Pharmaceutical compositions comprising mixed antibodies for Anti-CTLA4 and Anti-PD1

Abstract

The invention discloses a pharmaceutical composition containing an anti-CTLA4 and anti-PD1 mixed antibody, a pharmaceutical composition containing the mixed antibody and a chemotherapeutic drug, and an application of the pharmaceutical composition in treatment of non-small cell lung cancer.

Description

Pharmaceutical composition comprising mixed antibodies of anti-CTLA4 and anti-PD1 and therapeutic use thereof Technical Field

[0001] The present disclosure relates to the treatment of cancer, particularly non-small cell lung cancer, including immunotherapy and combination therapy. More specifically, the present disclosure relates to the use of a pharmaceutical composition comprising a mixed antibody against CTLA4 and anti-PD1 for the treatment of stage II-IIIB non-small cell lung cancer after complete surgical resection, or locally advanced or metastatic non-small cell lung cancer with negative PD-L1 expression (PDL1 TPS <1%), no epidermal growth factor receptor (EGFR) sensitive mutation or anaplastic lymphoma kinase (ALK) gene translocation change, and no systemic anti-tumor treatment. Background Art

[0002] According to GLOBOCAN 2020 data, lung cancer ranks second in incidence among cancers globally, with 2.2 million cases, and ranks first in mortality, with 1.8 million deaths worldwide. In China, both incidence and mortality rates rank first, with 816,000 new cases and 715,000 deaths, respectively.

[0003] Non-small cell lung cancer (NSCLC) accounts for approximately 85% of all lung cancers and includes two major pathological types: squamous cell carcinoma and non-squamous cell carcinoma. Non-squamous NSCLC accounts for approximately 60% of NSCLC, with lung adenocarcinoma being the most common type, accounting for approximately 40% of NSCLC. Among lung adenocarcinomas, approximately 10% of Caucasian patients and 40-60% of Asian patients carry EGFR-sensitive mutations, primarily including deletions in EGFR exon 19 and L858R mutations in exon 21. Mutations in other genes, such as ALK and ROS1, are also common.

[0004] Approximately 30% of all NSCLC patients are initially diagnosed with resectable early-stage NSCLC. For patients with early-stage NSCLC, surgery is the primary treatment; for patients with locally advanced NSCLC that is resectable, adjuvant chemotherapy is a common treatment. Among those who receive postoperative adjuvant therapy, 20%-30% of patients with stage I, 50% with stage II, and 60% with stage IIIA die within 5 years. However, the 5-year survival rate drops from 90% for resected stage IA1 to 41% for stage IIIA, indicating that some patients develop micrometastases during surgical resection, leading to treatment failure. Currently, only atezolizumab is approved in China for immunotherapy for this population, resulting in low patient accessibility and an urgent need to develop more effective drugs.

[0005] For patients whose tumors lack any genetic alterations (including EGFR mutations or ALK gene rearrangements, also known as ALK gene translocations), immune checkpoint inhibitors (ICIs) combined with platinum-based chemotherapy have become the standard first-line treatment. Data from studies such as Keynote-042 show that the PD-L1 expression rate is approximately 50-60%, with only one-third showing high expression. Patients with negative PD-L1 expression (defined as a tumor cell positive fraction [TPS] of less than 1%) account for approximately 40-50% of NSCLC patients in China, making them a relatively widely treated group.

[0006] Although immune checkpoint inhibitors combined with platinum-based chemotherapy have become the first-line standard of care for advanced, driver gene-negative NSCLC, there are significant differences in efficacy among patients with different PD-L1 expression patterns, with particularly low efficacy in patients with negative PD-L1 expression. Compared with patients with high PD-L1 expression (TPS ≥ 50%), the median PFS (mPFS) for the same treatment regimen is over 3 months lower, and the median OS (mOS) is over 6 months lower. For example, in the Keynote-189 study, the mPFS for patients with negative and high PD-L1 expression was 6.1 months and 9.4 months, respectively, and the mOS was 17.2 months and 27.7 months, respectively. Patients with both negative PD-L1 expression and negative driver gene NSCLC have a significant unmet treatment need.

[0007] Existing research results suggest that dual-drug therapy (combining two immune checkpoint inhibitors) may offer survival benefits for cancer patients. However, dual-drug therapy combined with chemotherapy is not currently approved in China. Approved immunotherapy drugs in China are intended for use in the general population or for patients with locally advanced or metastatic NSCLC who are PD-L1 positive. Therefore, a large, randomized, controlled study is urgently needed to validate the efficacy of dual-drug therapy combined with chemotherapy in patients with PDL1-negative, driver gene-negative, locally advanced or advanced metastatic NSCLC.

[0008] ZPML265 is a hybrid antibody drug formulation consisting of a recombinant humanized IgG1 monoclonal antibody targeting human CTLA4 and a recombinant humanized IgG4 monoclonal antibody targeting human PD1. These two different antibodies are produced by a single host cell. This hybrid antibody can simultaneously and specifically bind to CTLA4 and PD1, thereby blocking the immune checkpoint signaling pathways between CTLA4 and B7-1 / B7-2, and between PD-1 and PD-L1. This relieves the inhibitory effects of these two pathways on T lymphocytes, restoring their functional activity and anti-tumor immune responses, thereby enabling the body to fight and kill tumors.

[0009] Therefore, there is an unmet clinical need for patients with stage II-IIIB non-small cell lung cancer after complete surgical resection and locally advanced or metastatic non-small cell lung cancer with negative PDL1 expression and negative driver genes, and the development of more effective drugs is urgently needed.

[0010] SUMMARY OF THE INVENTION

[0011] The technical problem to be solved by the present disclosure is to provide an immunotherapy to fill the clinical gap in China where there are no dual-target immunotherapy drugs for non-small cell lung cancer, thereby solving such unmet clinical needs.

[0012] The present disclosure provides a pharmaceutical composition for treating non-small cell lung cancer, comprising an effective amount of a mixed antibody of anti-CTLA4 and anti-PD1.

[0013] At the same time, the present disclosure also provides a method for treating non-small cell lung cancer using a pharmaceutical composition comprising an effective amount of mixed antibodies of anti-CTLA4 and anti-PD1.

[0014] In some embodiments, the pharmaceutical composition further comprises a pharmaceutically acceptable carrier.

[0015] In some embodiments, the pharmaceutical composition further comprises an additional therapeutic agent, including but not limited to chemotherapeutic agents (chemotherapeutic drugs), cytotoxic agents, radiotherapeutic agents, cancer vaccines, tumor reducing agents, targeted anti-cancer agents, anti-angiogenic agents, biological response modifiers, cytokines, hormones, anti-metastatic agents, and immunotherapeutic agents.

[0016] In some embodiments, the additional therapeutic agent is a chemotherapy drug, preferably a chemotherapy drug is carboplatin / cisplatin, paclitaxel / vinorelbine, and / or pemetrexed / vinorelbine, wherein carboplatin and cisplatin can be used interchangeably, paclitaxel and vinorelbine can be used interchangeably, and pemetrexed and vinorelbine can be used interchangeably.

[0017] In some embodiments, the TNM stage of the non-small cell lung cancer described in the present disclosure is stage III or stage IV.

[0018] In other embodiments, the non-small cell lung cancer is stage II-IIIB non-small cell lung cancer after complete surgical resection.

[0019] In some other embodiments, the non-small cell lung cancer is locally advanced or metastatic non-small cell lung cancer that is negative for PD-L1 expression, has no epidermal growth factor receptor (EGFR) sensitive mutation or anaplastic lymphoma kinase (ALK) gene translocation change, and has not received systemic anti-tumor treatment.

[0020] In the present disclosure, non-small cell lung cancer includes squamous non-small cell lung cancer and non-squamous non-small cell lung cancer.

[0021] Preferably, when the non-small cell lung cancer is squamous non-small cell lung cancer, the dosage of the mixed antibody is 5 mg / kg, administered by intravenous infusion on the first day of each cycle, and a treatment cycle of 21 days. In addition, chemotherapy drugs can be additionally administered, and the chemotherapy drugs are carboplatin / cisplatin and paclitaxel / vinorelbine. Among them, the carboplatin dose is the target area under the curve (AUC) 5 mg / mL / min, or the cisplatin dose is 75 mg / m 2 and paclitaxel at a dose of 175 mg / m 2 , or vinorelbine at a dose of 25 mg / m 2 The drug is administered by intravenous infusion on the first day of each cycle, and a treatment cycle lasts for 21 days. If vinorelbine is selected, vinorelbine 25 mg / m2 is administered again on the eighth day of the treatment cycle. 2 After 2-4 cycles of combined antibody and chemotherapy, the combined antibody is maintained at a dose of 5 mg / kg, administered once every three weeks by intravenous infusion.

[0022] Preferably, when the non-small cell lung cancer is non-squamous non-small cell lung cancer, the dosage of the mixed antibody is 5 mg / kg, administered by intravenous infusion on the first day of each cycle, and 21 days is a treatment cycle. In addition, chemotherapy drugs can be additionally administered, and the chemotherapy drugs are carboplatin / cisplatin and pemetrexed / vinorelbine. Among them, the carboplatin dose is the target area under the curve (AUC) 5 mg / mL / min, or the cisplatin dose is 75 mg / m 2 ; and, pemetrexed at a dose of 500 mg / m 2 , or vinorelbine at a dose of 25 mg / m 2 The drug is administered by intravenous infusion on the first day of each cycle, and a treatment cycle lasts for 21 days. If vinorelbine is selected, vinorelbine 25 mg / m2 is administered again on the eighth day of the treatment cycle. 2 After 2-4 cycles of combined antibody and chemotherapy, the combined antibody is maintained at a dose of 5 mg / kg, administered once every three weeks by intravenous infusion.

[0023] In some embodiments, the hybrid antibody is produced by a single host cell that contains nucleic acids encoding two different antibodies, anti-CTLA4 and anti-PD1, wherein the sequences of the anti-CTLA4 antibody heavy chain HCDR1, HCDR2 and HCDR3 are shown in SEQ ID NOs: 1, 2, and 3, respectively, the sequences of the anti-CTLA4 antibody light chain LCDR1, LCDR2 and LCDR3 are shown in SEQ ID NOs: 4, 5, and 6, respectively, the sequences of the anti-PD1 antibody heavy chain HCDR1, HCDR2 and HCDR3 are shown in SEQ ID NOs: 9, 10, and 11, respectively, and the sequences of the anti-PD1 antibody light chain LCDR1, LCDR2 and LCDR3 are shown in SEQ ID NOs: 12, 13, and 14, respectively.

[0024] In some embodiments, the heavy chain variable region sequence of the anti-CTLA4 antibody is shown in SEQ ID NO: 7, the light chain variable region sequence of the anti-CTLA4 antibody is shown in SEQ ID NO: 8, the heavy chain variable region sequence of the anti-PD1 antibody is shown in SEQ ID NO: 15, and the light chain variable region sequence of the anti-PD1 antibody is shown in SEQ ID NO: 16.

[0025] In some embodiments, the heavy chain sequence of the anti-CTLA4 antibody is shown in SEQ ID NO: 17, the light chain sequence of the anti-CTLA4 antibody is shown in SEQ ID NO: 18, the heavy chain sequence of the anti-PD1 antibody is shown in SEQ ID NO: 19, and the light chain sequence of the anti-PD1 antibody is shown in SEQ ID NO: 20.

[0026] At the same time, the present disclosure also provides the use of a mixed antibody comprising an effective amount of anti-CTLA4 and anti-PD1 and a chemotherapy drug in the preparation of a pharmaceutical composition for treating stage II-IIIB non-small cell lung cancer after complete surgical resection.

[0027] Furthermore, the present disclosure provides the use of a mixed antibody comprising an effective amount of anti-CTLA4 and anti-PD1 and a chemotherapy drug in the preparation of a pharmaceutical composition for treating locally advanced or metastatic non-small cell lung cancer that is negative for PD-L1 expression, has no epidermal growth factor receptor (EGFR) sensitive mutation or anaplastic lymphoma kinase (ALK) gene translocation change, and has not received systemic anti-tumor treatment.

[0028] The results of a phase I clinical study of the hybrid antibody disclosed herein as a single drug showed that the enrolled patients with non-small cell lung cancer had a definite efficacy and good safety. At the same time, the clinical study of the hybrid antibody disclosed herein combined with chemotherapy in non-small cell lung cancer has also preliminarily shown a synergistic anti-tumor effect and demonstrated a relatively good clinical efficacy. Therefore, the hybrid antibody monotherapy or the combination of hybrid antibody + carboplatin / cisplatin + paclitaxel / vinorelbine or hybrid antibody + carboplatin / cisplatin + pemetrexed / vinorelbine provided by the present disclosure can be used for the medical treatment of stage III or IV non-small cell lung cancer, with expected controllable safety and good patient compliance, and can largely fill the gap in China where there are no dual-target immunotherapy drugs approved for non-small cell lung cancer, thereby solving the unmet urgent clinical needs. DETAILED DESCRIPTION

[0029] the term

[0030] All publications, patents, and patent applications mentioned in this specification are herein incorporated by reference to the same extent as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated by reference.

[0031] Before describing the present disclosure in detail below, it should be understood that the present disclosure is not limited to the specific methodologies, protocols and reagents described herein, as these may vary. It should also be understood that the terms used herein are only for the purpose of describing specific embodiments and are not intended to limit the scope of the present disclosure. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those of ordinary skill in the art to which the present disclosure belongs.

[0032] Certain embodiments disclosed herein include numerical ranges, and certain aspects of the present disclosure may be described in terms of ranges. Unless otherwise stated, it should be understood that numerical ranges or descriptions in terms of ranges are intended for simplicity and convenience and should not be considered as strict limitations on the scope of the present disclosure. Therefore, descriptions in terms of ranges should be considered to specifically disclose all possible subranges and all possible specific numerical points within the range, as these subranges and numerical points have been clearly stated herein. Regardless of the width of the numerical value, the above principles apply equally. When describing in terms of ranges, the range includes the endpoints of the range.

[0033] When referring to a measurable value such as an amount, a temporal duration, etc., the term "about" is meant to include variations of ±20%, or in some cases ±10%, or in some cases ±5%, or in some cases ±1%, or in some cases ±0.1% of the specified value.

[0034] As used herein, the three-letter and one-letter codes for amino acids are as described in J. Biol. Chem, 243, p3558 (1968).

[0035] The term "antibody" as used herein typically refers to a Y-shaped tetrameric protein comprising two heavy (H) polypeptide chains (HC) and two light (L) polypeptide chains (LC) held together by covalent disulfide bonds and non-covalent interactions. Natural IgG antibodies have this structure. Each light chain comprises a light chain variable domain (VL) and a light chain constant domain (CL). Each heavy chain comprises a heavy chain variable domain (VH) and a heavy chain constant domain (CH) (or heavy chain constant region (CH)).

[0036] Five major classes of antibodies are known in the art: IgA, IgD, IgE, IgG, and IgM. The corresponding heavy chain constant domains are called α, δ, ε, γ, and μ, respectively. IgG and IgA can be further divided into different subclasses, for example, IgG can be divided into IgG1, IgG2, IgG3, and IgG4, and IgA can be divided into IgA1 and IgA2. The light chains of antibodies from any vertebrate species can be assigned to one of two clearly distinct types, called κ and λ, based on the amino acid sequence of their constant domains.

[0037] The term "variable region" or "variable domain" shows significant changes in the amino acid composition from one antibody to another, and is primarily responsible for antigen recognition and binding. The variable region of each light chain / heavy chain pair forms an antibody binding site, so that a complete IgG antibody has two binding sites (i.e., it is bivalent). The variable region of the heavy chain (VH) and the variable region of the light chain (VL) each include three regions with extreme variability, referred to as hypervariable regions (HVRs), or more generally, referred to as complementary determining regions (CDRs). VH and VL each have four skeleton regions FR (or framework regions), represented by FR1, FR2, FR3, and FR4, respectively. Therefore, CDR and FR sequences typically appear in the following sequence of a heavy chain variable domain (VH) (or light chain variable domain (VL)): FR1-HCDR1 (LCDR1)-FR2-HCDR2 (LCDR2)-FR3-HCDR3 (LCDR3)-FR4.

[0038] The types of "antibodies" in a broad sense may include polyclonal antibodies, monoclonal antibodies, chimeric antibodies, humanized antibodies and primatized antibodies, CDR-grafted antibodies, human antibodies (including recombinantly produced human antibodies), recombinantly produced antibodies, intracellular antibodies, multispecific antibodies, bispecific antibodies, monovalent antibodies, multivalent antibodies, anti-idiotypic antibodies, synthetic antibodies (including muteins and variants thereof), etc.

[0039] The term "monoclonal antibody" (or "mAb") refers to a substantially homogeneous antibody produced by a single cell clone that is directed against a specific antigenic epitope. Monoclonal antibodies can be prepared using a variety of techniques known in the art, including hybridoma technology, recombinant technology, phage display technology, transgenic animals, synthetic technology, or a combination of these techniques.

[0040] It should be noted that the division of the CDRs and FRs in the antibody variable regions of the present disclosure is determined according to the Kabat definition. Other naming and numbering systems, such as Chothia, IMGT, or AHo, are also known to those skilled in the art. Therefore, humanized antibodies containing one or more CDRs derived from any naming system based on the antibody sequences of the present disclosure are clearly within the scope of the present disclosure.

[0041] The term "antigen" refers to a substance that is recognized and specifically bound by an antibody or antibody binding fragment. In a broad sense, an antigen can include any immunogenic fragment or determinant of a selected target, including a single epitope, multiple epitopes, a single domain, multiple domains, or a complete extracellular domain (ECD) or protein.

[0042] The term "epitope" refers to a site on an antigen to which an immunoglobulin or antibody specifically binds. An epitope can be formed by adjacent amino acids or non-adjacent amino acids juxtaposed by tertiary folding of a protein.

[0043] The terms "polypeptide," "peptide," and "protein" are used interchangeably herein to refer to polymers of amino acids of any length. The polymer may be linear, cyclic, or branched, it may contain modified amino acids, particularly conservatively modified amino acids, and it may be interrupted by non-amino acids. The term also includes amino acid polymers that have been modified, for example, by glycosylation, lipidation, acetylation, phosphorylation, methylation, and the like.

[0044] As used herein, the term "hybrid antibody" refers to an antibody containing a limited number of primary antibody species, optionally no more than two, three, four, five, six, seven, eight, nine, or ten, produced from host cells (optionally cells from a single host cell line) that have been transfected with DNA encoding at least two different antibodies (optionally full-length primate IgG antibodies) with different binding specificities. In some embodiments, DNA encoding at least two different heavy chains (HCs) and at least two different light chains (LCs) can be introduced into the same host cell. For example, a host cell can be transfected with DNA encoding at least two but no more than four different antibodies with different binding specificities. In some embodiments, the sequences of all transfected DNA encoding HCs and LCs can be mutated to alter the amino acid sequence of the antibody, thereby disfavoring non-homologous HC / LC pairing and strongly favoring homologous HC / LC pairing. In the case of introducing two different HCs into the host cell, one or both of the two different HCs can be optionally altered to disfavor heterodimer formation. In some embodiments, only one heavy chain is altered to prevent heterodimer formation. In some embodiments, when DNA encoding only two different antibodies is introduced into a host cell, only one of the antibodies encoded by the DNA comprises one or more partner-directed changes that favor cognate HC / LC pairing, while the other antibody does not comprise such changes. In some embodiments, the host cell produces only two major antibody species, wherein each HC predominantly pairs with its cognate LC, and most antibodies are tetramers containing two heavy chains having the same amino acid sequence and two light chains having the same amino acid sequence (see PCT / US2017 / 030676).

[0045] The term "pharmaceutical composition" refers to a formulation or combination of formulations containing one, two, or more active ingredients, which allows the active ingredients contained therein to exist in a biologically effective form and does not contain additional ingredients that are unacceptably toxic to the subject to which the formulation is administered. When a "pharmaceutical composition" is present as a combination of separate formulations containing two or more different active ingredients, they can be administered simultaneously, sequentially, separately, or at intervals, with the goal of exerting the biological activities of the multiple active ingredients for combined treatment of a disease.

[0046] The term "pharmaceutical carrier" or "pharmaceutically acceptable carrier" refers to a diluent, adjuvant (eg, Freund's adjuvant (complete and incomplete)), excipient, or vehicle with which the therapeutic agent is administered.

[0047] The term "effective amount" refers to a dose of a pharmaceutical formulation comprising an active ingredient of the present disclosure that, after administration to a patient in a single or multiple doses, produces the desired effect in the treated patient. The effective amount can be readily determined by the attending physician, who is skilled in the art, by considering a variety of factors such as: ethnic differences; weight, age, and health status; the specific disease involved; the severity of the disease; the response of the individual patient; the specific antibody administered; the mode of administration; the bioavailability characteristics of the administered formulation; the selected dosing regimen; and the use of any concomitant therapy.

[0048] The terms "host cell," "host cell line," and "host cell culture" are used interchangeably and refer to cells into which exogenous nucleic acid has been introduced, including the progeny of such cells. Host cells include "transformants" and "transformed cells," which include the primary transformed cell and progeny derived therefrom, without regard to the number of passages. Progeny may not be completely identical in nucleic acid content to the parent cell and may contain mutations. Mutant progeny that have the same function or biological activity as screened or selected for in the originally transformed cell are included herein.

[0049] The term "transfection" as used herein refers to the introduction of exogenous nucleic acid into eukaryotic cells. Transfection can be achieved by various means known in the art, including calcium phosphate-DNA coprecipitation, DEAE-dextran-mediated transfection, polybrene-mediated transfection, electroporation, microinjection, liposome fusion, lipofection, protoplast fusion, retroviral infection, and biolistics.

[0050] The term "isolated polynucleotide" or "isolated nucleic acid" refers to a nucleic acid molecule, DNA or RNA, that has been removed from its natural environment. For example, for the purposes of this disclosure, a recombinant polynucleotide encoding a polypeptide contained in a vector is considered isolated. Other examples of isolated polynucleotides include recombinant polynucleotides maintained in heterologous host cells or purified (partially or substantially) polynucleotides in solution. Isolated polynucleotides include polynucleotide molecules contained in cells that normally contain the polynucleotide molecule, but the polynucleotide molecule is present extrachromosomally or at a chromosomal location that is different from its natural chromosomal location. Isolated RNA molecules include in vivo or in vitro RNA transcripts of the present disclosure, as well as positive and negative strand forms and double-stranded forms. Isolated polynucleotides or nucleic acids of the present disclosure also include such molecules produced synthetically. In addition, the polynucleotide or nucleic acid may or may not include regulatory elements, such as promoters, ribosome binding sites, or transcription terminators.

[0051] The terms "nucleic acid molecule encoding," "coding DNA sequence," and "coding DNA" refer to the order of deoxyribonucleotides along a deoxyribonucleic acid chain. The order of these deoxyribonucleotides determines the order of amino acids along a polypeptide (protein) chain. Thus, a nucleic acid sequence encodes an amino acid sequence.

[0052] Methods for producing and purifying antibodies and antigen-binding fragments are well known and can be found in the prior art, such as the Cold Spring Harbor Manual of Antibody Laboratory Techniques, Chapters 5-8 and 15. The engineered antibodies or antigen-binding fragments thereof disclosed herein can be prepared and purified by conventional methods. For example, cDNA sequences encoding heavy and light chains can be cloned and recombined into expression vectors. The recombinant immunoglobulin expression vector can be stably transfected into CHO cells. Stable clones are obtained by expressing antibodies that specifically bind to human antigens. Positive clones are expanded in serum-free culture medium in a bioreactor to produce antibodies. The culture fluid that secretes the antibodies can be purified and collected by conventional techniques. The antibodies can be filtered and concentrated by conventional methods. Soluble mixtures and polymers can also be removed by conventional methods, such as molecular sieves and ion exchange.

[0053] As used herein, the term "individual" or "subject" refers to any animal, such as a mammal or marsupial. Individuals of the present disclosure include, but are not limited to, humans, non-human primates (e.g., cynomolgus or rhesus monkeys or other types of macaques), mice, pigs, horses, donkeys, cattle, sheep, rats, and any type of poultry.

[0054] As used herein, the terms "disease," "condition," or "disorder" refer to any change or disorder that damages or interferes with the normal function of a cell, tissue, or organ. For example, "disease" includes, but is not limited to, tumors, pathogen infection, autoimmune diseases, T-cell dysfunction, or immune tolerance deficiency (e.g., transplant rejection).

[0055] As used herein, the term "tumor" refers to a disease characterized by the pathological proliferation of cells or tissues, and their subsequent migration or invasion of other tissues or organs. Tumor growth is typically uncontrolled and progressive, without inducing or inhibiting normal cell proliferation. Tumor includes "cancer" and refers broadly to all malignant tumors.

[0056] As used herein, the term "treatment" refers to clinical intervention aimed at altering the course of a disease in an individual or cell, and can be either preventative or interventional in the clinical pathological process. Therapeutic effects include, but are not limited to, preventing the onset or recurrence of a disease, alleviating symptoms, reducing any direct or indirect pathological consequences of a disease, preventing metastasis, slowing the progression of a disease, improving or relieving the condition, and alleviating or improving the prognosis.

[0057] The term "combination" as used herein refers to a treatment regimen that provides at least two or more different therapies to achieve a specified therapeutic effect. The therapies can be physical, such as radiotherapy, or chemical, such as administering a drug to a subject, and the drug also includes a combination drug. "Combination drug" refers to a combination of two or more pharmaceutical preparations, each of which has an active ingredient, that need to be used in combination when administered to a subject. The active ingredients can be mixed together to form a single dosing unit, or they can be independently administered as dosing units and used separately; when administered, different pharmaceutical preparations can be administered essentially synchronously, simultaneously or sequentially.

[0058] Overall survival (OS) is the time from the start of study treatment to death from any cause. Patients who were still in follow-up at the cut-off date were censored based on the cut-off date. Subjects lost to follow-up were censored based on the date of last contact.

[0059] Progression-free survival (PFS) refers to the time from the start of study treatment to the first imaging assessment of disease progression or death from any cause (whichever occurs first).

[0060] The objective response rate (ORR) was defined as the best confirmed ORR during the study period, including both complete responses (CR) and partial responses (PR). According to RECIST v1.1, additional imaging was required to confirm the first occurrence of a PR or CR at ≥4 weeks.

[0061] Disease control rate (DCR) refers to the percentage of cases with the best efficacy evaluation as "CR+PR+SD".

[0062] Complete remission (CR): All target lesions disappear and the short diameter of all pathological lymph nodes (including target nodules and non-target nodules) must be reduced to <10mm.

[0063] Partial response (PR): The sum of the target lesion diameters decreased by at least 30% compared with the baseline level.

[0064] Disease progression (PD): The minimum value of the sum of all target lesion diameters measured during the entire experimental study is used as a reference, and the sum of diameters increases by at least 20% (if the baseline measurement value is the minimum, the baseline value is used as a reference); in addition, the absolute value of the sum of diameters must increase by at least 5 mm (the appearance of one or more new lesions is also considered as disease progression).

[0065] Stable disease (SD): The target lesion has not decreased to the level of PR, nor increased to the level of PD, but is somewhere in between. The minimum sum of the diameters can be used as a reference for research.

[0066] Duration of response (DOR) was defined as the time from the first tumor assessment as CR or PR (whichever was recorded first) to the first assessment as PD or death.

[0067] Treatment-emergent adverse events (TEAEs) refer to adverse events that occur during treatment.

[0068] Treatment-related adverse events (TRAEs) are adverse events related to study drugs that occur during treatment.

[0069] A serious adverse event (SAE) refers to an adverse medical event (AE) such as death, life-threatening, permanent or severe disability or loss of function, the need for hospitalization or prolonged hospitalization, and congenital abnormalities or birth defects that occurs after the subject receives the investigational drug.

[0070] Example

[0071] The present disclosure is further described below with reference to specific examples. It should be understood that these examples are only used to illustrate the present disclosure and are not intended to limit the scope of the present disclosure.

[0072] Example 1 Obtaining the hybrid antibody ZPML265

[0073] A hybrid antibody produced by a single host cell line was prepared using the method disclosed in PCT / US2017 / 030676. The hybrid antibody contains two active ingredients: a recombinant humanized IgG1 monoclonal antibody targeting human CTLA4 and a recombinant humanized IgG4 monoclonal antibody targeting human PD1. This hybrid antibody can specifically bind to both human CTLA4 and PD1. After the two antibodies are simultaneously expressed in a single host cell line, they are collected, purified, and combined with a pharmaceutically acceptable carrier to form a single hybrid antibody pharmaceutical formulation, ZPML265. The amino acid sequences of the anti-CTLA4 and anti-PD1 antibodies are shown in Table 1 below.

[0074] Table 1 Amino acid sequences of the components of the mixed antibody ZPML265

[0075] Example 2 A clinical study evaluating the safety and efficacy of ZPML265 combined with chemotherapy as a first-line treatment for patients with non-small cell lung cancer

[0076] In a Phase II clinical study conducted in China, 29 patients with squamous cell carcinoma and non-squamous cell carcinoma without EGFR / ALK / ROS1 gene mutations were enrolled. Patients were aged ≥18 years, had an Eastern Cooperative Oncology Group (ECOG) performance status of 0 or 1, had an expected survival of ≥3 months, had at least one measurable lesion according to RECIST v1.1, and had the following vital organ function levels before the first dose of the investigational drug:

[0077] 1) White blood cell count ≥3×10 9 / L;

[0078] 2) Absolute neutrophil count ≥1.5×10 9 / L;

[0079] 3) Platelets ≥100×10 9 / L;

[0080] 4) Hemoglobin ≥90 g / L;

[0081] 5) serum creatinine ≤1.5× upper limit of normal (ULN) or creatinine clearance (CLcr) calculated by the Cockcroft-Gault formula ≥50 mL / min;

[0082] 6) Total bilirubin ≤1.5×ULN (<3×ULN in patients with Gilbert's syndrome);

[0083] 7) Aspartate aminotransferase (AST) and alanine aminotransferase (ALT) ≤ 2.5 × ULN (≤ 5 × ULN is allowed when liver metastasis is present);

[0084] 8) Left ventricular ejection fraction (LVEF)>50%.

[0085] Dosage regimen:

[0086] The drugs used in this study are as follows: the treatment received by patients is divided into two stages, namely the lead-in period and the post-lead-in period, and the lead-in period includes 2 cycles. ZPML265 combined with chemotherapy is administered for 2 cycles in the lead-in period for squamous and non-squamous cell carcinoma, followed by ZPML265 maintenance treatment. ZPML265 is used until disease progression, intolerable toxicity, initiation of other anti-tumor treatments, loss of follow-up, death, withdrawal from the study, or 2 years of treatment (whichever occurs first). The dosing window period is ±3 days of the planned dosing date (calculated from the date after the first dose). If the dosing window period is exceeded, it is regarded as early or delayed administration, and the subsequent dosing date needs to be recalculated according to the last actual dosing date.

[0087] ZPML265: 5 mg / kg, administered once every three weeks (Q3W), intravenous infusion (IV) on the first day; the intravenous infusion time of ZPML265 is more than 30 minutes. After the infusion is completed, 20 ml of the same solvent is used to flush the tube. The entire infusion time including flushing is 60 minutes (±5 minutes); ZPML265 must not be injected by intravenous push.

[0088] Chemotherapy drug regimen:

[0089] Non-squamous cell carcinoma patients: pemetrexed (500 mg / m 2) + carboplatin (AUC 5 or 6 mg / mL / min) (according to Calvert's formula), administered once every three weeks as an intravenous infusion on day 1 of each cycle for two cycles.

[0090] Patients with squamous cell carcinoma: paclitaxel (175 mg / m 2 ) + carboplatin (AUC 5 or 6 mg / mL / min) (according to Calvert's formula), administered once every three weeks as an intravenous infusion on day 1 of each cycle for two cycles.

[0091] Effectiveness Results:

[0092] Efficacy data of the full analysis set: ORR was 58.6% (17 / 29), and DCR was 93.1% (27 / 29).

[0093] Analysis according to pathological type (squamous and non-squamous):

[0094] ① Seventeen patients with first-line wild-type squamous NSCLC were enrolled and treated with ZPML265 (5 mg / kg) + paclitaxel (175 mg / m 2 ) + carboplatin (AUC5 / 6), the ORR was 71% (12 / 17) and the DCR was 94% (16 / 17).

[0095] ② Twelve patients with first-line wild-type non-squamous NSCLC were enrolled and treated with ZPML265 (5 mg / kg) + pemetrexed (500 mg / m 2 ) + carboplatin (AUC5 / 6), the ORR was 42% (5 / 12) and the DCR was 92% (11 / 12).

[0096] The above results demonstrate that ZPML265 (5 mg / kg) combined with chemotherapy is effective for both squamous cell carcinoma and non-squamous cell carcinoma.

[0097] Safety results:

[0098] All 29 patients evaluable for safety experienced TRAEs. The incidence of grade ≥3 AEs was 13.8% (4 / 29), of which 2 (6.9%) were related to ZPML265. The incidence of infusion-related reactions was 67.0% (20 / 29). The incidence of immune-related adverse events was 75.9% (22 / 29). The incidence of SAEs was 24.1% (7 / 29). The incidence of SAEs related to study drug was 17.2% (5 / 29). The incidence of AEs leading to suspension of study drug was 41.4% (12 / 29). No AEs led to permanent discontinuation of study drug.

[0099] Preliminary conclusions:

[0100] ZPML265 combined with chemotherapy showed significant efficacy trends in patients with stage IIIB / C and stage IV non-small cell lung cancer, with overall safety controllable and good tolerability.

[0101] Example 3 Phase II clinical study evaluating the safety and efficacy of ZPML265 combined with chemotherapy and bevacizumab as first-line treatment for patients with non-small cell lung cancer

[0102] In a phase II clinical study conducted in China, a total of 31 patients with stage IIIB / C and stage IV EGFR wild-type non-small cell lung cancer (including squamous cell carcinoma and non-squamous cell carcinoma) and 31 patients with non-small cell lung cancer who failed EGFR-TKI treatment were enrolled.

[0103] Patients must be over 18 years old, have an Eastern Cooperative Oncology Group (ECOG) performance status score of 0 or 1, have an expected survival of ≥ 3 months, have at least one measurable lesion according to RECIST v1.1, and have the following vital organ function level before the first use of the investigational drug:

[0104] 1) White blood cell count ≥3×10 9 / L;

[0105] 2) Absolute neutrophil count ≥1.5×10 9 / L;

[0106] 3) Platelets ≥100×10 9 / L;

[0107] 4) Hemoglobin ≥90 g / L;

[0108] 5) serum creatinine ≤1.5× upper limit of normal (ULN) or creatinine clearance (CLcr) calculated by the Cockcroft-Gault formula ≥50 mL / min;

[0109] 6) Total bilirubin ≤1.5×ULN (<3×ULN in patients with Gilbert's syndrome);

[0110] 7) AST and ALT ≤ 2.5 × ULN (≤ 5 × ULN is allowed when liver metastasis is present);

[0111] 8) INR or APTT ≤ 1.5 × ULN;

[0112] 9) Left ventricular ejection fraction (LVEF)>50%.

[0113] Dosage regimen:

[0114] The drugs used in this study are as follows: the treatment received by patients is divided into two stages, namely the lead-in period and the post-lead-in period, and the lead-in period includes 4 cycles. During the lead-in period for non-squamous cell carcinoma, ZPML265 combined with chemotherapy and bevacizumab is administered for 4 cycles, followed by maintenance treatment with ZPML265 combined with pemetrexed and bevacizumab. During the lead-in period for squamous cell carcinoma, ZPML265 combined with chemotherapy is administered for 4 cycles, followed by maintenance treatment with ZPML265. ZPML265 is administered until disease progression, intolerable toxicity, initiation of other anti-tumor treatments, loss to follow-up, death, withdrawal from the study, or 2 years of treatment (whichever occurs first). The dosing window period is ±3 days of the planned dosing date (calculated from the date after the first dose). If the dosing window period is exceeded, it will be regarded as early or delayed administration, and the subsequent dosing date must be recalculated according to the last actual dosing date.

[0115] ZPML265: 5 mg / kg, administered once every three weeks, via intravenous infusion on the first day; the intravenous infusion time of ZPML265 is more than 30 minutes. After the infusion is completed, 20 ml of the same solvent is used to flush the tube. The entire infusion time, including the flushing, is 60 minutes (± 5 minutes); ZPML265 must not be injected by intravenous push.

[0116] Chemotherapy drug regimen:

[0117] Non-squamous cell carcinoma patients: Lead-in phase: ZPML265 (5 mg / kg) + pemetrexed (500 mg / m 2 ) + carboplatin (AUC5 / 6) + bevacizumab (15 mg / kg), administered once every three weeks, intravenous infusion on day 1, for 4 cycles. After the lead-in period: ZPML265 (5 mg / kg) + pemetrexed (500 mg / m 2 ) + bevacizumab (15 mg / kg), administered once every three weeks, intravenous infusion on day 1.

[0118] Squamous cell carcinoma patients: Lead-in phase: ZPML265 (5 mg / kg) + paclitaxel (175 mg / m 2 ) + carboplatin (AUC5 / 6), administered once every three weeks by intravenous infusion on day 1 for 4 cycles. After the lead-in phase: ZPML265 (5 mg / kg), administered once every three weeks by intravenous infusion on day 1.

[0119] Efficacy results (EGFR wild-type NSCLC data):

[0120] (1) Effectiveness for the entire population:

[0121] There were 31 cases of EGFR wild-type NSCLC, with an ORR of 45.2% (14 / 31), a DCR of 83.9 (26 / 31), and a PFS of 5.45 (4.271, 11.236).

[0122] Analysis according to pathological type (squamous and non-squamous):

[0123] ① Eleven patients with first-line wild-type squamous NSCLC were enrolled and treated with ZPML265 (5 mg / kg) + paclitaxel (175 mg / m 2 ) + carboplatin (AUC5 / 6) for 4 cycles, the ORR was 36.4% (4 / 11), the DCR was 90.9% (10 / 11), and the mPFS was 6.97 (4.107, -) months.

[0124] ② Twenty patients with first-line wild-type non-squamous NSCLC were enrolled and treated with ZPML265 (5 mg / kg) + pemetrexed (500 mg / m 2 ) + carboplatin (AUC5 / 6) + bevacizumab (15 mg / kg) for 4 cycles, the ORR was 40% (8 / 20), the DCR was 94.1% (16 / 20), and the mPFS was 7.69 (4.271, -) months.

[0125] The above results demonstrate that the combined treatment of ZPML265 (5 mg / kg) and chemotherapy is effective for both non-squamous cell carcinoma and squamous cell carcinoma.

[0126] (2) Preliminary efficacy in PD-L1 negative subjects—the target study population

[0127] Based on Example 2 and Example 3, there were a total of 12 first-line NSCLC patients with negative PD-L1 expression and negative driver genes. The initial response rate of ZPML265 combined with platinum-containing chemotherapy ± bevacizumab treatment was 50% (6 / 12), the DCR was 91.7% (11 / 12), and the mPFS was 6.14 (2.103, -) months, indicating that ZPML265 combination therapy has good therapeutic potential for first-line NSCLC patients with negative PD-L1 expression and negative driver genes, with good response and tolerable adverse reactions.

[0128] Safety results:

[0129] The following are the safety results of a comprehensive analysis of Example 2 (29 cases of wild-type NSCLC) and Example 3 (31 cases of EGFR wild-type NSCLC):

[0130] Among the 60 safety-evaluable patients, 59 experienced TRAEs. The incidence of grade ≥3 AEs was 40% (24 / 60), of which 13 (21.7%) were related to ZPML265, a lower incidence compared to comparable studies (47% in the CM-9LA study and 51.8% in the POSEIDON study). Immune-related adverse events occurred in 41 patients (68.3%), of which 35 (58.3%) were related to ZPML265. TEAEs led to drug discontinuation in 34 patients (56.7%), of which 30 (50.0%) were related to ZPML265. Three patients (5.0%) experienced TEAEs, of which two (3.3%) were related to ZPML265, a lower incidence compared to comparable studies (19% in the CM-9LA study and 15.1% in the POSEIDON study). No adverse reactions resulted in death in the study.

[0131] Preliminary conclusions:

[0132] ZPML265 combined with chemotherapy showed a significant efficacy trend in patients with stage IIIB / C and stage IV non-small cell lung cancer (including NSCLC with negative driver genes and negative PD-L1 expression), with controllable overall safety and good tolerability.

[0133] Example 4 Phase III clinical study to evaluate the efficacy and safety of ZPML265 combined with chemotherapy as a first-line treatment for patients with PD-L1 negative, locally advanced or metastatic non-small cell lung cancer

[0134] In a Phase III clinical study conducted in China, 650 patients with PD-L1 expression-negative, locally advanced or metastatic non-small cell lung cancer, including squamous cell carcinoma and non-squamous cell carcinoma, were planned to be enrolled, excluding NSCLC patients with EGFR and ALK mutations.

[0135] Negative PD-L1 expression was defined as TPS < 1% (TPS: the percentage of tumor cells with PD-L1 membrane staining of any intensity among tumor cells).

[0136] Patients must be over 18 years old, have an Eastern Cooperative Oncology Group (ECOG) performance status of 0 or 1, have an expected survival of ≥ 3 months, have at least one measurable lesion according to RECIST v1.1, have not received systemic treatment for advanced or metastatic NSCLC, and have the following vital organ function levels before the first use of the investigational drug:

[0137] (1) Absolute neutrophil count ≥1.5×10 9 / L;

[0138] (2) Platelet count ≥100×10 9 / L;

[0139] (3) Hemoglobin ≥90 g / L;

[0140] (4) serum creatinine ≤1.5× upper limit of normal (ULN) or creatinine clearance (CLcr) calculated by the Cockcroft-Gault formula ≥50 mL / min;

[0141] (5) Total bilirubin ≤1.5×ULN (≤3×ULN in patients with Gilbert syndrome);

[0142] (6) AST and ALT ≤ 2.5 × ULN (≤ 5 × ULN for patients with liver metastasis);

[0143] (7) International normalized ratio (INR) or activated partial thromboplastin time (APTT) ≤ 1.5 × ULN, unless the subject is receiving anticoagulant therapy;

[0144] (8) Left ventricular ejection fraction (LVEF) ≥ 50%.

[0145] Dosage regimen:

[0146] The medications used in this study are as follows: Treatment for subjects is divided into two phases: a run-in phase and a maintenance phase. The run-in phase consists of four cycles. Subjects receive the treatment regimens shown in the table below, according to study group and tissue typing, until disease progression, intolerable toxicity, withdrawal of informed consent, non-compliance with study treatment or procedures, investigator's decision to withdraw the subject from the study, or other reasons specified in the protocol (the maximum duration of ZPML265 treatment is 2 years [35 cycles]).

[0147] Table 2 Phase III clinical study plan for the efficacy and safety of ZPML265 combined with chemotherapy as first-line treatment for patients with PD-L1 negative, locally advanced or metastatic non-small cell lung cancer

[0148] Example 5: A randomized, double-blind, multicenter phase III clinical study evaluating ZPML265 combined with platinum-containing chemotherapy versus placebo combined with platinum-containing chemotherapy in the adjuvant treatment of stage II-IIIB non-small cell lung cancer after complete surgical resection

[0149] This randomized, double-blind, multicenter Phase III trial of ZPML265 plus platinum-based chemotherapy versus placebo plus platinum-based chemotherapy as adjuvant therapy for stage II-IIIB non-small cell lung cancer following complete surgical resection enrolled patients with pathologically confirmed squamous or non-squamous NSCLC. Patients were eligible and age ≥18 years, regardless of gender. ECOG performance status was 0-1. Patients were eligible for stage II-IIIB according to the 8th edition of the American Joint Committee on Cancer (AJCC) staging system and had undergone curative resection (R0). Disease stages included: T2b-T4N0, T1-T4N1, T1-T2bN2, and curatively resectable T3-T4N2. Resection procedures included: lobectomy, sleeve lobectomy, bilobectomy, or pneumonectomy. Note: Segmentectomy or wedge resection was not permitted. Patients enrolled in the study must have received adjuvant therapy within 8 weeks (≤56 days) of surgery and fully recovered from surgery to be able to receive the study drug. Non-squamous NSCLC subjects must not have EGFR-sensitive mutations or ALK fusion genes (test results from local medical institutions must be accepted; if no test results are available, tumor tissue specimens must be provided for central laboratory testing before randomization). They must have adequate organ function before the first dose of study treatment (no blood components, leukocyte-raising drugs, or platelet-raising drugs were allowed within 7 days before obtaining laboratory tests).

[0150] 1) White blood cell count ≥4.0×10 9 / L;

[0151] 2) Absolute neutrophil count ≥1.5×10 9 / L;

[0152] 3) Platelet count ≥100×10 9 / L;

[0153] 4) Hemoglobin ≥90 g / L;

[0154] 5) serum creatinine ≤1.5×upper limit of normal (ULN) or creatinine clearance (CrCL) calculated by the Cockcroft-Gault formula ≥50 mL / min;

[0155] 6) Total bilirubin ≤1.5×ULN (≤3×ULN for patients with Gilbert syndrome);

[0156] 7) AST and ALT ≤ 2.5 × ULN;

[0157] 8) INR or APTT ≤ 1.5 × ULN;

[0158] 9) Left ventricular ejection fraction (LVEF) ≥ 50%.

[0159] Dosage regimen:

[0160] ZPML265: 5 mg / kg, administered once every three weeks (Q3W), i.e., three weeks as a dosing cycle, intravenous infusion (IV) on the first day (D1) of each cycle.

[0161] Placebo: A preparation for the control group that does not contain the active ingredient ZPML265 but has the same excipients as ZPML265 and is administered intravenously.

[0162] Chemotherapy drugs, including paclitaxel, pemetrexed, vinorelbine, cisplatin, and carboplatin: intravenous infusion.

[0163] 1) Squamous cell NSCLC subjects were randomized to receive one of the following treatments:

[0164] Experimental group: ZPML265 5mg / kg (D1) + vinorelbine 25mg / m 2 (D1, D8) or paclitaxel 175 mg / m 2 (D1) + cisplatin 75 mg / m 2 (D1) or carboplatin AUC = 5 mg / mL / min (D1), with one dosing cycle every three weeks, for a total of 2-4 dosing cycles; then ZPML265 5 mg / kg (D1) was maintained, administered once every three weeks until 16 cycles of ZPML265 were completed.

[0165] Control group: placebo 5 mg / kg (D1) + vinorelbine 25 mg / m 2 (D1, D8) or paclitaxel 175 mg / m 2 (D1) + cisplatin 75 mg / m 2 (D1) or carboplatin AUC = 5 mg / mL / min (D1), with one dosing cycle every three weeks, for a total of 2-4 dosing cycles; then ZPML265 5 mg / kg (D1) was maintained, administered once every three weeks, until 16 cycles of placebo were completed.

[0166] 2) Non-squamous NSCLC subjects were randomized to receive one of the following treatments:

[0167] Experimental group: ZPML265 5mg / kg (D1) + pemetrexed 500mg / m 2 (D1) or vinorelbine 25 mg / m 2 (D1, D8) + cisplatin 75 mg / m 2(D1) or carboplatin AUC = 5 mg / mL / min (D1), with one dosing cycle every three weeks, for a total of 2-4 dosing cycles; then ZPML265 5 mg / kg (D1) was maintained, administered once every three weeks until 16 cycles of ZPML265 were completed.

[0168] Control group: placebo 5 mg / kg (D1) + pemetrexed 500 mg / m 2 (D1) or vinorelbine 25 mg / m 2 (D1, D8) + cisplatin 75 mg / m 2 (D1) or carboplatin AUC = 5 mg / mL / min (D1), with one dosing cycle every three weeks, for a total of 2-4 cycles; followed by maintenance with placebo 5 mg / kg (D1), administered once every three weeks until 16 cycles of placebo are completed.

Claims

1. A pharmaceutical composition for treating non-small cell lung cancer, comprising an effective amount of a mixed antibody of anti-CTLA4 and anti-PD1, wherein the mixed antibody is produced by a single host cell containing nucleic acids encoding two different antibodies, anti-CTLA4 and anti-PD1, wherein: The sequences of the anti-CTLA4 antibody heavy chain HCDR1, HCDR2 and HCDR3 are shown in SEQ ID NOs: 1, 2 and 3, respectively; the sequences of the anti-CTLA4 antibody light chain LCDR1, LCDR2 and LCDR3 are shown in SEQ ID NOs: 4, 5 and 6, respectively; the sequences of the anti-PD1 antibody heavy chain HCDR1, HCDR2 and HCDR3 are shown in SEQ ID NOs: 9, 10 and 11, respectively; and the sequences of the anti-PD1 antibody light chain LCDR1, LCDR2 and LCDR3 are shown in SEQ ID NOs: 12, 13 and 14, respectively.

2. The pharmaceutical composition according to claim 1, wherein The heavy chain variable region sequence of the anti-CTLA4 antibody is shown in SEQ ID NO: 7, the light chain variable region sequence of the anti-CTLA4 antibody is shown in SEQ ID NO: 8, the heavy chain variable region sequence of the anti-PD1 antibody is shown in SEQ ID NO: 15, and the light chain variable region sequence of the anti-PD1 antibody is shown in SEQ ID NO:

16.

3. The pharmaceutical composition according to claim 1, wherein The heavy chain sequence of the anti-CTLA4 antibody is shown in SEQ ID NO: 17, the light chain sequence of the anti-CTLA4 antibody is shown in SEQ ID NO: 18, the heavy chain sequence of the anti-PD1 antibody is shown in SEQ ID NO: 19, and the light chain sequence of the anti-PD1 antibody is shown in SEQ ID NO:

20.

4. The pharmaceutical composition according to claim 1, wherein the non-small cell lung cancer has a TNM stage of III or IV. The pharmaceutical composition according to claim 1 , wherein the non-small cell lung cancer is stage II-IIIB non-small cell lung cancer after complete surgical resection.

6. The pharmaceutical composition according to claim 1, wherein the non-small cell lung cancer is locally advanced or metastatic non-small cell lung cancer that is negative for PD-L1 expression, has no EGFR sensitive mutation or ALK gene translocation change, and has not received systemic anti-tumor treatment.

7. The pharmaceutical composition according to any one of claims 4 to 6, wherein the non-small cell lung cancer is squamous non-small cell lung cancer.

8. The pharmaceutical composition according to claim 7, wherein the dosage of the mixed antibody is 5 mg / kg, administered by intravenous infusion on the first day of each cycle, and a treatment cycle of 21 days.

9. The pharmaceutical composition of claim 8, further comprising a chemotherapeutic drug.

10. The pharmaceutical composition according to claim 9, wherein the chemotherapy drugs are carboplatin / cisplatin and paclitaxel / vinorelbine.

11. The pharmaceutical composition according to claim 10, wherein Carboplatin dose to target area under the curve (AUC) 5 mg / mL / min or cisplatin dose to 75 mg / m 2 ; and, paclitaxel at a dose of 175 mg / m 2 , or vinorelbine at a dose of 25 mg / m 2 The drug is administered by intravenous infusion on the first day of each cycle, and a treatment cycle lasts for 21 days. If vinorelbine is selected, vinorelbine 25 mg / m2 is administered again on the eighth day of the treatment cycle. 2 .

12. The pharmaceutical composition according to claim 11, wherein after 2-4 cycles of administration of the mixed antibody and the chemotherapy drug, the mixed antibody is used as maintenance therapy at a dose of 5 mg / kg, administered once every three weeks by intravenous infusion.

13. The pharmaceutical composition according to any one of claims 4 to 6, wherein the non-small cell lung cancer is non-squamous non-small cell lung cancer.

14. The pharmaceutical composition according to claim 13, wherein the dosage of the mixed antibody is 5 mg / kg, administered by intravenous infusion on the first day of each cycle, and a treatment cycle of 21 days.

15. The pharmaceutical composition of claim 14, further comprising a chemotherapeutic drug. The pharmaceutical composition according to claim 15 , wherein the chemotherapy drugs are carboplatin / cisplatin and pemetrexed / vinorelbine.

17. The pharmaceutical composition according to claim 16, wherein Carboplatin dose to target area under the curve (AUC) 5 mg / mL / min or cisplatin dose to 75 mg / m 2 ; and, pemetrexed at a dose of 500 mg / m 2 , or vinorelbine at a dose of 25 mg / m 2 The drug is administered by intravenous infusion on the first day of each cycle, and a treatment cycle lasts for 21 days. If vinorelbine is selected, vinorelbine 25 mg / m2 is administered again on the eighth day of the treatment cycle. 2 .

18. The pharmaceutical composition according to claim 17, wherein after 2-4 cycles of administration of the mixed antibody and the chemotherapy drug, the mixed antibody is used as maintenance therapy at a dose of 5 mg / kg, administered once every three weeks by intravenous infusion.

19. A method for treating non-small cell lung cancer, comprising administering the pharmaceutical composition of any one of claims 1 to 18 to a subject.

20. Use of a mixed antibody comprising an effective amount of anti-CTLA4 and anti-PD1 and a chemotherapeutic drug in the preparation of a pharmaceutical composition for treating non-small cell lung cancer, wherein the mixed antibody is produced by a single host cell containing nucleic acids encoding two different antibodies, anti-CTLA4 and anti-PD1, wherein: The heavy chain sequence of the anti-CTLA4 antibody is shown in SEQ ID NO: 17, the light chain sequence of the anti-CTLA4 antibody is shown in SEQ ID NO: 18, the heavy chain sequence of the anti-PD1 antibody is shown in SEQ ID NO: 19, and the light chain sequence of the anti-PD1 antibody is shown in SEQ ID NO: 20; the non-small cell lung cancer is stage II-IIIB non-small cell lung cancer after complete surgical resection; or, the non-small cell lung cancer is locally advanced or metastatic non-small cell lung cancer with negative PD-L1 expression, no EGFR sensitive mutation or ALK gene translocation change, and no prior systemic anti-tumor treatment.