims 2-46, wherein the anti-CTLA4 antibody comprises a constant region preferably derived from a human antibody, preferably the constant region is selected from the constant region of human IgGl, IgG2, IgG3 or IgG4.

48. The anti-CTLA4 antibody or fragment thereof for use according to claim 47 or the chemotherapy agent for use according to claim 47, wherein the anti-CTLA4 antibody comprises an IgGl Fc, preferably comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 17 or 18 or an amino acid sequence comprising at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 17 or 18.

49. The anti-CTLA4 antibody or fragment thereof for use according to claims 1, 3-48 or the chemotherapy agent for use according to claims 2-48, wherein the anti-CTLA4 antibody comprises: a) a heavy chain comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 19, 21, or 23 or an amino acid sequence comprising at least 80%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 19, 21, or 23; andBNT ref [P1819WO01] / / C&F ref. [240673WO]b) a light chain comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 25, 27, or 29 or an amino acid sequence comprising at least 80%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 25, 27, or 29.

50. The anti-CTLA4 antibody or fragment thereof for use according to claim 49 or the chemotherapy agent for use according to claim 49, wherein the anti-CTLA4 antibody comprises: a) a heavy chain comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 19 or an amino acid sequence comprising at least 80%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 19; and b) a light chain comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 27 or an amino acid sequence comprising at least 80%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 27.

51. The anti-CTLA4 antibody or fragment thereof for use according to claim 49 or the chemotherapy agent for use according to claim 49, wherein the anti-CTLA4 antibody comprises: a) a heavy chain comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 23 or an amino acid sequence comprising at least 80%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 23; and b) a light chain comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 27 or an amino acid sequence comprising at least 80%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 27.BNT ref [P1819WO01] / / C&F ref. [240673WO]52. The anti-CTLA4 antibody or fragment thereof for use according to claim 49 or the chemotherapy agent for use according to claim 49, wherein the anti-CTLA4 antibody comprises: a) a heavy chain comprising or consisting of an amino acid sequence set forth in SEQ ID NO: 23 or an amino acid sequence comprising at least 80%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 23; and b) a light chain comprising or consisting of amino acid sequence set forth in SEQ ID NO: 29 or an amino acid sequence comprising at least 80%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 29.

53. The anti-CTLA4 antibody or fragment thereof for use according to claims 1, 3-52 or the chemotherapy agent for use according to claims 2-52, wherein the anti-CTLA4 antibody or fragment thereof is capable of binding to human CTLA4.

54. The anti-CTLA4 antibody or fragment thereof for use according to claims 1, 3-53 or the chemotherapy agent for use according to claims 2-53, wherein the anti CTLA4 antibody or fragment thereof is a humanized anti-CTLA4 antibody or fragment thereof.

55. The anti-CTLA4 antibody or fragment thereof for use according to claims 1, 3-46, 53, 54, or the chemotherapy agent for use according to claims 2-46, 53, 54, wherein the anti-CTLA4 antibody fragment is an antigen-binding fragment or a variant thereof.

56. The anti-CTLA4 antibody or fragment thereof for use according to claim 55 or the chemotherapy agent for use according to claim 55, wherein the antigen binding fragment or variant thereof is selected from the group consisting of a Fab, a Fab’, aBNT ref [P1819WO01] / / C&F ref. [240673WO]F(ab’)2, a scFV, a diabody, a triabody, a minibody, and a single-domain antibody (sdAB), and variants thereof.

57. An anti-CTLA4 antibody or fragment thereof and a chemotherapy agent for use in a method of treating cancer in a subject in need thereof, wherein the method comprises administering to the subject: a) the anti-CTLA4 antibody or fragment thereof; and b) the chemotherapy agent.

58. A composition comprising: a) an anti-CTLA4 antibody or fragment thereof; and b) a chemotherapy agent.

59. The composition according to claim 58, comprising the anti-CTLA4 antibody or fragment thereof in a range from about 0.1 mg / kg to about 45 mg / kg body weight, preferably from about 1 mg / kg to about 30 mg / kg body weight, more preferably from about 2 mg / kg to about 20 mg / kg body weight, even more preferably from about 4 mg / kg to about 15 mg / kg body weight, and most preferably from about 5 mg / kg to about 12 mg / kg body weight.

60. The composition according to claim 58 or 59, comprising the chemotherapy agent in a range from about 0.1 to about 300 mg / m2body surface area, preferably from about 1 to about 200 mg / m2body surface area, more preferably from about 10 to about 150 mg / m2body surface area, even more preferably from about 25 to about 100 mg / m2body surface area, and most preferably from about 30 to about 80 mg / m2body surface area.BNT ref [P1819WO01] / / C&F ref. [240673WO]61. The composition according to claim 60, comprising about 75 mg / m2body surface area, 55 mg / m2body surface area, or 37.5 mg / m2body surface area of the chemotherapy agent.

62. The composition according to claims 58-61, comprising one or more pharmaceutically acceptable carriers, diluents and / or excipients.

63. The composition according to claims 58-62, wherein the composition is a pharmaceutical composition.

64. A method of treating cancer in a subject in need thereof, wherein the method comprises administering to the subject: a) an anti-CTLA4 antibody or fragment thereof; and b) a chemotherapy agent.

66. A combination of an anti-CTLA4 antibody or fragment thereof and a chemotherapy agent.BNT ref [P1819WO01] / / C&F ref. [240673WO]