WO2023185720A1

WO2023185720A1 - Pharmaceutical composition containing mixed antibody of anti-ctla4 and anti-pd1 and therapeutic use thereof

Info

Publication number: WO2023185720A1
Application number: PCT/CN2023/084041
Authority: WO
Inventors: 房良华; 汪华媛
Original assignee: 齐鲁制药有限公司
Priority date: 2022-03-28
Filing date: 2023-03-27
Publication date: 2023-10-05

Abstract

Provided is a use of a combination chemotherapy agent containing a mixed antibody of anti-CTLA4 and anti-PD1, for example, carboplatin/cisplatin and etoposide, in treating small cell lung cancer.

Description

Pharmaceutical compositions containing mixed antibodies against CTLA4 and anti-PD1 and their therapeutic uses

Technical field

The present disclosure relates to the treatment of cancer, particularly 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 to treat small cell lung cancer.

Background technique

According to GLOBOCAN 2020 data, lung cancer ranks second in cancer incidence globally, with 2.2 million cases, and ranks first in death incidence, with 1.8 million deaths from lung cancer worldwide. In China, both the morbidity and mortality rates rank first, with the number of new lung cancer cases and deaths reaching 820,000 and 710,000 respectively.

Small cell lung cancer (SCLC) accounts for approximately 15% of all lung cancer patients, and approximately 2/3 of SCLC patients are in extensive-stage SCLC (ES-SCLC) at the time of onset. There are approximately 110,000 new cases of small cell lung cancer in China every year, including more than 60,000 new cases of extensive-stage small cell lung cancer.

ES-SCLC is sensitive to chemotherapy, with an objective response rate as high as 40-70%. However, the duration of response is short, with a median PFS of less than 6 months, an mOS of less than 12 months, a five-year survival rate of no more than 6%, and a very poor prognosis. Although the combined use of immunotherapy on the basis of chemotherapy can prolong the overall survival of ES-SCLC, the efficacy is still poor. Currently, the immunosuppressive drugs already on the market in China for the first-line treatment of extensive-stage small cell lung cancer include atezolizumab, durvalumab, and slulimumab, but atezolizumab and durvalumab are Rilumab is expensive and has poor drug accessibility. Pharmacoeconomic analysis shows that atezolizumab combined with chemotherapy or durvalumab combined with chemotherapy is not cost-effective compared with chemotherapy alone for the first-line treatment of ES-SCLC. Advantages: There are still few first-line treatments for ES-SCLC available in China, and there is an urgent need to develop drugs with better efficacy to enrich the treatment landscape and prolong the survival of patients.

ZPML265 is a mixed antibody pharmaceutical preparation, which is a dual-target immunity (referred to as dual-immunity) therapeutic drug. It consists of a recombinant humanized IgG1 monoclonal antibody targeting human CTLA4 and a recombinant humanized IgG4 monoclonal antibody targeting human PD1. Composed of two different antibodies produced by a single host cell. This mixed antibody can specifically bind to CTLA4 and PD1 at the same time, thereby blocking the two immune checkpoint signaling pathways of CTLA-4 and B7-1/B7-2, as well as PD-1 and PD-L1, and releasing the impact of the two pathways on T lymphocytes. The inhibitory effect of cells restores their functional activity and anti-tumor immune response, thereby enabling the body to achieve the purpose of fighting and killing tumors.

Although combined with anti-PD-(L)1 immunotherapy on the basis of chemotherapy, the overall survival of ES-SCLC can be prolonged, but the efficacy is still poor, and there is still a very urgent clinical need. It is expected that more and better drugs will be used for the treatment of advanced stage patients. Longer survival benefit for SCLC patients.

Summary of the invention

The technical problem to be solved by this disclosure is to provide an immunotherapy to fill the gap in the international dual-target immunity (double immunity) Therapeutics address clinical gaps in small cell lung cancer, thereby addressing such unmet clinical needs.

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

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

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

In some embodiments, the additional therapeutic agent is a chemotherapeutic agent, preferably a combination of carboplatin/cisplatin and etoposide.

Therefore, the present disclosure also preferably provides a pharmaceutical composition for treating small cell lung cancer, which contains an effective amount of an anti-CTLA4 and anti-PD1 mixed antibody and a chemotherapeutic drug.

The present disclosure also particularly preferably provides a pharmaceutical composition for treating small cell lung cancer, which contains an effective amount of a mixed antibody against CTLA4 and anti-PD1, carboplatin/cisplatin, and etoposide.

In some embodiments, the mixed antibodies are produced from 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 chains HCDR1, HCDR2 and HCDR3 are respectively SEQ. ID NO: 1, 2, and 3, the sequences of the anti-CTLA4 antibody light chain LCDR1, LCDR2, and LCDR3 are respectively SEQ ID NO: 4, 5, and 6, and the sequences of the anti-PD1 antibody heavy chain HCDR1, HCDR2, and HCDR3 are respectively They are shown in SEQ ID NO: 9, 10, and 11, and the sequences of the anti-PD1 antibody light chain LCDR1, LCDR2, and LCDR3 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 set forth in SEQ ID NO:7, the light chain variable region sequence of the anti-CTLA4 antibody is set forth in SEQ ID NO:8, and the heavy chain sequence of the anti-PD1 antibody is set forth in SEQ ID NO:7. 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 SEQ ID NO: 17, the light chain sequence of the anti-CTLA4 antibody is 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 small cell lung cancer is extensive stage small cell lung cancer.

Wherein, the dosage of the mixed antibody is 5 mg/kg, administered once every three weeks, administered by intravenous infusion on the first day, and 21 days constitute a cycle.

And, the dose of chemotherapy drug carboplatin is the target AUC 5mg/min/mL, or the dose of cisplatin is 75mg/m ² , administered by intravenous infusion on the first day; the dose of etoposide is 100mg/ ^m2 , administered on the 1st and 2nd days. , intravenous infusion for 3 days, with a cycle of 21 days.

Furthermore, after 4-6 treatment cycles, the mixed antibody is still administered as a maintenance treatment once every 3 weeks at a dose of 5 mg/kg, administered by intravenous infusion on the first day, with a cycle of 21 days until disease progression.

At the same time, the present disclosure also provides a method for treating small cell lung cancer, which method includes administering a pharmaceutical composition to a subject, and the pharmaceutical composition is the aforementioned mixed antibody containing an effective amount of anti-CTLA4 and anti-PD1.

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

The present disclosure also particularly preferably provides a method for treating small cell lung cancer, which method includes administering to a subject a pharmaceutical composition, the pharmaceutical composition being the aforementioned mixed antibody containing an effective amount of anti-CTLA4 and anti-PD1, Carboplatin/cisplatin and etoposide.

The results of a phase I clinical study of the disclosed mixed antibody as a single drug showed that the 26 patients with small cell lung cancer enrolled in the group had accurate efficacy and good safety. At the same time, the clinical research based on the mixed antibody combined with chemotherapy of the present disclosure in small cell lung cancer has also initially shown a synergistic anti-tumor effect and demonstrated superior clinical efficacy. Therefore, the combination of mixed antibodies or mixed antibodies and chemotherapy drugs (such as carboplatin/cisplatin + etoposide) provided by the present disclosure is expected to have controllable safety and patient compliance for medical treatment of extensive-stage small cell lung cancer. It is good and can fill the gap in the world that no dual-target immunotherapy drugs have been approved for small cell lung cancer to a large extent, and solve the urgent unmet clinical needs.

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. Enter the general.

Before the present disclosure is described in detail below, it is to be understood that this disclosure is not limited to the specific methodologies, protocols, and reagents described herein, as these may vary. It should also 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 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 include numerical ranges, and certain aspects of the disclosure may be described in terms of ranges. Unless otherwise stated, it should be understood that numerical ranges or range descriptions are only for the purpose of simplicity and convenience and should not be considered to strictly limit the scope of the present disclosure. Accordingly, descriptions expressed in range terms should be considered to specifically disclose all possible subranges and all possible specific numerical points within such ranges, as if such subranges and numerical points were expressly written herein. The above principles apply equally regardless of the width of the values stated. When a range description is used, the range includes the endpoints of the range.

When referring to measurable values such as amounts, temporary durations, etc., the term "about" is meant to include ±20%, or in some cases ±10%, or in some cases ±5%, or in some cases ±20% of the specified value. A variation of ±1% in some cases, or ±0.1% in some cases.

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

As used herein, the term "antibody" typically refers to an antibody containing two heavy (H) polypeptide chains (HC) and two light (L) polypeptide chains (LC) held together by covalent disulfide bonds and non-covalent interactions. ) Y-type 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 contains a variable knot domain (VH) and constant region (CH).

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, such as IgG can be divided into IgG1, IgG2, IgG3, IgG4, 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 types, termed kappa and lambda, based on the amino acid sequence of their constant domains.

The term "variable region" or "variable domain" shows significant variation in the amino acid composition from one antibody to another and is primarily responsible for antigen recognition and binding. The variable regions of each light/heavy chain pair form the antibody binding site, such that a complete IgG antibody has two binding sites (i.e. 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 the hypervariable region (HVR), or more commonly, as Complementarity determining region (CDR), VH and VL each have 4 framework regions FR (or framework regions), represented by FR1, FR2, FR3, and FR4 respectively. Therefore, CDR and FR sequences typically occur in the following sequence of the heavy chain variable domain (VH) (or light chain variable domain (VL)): FR1-HCDR1(LCDR1)-FR2-HCDR2(LCDR2)-FR3 -HCDR3(LCDR3)-FR4.

Types of "antibodies" in a broad sense can 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 Mutated proteins and their variants), etc.

The term "monoclonal antibody" (or "monoclonal antibody") refers to a substantially homogeneous antibody produced by a single cell clone and directed only 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 the above technologies.

It should be noted that the division of CDRs and FRs in the antibody variable region of the present disclosure is determined based on 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 nomenclature system based on the antibody sequences of the disclosure are expressly within the scope of the disclosure.

The term "antigen" refers to a substance 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 the selected target, including single epitopes, multiple epitopes, and single structures. 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 contain 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 antibodies" refers to cells containing DNA from a host that has been transfected with DNA encoding at least two different antibodies with different binding specificities, optionally full-length primate IgG antibodies A limited number of major 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, the host cell can be genetically modified with DNA encoding at least two but no more than four different light chains (LC). 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 changing the amino acid sequence of the antibody to disfavor non-homologous HC/LC pairing and strongly favor homologous HC/LC pairing. Where two different HCs are introduced into a host cell, one or both of the two different HCs can optionally be altered such that formation of heterodimers is not favored. 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 homology is favored. HC/LC pairing, whereas the other antibody did not contain such changes. In some embodiments, the host cell produces only two major antibody species, where each HC primarily pairs with its cognate LC, and most antibodies are those containing two heavy chains with the same amino acid sequence and two heavy chains with the same amino acid sequence. Tetramer of light chains (see PCT/US2017/030676).

The term "pharmaceutical composition" refers to a preparation or a combination of preparations containing one, two or more active ingredients, which allows the active ingredients contained therein to be present in a biologically active form and which does not contain any chemicals necessary for administration. Additional components of the formulation have unacceptable toxicity to subjects. When a "pharmaceutical composition" exists in the form of a combination of separate preparations containing two or more active ingredients, it can be administered simultaneously, sequentially, separately or at intervals. The purpose is to exert the biological activity of the multiple active ingredients and use them together. For treating diseases.

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 that dose of a pharmaceutical formulation containing an active ingredient of the present disclosure that produces the desired effect in the patient being treated when administered to the patient in single or multiple doses. The effective amount can be readily determined by the attending physician, who is one of skill in the art, by considering 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 dosing regimen selected; and the use of any concomitant therapy.

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

The term "transfection" as used herein refers to the introduction of exogenous nucleic acid into a eukaryotic cell. Transfection can be achieved by various means known in the art, including electroporation, microinjection, liposome fusion, etc.

The terms "nucleic acid molecule encoding", "encoding DNA sequence" and "encoding DNA" refer to the sequence of deoxyribonucleotides along a deoxyribonucleic acid chain. The order of these deoxyribonucleotides determines the order of the 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 available in the art, such as Cold Spring Harbor's Antibody Experimentation Technical Guide, Chapters 5-8 and 15. Engineered antibodies of the present disclosure, or antigen-binding fragments thereof, may 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 were expanded in serum-free medium in bioreactors to produce antibodies. The culture medium secreting antibodies can be purified and collected using conventional techniques. Antibodies can be filtered and concentrated using conventional methods. Soluble mixtures and polymers can also be removed by conventional methods, such as molecular sieves and ion exchange.

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

As used herein, the terms "disease" or "disorder" or "disorder" or the like refer to any change or disorder that impairs or interferes with the normal function of a cell, tissue or organ. For example, the "disease" includes but is not limited to: tumors, pathogenic infections, autoimmune diseases, T cell dysfunction diseases, or immune tolerance defects (such as transplant rejection), etc.

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

The term "treatment" as used herein refers to clinical intervention in an attempt to modify an individual or to treat a cell-induced disease process, either prophylactically or in the clinical pathological process. Therapeutic effects include, but are not limited to, preventing the occurrence or recurrence of the disease, alleviating symptoms, reducing the direct or indirect pathological consequences of any disease, preventing metastasis, slowing down the progression of the disease, improving or alleviating the condition, alleviating or improving the prognosis, etc.

As used herein, the term "combination" refers to a treatment regimen that provides at least two or more different therapies to achieve a specified therapeutic effect, and the therapies may be either physical, such as radiation therapy, or chemical, such as administration of Subject drugs, the drugs also include combination drugs. "Combination drug" refers to a combination of two or more pharmaceutical preparations each having active ingredients, which need to be used together when administered to a subject. The active ingredients can be mixed together to form a single dosing unit, or they can be formed into separate dosing units and used separately; during administration, different pharmaceutical preparations can be administered substantially simultaneously, simultaneously or sequentially.

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

Progression-free survival (PFS) refers to the time from the start of study treatment to the first imaging evaluation of 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, the first occurrence of PR or CR requires additional imaging to confirm the lesion at ≥4 weeks.

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

Complete response (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 <10 mm.

Partial response (PR): The sum of target lesion diameters is reduced by at least 30% from baseline.

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

Stable disease (SD): The reduction of the target lesion does not reach the PR level, and the increase does not reach the PD level. It is somewhere 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 from when the tumor was first assessed as CR or PR (whichever was recorded first) to when the tumor was first assessed as PD or death.

Treatment-emergent adverse events (TEAE) refer to adverse events that occur during treatment.

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

Serious adverse events (SAE) refer to death, life-threatening, permanent or severe disability or loss of function after the subject receives the investigational drug, the subject needs hospitalization or prolonged hospitalization, and adverse events such as congenital anomalies or birth defects. Medical events.

Example

The present disclosure will be further described below with reference to specific embodiments. 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.

Example 1 Obtaining Mixed Antibody ZPML265

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

Table 1 Amino acid sequence of each component of mixed antibody ZPML265

Example 2 Effectiveness and safety of ZPML265 monotherapy in the treatment of extensive-stage small cell lung cancer

Administration:

In a phase I clinical study conducted in China, 26 patients with pathologically confirmed extensive-stage small cell lung cancer were enrolled. Over 18 years of age, Eastern Cooperative Oncology Group (ECOG) performance status score of 0 or 1, expected survival ≥3 months, functional level of vital organs before first use of 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) (in patients with liver cancer or liver metastasis, ≤ 5 × ULN is allowed);

6) Total bilirubin ≤1.5×ULN (patients with Gilbert syndrome are allowed ≤3×ULN);

7) International normalized ratio (INR) ≤ 1.5; activated partial thromboplastin time (APTT) ≤ 1.5 × ULN;

8) Serum creatinine ≤1.5×ULN. If the subject’s creatinine level is >1.5×ULN, the creatinine clearance (CrCl) must be ≥50mL/min (calculated according to the Cockcroft-Gault formula);

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

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

Effectiveness results:

Efficacy data obtained from ZPML265 monotherapy in 26 patients with extensive-stage small cell lung cancer: the confirmed ORR was 23.1% and the DCR was 34.6%. There were 12 cases of SCLC who had received ≥1 line of treatment and had not received immunotherapy. Among them, the best efficacy evaluation result of 3 subjects was PR and 2 cases were SD. The confirmed ORR was 25.0% and the confirmed DCR was 41.7%. Among the 11 cases of SCLC who had received ≥1 line of treatment and immunotherapy in the past, the best efficacy evaluation results of 2 subjects were PR and 1 case was SD. The confirmed ORR was 18.2% and the confirmed DCR was 27.3%. This study shows the promising therapeutic effect of ZPML265 single agent in the treatment of SCLC.

Safety results:

Safety data summarized data from 609 subjects treated with ZPML265. As of May 31, 2022, 546 (89.7%) patients had experienced TEAEs, and 451 (74.1%) patients had experienced TEAEs related to ZPML265. Grade ≥3 TEAEs and SAEs were reported in 187 (30.7%) and 156 (25.6%) patients, respectively, and grade ≥3 TEAEs and SAEs related to ZPML265 were reported in 102 (16.7%) and 75 (12.3%) patients. SAE related to ZPML265.

Among them, TRAEs with an incidence of ≥10% include: rash (17.6%), pruritus (12.3%), hypothyroidism (11.3%), hyperthyroidism (10.3%), anemia (10%), and aspartate Acid aminotransferase was elevated (10%). The incidence of grade ≥3 TRAEs was 16.7%, and TRAEs with an incidence ≥1% included anemia (2.3%), increased aspartate aminotransferase (1.1%), and increased alanine aminotransferase (1.0% ).

Compared with ZPML265 monotherapy, another meta-analysis that summarized 2664 cases of Nivolumab combined with Ipilimumab showed that the overall treatment-related serious adverse event rate was 29.6%, and the incidence of grade ≥3 TRAE was 39.9%, which was higher than Data related to ZPML265 monotherapy.

It can be seen from the existing data that the safety of the treatment is acceptable, and no new safety signals were found compared with other immunotherapy drugs.

in conclusion:

Overall, ZPML265 monotherapy has shown promising therapeutic effects in the treatment of extensive-stage small cell lung cancer. In terms of safety, the incidence of TRAEs is similar to that of anti-PD-(L)1 monotherapy and lower than that of Nivolumab+Ipilimumab. The treatment, that is, the safety is better than the existing clinical solutions in the current technology while ensuring the effectiveness of the drug.

Example 3 A phase II clinical study evaluating ZPML265 combined with carboplatin and etoposide in the first-line treatment of extensive-stage small cell lung cancer

In the Phase II clinical study of ZPML265 combined with carboplatin and etoposide as first-line treatment of extensive-stage small cell lung cancer, patients with histologically or cytologically confirmed extensive-stage small cell lung cancer were enrolled. Patients are aged ≥18 years and ≤75 years old, regardless of gender. ECOG performance status score 0-1. Expected survival ≥12 weeks. The functional level of vital organs must meet the following requirements before using the investigational drug for the first time:

1) Blood routine (no blood transfusion or correction with hematopoietic stimulating factor drugs within 14 days before obtaining the laboratory test): white blood cell count ≥3.0×10 ⁹ /L; absolute neutrophil count ≥1.5×10 ⁹ /L; platelets ≥100×10 ⁹ /L; hemoglobin ≥90g/L;

2) Liver function: aspartate aminotransferase ≤2.5×ULN in subjects without liver metastasis; alanine liver aminotransferase ≤2.5×ULN; ALT and AST in subjects with liver metastasis ≤5×ULN; serum Total bilirubin ≤1.5×ULN (if Gilbert syndrome, total bilirubin ≤3.0 mg/dL);

3) Renal function: serum creatinine ≤1.5×ULN or creatinine clearance ≥50mL/min;

4) Coagulation function: international normalized ratio ≤ 1.5 × ULN, activated partial thromboplastin time ≤ 1.5 × ULN (only applicable to patients who are not currently receiving anticoagulant therapy. Patients who are currently receiving anticoagulant therapy should receive a stable dose of anticoagulant therapy);

5) Others: lipase ≤ 1.5 × ULN (if lipase is > 1.5 × ULN without clinical or imaging evidence of pancreatitis, you can be included); amylase ≤ 1.5 × ULN ( if amylase > 1.5 × ULN without clinical or imaging evidence of pancreatitis, you can be included) Patients with pancreatitis confirmed by imaging studies can be enrolled); alkaline phosphatase ≤2.5×ULN, subjects with liver metastasis or bone metastasis, ALP ≤5×ULN.

Dosing regimen:

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

The dose of carboplatin is the target AUC of 5mg/min/mL, administered by intravenous infusion on the 1st day, and the dose of etoposide is 100mg/m ² , administered by intravenous infusion on the 1st, 2nd and 3rd days, with a cycle of 21 days. Dosing for 4-6 cycles.

Effectiveness results:

ZPML265 single agent combined with chemotherapy was used to treat 40 patients with extensive-stage small cell lung cancer. The efficacy analysis was based on 39 evaluable patients. As of January 16, 2023, the confirmed ORR was 89.7% (35/39) and 35 patients (92.1%). The best overall tumor assessment of the subjects was PR, 3 subjects (7.7%) subjects had the best overall tumor assessment of SD, 1 subject (2.6%) was PD, the overall DCR was 97.4% (38/39), 3 subjects The monthly PFS rate was 94.8%, and the median PFS was 5.7 months (95% confidence interval: 5.4-7.1 months).

The current ORR of ZPML265 combined with chemotherapy is better than the PD-(L)1 inhibitor + chemotherapy data in similar studies (atezolizumab vs. chemotherapy ORR: 60.2%, durvalumab vs. chemotherapy ORR: 68%).

Safety results:

All 40 subjects received at least one dose of ZPML265 treatment. As of January 16, 2023, the median treatment duration of ZPML265 was 25.64 weeks (range: 3.0-38.9 weeks). 40 subjects entered the safety analysis set. (100%) all experienced at least one TEAE, with 80% of patients experiencing an adverse event related to ZPML265. TRAEs with an incidence of ≥30% were anemia (45%), decreased platelet count (35.0%), decreased neutrophil count (30.0%), decreased white blood cell count (30.0%), and nausea (30.0%). Among them, 15 patients (37.5%) experienced grade ≥3 TEAEs, of which TRAEs with an incidence rate of ≥10% were decreased neutrophil count (20.0%) and platelet count (10.0%). There were no TRAEs resulting in death.

No TEAEs occurred that resulted in permanent discontinuation of ZPML265 or chemotherapy.

The safety data are comparable to those of single-target immune (single-immune) drugs combined with chemotherapy. Common adverse events are hematological toxicity, which is a common toxicity of the chemotherapy drugs etoposide and carboplatin. The grade 3-4 event rate was 90%, similar to the rate with slulimab in the ASTRUM-005 study (82.5%).

in conclusion:

Overall, ZPML265 combined with chemotherapy has shown promising therapeutic effects in the first-line treatment of extensive-stage small cell lung cancer. In terms of safety, the incidence of grade 3-4 TRAEs is similar to that of anti-PD-)L)1 monotherapy, that is, it ensures that the drug The efficacy and safety profile are comparable to existing clinical protocols in the state of the art.

Example 4 Phase III clinical study evaluating ZPML265 combined with carboplatin/cisplatin and etoposide in the first-line treatment of extensive-stage small cell lung cancer

In the Phase III clinical study of ZPML265 combined with carboplatin/cisplatin and etoposide as first-line treatment of extensive-stage small cell lung cancer, patients with histologically or cytologically confirmed extensive-stage small cell lung cancer were enrolled. Patients are aged 18-75 years old, regardless of gender. ECOG performance status score 0-1. Expected survival ≥12 weeks. The functional level of vital organs must meet the following requirements before using the investigational drug for the first time:

(1) Blood routine (no blood transfusion or correction with hematopoietic stimulating factor drugs within 14 days before the laboratory test during the screening period):

a) White blood cell count (white blood cell, WBC) ≥ 4.0×10 ⁹ /L;

b) Absolute neutrophil count (ANC) ≥ 2.0×10 ⁹ L;

c) Platelet (PLT) ≥100×10 ⁹ /L;

d) Hemoglobin content (hemoglobin, HGB) ≥ 9.0g/dL;

(2) Liver function: aspartate aminotransferase (AST) ≤2.5×ULN in patients without liver metastasis; alanine aminotransferase (ALT) ≤2.5×ULN in patients with liver metastasis. , AST ≤ 5 × ULN; serum total bilirubin (TBIL) ≤ 1.5 × ULN (except for Gilbert syndrome, total bilirubin ≤ 3.0 mg/dL);

(3) Renal function: serum creatinine ≤1.5×ULN or creatinine clearance rate (CrCl) ≥50mL/minute, and urine protein <2+. If the urine protein is 2+, you need to additionally test the 24-hour urine protein quantification or collect random urine samples to evaluate the urine protein/creatinine ratio. If the 24-hour urine protein quantification is <1g or the urine protein/creatinine ratio is <2, you can be included in the group;

(4) Coagulation function: international normalized ratio (INR) ≤ 1.5 × ULN, activated partial thromboplastin time (APTT) ≤ 1.5 × ULN (only applicable to those who are not currently receiving anticoagulation therapy Patients (those currently receiving anticoagulant therapy should receive a stable dose of anticoagulant therapy);

(5) Others: alkaline phosphatase (ALP) ≤ 2.5 × ULN. For patients with liver metastasis or bone metastasis, ALP ≤ 5 × ULN.

Dosing regimen:

1) Experimental group: ZPML265+carboplatin/cisplatin+etoposide:

The intensive treatment period is 21 days as a treatment cycle, and the treatment is continued for 4-6 cycles;

ZPML265, 5mg/kg, day 1 (D1), intravenous (IV) infusion, Q3W;

Carboplatin AUC 5 or cisplatin 75 mg/m ² , D1, IV infusion, Q3W;

Etoposide 100 mg/m ² , D1, 2, 3, IV infusion, Q3W;

Patients who did not experience disease progression received ZPML265 single-agent maintenance therapy, 5 mg/kg, D1, IV infusion, Q3W.

2) Control group: adebelimab + carboplatin + etoposide:

Adebelimab, 20 mg/kg, D1, IV infusion, Q3W;

Carboplatin AUC 5, D1, IV infusion, Q3W;

Etoposide 100 mg/m ² , D1, 2, 3, IV infusion, Q3W;

Patients without disease progression received aderbelimab single-agent maintenance therapy, 20 mg/kg, D1, IV infusion, Q3W.

In addition, after the disease progresses, for patients in the experimental group, if the researchers believe that there may still be clinical benefit from continued treatment with ZPML265, ZPML265 treatment can be combined with second-line chemotherapy. That is, patients can choose to enter the following second-line treatments:

Experimental group: chemotherapy + ZPML265, 21 days as a treatment cycle

Control group: chemotherapy, 21 days as a treatment cycle

Chemotherapy regimens were selected according to the latest CSCO guidelines.

The above-described embodiments of the present disclosure are illustrative only, and any person skilled in the art will recognize, or be able to ascertain, numerous equivalents to the specific compounds, materials, and operations without the need for more than routine experimentation. All such equivalents are within the scope of the disclosure and included by the claims.

Claims

A pharmaceutical composition for treating small cell lung cancer, which contains an effective amount of anti-CTLA4 and anti-PD1 mixed antibodies and chemotherapy drugs. The mixed antibody is composed of nucleic acids encoding two different anti-CTLA4 and anti-PD1 antibodies. Produced by a single host cell, in which 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 sequences of the anti-CTLA4 antibody light chain LCDR1, LCDR2 and LCDR3 are respectively SEQ ID NO: 1, 2 and 3. ID NO: 4, 5, and 6, the sequences of the anti-PD1 antibody heavy chain HCDR1, HCDR2, and HCDR3 are respectively SEQ ID NO: 9, 10, and 11, and the sequences of the anti-PD1 antibody light chain LCDR1, LCDR2, and LCDR3 are respectively Shown as SEQ ID NO: 12, 13, 14.
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.
The pharmaceutical composition of 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, and the heavy chain sequence of the anti-PD1 antibody is shown in SEQ ID NO: 17. 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.
The pharmaceutical composition according to any one of claims 1 to 3, wherein the small cell lung cancer is extensive stage small cell lung cancer.
The pharmaceutical composition according to claim 4, wherein the chemotherapy drug is carboplatin/cisplatin and etoposide.
The pharmaceutical composition according to claim 5, during the treatment period: the dosage of the mixed antibody is 5 mg/kg, administered once every three weeks, administered by intravenous infusion on the first day, and 21 days constitute a cycle.
The pharmaceutical composition according to claim 5, during the treatment period: the dose of carboplatin is the target AUC 5mg/min/mL, or the dose of cisplatin is 75mg/ m2 , administered by intravenous infusion on the first day; the dose of etoposide is 100 mg/m 2 , administered by intravenous infusion on days 1, 2, and 3, with a cycle of 21 days.
The pharmaceutical composition according to claim 6 or 7, after 4-6 treatment periods, the mixed antibody maintenance treatment is continued, and the dosage of the mixed antibody is 5 mg/kg, administered by intravenous infusion on the first day, every three Administer once a week until disease progression.
A method for treating small cell lung cancer, the method comprising administering to a subject a pharmaceutical composition containing an effective amount of a mixed antibody against CTLA4 and anti-PD1 and a chemotherapeutic drug, the mixed antibody is composed of a compound encoding an anti-CTLA4 and an anti-PD1 The nucleic acids of two different PD1 antibodies are produced by a single host cell. Among them, the sequences of the anti-CTLA4 antibody heavy chains HCDR1, HCDR2 and HCDR3 are shown in SEQ ID NO: 1, 2 and 3 respectively, and the anti-CTLA4 antibody light chains LCDR1 and LCDR2 The sequences of the anti-PD1 antibody heavy chain HCDR1, HCDR2 and HCDR3 are shown in SEQ ID NO: 9, 10 and 11 respectively. The anti-PD1 antibody light chain LCDR1 The sequences of LCDR2 and LCDR3 are shown in SEQ ID NO: 12, 13 and 14 respectively.
The method of claim 9, 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 is shown in SEQ ID NO: 8. The heavy chain variable region sequence of the PD1 antibody is SEQ ID NO: 15 is shown, and the light chain variable region sequence of the anti-PD1 antibody is shown in SEQ ID NO: 16.
The method of claim 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 It 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 method according to any one of claims 9-11, wherein the small cell lung cancer is extensive-stage small cell lung cancer.
The method of claim 12, wherein the chemotherapy drugs are carboplatin/cisplatin and etoposide.
The method according to claim 13, during the treatment period: the dosage of the mixed antibody is 5 mg/kg, administered once every three weeks, administered by intravenous infusion on the first day, and 21 days constitute a cycle.
The method according to claim 13, during the treatment period: the dose of carboplatin is the target AUC 5mg/min/mL, or the dose of cisplatin is 75mg/ m2 , administered by intravenous infusion on the first day; the dose of etoposide is 100mg/ m 2 , intravenous infusion on days 1, 2, and 3, with a cycle of 21 days.
The method according to claim 14 or 15, after 4-6 treatment periods, the mixed antibody maintenance treatment is continued, and the dosage of the mixed antibody is 5 mg/kg, administered by intravenous infusion on the first day, and administered every three weeks. The drug is given once until the disease progresses.
A pharmaceutical composition for treating extensive-stage small cell lung cancer, which contains an effective amount of a mixed antibody against CTLA4 and anti-PD1. The mixed antibody is composed of nucleic acids encoding two different antibodies against CTLA4 and anti-PD1. Produced by a single host cell, 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 is SEQ ID NO: 19 is shown, and the light chain sequence of the anti-PD1 antibody is SEQ ID NO: 20.