WO2023160517A1 - Pharmaceutical composition comprising mixed antibodies anti-ctla4 and anti-pd1 and therapeutic use thereof - Google Patents

Abstract

The present disclosure relates to the use of a pharmaceutical composition comprising mixed antibodies anti-CTLA4 and anti-PD1 in combination with lenvatinib to treat kidney cancer.

Description

Pharmaceutical composition comprising mixed antibody against CTLA4 and anti-PD1 and therapeutic use thereof technical field

The present disclosure relates to the treatment of cancer, particularly renal 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 in the treatment of renal cancer.

Background technique

Worldwide, the incidence of renal cancer accounts for about 3% of adult malignant tumors, and its distribution has obvious regional differences. The incidence rate is the highest in Western developed countries such as North America and Western Europe, while the incidence rate in developing countries such as Africa and Asia is the lowest. Worldwide, the incidence of renal cancer is currently ranked 9th among male malignancies (214,000 new cases) and 14th among women (124,000 new cases). In recent decades, the incidence of RCC has shown a continuous increase in most countries and regions, including North America, parts of Europe, Asia, Oceania and parts of Latin America, but the mortality rate of RCC tends to be stable or low in developed countries. decline. According to the data from the China Cancer Registry Annual Report, from 1988 to 2014, the incidence of RCC in China showed an upward trend. From 2005 to 2009, the incidence rates of renal cancer were 3.96/100,000, 4.44/100,000, 4.64/100,000, 5.08/100,000, and 4.5/100,000, respectively. In February 2018, the National Cancer Center of China compiled data from 339 cancer registries, covering a total of 280 million people in China. Statistics show that the incidence of kidney cancer in China in 2014 was 4.99/100,000.

Among the treatment options for renal cancer, the 5-year survival rate of surgical treatment of localized and locally advanced renal cancer can reach 93%, but about 20%-40% of early renal cancer will recur within 5 years after radical surgery.

Advanced RCC is mainly treated with medical treatment. In 2005, the FDA approved the targeted drug Sorafenib for the treatment of advanced renal cancer. Since then, first-line treatment combined with immunotherapy has become an important development trend. At present, a total of 6 immunotherapy programs have been approved for marketing: 1) FDA approved Nivolumab for second-line treatment of advanced renal cancer in 2015; 2) FDA approved it in April 2018 Nivolumab combined with Ipilimumab for first-line treatment of advanced renal cancer; 3) FDA approved Pembrolizumab + axitinib in April and May 2019 and 4) Avelumab (anti-PD-L1 monoclonal antibody) + axitinib for advanced renal cancer First-line treatment; 5) In January 2021, the FDA approved Nivolumab + cabozantinib for the first-line treatment of advanced renal cancer; 6) In August 2021, the FDA approved pembrolizumab + lenvatinib for the first-line treatment of advanced renal cancer. However, the combination of double immunity (combination of two immune checkpoint inhibitors) and targeted drugs for the treatment of renal cancer has not yet been marketed.

ZPML265 is a mixed antibody drug formulation consisting of a recombinant humanized IgG1 monoclonal antibody targeting human CTLA4 and a recombinant humanized IgG4 monoclonal antibody targeting human PD1, two different antibodies produced by a single host cell . The mixed antibody can specifically bind CTLA4 and PD1 at the same time, thereby blocking the two immune checkpoint signaling pathways of CTLA4 and B7-1/B7-2 and PD1 and PDL1, releasing the inhibitory effect of the two pathways on T lymphocytes, and restoring Its functional activity and anti-tumor immune response enable the body to fight and kill tumors.

Lenvatinib is a multi-target tyrosine kinase inhibitor (TKI) with Has the following structural formula:

Lenvatinib has a novel binding mode, in addition to inhibiting other pro-angiogenic and oncogenic signaling pathway-related tyrosine kinases involved in tumor proliferation, it can also selectively inhibit the kinase activity of vascular endothelial growth factor (VEGF) receptors, such as VEGF-1, VEGF-2 and VEGF-3 etc. In 2016, the FDA and EMA successively approved the drug combined with everolimus for the treatment of advanced renal cell carcinoma; in August 2021, the FDA approved the drug combined with pembrolizumab for the first-line treatment of adult patients with advanced renal cell carcinoma.

However, as a new treatment plan, double-immunization plus targeting is still a blank in the clinical field of renal cancer in the world. There is no such treatment plan approved for marketing, and there is a wide space for exploration and urgent unmet clinical needs.

Summary of the invention

The technical problem to be solved by the present disclosure is to provide a combination therapy of double immunity and targeting to fill the clinical gap in which combination therapy of double immunity and targeting has not yet been applied to kidney cancer, so as to solve such unmet clinical needs .

The present disclosure provides a pharmaceutical composition for treating renal cancer, comprising an effective amount of a mixed antibody of anti-CTLA4 and anti-PD1.

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

In some embodiments, the pharmaceutical composition further comprises additional therapeutic agents, including but not limited to chemotherapeutics, cytotoxic agents, radiotherapeutics, cancer vaccines, tumor reducing agents, targeted anticancer agents, antivascular Genetic agents, biological response modifiers, cytokines, hormones, anti-metastatic and immunotherapeutic agents.

In some embodiments, the additional therapeutic agent is preferably lenvatinib or a pharmaceutically acceptable salt thereof, more preferably lenvatinib mesylate.

Therefore, the present disclosure preferably also provides a pharmaceutical composition for treating renal cancer, which contains an effective amount of a mixed antibody against CTLA4 and anti-PD1 and lenvatinib.

In some embodiments, the mixed antibody is produced by a single host cell containing nucleic acids encoding two different anti-CTLA4 and anti-PD1 antibodies, wherein the sequences of the heavy chains HCDR1, HCDR2 and HCDR3 of the anti-CTLA4 antibody are SEQ ID NO: ID NO: 1, 2, 3, the sequences of anti-CTLA4 antibody light chain LCDR1, LCDR2 and LCDR3 are respectively shown in SEQ ID NO: 4, 5, 6, and the sequences of anti-PD1 antibody heavy chain HCDR1, HCDR2 and HCDR3 are respectively The sequences of the anti-PD1 antibody light chains LCDR1, LCDR2 and LCDR3 are shown in SEQ ID NO: 9, 10 and 11, and the sequences are shown in SEQ ID NO: 12, 13 and 14, respectively.

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, and the heavy chain of the anti-PD1 antibody The variable region sequence 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.

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, and the heavy chain sequence of the anti-PD1 antibody is SEQ ID NO: As shown in 19, the light chain sequence of the anti-PD1 antibody is shown in SEQ ID NO: 20.

Preferably, the renal cancer is clear cell renal carcinoma.

Preferably, the renal cancer is metastatic or recurrent renal cell carcinoma.

The dosage of the mixed antibody is 5 mg/kg, administered once every three weeks, by intravenous infusion; the initial dosage of lenvatinib mesylate is 20 mg or 14 mg or 10 mg, administered orally once a day.

The present disclosure also provides a method for treating renal cancer, the method comprising administering to a subject a pharmaceutical composition, the pharmaceutical composition being the aforementioned pharmaceutical composition comprising an effective amount of a mixed antibody of anti-CTLA4 and anti-PD1.

And, the present disclosure also provides a method for treating kidney cancer, the method comprising administering to a subject a pharmaceutical composition, the pharmaceutical composition being the aforementioned mixed antibody comprising an effective amount of anti-CTLA4 and anti-PD1 and lenval The pharmaceutical composition of Tini.

The present disclosure also provides the use of the pharmaceutical composition comprising an effective amount of the anti-CTLA4 and anti-PD1 mixed antibody and lenvatinib in the preparation of a drug for treating renal cancer.

The result of a phase I clinical study of the mixed antibody of the present disclosure as a single drug shows that the curative effect is definite and the safety is good in 18 patients with renal cancer enrolled. At the same time, the clinical research based on the combination of the mixed antibody of the present disclosure combined with lenvatinib also shows that immune checkpoint inhibitors combined with TKI can play a synergistic anti-tumor effect in advanced renal cancer. Therefore, the mixed antibody or the combined regimen of mixed antibody and lenvatinib provided by the present disclosure is expected to have controllable safety and good patient compliance for the medical treatment of advanced renal cell carcinoma, and can largely fill the dual There is a clinical gap in the treatment of kidney cancer with the combination of immunotherapy and targeted therapy, and it solves the urgent clinical needs that have not been met.

Detailed ways

the term

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 and In general.

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

Certain embodiments disclosed herein encompass numerical ranges, and certain aspects of the disclosure can be described in terms of ranges. Unless otherwise stated, it should be understood that the numerical range or the way described in the range is only for the purpose of brevity and convenience, and should not be regarded as are intended to be strict limitations on the scope of the present disclosure. Accordingly, description in range form should be considered to specifically disclose all possible subranges and all possible specific numerical points within that range, as if such subranges and numerical points had been expressly written herein. The above principles apply equally regardless of the breadth or narrowness of the numerical values stated. When a range is used, that range includes the range endpoints.

When referring to measurable values such as amounts, temporal durations, etc., the term "about" is meant to include ±20%, or in some cases ±10%, or in some cases ±5%, or within ±1% in some cases, or ±0.1% in some cases.

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

The term "antibody", as used herein, typically refers to a 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. ) Y-tetrameric protein. Natural IgG antibodies have such a structure. Each light chain consists of a variable domain (VL) and a constant domain (CL). Each heavy chain comprises a variable domain (VH) and constant region (CH).

Five major classes of antibodies are known in the art: IgA, IgD, IgE, IgG, and IgM, and the corresponding heavy chain constant domains are called α, δ, ε, γ, and μ, respectively. IgG and IgA can be further divided into different For example, IgG can be divided into IgG1, IgG2, IgG3, 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 distinct classes, called kappa and lambda, based on the amino acid sequence of their constant domains.

The term "variable region" or "variable domain" exhibits significant variation in amino acid composition from one antibody to another and is primarily responsible for antigen recognition and binding. The variable regions of each light chain/heavy chain pair form the antibody combining site such that an intact IgG antibody has two binding sites (ie it is bivalent). The variable region (VH) of the heavy chain and the variable region (VL) of the light chain each contain three regions of extreme variability known as hypervariable regions (HVR) or, more commonly, Complementarity-determining regions (CDR), VH and VL each have four framework regions FR (or called framework regions), which are represented by FR1, FR2, FR3, and FR4, respectively. Thus, the CDR and FR sequences typically appear in the following sequence of the heavy chain variable domain (VH) (or the light chain variable domain (VL)): FR1-HCDR1(LCDR1)-FR2-HCDR2(LCDR2)-FR3 -HCDR3 (LCDR3)-FR4.

Types of "antibodies" in a broad sense 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, intrabodies, multispecific antibodies, bispecific antibodies, single chain antibodies, monovalent antibodies, multivalent antibodies, single domain antibodies, nanobodies, synthetic antibodies (including Mutant proteins and their variants) and so on.

The term "monoclonal antibody" (or "monoclonal antibody") refers to a substantially homogeneous antibody produced by a single cell clone that only targets a specific epitope. Monoclonal antibodies can be prepared using various techniques known in the art, including hybridoma technology, recombinant technology, phage display technology, transgenic animals, synthetic technology or a combination of the above technologies, etc.

It should be noted that the division of CDRs and FRs of antibody variable regions in the present disclosure is determined according to the Kabat definition. However, other naming and numbering systems, such as Chothia, IMGT or AHo etc., are also known to those skilled in the art. Therefore, with the antibody of the present disclosure On a sequence basis, humanized antibodies comprising one or more CDRs derived from any nomenclature system expressly remain within the scope of the present disclosure.

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

The terms "polypeptide", "peptide" and "protein" are used interchangeably herein to refer to a polymer of amino acids of any length. The polymer may be linear, cyclic or branched, it may comprise modified amino acids, especially 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.

As used herein, the term "mixed antibody" refers to a mixture of antibodies from a host cell (any antibody) that has been transfected with DNA encoding at least two different antibodies with different binding optionally cells from a single host cell line) produce a limited number of primary antibody species, optionally no more than two, three, four, five, six, seven, eight, nine or ten kinds. In some embodiments, DNA encoding at least two different heavy chains (HC) and at least two different light chains (LC) can be introduced into the same host cell, for example, a host cell can be encoded with at least two but no more than four DNA transfection of different antibodies with different binding specificities. In some embodiments, the sequences of all transfected DNA encoding HC and LC can be mutated, thereby altering the amino acid sequence of the antibody to disfavor non-cognate HC/LC pairing and strongly favor homologous HC/LC pairing. Where two different HCs are introduced into the host cell, one or both of the two different HCs may optionally be altered so as to disfavor heterodimer formation. In some embodiments, only one heavy chain is altered to prevent heterodimer formation. In some embodiments, where DNA encoding only two different antibodies is introduced into a host cell, only one of the antibodies encoded by the DNA contains one or more partner orientation changes such that homologous HC/LC pairing, while the other antibody does not contain such changes. In some embodiments, the host cell produces only two major antibody species, where each HC is predominantly paired with its cognate LC, and the majority of the antibodies are two heavy chains with the same amino acid sequence and two heavy chains with the same amino acid sequence. Tetramers of light chains (see PCT/US2017/030676).

The term "pharmaceutically acceptable salt" refers to a salt of a compound, prepared from a compound having specified substituents and a relatively non-toxic acid or base. In general, such salts are prepared by reacting the free acid or base form of these compounds with a stoichiometric amount of the appropriate base or acid in water or an organic solvent or a mixture of both.

The term "pharmaceutical composition" refers to a preparation or combination of preparations containing one, two or more active ingredients, which allows the active ingredients contained therein to exist in a form effective for biological activity, and does not contain any necessary ingredients for administration. Subjects of the formulation described above had unacceptably toxic additional ingredients. When the "pharmaceutical composition" exists in the form of a combination of different separate preparations containing two or more active ingredients, they can be administered simultaneously, sequentially, separately or at intervals, the purpose of which is to exert the biological activities of multiple active ingredients and to use them together. Yu Zhi cure disease.

The term "pharmaceutically acceptable 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.

The term "effective amount" refers to the dose of a pharmaceutical formulation comprising an active ingredient of the present disclosure which, when administered to a patient in single or multiple doses, produces the desired effect in a treated patient. An effective amount can be readily determined by the attending physician, who is skilled in the art, by considering various factors such as ethnic differences; body weight, age and health; the particular 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 formulation administered; the chosen dosing regimen; and the use of any concomitant therapy.

The terms "host cell", "host cell line" and "host cell culture" are used interchangeably and refer to a cell into which exogenous nucleic acid has been introduced, including the progeny of such a cell. Host cells include "transformants" and "transformed cells," which include the primary transformed cell and progeny derived therefrom, regardless of the number of passages. Progeny may not be identical in nucleic acid content to the parent cell, but may contain mutations. Included herein are mutant progeny that have the same function or biological activity as screened or selected for in the originally transformed cell.

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 electroporation, microinjection, liposome fusion, and the like.

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

Methods for producing and purifying antibodies and antigen-binding fragments are well known and can be found in the art, eg, Cold Spring Harbor's Antibody Laboratory Technique Guide, Chapters 5-8 and 15. Antibodies or antigen-binding fragments thereof engineered in the present disclosure can be prepared and purified using 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 stably transfect CHO cells. Stable clones are obtained by expressing antibodies that specifically bind to human antigens. Positive clones are expanded in serum-free medium in bioreactors for antibody production. The culture fluid that secretes the antibody can be purified and collected using conventional techniques. Antibodies can be concentrated by filtration using conventional methods. Soluble mixtures and aggregates can also be removed by conventional methods such as molecular sieves and ion exchange.

The term "individual" or "subject" as used herein refers to any animal, such as a mammal or a marsupial. Subjects of the present disclosure include, but are not limited to, humans, non-human primates (such as cynomolgus or rhesus or other types of rhesus monkeys), mice, pigs, horses, donkeys, cows, sheep, rats, and any species poultry.

As used herein, the term "disease" or "condition" or "disorder" and the like refers to any change or disorder that damages or interferes with the normal function of a cell, tissue or organ. For example, the "disease" includes but is not limited to: tumor, pathogenic infection, autoimmune disease, T cell dysfunction disease, or immune tolerance defect (such as transplant rejection), etc.

The term "neoplastic" as used herein 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 usually uncontrolled and progressive, without inducing or inhibiting normal cell proliferation. Tumor includes "cancer", which generally refers to all malignant tumors.

The term "kidney cancer" used in this article is also called "renal cell carcinoma", which is a malignant tumor originating from the renal tubular epithelium, accounting for 80% to 90% of renal malignant tumors. The most common histopathological type of RCC is clear cell RCC.

The term "treatment" as used herein refers to clinical intervention in an attempt to alter the course of a disease caused by an individual or a cell, either for prevention or for intervention in the course of clinical pathology. Therapeutic effects include, but are not limited to, preventing the occurrence or recurrence of the disease, relieving symptoms, reducing any direct or indirect pathological consequences of the disease, preventing metastasis, slowing down the progression of the disease, improving or relieving the disease, remission or improving the prognosis, etc.

As used herein, the term "combination" relates to a treatment regimen that provides at least two or more different therapies, either physical, such as radiotherapy, or chemical, such as administration of Drugs for the subject, which also include combination drugs. "Combined drug" refers to a combination comprising two or more pharmaceutical preparations each having an active ingredient, which need to be used in combination when administered to a subject. The active ingredients can be mixed together to form a single administration unit, or they can be independently used as administration units and used separately; when administered, different pharmaceutical preparations can be administered substantially synchronously, simultaneously or sequentially.

Overall survival (OS) refers to the time between the start of study treatment and death from any cause. As of the analysis cut-off date (Cut-off), the patients who are still being followed up will use the cut-off date as the censoring time, and the subjects lost to follow-up will use the date of the last contact with them as the censoring time.

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

Objective response rate (ORR) was defined as the best confirmed ORR during the study period, including complete response (CR) and partial response (PR) cases. According to RECIST v1.1, when PR or CR occurs for the first time, additional imaging examination lesions need to be confirmed at ≥ 4 weeks.

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

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

Partial Response (PR): The sum of the target lesion diameters is reduced by at least 30% compared with the baseline level.

Progression of disease (PD): taking the minimum value of the sum of the diameters of all target lesions measured throughout the experimental study as a reference, the sum of the diameters increased by at least 20% (if the baseline measurement value is the smallest, the baseline value is used as a reference); otherwise In addition, the absolute value of the sum of diameters must increase by at least 5mm (the appearance of one or more new lesions is also considered as disease progression).

Stable disease (SD): The degree of reduction of the target lesion does not reach the PR level, and the degree of increase does not reach the PD level, in between, The minimum value of the sum of diameters can be used as a reference during research.

Duration of response (DOR) was defined as the time between when the tumor was first assessed as CR or PR (whichever was recorded first) and when it was first assessed as PD or death.

Adverse events during treatment (TEAE) refer to adverse events that occurred during treatment.

Treatment-related adverse events (TRAEs) refer to adverse events related to the study drug that occurred during treatment.

Serious adverse event (SAE) refers to the death, life-threatening, permanent or severe disability or loss of function of the subject after receiving the test drug, the subject's need for hospitalization or prolonged hospitalization, and congenital abnormalities or birth defects. medical event.

Example

The present disclosure will be further elaborated below in conjunction with specific embodiments. It should be understood that these examples are only for illustrating the present disclosure and are not intended to limit the scope of the present disclosure.

Example 1 Obtaining of Mixed Antibody ZPML265

Prepare a mixed antibody produced by a single host cell line (see PCT/US2017/030676), which contains two active ingredients, namely a recombinant humanized IgG1 monoclonal antibody targeting human CTLA4 and a human PD1 targeting Recombinant humanized IgG4 monoclonal antibody. This hybrid antibody can specifically bind human CTLA4 and PD1 at the same time. After the two antibodies are expressed simultaneously by a single host cell line, they are collected, purified, and combined with a pharmaceutically acceptable carrier to form a single mixed antibody pharmaceutical preparation ZPML265. The respective amino acid sequences of the anti-CTLA4 antibody and the anti-PD1 antibody are shown in Table 1 below.

Table 1 Amino acid sequences of components of the mixed antibody ZPML265

Example 2 Efficacy and safety of ZPML265 monotherapy in advanced renal cell carcinoma

Administration:

In a phase I clinical study conducted in China, 18 patients with pathologically confirmed metastatic or recurrent renal cell carcinoma were enrolled. The patients were over 18 years old, and the Eastern Cooperative Oncology Group (ECOG) physical status score was 0 or 1. The expected survival time is ≥ 3 months, and the functional level of vital organs before the first use of the investigational drug:

1) Absolute neutrophil count ≥ 1.5×10 ⁹ /L;

2) Platelets ≥ 75×10 ⁹ /L;

3) Hemoglobin ≥ 90g/L;

4) Serum albumin ≥ 30g/L;

5) Alanine aminotransferase (AST) and aspartate aminotransferase (ALT) ≤ 2.5 × upper limit of normal reference value (ULN) (for liver cancer or liver metastasis, ≤ 5 × ULN is allowed);

6) Total bilirubin ≤ 1.5×ULN (≤3×ULN for Gilbert syndrome);

7) International normalized ratio (INR) ≤ 1.5 [INR ≤ 2.3 or prothrombin time (PT) prolongation ≤ 6 seconds in subjects with liver cancer]; activated partial thromboplastin time (APTT) ≤ 1.5 × ULN;

8) Serum creatinine ≤ 1.5 × ULN, if the subject’s creatinine level > 1.5 × ULN, creatinine clearance (CrC1) ≥ 50 mL/min (calculated according to the Cockcroft-Gault formula);

9) Cardiac left ventricular ejection fraction (LVEF) > 50%.

Dosing regimen: The dose of ZPML265 is 5mg/kg, administered by intravenous infusion once every three weeks.

Validity results:

Efficacy data obtained in 18 cases of advanced renal cell carcinoma treated with ZPML265 monotherapy: ORR was 33.3%, DCR was 72.2%, and the trend of efficacy was significant.

Security Results:

Among the 18 patients with advanced RCC, 11 (61.1%) subjects had TRAEs, most of which were grade 1-2 (50.0%); 2 (11.1%) subjects had ≥3 grade TRAEs.

Refer to the treatment plan of drugs similar to ZPML265, for example, in a clinical study of Nivolumab (anti-PD1 antibody) combined with Ipilimumab (anti-CTLA4 antibody) in the first-line treatment of advanced renal cancer, among 547 patients in the Nivolumab combined with Ipilimumab group, 509 patients (93 %) had any grade TRAE, with 250 (46%) grade 3-4 TRAEs. There were 118 (22%) TRAEs leading to discontinuation; 8 patient deaths were treatment-related.

Table 2 lists the comparison of Nivolumab combined with Ipilimumab in the Checkmate214 study and ZPML265 monotherapy in advanced renal cancer with an incidence of ≥10% of TRAEs.

Table 2 TRAEs with an incidence of ≥10% in the Nivolumab combined with Ipilimumab group and ZPML265 monotherapy in advanced renal cancer in the Checkmate214 study

in conclusion:

Overall, the efficacy of ZPML265 monotherapy in the treatment of advanced renal cancer is similar to that of similar drugs; however, in terms of safety, all The incidence of TRAE is lower than that of similar drugs, that is, the safety of the drug is better than that of existing clinical solutions in the prior art while ensuring the effectiveness of the drug.

Example 3 An evaluation of the tolerance, safety, and pharmacokinetics of ZPML265 combined with lenvatinib in patients with advanced renal cell carcinoma Phase Ib clinical study of preliminary efficacy

In the phase Ib clinical study of ZPML265 combined with lenvatinib in the treatment of advanced renal cell carcinoma, patients with advanced renal cell carcinoma (metastatic or unresectable) mainly clear cell type confirmed by pathology were selected. The age of the patient is ≥18 years old, male or female. Eastern Cooperative Oncology Group (ECOG) performance status score 0-1. Expected survival period ≥ 6 months. Before the first use of the test drug, the functional level of vital organs must meet the following requirements (no blood components, hematopoietic stimulating factors, cell growth factors, whitening drugs, platelet increasing drugs, etc. are allowed to be used within 7 days before obtaining laboratory tests):

1) Absolute neutrophil count ≥ 1.5×10 ⁹ /L;

2) Platelets ≥ 100×10 ⁹ /L;

3) Hemoglobin ≥ 90g/L;

4) Serum albumin ≥ 30g/L;

5) AST and ALT ≤ 2.5 × upper limit of normal reference value (ULN); if accompanied by liver metastasis, ALT and AST ≤ 5 × ULN;

6) Total bilirubin ≤ 1.5×ULN (≤3×ULN for Gilbert syndrome);

7) Serum creatinine ≤ 1.5 × ULN, if the patient’s creatinine level > 1.5 × ULN, the creatinine clearance rate (CLcr) calculated by the Cockcroft-Gault equation is ≥ 50 mL/min;

8) Cardiac left ventricular ejection fraction (LVEF) > 50%;

9) Proteinuria <2+ (when urine protein is ≥2+, 24h urine protein quantification should be performed, and <1g can be selected);

10) International normalized ratio (INR) ≤ 1.5 × ULN or prothrombin time (PT) prolongation ≤ 1.5 × ULN; activated partial thromboplastin time (APTT) ≤ 1.5 × ULN.

Dosing regimen: The initial dose of lenvatinib mesylate is 20 mg or 14 mg or 10 mg, administered orally once a day; the dose of ZPML265 is 5 mg/kg, administered by intravenous infusion once every three weeks.

Evaluation indicators: The endpoints of this study are safety and efficacy indicators and other indicators.

Validity results:

Eleven subjects who received ZPML265 combined with lenvatinib received at least one efficacy evaluation. In the best efficacy evaluation results, ORR was 54.5%, DCR was 100%, and the efficacy trend was significant.

Security Results:

Among the 13 patients with advanced RCC who received ZPML265 combined with lenvatinib, 11 (84.6%) subjects had TEAEs, and 8 (61.5%) subjects had ≥ grade 3 TEAEs. One TEAE (7.7%) led to drug discontinuation.

Refer to the treatment plan of similar drugs, for example, in a clinical study of pembrolizumab (anti-PD1 antibody) combined with lenvatinib in the first-line treatment of advanced renal cancer, among 352 patients in the pembrolizumab combined with lenvatinib group, 351 cases (99.7% ) had any grade TEAE, and 290 cases (82.4%) had ≥3 grade TEAE. There were 131 (37.2%) TEAEs leading to drug discontinuation. in conclusion:

Overall, the efficacy of ZPML265 combined with lenvatinib in the first-line treatment of advanced RCC is similar to that of similar drugs combined with lenvatinib; in terms of safety, the incidence of all TEAEs is lower than that of similar drugs combined with lenvatinib Incidence of TEAEs in the treatment regimen. That is, while ensuring the effectiveness of the drug, the safety is better than that of similar clinical solutions in the prior art.

The above-described embodiments of the present disclosure are illustrative only, and any skilled in the art will recognize, or can ascertain, numerous equivalents to the specific compounds, materials, and procedures without undue undue experimentation. All such equivalents are within the scope of this disclosure and are encompassed by the claims.

Claims (14)

1. A pharmaceutical composition for treating kidney cancer, which contains an effective amount of anti-CTLA4 and anti-PD1 mixed antibody and lenvatinib, the mixed antibody is composed of two different antibodies encoding anti-CTLA4 and anti-PD1 The nucleic acid is produced by a single host cell, wherein the sequences of the heavy chain HCDR1, HCDR2 and HCDR3 of the anti-CTLA4 antibody are shown in SEQ ID NO: 1, 2 and 3 respectively, and the sequences of the light chain LCDR1, LCDR2 and LCDR3 of the anti-CTLA4 antibody are respectively The sequences of anti-PD1 antibody heavy chain HCDR1, HCDR2 and HCDR3 shown in SEQ ID NO: 4, 5 and 6 are respectively shown in SEQ ID NO: 9, 10 and 11, the sequences of anti-PD1 antibody light chain LCDR1, LCDR2 and LCDR3 Shown in SEQ ID NO: 12, 13, 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, and 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 or 2, 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, and the anti-PD1 antibody 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 or 2, wherein lenvatinib is lenvatinib mesylate.
5. The pharmaceutical composition according to claim 1 or 2, wherein the renal cancer is clear cell renal carcinoma.
6. The pharmaceutical composition according to claim 1 or 2, wherein the renal cancer is metastatic or recurrent renal cell carcinoma.
7. The pharmaceutical composition according to claim 4, the dosage of the mixed antibody is 5mg/kg, administered once every three weeks, administered by intravenous infusion; the initial dosage of lenvatinib mesylate is 20mg or 14mg or 10mg, once a day, orally.
8. A method for treating renal cancer, the method comprising administering to a subject a pharmaceutical composition comprising an effective amount of a mixed antibody of anti-CTLA4 and anti-PD1 and lenvatinib, the mixed antibody is composed of an anti-CTLA4 and anti-PD1 encoding Produced by a single host cell of the nucleic acids of two different anti-PD1 antibodies, wherein the sequences of the anti-CTLA4 antibody heavy chain HCDR1, HCDR2 and HCDR3 are shown in SEQ ID NO: 1, 2, and 3, respectively, and the anti-CTLA4 antibody light chain LCDR1, The sequences of LCDR2 and LCDR3 are shown in SEQ ID NO: 4, 5, and 6, respectively, and the sequences of the anti-PD1 antibody heavy chain HCDR1, HCDR2, and HCDR3 are shown in SEQ ID NO: 9, 10, and 11, respectively, and the anti-PD1 antibody light chain The sequences of LCDR1, LCDR2 and LCDR3 are shown in SEQ ID NO: 12, 13 and 14, respectively.
9. The method according to claim 8, 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, and the anti-CTLA4 antibody The heavy chain variable region sequence of the 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.
10. The method according to claim 8 or 9, 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, and the heavy chain sequence of the anti-PD1 antibody The chain sequence is shown in SEQ ID NO: 19, and the light chain sequence of the anti-PD1 antibody is shown in SEQ ID NO: 20.
11. The method according to claim 8 or 9, wherein lenvatinib is lenvatinib mesylate.
12. The method according to claim 8 or 9, wherein the renal cancer is clear cell renal carcinoma.
13. The method according to claim 8 or 9, wherein the renal cancer is metastatic or recurrent renal cell carcinoma.
14. The method according to claim 11, the dosage of the mixed antibody is 5mg/kg, administered once every three weeks, administered by intravenous infusion; the initial dosage of lenvatinib mesylate is 20mg or 14mg or 10mg, Once a day, oral administration.