WO2023138574A1 - Drug composition of spiro aryl phosphine oxide and anti-vegf antibody - Google Patents

Abstract

Disclosed in the present invention is a drug composition of an ALK inhibitor and an antibody, the drug composition comprising a spiro aryl phosphine oxide and an anti-VEGF antibody, and the ALK inhibitor being a compound of formula (I) or a pharmaceutically acceptable salt thereof. The drug composition disclosed by the present invention is used for treating lung cancer and shows high anti-tumor activity.

Description

Drug Combination of Spiroaryl Phosphate Oxide and Anti-VEGF Antibody

This application claims the priority of the Chinese patent application with the application number 202210052516.5 filed on January 18, 2022, entitled "Drug Combination of Spiroaryl Phosphate Oxide and Anti-VEGF Antibody", the entire content of which is incorporated in this application by reference.

technical field

The invention belongs to the field of medicinal chemistry and relates to a combined pharmaceutical composition for treating tumors. Specifically, the present invention relates to the drug combination of spirocyclic aryl phosphorus oxide and antibody and its use in the preparation of antitumor drugs.

Background technique

Anaplastic lymphoma kinase (ALK) is a receptor tyrosine kinase (RTK) and a member of the insulin receptor (IR) superfamily. In 1994, ALK was first discovered in 60-80% of anaplastic large cell lymphoma (ALCLS) cell lines in the form of fusion protein NPM (nucleophosmin)-ALK. NPM-ALK was caused by t(2;5) chromosome translocation. Although the physiological function of ALK in cancer is unclear, ALK fusion proteins have been identified in various human cancers such as breast cancer, colorectal cancer, inflammatory myofibroblastic tumor (IMT), diffuse large B-cell lymphoma (DLBCL), and most notably in non-small cell lung cancer (NSCLC). Therefore, the kinase activity associated with ALK fusion proteins is considered to play a very important role in the survival and proliferation of human cancer cells.

The ALK gene provides a signaling instruction for receptor tyrosine kinases to transmit signals from the cell surface into the cell. This process begins when a kinase is stimulated at the cell surface and then attaches to a similar kinase (dimerization). After dimerization, the kinase is tagged with a phosphate group, a process called phosphorylation. Phosphorylation-activated kinase, an activated kinase is another protein capable of transferring a phosphate group into the cell, activating a series of proteins that continue through the signaling pathway. These signaling pathways are important for many cellular processes, such as cell growth and division (proliferation) or maturation (differentiation).

Often ALK is rearranged such that the tyrosine kinase domain of ALK is fused to the 5′-terminal domain of another protein, such as echinoderm microtubule-associated protein-like 4 (EML4) in NSCLC or nucleophosphoprotein (NPM) in anaplastic large cell lymphoma. The breakpoint of all ALK fusion genes is very conserved, located in the upstream kinase domain of the exon-encoded intragenic region. Since the part of the rearrangement involved in ALK does not include the transmembrane domain, the resulting fusion protein re-migrates from the cell membrane to the cytoplasm. The 5'-end of the fusion protein usually contains a coiled-coil or leucine zipper domain, which oligomerizes the fusion protein and leads to ligand-dependent activation of the ALK tyrosine kinase. This in turn constitutively activates downstream signaling pathways such as Ras/MAPK, PI3K/AKT, and JAK/STAT. ALK-driven lung cancers respond and regress following treatment with ALK small-molecule tyrosine kinase inhibitors. This finding demonstrates "oncogene dependency," whereby cancer cells become dependent on oncogenic driver genes and are thus highly sensitive to suppressing oncogenes.

Crizotinib is the first ALK inhibitor approved by the FDA for the treatment of ALK-positive lung cancer. Although patients initially responded very well to crizotinib therapy, most patients relapsed during the first year of treatment due to the development of drug resistance. In April 2014, the FDA approved Ceritinib for the treatment of anaplastic lymphoma kinase (ALK)-positive metastatic non-small cell lung cancer (NSCLC), including patients who are effective and resistant to crizotinib. However, drug resistance always occurs with the prolongation of treatment time, so that the drug loses its effectiveness.

Spirocyclic aryl phosphine oxide, chemical name: (2-((5-chloro-2-((2-methoxy-4-(9-methyl-3,9-diazaspiro[5.5]undec-3-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)dimethylphosphine oxide, is a novel ALK/ROS1 inhibitor for the treatment of ALK/ROS1 positive non-small cell lung cancer. It has the following structural formula:

The compound of formula I inhibits the phosphorylation of ALK by inhibiting the activation of ALK kinase, thereby preventing the phosphorylation of downstream signaling molecules, Such as ERK, STAT5 and AKT. In addition, the compound of formula I inhibits the activation of ROS1 by inhibiting the phosphorylation of ROS1 fusion kinase, thereby preventing the phosphorylation of downstream signaling molecules such as ERK or AKT. Ultimately inhibits ALK or ROS1 kinase-driven tumor growth by inhibiting kinase phosphorylation. Related patent documents include: patent WO2016/000581 discloses spirocyclic aryl phosphorus oxides as ALK inhibitors; patent CN106928275 discloses the preparation method and intermediates of spirocyclic aryl phosphorus oxides; patent CN110407877 discloses polymorphic forms of spirocyclic aryl phosphorus oxides.

As early as 1971, it was discovered that tumor angiogenesis plays a vital role in the growth, development and metastasis of tumors. Therefore, the inhibition of abnormal angiogenesis has become a new focus of cancer treatment, and its generation is the result of the interaction between the tumor and the environment. There are many influencing factors, among which vascular endothelial growth factor (VEGF) is the main relevant mediator. VEGF promotes the proliferation and migration of abnormal tumor vascular endothelial cells and increases the permeability of abnormal blood vessels by binding to VEGF receptor (VEGFR), while anti-angiogenic agents can prune and normalize tumor neovascularization. Angiogenesis is an extremely complex process that requires the participation of vasculature, circulating endothelial cells, and pro-angiogenic mediators including VEGF. Among them, VEGF/VEGFR is an important signal transduction pathway. VEGF combines with VEGFR-1 and VEGFR-2 expressed in tissue epithelial cells to activate intracellular signal transduction pathways, leading to vascular endothelial cell proliferation and neovascularization. The expression of VEGF is related to the progression, stage, ascites formation, shortened disease-free survival time and poor prognosis of malignant tumors. Immunohistochemical examination found that VEGF was expressed in the tumor and its metastatic tissues, and could be detected in malignant ascites and serum. The expression of VEGF and VEGFR detected in tumor patients is an independent prognostic factor.

Bevacizumab (AVASTIN ^® ) is a recombinant humanized immunoglobulin G1 (IgG1) monoclonal antibody that can bind to VEGF-A, inhibit its binding to VEGF receptor-2 (VEGFR-2), and then inhibit the biological effects of VEGF, including affecting the permeability and proliferation of blood vessels, as well as the migration and survival of endothelial cells, so as to inhibit tumor angiogenesis, growth and metastasis. In addition, some other possible mechanisms of action of bevacizumab are being extensively investigated. Compared with single administration, the combination of bevacizumab and chemotherapy drugs can improve the anti-tumor efficacy, which may benefit from the fact that bevacizumab can reduce the pressure of intratumoral tissue space, thereby enhancing the penetration of chemotherapy drugs inside the tumor. At present, bevacizumab has been widely used in the treatment of colorectal cancer, lung cancer, ovarian cancer, cervical cancer, glioblastoma and other tumors worldwide.

The biggest challenge encountered in the process of tumor treatment is the poor efficacy of drugs due to tumor immune tolerance and escape. Therefore, the combination of small molecule anti-tumor compounds and anti-VEGF antibodies to treat tumors has important application value.

Contents of the invention

The purpose of the present invention is to provide a combination pharmaceutical composition, which comprises ALK inhibitor and anti-VEGF antibody. Specifically, the combined pharmaceutical composition of the present invention comprises a compound of formula (I) or a pharmaceutically acceptable salt thereof as an ALK inhibitor, and an anti-VEGF antibody.

In some aspects of the present invention, the anti-VEGF antibody in the above combined pharmaceutical composition is bevacizumab.

In some schemes of the present invention, the bevacizumab in the above-mentioned combination pharmaceutical composition may be AVASTIN ^® , Agoda ^® .

In some aspects of the present invention, the above-mentioned combined pharmaceutical composition is a non-fixed combination. In some aspects, the anti-VEGF antibody and the compound of Formula I, or a pharmaceutically acceptable salt thereof, in the non-fixed combination are each in the form of a pharmaceutical composition. In some aspects, there is also provided a kit of a combined pharmaceutical composition for treating lung tumors, which contains (a) a first pharmaceutical composition comprising an anti-VEGF antibody; and (b) a second pharmaceutical composition comprising a compound of formula I as an active ingredient.

The present invention also provides an application of a combined pharmaceutical composition in the preparation of a drug for treating and/or preventing lung cancer, the pharmaceutical composition comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof as an ALK inhibitor, and an anti-VEGF antibody.

In some aspects of the invention, the anti-VEGF antibody in the above use is bevacizumab.

In some schemes of the present invention, bevacizumab (bevacizumab) in the above-mentioned application can be

In some aspects of the present invention, the anti-VEGF antibody and the compound of formula I or a pharmaceutically acceptable salt thereof in the above application are each in the form of a pharmaceutical composition.

In some aspects of the present invention, the lung cancer in the above application is positive for ALK and/or ROS1 fusion gene.

In some aspects of the present invention, the lung cancer mentioned in the above application is early-middle stage, locally advanced, advanced metastatic non-small cell lung cancer.

In some aspects of the present invention, the lung cancer in the above application is early-middle stage, locally advanced, advanced metastatic non-small cell lung cancer that is positive for ALK and/or ROS1 fusion gene.

In some aspects of the present invention, the lung cancer in the above application is non-small cell lung cancer that has not been treated with ALK and/or ROS1 inhibitors or failed to be treated with ALK and/or ROS1 inhibitors.

In some aspects of the present invention, the non-small cell lung cancer in the above application is adenocarcinoma, squamous cell carcinoma, adenosquamous cell carcinoma or large cell carcinoma.

In some aspects of the present invention, the anti-VEGF antibody and the compound of formula I or a pharmaceutically acceptable salt thereof in the above application are each in the form of a pharmaceutical composition, and can be administered simultaneously, sequentially or at intervals.

The invention also provides a method for treating a subject suffering from cancer or tumor comprising administering to said subject a therapeutically effective amount of an ALK inhibitor and a therapeutically effective amount of an anti-VEGF antibody.

In some aspects of the present invention, the ALK inhibitor in the above treatment method is a spirocyclic aryl phosphorus oxide represented by formula I.

In some aspects of the invention, the anti-VEGF antibody in the above method of treatment is bevacizumab.

In some schemes of the present invention, bevacizumab (bevacizumab) in the above-mentioned treatment method can be

In some aspects of the present invention, the anti-VEGF antibody and the compound of formula I or a pharmaceutically acceptable salt thereof in the above treatment methods are each in the form of a pharmaceutical composition.

In some aspects of the present invention, the lung cancer in the above treatment method is positive for ALK and/or ROS1 fusion gene.

In some schemes of the present invention, the lung cancer mentioned in the above treatment method is early-middle stage, locally advanced, advanced metastatic non-small cell lung cancer.

In some schemes of the present invention, the lung cancer described in the above treatment method is ALK and/or ROS1 fusion gene-positive early-middle stage, locally advanced, advanced metastatic non-small cell lung cancer.

In some aspects of the present invention, the lung cancer in the above treatment method is non-small cell lung cancer that has not been treated with an ALK/ROS1 inhibitor or failed to be treated with an ALK/ROS1 inhibitor.

In some aspects of the present invention, the non-small cell lung cancer in the above treatment method is adenocarcinoma, squamous cell carcinoma, adenosquamous cell carcinoma or large cell carcinoma.

In some aspects of the present invention, the anti-VEGF antibody and the compound of formula I or a pharmaceutically acceptable salt thereof in the above treatment method are each in the form of a pharmaceutical composition, and can be administered simultaneously, sequentially or at intervals.

The present invention also provides a use of a compound of formula I or a pharmaceutically acceptable salt thereof for treating lung cancer, wherein the compound of formula I or a pharmaceutically acceptable salt thereof is used in combination with an anti-VEGF antibody.

The present invention also provides the use of an anti-VEGF antibody for treating lung cancer. The anti-VEGF antibody is used in combination with the compound of formula I or a pharmaceutically acceptable salt thereof.

In some aspects, the anti-VEGF antibody for the above uses is selected from bevacizumab.

In some schemes of the present invention, the bevacizumab used above can be

The present invention also provides a pharmaceutical pack comprising a single packaged pharmaceutical composition in separate containers, comprising a pharmaceutical composition comprising a compound of formula I or a pharmaceutically acceptable salt thereof in one container, and comprising a pharmaceutical composition comprising an anti-VEGF antibody in a second container.

Pharmaceutical compositions of compounds of formula I

Pharmaceutical compositions of compounds of formula I can be formulated for specific routes of administration, such as oral administration, parenteral administration, rectal administration, and the like. Oral administration is preferred, eg as a tablet.

In some embodiments of the present invention, the dosage of the pharmaceutical composition of the compound of formula I is 60 mg/time to 180 mg/time, or the dosage is 90 mg/time to 180 mg/time.

In some embodiments of the present invention, according to once-daily administration, the pharmaceutical composition of the compound of formula I is administered about 60 mg-180 mg; or 90 mg-180 mg each time.

In some embodiments of the present invention, the pharmaceutical composition of the compound of formula I is about 60 mg to 180 mg; or 90 mg to 180 mg.

Compound of formula I

As used in the present invention, the compound of formula I is a spirocyclic aryl phosphorus oxide with a chemical name of (2-((5-chloro-2-((2-methoxy-4-(9-methyl-3,9-diazaspiro[5.5]undec-3-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)dimethylphosphine oxide, which is a newly developed highly selective anaplastic lymphoma kinase (ALK) inhibitor, which has the following structural formula:

Related patent documents include: patent WO2016/000581 discloses spirocyclic aryl phosphorus oxides as ALK inhibitors; patent CN106928275 discloses the preparation method and intermediates of spirocyclic aryl phosphorus oxides; patent CN110407877 discloses polymorphic forms of spirocyclic aryl phosphorus oxides.

Bevacizumab

As used in the present invention, bevacizumab is an anti-VEGF antibody, and its sequence and structure can be found in document WO1998/045331. In December 2019, the first domestic bevacizumab produced by Qilu Pharmaceutical Officially approved by the National Medical Products Administration (NMPA) of China for the treatment of patients with advanced, metastatic or recurrent non-small cell lung cancer (NSCLC) and metastatic colorectal cancer. The amino acid sequence of bevacizumab is as follows:

Heavy chain:

Light chain:

Definition and Description

Unless otherwise stated, the following terms and phrases used herein are intended to have the following meanings. A specific term or phrase should not be considered indeterminate or unclear if it is not specifically defined, but should be understood according to its ordinary meaning. When a trade name appears herein, it is intended to refer to its corresponding trade name or its active ingredient.

The term "pharmaceutically acceptable" as used herein refers to those compounds, materials, compositions and/or dosage forms, which are suitable for use in contact with human and animal tissues within the scope of sound medical judgment, without excessive toxicity, irritation, allergic reaction or other problems or complications, commensurate with a reasonable benefit/risk ratio.

The term "pharmaceutically acceptable salt" refers to a salt of a compound of the present invention, which is prepared from a compound having a specific substituent found in the present invention and a relatively non-toxic acid or base. When compounds of the present invention contain relatively acidic functional groups, base addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of base, either neat solution or in a suitable inert solvent. When compounds of the present invention contain relatively basic functionalities, acid addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of the acid, either neat solution or in a suitable inert solvent. The pharmaceutically acceptable salts of the present invention can be synthesized from the parent compound containing acid groups or bases by conventional chemical methods. 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 "antibody" refers to a binding protein having at least one antigen binding domain. Antibodies and fragments thereof of the present invention may be whole antibodies or any fragments thereof. Accordingly, the antibodies and fragments of the invention include monoclonal antibodies or fragments thereof and antibody variants or fragments thereof, as well as immunoconjugates. Examples of antibody fragments include Fab fragments, Fab' fragments, F(ab)' fragments, Fv fragments, isolated CDR regions, single chain Fv molecules (scFv), and other antibody fragments known in the art. Antibodies and fragments thereof may also include recombinant polypeptides, fusion proteins and bispecific antibodies. The anti-PD-L1 antibodies and fragments thereof disclosed herein can be of the IgG1, IgG2, IgG3 or IgG4 isotype.

The term "humanized antibody" refers to an antibody in which the antigen-binding site is derived from a non-human species and the variable region framework is derived from human immunoglobulin sequences. Humanized antibodies may contain substitutions in the framework regions such that the framework may not be an exact copy of expressed human immunoglobulin or germline gene sequences.

The term "monoclonal antibody" ("mAb") refers to an antibody molecule of single molecular composition. Monoclonal antibody compositions exhibit a single binding specificity and affinity for a particular epitope, or, in the case of bispecific monoclonal antibodies, dual binding specificities for two different epitopes. A mAb is an example of an isolated antibody. mAbs can be produced by hybridoma, recombinant, transgenic or other techniques known to those skilled in the art.

The term "VEGF" or "VEGF-A" refers to the 165 amino acid human vascular endothelial growth factor and related 121, 145, 189, and 206 amino acid human vascular endothelial growth factors, e.g., as described by Leung et al. Science, 246:1306 (1989), and Houck et al., Mol. Its naturally occurring allelic and processed forms. VEGF-A is part of a gene family that includes VEGF-B, VEGF-C, VEGF-D, VEGF-E, VEGF-F, and P1GF. VEGF-A mainly binds to two high-affinity receptor tyrosine kinases VEGFR-1 (F1t-1) and VEGFR-2 (F1k-1/KDR), which is the main transmitter of VEGF-A mitotic signal in vascular endothelial cells. Additionally, neuropilin-1 has been identified as a receptor for the heparin-binding isoform of VEGF-A and may play a role in vascular development. The term "VEGF" or "VEGF-A" also refers to VEGFs from non-human species such as mice, rats or primates. Sometimes, VEGF from a particular species is indicated by a term, such as hVEGF for human VEGF or mVEGF for murine VEGF. Typically, VEGF refers to human VEGF. The term "VEGF" is also used to refer to truncated forms or fragments of polypeptides comprising amino acids 8-109 or 1-109 of the 165 amino acid human vascular endothelial growth factor. References to any of said forms of VEGF can be identified in the present invention, for example, by "VEGF(8-109)", "VEGF(1-109)" or "VEGF165". Amino acid positions for "truncated" native VEGF are numbered as indicated in the native VEGF sequence.

The term "anti-VEGF antibody" refers to an antibody that binds VEGF with sufficient affinity and specificity. The antibody selected typically has a binding affinity for VEGF, eg, the antibody can bind hVEGF with a Kd value from 1 pM to 100 nM. For example, antibody affinity can be determined by surface plasmon resonance-based assays such as the BIAcore assay described in PCT Publication WO2005/012359; by enzyme-linked immunosorbent assays (ELISA); and competition assays such as RIA's. In certain embodiments, the anti-VEGF antibodies of the invention can be used as therapeutic agents in targeting and interfering with diseases or disorders in which VEGF activity is implicated. In addition, the antibodies can be tested for other biological activities, eg, to assess their efficacy as therapeutic agents. Such assays are known in the art and depend on the target antigen and the intended application of the antibody. Examples include HUVEC inhibition assays; Tumor cell growth inhibition assays (eg, described in WO89/06692); antibody-dependent cellular cytotoxicity (ADCC) and complement-mediated cytotoxicity (CDC) assays (eg, patent US 5,500,362); and agonist activity or hematopoietic assays (see WO95/27062). Anti-VEGF antibodies generally do not bind other VEGF homologues, such as VEGF-B or VEGF-C, nor do they bind other growth factors, such as P1GF, PDGF, or bFGF.

The term "treatment" generally refers to obtaining a desired pharmacological and/or physiological effect. The effect may be prophylactic in terms of complete or partial prevention of the disease or its symptoms; and/or therapeutic in terms of partial or complete stabilization or cure of the disease and/or side effects due to the disease. "Treatment" as used herein encompasses any treatment of a disease in a patient, including: (a) preventing the disease or symptom in a patient susceptible to the disease or symptom but not yet diagnosed with the disease; (b) inhibiting the symptom of the disease, i.e. preventing its development; or (c) relieving the symptom of the disease, i.e. causing regression of the disease or symptom.

As used herein, the term "subject" refers to mammals, such as rodents, felines, canines, and primates. Preferably, the subject of the invention is a human.

As used herein, the term "administering" means physically introducing a composition comprising a therapeutic agent into a subject using any of a variety of methods and delivery systems known to those skilled in the art. Routes of administration for anti-VEGF antibodies include intravenous, intramuscular, subcutaneous, intraperitoneal, spinal or other parenteral routes of administration, eg, by injection or infusion. As used herein, "parenteral administration" refers to modes of administration other than enteral and topical administration, typically by injection, and includes, but is not limited to, intravenous, intramuscular, intraarterial, intrathecal, intralymphatic, intralesional, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcutaneous, intraarticular, subcapsular, subarachnoid, intraspinal, epidural, and intrasternal injection and infusion, and in vivo electroporation. In certain embodiments, the anti-VEGF antibody is administered non-parenterally, and in certain embodiments, orally. Other non-parenteral routes include topical, epidermal or mucosal routes of administration, eg, intranasal, vaginal, rectal, sublingual or topical. Administration can also be performed, eg, once, multiple times, and/or over one or more extended periods of time.

The term "subject" includes any human or non-human animal. The term "non-human animal" includes, but is not limited to, vertebrates such as non-human primates, sheep, dogs, and rodents such as mice, rats, and guinea pigs. In certain embodiments, the subject is a human. The terms "subject" and "patient" are used interchangeably in certain contexts herein.

A "therapeutically effective amount" or "therapeutically effective dose" of a drug or therapeutic agent is any amount of the drug which, when used alone or in combination with another therapeutic agent, protects a subject from disease onset or promotes regression of disease as evidenced by a reduction in the severity of disease symptoms, an increase in the frequency and duration of disease-free periods, or prevention of impairment or disability resulting from disease affliction. The ability of a therapeutic agent to promote disease regression can be evaluated using a variety of methods known to the skilled practitioner, such as in human subjects during clinical trials, in animal model systems predicting efficacy in humans, or by measuring the activity of the agent in in vitro assays.

A therapeutically effective amount of a drug includes a "prophylactically effective amount," which is any amount of drug that inhibits the onset or recurrence of cancer when administered alone or in combination with an antineoplastic agent to a subject at risk of developing cancer (e.g., a subject with a premalignant condition) or a subject at risk of cancer recurrence. In certain embodiments, the prophylactically effective amount completely prevents the occurrence or recurrence of cancer. "Inhibiting" the occurrence or recurrence of cancer means reducing the likelihood of the occurrence or recurrence of cancer, or preventing the occurrence or recurrence of cancer altogether.

The term "recurrent" cancer is cancer that regenerates at the original site or at a distant site after a response to initial treatment (eg, surgery). A "locally recurrent" cancer is one that, after treatment, arises in the same location as a previously treated cancer.

The term "unresectable" cancer is one that cannot be removed by surgery.

The term "metastatic" cancer refers to cancer that has spread from one part of the body, such as the lungs, to another part of the body.

The term "fixed combination" means that the active ingredients (eg, anti-VEGF antibody or compound of formula I) are administered to a subject simultaneously in a fixed total dose or dose ratio, or in the form of a single entity, pharmaceutical composition or formulation.

The term "non-fixed combination" means that two or more active components are administered to a subject simultaneously, concurrently or sequentially as independent entities (eg, pharmaceutical composition, preparation) without specific time limit, wherein the active components administered to the subject reach a therapeutically effective level. Examples of non-fixed combinations are cocktail therapy, eg administration of 3 or more active ingredients. In non-fixed combinations, the individual active ingredients may be packaged, sold or administered as entirely separate pharmaceutical compositions. The "non-fixed combination" also includes the combined use of "fixed combinations" or independent entities of any one or more active components.

The term "combination" or "combined use" means that two or more active substances can be administered to a subject together in a mixture, simultaneously as a single formulation, or sequentially as a single formulation in any order.

The term "pharmaceutical composition" refers to one or more active ingredients of the present application (such as anti-VEGF antibody or compound of formula I) or its drug A mixture of drug combination and pharmaceutically acceptable excipients. The purpose of the pharmaceutical composition is to facilitate administration of a compound of the present application, or a pharmaceutical combination thereof, to a subject.

The term "synergistic effect" means that two or more ingredients (eg, anti-VEGF antibody or compound of formula I) produce an effect (eg, inhibition of lung cancer growth) that is greater than the simple additive effect of the ingredients administered alone.

Application method

The following content does not limit the administration mode of the pharmaceutical combination of the present invention.

The components in the combined drug combination of the present invention can be formulated separately. In one embodiment, the components in the combination pharmaceutical composition of the present invention can be formulated into a pharmaceutical composition suitable for single or multiple administration.

The components in the combined pharmaceutical composition of the present invention can be administered alone, or part or all of them can be administered together. The components in the combined pharmaceutical composition of the present invention may be administered substantially differently, or part or all of them may be administered substantially simultaneously.

The components in the combined pharmaceutical composition of the present invention can be administered independently, or part or all of them together, in various suitable routes, including, but not limited to, oral or parenteral (by intravenous, intramuscular, topical or subcutaneous routes). In some embodiments, the components of the combined pharmaceutical composition of the present invention can be administered independently, or part or all of them can be administered together orally or by injection, such as intravenous injection or intraperitoneal injection.

The components in the combined pharmaceutical composition of the present invention can be each independently, or some or all of them can be suitable dosage forms together, including, but not limited to, tablets, buccal tablets, pills, capsules (such as hard capsules, soft capsules, enteric-coated capsules, microcapsules), elixirs, granules, syrups, injections (intramuscular, intravenous, intraperitoneal), granules, emulsions, suspensions, solutions, dispersions and dosage forms of sustained release preparations for oral or parenteral administration.

The components in the combined pharmaceutical composition of the present invention may be each independently, or part or all of them together contain pharmaceutically acceptable carriers and/or excipients.

The combination pharmaceutical compositions of the present invention may also contain additional therapeutic agents. In one embodiment, the additional therapeutic agent may be a cancer therapeutic agent known in the art, preferably a lung cancer therapeutic agent.

In some specific schemes of the present invention, the curative effect of the compound of formula I and anti-VEGF antibody alone or in combination on lung tumors was also investigated. As a result of the experiment, it was surprisingly found that the compound of formula I had an obvious synergistic effect with the anti-VEGF antibody, breaking the established immune tolerance of the body to tumor cells.

Description of drawings

The drawings described here are used to provide a further understanding of the application and constitute a part of the application. The schematic embodiments and descriptions of the application are used to explain the application and do not constitute an improper limitation to the application.

Figure 1 shows the human lung cancer LU-01-1443 xenograft model tumor-bearing mice administered formula I compound, bevacizumab And formula I compound and bevacizumab Weight change after joint;

Figure 2 shows the human lung cancer LU-01-1443 xenograft model tumor-bearing mice administered formula I compound, bevacizumab And formula I compound and bevacizumab Relative body weight change after combination;

Figure 3 shows the human lung cancer LU-01-1443 xenograft model tumor-bearing mice administered formula I compound, bevacizumab And formula I compound and bevacizumab Tumor growth curve after combination;

Figure 4 shows the human lung cancer LU-01-1443 xenograft model tumor-bearing mice administered formula I compound, bevacizumab And formula I compound and bevacizumab Relative tumor growth curves after combination.

Detailed ways

The present invention will be described in detail through examples below, but these examples are only for illustration and do not limit any scope of the present invention. Likewise, the invention is not limited to any specific preferred embodiments described herein. Those skilled in the art should understand that equivalent replacements or corresponding improvements to the technical features of the present invention still fall within the protection scope of the present invention.

Table 1. List of abbreviations

Example 1 In vivo efficacy of spirocyclic aryl phosphine oxides in combination with bevacizumab

The purpose of this experiment is to evaluate the in vivo efficacy of the compound of formula I combined with bevacizumab on the subcutaneous xenograft tumor BALB/c nude mouse model of human lung cancer LU-01-1443.

Experimental program

Table 2. Animal grouping and dosing regimen for in vivo efficacy experiments

Note: a. N: the number of mice in each group; b. When the body weight dropped by more than 15%, the mice were stopped for observation and treatment until their body weight recovered to more than 10%; c. The vehicle was a mixed solution of 5% absolute ethanol and polyoxyethylene 40 hydrogenated castor oil (1:1) + 95% sterile water for injection.

Experimental sample

Name: compound of formula I

Provider: Qilu Pharmaceutical Co., Ltd.

Batch number: T9001L51N

Property description: off-white powder

Purity: 99.6% Molecular Weight: 569.08

Storage conditions: Store at room temperature and avoid light

Name: Bevacizumab

Provider: Qilu Pharmaceutical Co., Ltd.

Batch number: 2020040114KEA

Description of properties: colorless transparent liquid

Concentration: 25mg/mL

Storage conditions: 4°C protected from light

Experimental method and steps

PDX model building

The establishment of the human lung cancer LU-01-1443 model was initially derived from surgically resected clinical samples, which were defined as the P0 generation after implantation into mice. Implantation of tumor tissue from generation P0 into the next generation is referred to as generation P1. Continue to implant in nude mice by analogy. Among them, FP3 tumors were resuscitated through P2 generation. The next generation generated by the FP3 generation is defined as FP4, and so on. The model used in this experiment is the FP5 generation. The LU-01-1443 model is EML4-ALK Fusion, VEGFA/B/C secreted.

tumor inoculation

LU-01-1443 tumor tissue pieces of 20-30 mm ³ were subcutaneously inoculated in the right forelimb axilla of each mouse. On day 13 after inoculation, when the average tumor volume of the enrolled animals reached 115 mm ³ , group administration began. The experimental grouping and dosing regimen are shown in Table 2.

Preparation of test substance

Table 3. Test substance preparation method

Note: The drug is prepared and used now, and the drug needs to be mixed gently before administration.

Daily observation of laboratory animals

The health status and death of the animals were monitored every day. Routine inspections included observing the effects of tumor growth and drug treatment on the daily behavior of the animals, such as behavioral activities, food and water intake (visual inspection only), changes in body weight (body weight was measured once a day), appearance signs or other abnormal conditions. Animal deaths and side effects within groups were recorded based on the number of animals in each group.

Tumor Measurements and Experimental Indicators

The experimental index is to investigate whether tumor growth is inhibited, delayed or cured. Tumor diameters were measured twice a week with vernier calipers. The formula for calculating the tumor volume is: V=0.5a×b ² , where a and b represent the long diameter and short diameter of the tumor, respectively.

The antitumor efficacy of compounds was evaluated by TGI (%) or relative tumor proliferation rate T/C (%). TGI (%) reflects tumor growth inhibition rate. Calculation of TGI (%): TGI (%)=[1-(average tumor volume at the end of administration of a treatment group-average tumor volume at the beginning of administration of this treatment group)/(average tumor volume at the end of treatment of the vehicle control group-average tumor volume at the beginning of treatment of the vehicle control group)]×100%.

Relative tumor proliferation rate T/C (%): the calculation formula is as follows: T/C%=TRTV/CRTV×100% (TRTV: RTV of the treatment group; CRTV: RTV of the negative control group). The relative tumor volume (RTV) was calculated according to the results of tumor measurement, and the calculation formula was RTV=Vt/V0, where V0 was the average tumor volume measured during group administration (i.e. d0), Vt was the average tumor volume during a certain measurement, and TRTV and CRTV took the data on the same day.

Statistical Analysis

The mean and standard error (SEM) of the relative tumor volume at each time point of each group were included (see Table 4 for specific data). The experiment ended on day 14 after dosing, so statistical analysis was performed based on this data to evaluate differences between groups. The comparison between the two groups was analyzed by T-test, and SPSS 17.0 was used for all data analysis. p<0.05 considered significant difference.

Experimental results

Mortality, morbidity, and weight change

In this model, vehicle group, formula I compound single drug group, bevacizumab Group and formula I compound + bevacizumab The body weight of the animals in the combined group remained stable. No animals became ill or died. Formula I compound, bevacizumab And formula I compound and bevacizumab The effect of the combination on the body weight of the human lung cancer LU-01-1443 xenograft tumor female BALB/c nude mouse model is shown in Figure 1 and Figure 2.

tumor volume

The human lung cancer LU-01-1443 xenograft tumor female BALB/c nude mouse model was given the compound of formula I and bevacizumab And formula I compound and bevacizumab The changes in tumor volume in each group after combined treatment are shown in Table 4.

Table 4. Tumor volume at different time points in each group

Note: a. Mean ± SEM; b. Days after administration.

tumor growth curve

The human lung cancer LU-01-1443 xenograft tumor female BALB/c nude mouse model was given the compound of formula I and bevacizumab And formula I compound and bevacizumab The tumor growth curves and relative tumor growth curves of each group after combined treatment are shown in Fig. 3 and Fig. 4 .

Anti-tumor drug efficacy evaluation index

Table 5. Evaluation of antitumor efficacy of human lung cancer LU-01-1443 xenograft model (calculated based on the tumor volume on the 14th day after administration)

Note:

a. Mean ± SEM;

b. For the specific calculation of tumor growth inhibition T/C and TGI, please refer to the "tumor measurement and experimental indicators" part of the instructions;

c.p value is analyzed by T-test according to the relative tumor volume of each mouse in different groups and the vehicle group as the control;

The dp value is according to the relative tumor volume of each mouse in different groups with formula I compound group as contrast, and formula I compound+bevacizumab Groups were compared using T-test;

The ep value is according to the relative tumor volume of each mouse in different groups with bevacizumab group as contrast, and formula I compound+bevacizumab Groups were compared using T-test.

Table 6. Antitumor efficacy evaluation of human lung cancer LU-01-1443 xenograft tumor model (calculated based on tumor weight on the 14th day after administration)

Note:

a. Mean ± SEM;

b. The p value is analyzed by T-test according to the tumor weight of each mouse in different groups and the vehicle group as a control;

The cp value is based on the tumor weight of each mouse in different groups with formula I compound group as contrast, and formula I compound+bevacizumab Groups were compared using T-test;

The dp value was compared with bevacizumab according to the tumor weight of each mouse in different groups Group is control, with formula I compound+bevacizumab Groups were compared using T-test.

Experimental results

In this experiment, we evaluated the in vivo efficacy of the test drug compound of formula I, bevacizumab and the combination of compound of formula I and bevacizumab in the xenograft tumor model of human lung cancer LU-01-1443. The animal body weight and tumor volume of each group at different time points are shown in Table 4, Table 5, Table 6 and Figure 3 and Figure 4 .

On the 14th day after administration, the end point of the experiment, the tumor volume of the tumor-bearing mice in the vehicle control group reached 2,623 mm ³ . Compared with the vehicle control group, the test substance bevacizumab Single use has a certain delay effect on tumor growth, and the tumor volume is 1,722mm ³ (T/C=63.93%, TGI=35.92%, p=0.044). Compound of formula I alone and compound of formula I + bevacizumab The combination has a significant anti-tumor effect, and the tumor volumes are 890mm ³ (T/C=35.28%, TGI=69.11%, p=2.10E-04) and 311mm ³ (T/C=11.81%, TGI=92.19%, p=8.69E-05). Formula I compound and bevacizumab The anti-tumor effect after combined use is significantly better than that of the compound of formula I or bevacizumab Single use (p=0.004, 0.003).

Test drug formula I compound and bevacizumab Figure 1 and Figure 2 show the effects on body weight changes in tumor-bearing mice. During the experiment, vehicle group, formula I compound single drug group, bevacizumab Single drug group, compound of formula I + bevacizumab The body weight of the animals in the combined group remained stable.

in conclusion

In summary, in this experiment, the human lung cancer LU-01-1443 model tumor-bearing mice had no effect on the tested drug formula I compound, bevacizumab Both alone and in combination were well tolerated. And, formula I compound and bevacizumab The combination therapy has an obvious synergistic effect, and the tumor inhibitory effect is remarkable, and it is significantly better than the compound of formula I or bevacizumab at the same dose single-use effect.

The above descriptions are only preferred embodiments of the present invention, and are not intended to limit the present invention. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principles of the present invention shall be included within the protection scope of the present invention.

Claims (10)

1. A combined pharmaceutical composition comprising an anti-VEGF antibody and a compound represented by formula I or a pharmaceutically acceptable salt thereof
   
2. The combined pharmaceutical composition according to claim 1, wherein the anti-VEGF antibody is bevacizumab.
3. According to the combined pharmaceutical composition of claim 1 or 2, the anti-VEGF antibody and the compound of formula I or a pharmaceutically acceptable salt thereof are each in the form of a pharmaceutical composition.
4. Application of the combination pharmaceutical composition described in any one of claims 1-3 in the preparation of drugs for treating and/or preventing lung cancer.
5. The use according to claim 4, wherein the lung cancer is ALK and/or ROS1 fusion gene positive.
6. The use according to any one of claims 4-5, wherein the lung cancer is early-middle stage, locally advanced, advanced metastatic non-small cell lung cancer.
7. The use according to any one of claims 4-6, wherein the lung cancer is early-middle stage, locally advanced, advanced metastatic non-small cell lung cancer positive for ALK and/or ROS1 fusion gene.
8. The use according to any one of claims 4-7, wherein the lung cancer is non-small cell lung cancer that has not been treated with ALK and/or ROS1 inhibitors or failed to be treated with ALK and/or ROS1 inhibitors.
9. The use according to any one of claims 6-8, wherein the non-small cell lung cancer is adenocarcinoma, squamous cell carcinoma, adenosquamous cell carcinoma or large cell carcinoma.
10. The use according to any one of claims 4-9, wherein the anti-VEGF antibody and the compound of formula I or a pharmaceutically acceptable salt thereof are each in the form of a pharmaceutical composition and can be administered simultaneously, sequentially or at intervals.