HK40040200A

HK40040200A - Combination of poziotinib with cytotoxic agent and/or other molecularly targeted agent and use thereof

Info

Publication number: HK40040200A
Application number: HK62021029711.0A
Authority: HK
Inventors: 古鲁·雷迪
Original assignee: 光谱制药有限公司
Priority date: 2018-06-25
Filing date: 2019-06-25
Publication date: 2021-07-30

Description

Composition of bosutinib and cytotoxic agent and/or other molecular targeting agent and application thereof

Cross Reference to Related Applications

This application claims the benefit of U.S. provisional application No. 62/689,530 filed 2018, month 6, 25, the entire disclosure of which is incorporated herein by reference in its entirety.

Technical Field

One or more embodiments relate to a combination medicament comprising bosutinib and a cytotoxic agent and/or a molecular targeting agent and uses of the medicament.

Background

Pozitinib is a low molecular weight compound that selectively and irreversibly inhibits the Epidermal Growth Factor Receptor (EGFR) family (including Her1, Her2, and Her 4). Bosutinib is also a pan-Her inhibitor that is very effective in inhibiting EGFR and Her2 activation and resistance mutations. The activity of poqitinib is disclosed in US 8188102B and US 2013/0071452a1, which are hereby incorporated by reference. In US 8188102B, the compound of formula 1 is the compound of example 36.

EGFR is the first known growth factor receptor in protein tyrosine kinases, a protein consisting of a receptor moiety and a tyrosine kinase moiety, which transmits extracellular signals across the cell membrane into the cell. The EGFR tyrosine kinase family has four subtypes of receptors EGFR/ErbB1, Her2/ErbB2, Her3/ErbB3 and Her4/ErbB4 (hereinafter also referred to as "Her 1, Her2, Her3 and Her 4", respectively). All of these can form a signaling complex comprising two different members of the family, either as homodimers or heterodimers. EGFR plays an important role in normal cell regulation through intracellular signal transduction, but when signals transmitted through receptors cannot be controlled due to overexpression or activation mutations, EGFR activates a cell signaling system to induce growth or differentiation of tumor cells, neovascularization, metabolism, and drug resistance expression (resistance expression). (WellsA. int J Biochem Cell Bio1.,1999,31,637 and Nancy E.Hynes and Heidi A.Lane, Nature Reviews Cancer 5,341,2005). EGFR has been reported to be abnormally overexpressed or frequently mutated in most solid cancer cells, which is associated with poor prognosis. Therefore, research for developing anticancer agents targeting EGFR has been actively conducted, and it is expected that anticancer effects will be very good if cancer cells are blocked from signaling through EGFR.

EGFR targeted anticancer agents are classified into monoclonal antibody drugs targeting the extracellular domain of the receptor and low molecular weight drugs targeting intracellular tyrosine kinase. Examples of EGFR-targeted antibody drugs include trastuzumab (trastuzumab) which is a Her 2-specific humanized monoclonal antibody and Cetuximab (Cetuximab) which is a Her 1-specific chimeric (mouse/human) monoclonal antibody. Trastuzumab and cetuximab are used as sole or combined therapeutics for Her2 positive advanced breast and gastric cancers, and Her1 positive metastatic colorectal cancers and head and neck cancers, respectively. Examples of EGFR-targeted low molecular weight drugs include gefitinib (gefitinib) and erlotinib (erlotinib), which are selective inhibitors of EGFR/ErbB1, and lapatinib (lapatinib), which is an inhibitor of EGFR and Her 2. Gefitinib and erlotinib are used as therapeutics for lung cancer, lapatinib is used as a therapeutic for Her2 positive advanced breast cancer, and clinical trials are underway to expand the indications for treatment of other solid cancers.

However, in recent years, it has been reported that the expression of resistance in EGFR-targeted therapies reduces the response time of the drugs used. Non-small cell lung cancer (NSCLC) patients with EGFR-activated mutants treated with gefitinib or erlotinib were reported to develop resistance to the drug after about 8 to 16 months, and about 60% developed resistance due to the EGFRT790M mutation (helenaa. yu et al, clin. cancer res.19(8),2240,2013). Furthermore, in the case of Her2 positive metastatic breast Cancer patients treated with the antibody drug trastuzumab, 66% to 88% of patients are known to exhibit new resistance (de novo resistance) or acquired resistance (Alice Chung et al, clin. Breast Cancer 13(4),223,2013) due to various mechanisms.

In this regard, although EGFR-targeted therapeutics have considerable efficacy in the treatment of EGFR/Her2 overexpressing or mutated solid cancers, the development of EGFR-targeted therapeutics has limitations in that the efficacy cannot be maintained for a long time due to the development of primary and secondary resistance. Therefore, there is a pressing need for an effective treatment method that can improve the efficacy and overcome drug resistance in the treatment of solid cancers that overexpress or mutate EGFR/Her 2.

Disclosure of Invention

The present invention addresses this need. According to one embodiment, there is provided a combination comprising as active ingredients a composition of bozitinib and at least one agent selected from the group consisting of cytotoxic agents (cytotoxic agents) and molecular targeting agents (molecular targeted agents) for the treatment of a tumor associated with overexpression or amplification of at least one gene of HER1, HER2 and HER4 or a mutant of HER1, HER2 or HER4 in a subject.

According to another embodiment, there is provided a use of a combination medicament for the manufacture of a medicament for the treatment of a tumor associated with overexpression or amplification of at least one gene of HER1, HER2 and HER4 or a mutant of HER1, HER2 or HER4 in a subject.

According to another embodiment, there is provided a method of treating a tumor associated with overexpression or amplification of HER1, HER2 or HER4 or a mutant of HER1, HER2 or HER4 in a subject, the method comprising administering a composition comprising a plurality of active ingredients comprising a therapeutically effective amount of bosutinib and at least one agent selected from cytotoxic agents and molecular targeting agents.

According to another embodiment, there is provided a kit for treating a tumor associated with overexpression or amplification of HER1, HER2 or HER4 or a mutant of HER1, HER2 or HER4 in a subject, the kit comprising a first part and a second part, wherein the first part comprises bosutinib and the second part comprises at least one active ingredient selected from a cytotoxic agent and a molecular targeting agent. The kit may further comprise a package insert comprising instructions for treating a tumor associated with overexpression or amplification of HER1, HER2, or HER4, or a mutant thereof, in a subject.

Drawings

These and/or other aspects will become apparent and more readily appreciated from the following description of the embodiments, taken in conjunction with the accompanying drawings of which:

fig. 1A to 1D show an example of synergy in BT-474 cells (a breast cancer cell line) that are Her2 overexpressed and sensitive to trastuzumab. FIG. 1A shows dose response curves for 10 μ M cisplatin (cissplatin) and bozitinib concentrations ranging from 0.25nM to 2nM, administered alone and in combination. Figure 1B shows dose response curves for paclitaxel (paclitaxel) and boresinib administered alone and in combination at a ratio of paclitaxel to boresinib 1:1 at concentrations ranging from 0.25nM to 2 nM. FIG. 1C shows dose response curves for 5 μ M5-FU and a concentration range of 0.25nM to 2nM of Boletinib alone and in combination. Figure 1D shows dose response curves for trastuzumab at concentrations ranging from 5nM to 100nM and pofzetinib at concentrations ranging from 0.05nM to 1nM administered alone and in combination at a ratio of trastuzumab pozetinib 100: 1;

figures 2A to 2C show examples of synergy in MDA-MB-453 cells (a breast cancer cell line) that are HER2 overexpressed, trastuzumab-resistant, and harboring a PIK3CA mutant. Figure 2A shows dose response curves for paclitaxel and bociclib alone and in combination in a paclitaxel to bociclib 1:1 ratio, both at concentrations ranging from 0.3nM to 2.5 nM. Figure 2B shows dose response curves of 5-FU at a concentration ranging from 60nM to 500nM and pozzatinib at a concentration ranging from 0.3nM to 2.5nM administered alone and/or in combination with 5-FU: pozzatinib at a ratio of 200: 1. Figure 2C shows dose response curves for trastuzumab at a concentration ranging from 0.5nM to 25nM and pofzetinib at a concentration ranging from 0.05nM to 2.5nM administered alone and in combination at a trastuzumab pozetinib ratio of 10: 1;

FIG. 3 shows an example of synergy in MCF-7 cells, an estrogen receptor positive breast cancer cell line, where Her1 and Her2 are not overexpressed. FIG. 3 shows dose response curves for vinorelbine (vinorelbine) at 2nM and bovatinib alone and in combination at concentrations ranging from 0.1nM to 10 nM;

figure 4 shows an example of synergy in MDA-MB-361 cells (a breast cancer cell line) that are Her2 overexpressed, trastuzumab-resistant, and with PIK3CA mutant. FIG. 4 shows dose response curves for 1nM vinorelbine and a concentration range of 1 μ M to 10 μ M of bocetitinib administered alone and in combination;

FIG. 5 shows an example of synergy in TE-4 cells (an esophageal cancer cell line), which are Her2 overexpressed. FIG. 5 shows dose-response curves for 5-FU and 0.5nM of polazitinib alone and in combination at concentrations ranging from 10nM to 50000 nM;

fig. 6A and 6B show examples of synergy in HCC827 cells, a NSCLC (non-small cell lung cancer) cell line, with Her1L858R mutants. Figure 6A shows dose response curves for paclitaxel at a concentration ranging from 0.5nM to 20nM and for polazinib at a concentration ranging from 0.05nM to 2nM administered alone in combination with paclitaxel: polazinib: 10: 1. Figure 6B shows dose response curves for cisplatin at a concentration ranging from 0.1nM to 4 μ Μ and pozitinib at a concentration ranging from 0.05nM to 2nM administered alone and/or in combination at a cisplatin to pozitinib ratio of 2000: 1; and

FIGS. 7A and 7B show examples of synergy in H1975 cells (a NSCLC cell line carrying the Her1L858R/T790M mutant). Figure 7A shows dose response curves for paclitaxel at a concentration ranging from 1.5nM to 50nM and for polazinib at a concentration ranging from 0.15nM to 5nM administered alone in combination with paclitaxel: polazinib: 10: 1. Figure 7B shows dose response curves for cetuximab at a concentration ranging from 80nM to 5000nM and for combined administration of bocetirizine alone and/or in a ratio of cetuximab to bocetirizine ═ 200:1 at a concentration ranging from 0.4nM to 25 nM.

Figures 8A and 8B show the effect of combined administration of bosutinib and Pemetrexed (Pemetrexed) or Letrozole (Letrozole) on Calu3 cell line.

Detailed Description

Reference will now be made in detail to embodiments, examples of which are illustrated in the accompanying drawings, wherein like reference numerals refer to the like elements throughout. In this regard, embodiments of the present application may take different forms and should not be construed as limited to the descriptions set forth herein. Accordingly, the embodiments are described below in order to explain aspects of the present specification by referring to the figures only. As used herein, the term "and/or" includes any and all combinations of one or more of the associated listed items. Expressions such as "at least one of" when preceding a list of elements modify the entire list of elements without modifying individual elements in the list.

According to one embodiment, a combination medicament for the treatment of a tumor is provided. In a more specific embodiment, the tumor is associated with overexpression or amplification of at least one gene of HER1, HER2 and HER4 or a mutant of HER1, HER2 or HER4 in a subject, said combination comprising as active ingredients a composition of bosutinib and at least one cytotoxic agent and/or at least one molecular targeting agent.

The bosutinib, namely 1- [4- [4- (3,4-dichloro-2-fluoroanilino) -7-methoxyquinazolin-6-yl ] oxypiperidin-1-yl ] prop-2-en-1-one (1- [4- [4- (3, 4-dichoro-2-fluoroanilino) -7-methoxyquinazolin-6-yl ] oxyperidin-1-yl ] prop-2-en-1-one), or a pharmaceutically acceptable hydrate and/or salt thereof has a structure shown in formula 1.

The pharmaceutically acceptable salt may be an inorganic acid salt, an organic acid salt or a metal salt. The inorganic acid salt may be a salt of hydrochloric acid, phosphoric acid, sulfuric acid or disulfonic acid. The organic acid salt may be a salt of malic acid, maleic acid, citric acid, fumaric acid, benzenesulfonic acid (besylic acid), camphorsulfonic acid (camsylic acid), or 1, 2-ethanedisulfonic acid (edisylic acid). The metal salt may be a calcium, sodium, magnesium, strontium or potassium salt. In one embodiment, the brigatinib may be a hydrochloride salt in tablet form. The bovatinib may be administered in an amount of 0.1mg to 50 mg.

Bosutinib is a low molecular weight compound that selectively and irreversibly inhibits the Epidermal Growth Factor Receptor (EGFR) family (including Her1, Her2, and Her 4). Bosutinib is also a very potent pan-Her inhibitor in inhibiting EGFR and Her2 activation and drug resistant mutants. The activity of poqitinib is disclosed in US 8188102B and US 2013/0071452a1, the entire contents of which are incorporated herein by reference. In US 8188102B, the compound of formula 1 is the compound of example 36. Poecitinib can inhibit the growth of tumor cells in various cancers with Her1 or Her2 overexpression or activation mutants in vitro, and can effectively inhibit the growth of lung cancer cells with gefitinib or erlotinib resistance. In addition, poecitinib effectively prevented tumor growth in xenograft animal models (heter-transplantation animal models), which are animal bodies transplanted with such tumor cells. In addition, bosutinib has a broad and excellent inhibitory effect on EGFR and mutants thereof, and the therapeutic fields including the field of resistance to other known EGFR-targeting antibody drugs and low molecular weight drugs can be broader and more effective. Combination therapy with other drugs on this basis may also improve the resistance effect against various solid cancers, and may improve the remission rate (survival) and prolong survival time, compared to the effects produced using conventional therapeutic agents.

Cytotoxic agents are agents that have a cytotoxic effect on cells. Cytotoxic effects refer to the depletion, elimination and/or death of target cells (i.e., tumor cells). The cytotoxic agent may be at least one selected from the group consisting of: antimetabolites, mitotic inhibitors, alkylating agents, platinum-based antineoplastic drugs (platinum-based anticancer), antibody-based EGFR inhibitors, antibody-based HER2/3 inhibitors, angiogenesis inhibitors, mTOR inhibitors, CDK4 and CDK6 inhibitors, or aromatase inhibitors. The composition may comprise at least two cytotoxic agents. For example, the composition may comprise at least 2, at least 3 or at least 4 or all selected from the group consisting of antimetabolites, mitotic inhibitors, alkylating agents, angiogenesis inhibitors and platinum-based antineoplastic agents.

An antimetabolite may be an agent that inhibits DNA synthesis in a cell by inhibiting the formation of the base purine or pyrimidine of a nucleotide. In one embodiment, the antimetabolite may be selected from the group consisting of Capecitabine (Capecitabine), 5-Fluorouracil (5-fluoroouracil), Gemcitabine (Gemcitabine), pemetrexed, Methotrexate (Methotrexate), 6-Mercaptopurine (6-mercaptoprine), Cladribine (Cladribine), Cytarabine (Cytarabine), doxifluridine (doxifrudine), Floxuridine (Floxuridine), Fludarabine (Fludarabine), hydroxyurea (hydroxyycarbamide), dacarbazine (decazine), hydroxyurea (hydroxyuride), and asparaginase (asparaginase). In a more specific embodiment, the antimetabolite is a base analog, and the term "base analog" herein includes nucleotide and nucleoside analogs in addition to purine base analogs (e.g., 5-fluorouracil).

The mitotic inhibitor may be a microtubule destabilizer, a microtubule stabilizer, or a combination thereof. The mitotic inhibitor can be a taxane (taxanes), vinca alkaloids (vinca alkaloids), epothilone (epothilone), or combinations thereof. In a particular embodiment, the mitotic inhibitor is a taxane, such as including but not limited to paclitaxel, docetaxel, and cabazitaxel. In another specific embodiment, the mitotic inhibitor is a vinca alkaloid or derivative thereof, including, for example, but not limited to, vinblastine (vinblastine), vincristine (vinchristine), vinflunine (vinflunine), vinorelbine, vincaminol (vinaminol), vinbuxine (vinburnine), vinendosidine (vinderidine), and vindesine (vindesine).

The mitotic inhibitor is selected from BT-062, HMN-214, eribulin mesylate (eribulin mesylate), vindesine, EC-1069, EC-1456, EC-531, vintafolide, 2-methoxyestradiol (2-methoxythioridol), GTx-230, trastuzumab emtansine (T-DM1), crolizin, D1302A-maytansine conjugate IMGN-529 (D1302A-maytansinoids IMGN-529), moxon-lovotuzumab mertansine (LORTUzumab mertansine), SAR-3419, su-566658, IMP-03138, topotecan/vincristine combination, BPH-8, sburethrin tromethamine, suturamine (RX), sulbactam), sodium phosphate (RX 21101), vinblastine A, docetaxel A, vinblastine A, vinblastine acetate A, vinblastine A, vinblastine, vinblasti, Paclitaxel (patupilone), ixabepilone (ixabepilone), epothilone D, paclitaxel, docetaxel, DJ-927, discodermolide (discodermolide), escarginol (eleutherobin), and pharmaceutically acceptable salts or combinations thereof.

As used herein, an "alkylating agent" is the addition of one or more alkyl groups (C) to a nucleic acid_nH_mWhere n and m are integers). In the present invention, the alkylating agent is selected from the group consisting of nitrogen mustards, nitrosoureas, alkyl sulfonates, triazines, ethyleneimines, and combinations thereof. Non-limiting examples of nitrogen mustardsExamples include dichloromethyldiethanamine (mechlororethamine), chlorambucil (chlorambucil), cyclophosphamide (cyclophosphamide), bendamustine (bendamustine), ifosfamide (ifosfamide), melphalan (melphalan), melphalan flufenamide (melphalan flufenamide), and pharmaceutically acceptable salts thereof. Non-limiting examples of nitrosoureas include streptozotocin (streptozocin), carmustine (carmustine), lomustine (lomustine), and pharmaceutically acceptable salts thereof. Non-limiting examples of alkyl sulfonates include busulfan and pharmaceutically acceptable salts thereof. Non-limiting examples of triazines include dacarbazine, temozolomide (temozolomide), and pharmaceutically acceptable salts thereof. Non-limiting examples of ethyleneimine include thiotepa (thiotepa), hexamethylmelamine (altretamine), and pharmaceutically acceptable salts thereof. Other alkylating agents include ProLindac^TMAc-225 BC-8, ALF-2111, trofosfamide (trofosfamide), MDX-1203, thioureidobutyronitrile (thioureidobutyronitrile), mitobronitol (mitobronitol), dibromodulcitol (mitolactol), nimustine (nimustine), glufosfamide (glufosfamide), HuMax-TAC and PBDADC combinations, BP-C1, troosulfan (treosulfan), nifurolimus (nifurtimox), propenesulfonate P-toluenesulfonate (impron tosilate), ramustine (ranimustine), ND-01, HH-1, 22P1G cells and ifosfamide (ifosfamide) combinations, estramustine phosphate (ramustine phosphate), ranimustine (prednimustine), pyributidine (tripterine), tolidine (tlutinine), melphalan (tluamin), melphalan (platinum), melphalan (SG33), carboplatin (platinum), and pharmaceutically acceptable salts thereof.

Angiogenesis inhibitors are substances that inhibit the growth of new blood vessels (angiogenesis). Some angiogenesis inhibitors are endogenous and are a normal part of the body's control, others are obtained exogenously through drugs or diets. In at least one embodiment, the angiogenesis inhibitor comprises bevacizumab (bevcizumab), sunitinib (sunitinib), sorafenib (sorafenib), or pazopanib (pazopatinib).

The platinum antineoplastic agent can be selected from cisplatin, Carboplatin (Carboplatin), Dicycloplatin (Dicycloplatin), Eptaplatin (Eptaplatin), Lobaplatin (Lobaplatin), miiplatin (Miriplatin), nedaplatin, Oxaliplatin (Oxaliplatin), Picoplatin (Picoplatin) and Satraplatin (Satraplatin).

As used herein, a "molecular targeting agent" is a substance that, when administered to a subject, interferes with the function of a single molecule or group of molecules, preferably those involved in tumor growth and progression. Non-limiting examples of the molecular targeting agents of the present invention include signal transduction inhibitors, modulators of gene expression and other cellular functions, immune system modulators, antibody-drug conjugates, and compositions thereof.

The molecular targeting agent may be selected from the group consisting of epidermal growth factor receptor family inhibitors (EGFRI), mammalian target of rapamycin (mTOR) inhibitors, immune checkpoint inhibitors, Anaplastic Lymphoma Kinase (ALK) inhibitors, B-cell lymphoma-2 (BCL-2) inhibitors, B-Raf inhibitors, Cyclin Dependent Kinase Inhibitors (CDKi) (such as CDK4/CDK6 inhibitors), palbociclib (palbociclib), ERK inhibitors, histone deacetylase inhibitors (HDACI), heat shock protein-90 inhibitors (HSP90i), Janus kinase inhibitors, mitogen-activated protein kinase (MAPK) inhibitors, MEK inhibitors (such as MEK1/MEK2 inhibitors), trimiprtinib (trametinib), poly-ADP ribose polymerase (PARP) inhibitors, phosphoinositide 3-kinase inhibitors (PI3Ki), Ras inhibitors, sodium-glucose cotransporter (SGR) inhibitors and groups thereof A compound (I) is provided.

Suitable sodium-glucose cotransporter (SGLT) inhibitors, also known as sodium-dependent glucose cotransporter inhibitors, include inhibitors of sodium/glucose cotransporter 1(SGLT 1).

The molecular targeting agent may be selected from ado-trastuzumab (T-DM1), alemtuzumab (alemtuzumab), cetuximab, lapinimumab (ipilimumab), ofatumumab (ofatumumab), panitumumab (panitumumab), pertuzumab (pertuzumab), rituximab (rituximab), tositumomab (tositumomab), 131I-tositumomab (131I-tositumomab), trastuzumab, vetuzumab (brentuximab vedotin), dinil interleukin (denileukin diftitox), ibritumomab (ibritumomab tix), axitinib (axitinib), bortezomib (bortezomib), sultinib (bostinib), carboplatin (bortetinib), carbotinib (bortetinib), zotinib (azatinib), erlotinib (zotinib (erlotinib), erlotinib (lapatinib), erlotinib (lapatinib), erlotinib (, Regorafenib, ruxolitinib, sorafenib, sunitinib, tofacitinib, vandetanib, vemurafenib, alitretinoin (alitatentoin), bexarotene (bexarotene), everolimus (everolimus), romidepsin (romidepsin), temsirolimus (temsirolimus), tretinoin (tretinoin), vorinostat (vorinostat), and pharmaceutically acceptable salts thereof or compositions thereof. Molecular targeting agents may include antibodies, antibody moieties or antibody-conjugated drugs.

The EGFR inhibitor may be selected from erlotinib, gefitinib, lapatinib, canertinib, pelitinib, neratinib, R, E-N- (7-chloro-1- (1- (4- (dimethylamino) but-2-enyl) azepin-3-yl) -1H-benzo [ d ] imidazol-2-yl) -2-methylisonicotinamide ((R, E) -N- (7-chloro-1- (1- (4- (dimethylamino) but-2-enyl) azepan-3-yl) -1H-benzol [ d ] imidizol-2-yl) -2-methylisonicotinamide), trastuzumab, panitumumab, matuzumab, Rituximab (necitumumab), pertuzumab, nimotuzumab (nimotuzumab), zalutumumab (zalutumumab), cetuximab, icotinib (icotinib), afatinib (affatinib), and pharmaceutically acceptable salts thereof. In one embodiment, the EGFR inhibitor may be an antibody-based EGFR inhibitor, such as cetuximab, in another embodiment the EGFR inhibitor is nixituzumab, and in another embodiment the EGFR inhibitor is panitumumab. The molecular targeting agent may be an anti-EGFR family antibody or a complex (complex) comprising an anti-EGFR family antibody. The anti-EGFR family antibody may be an anti-HER 1 antibody, an anti-HER 2 antibody, or an anti-HER 4 antibody.

A tumor associated with overexpression or amplification of at least one of HER1, HER2, and HER4, or a mutant thereof, can be an abnormal growth of tissue, which if it forms a mass, is generally referred to as a tumor with overexpression of at least one of HER1, HER2, HER4, and a mutant thereof, or amplification encoded by at least one of HER1, HER2, HER4, or a mutant thereof. In one embodiment, the mutant may be present in any one of exons 18, 19, 20 and 21, or any combination thereof. For example, the mutant may be HER1 having an exon 19 deletion, a T790M substitution, a L828R substitution, or a combination thereof. In another embodiment, the mutant may be at exon 20 of HER2, e.g. an exon 20 insertion mutant. In another embodiment, one or more mutations are present in either exon 19 or 20 or both. For example, one or more EGFR exon 20 mutants include one or more point mutants, insertions and/or deletions between amino acids 763-778 and 3-18 nucleotides.

In other embodiments, the subject may be determined to have 2, 3 or 4 EGFR exon 20 mutants at one or more residues selected from the group consisting of a763, a767, S768, V769, D770, N771, P772, and H773. In another embodiment, the subject may be determined to be free of EGFR mutants at residue C797. In some embodiments, the one or more EGFR mutants comprise substitutions and/or deletions at a763, a767, S768, V769, D770, N771, P772, and H773in exon 20. In some embodiments, the one or more exon 20 mutants are selected from the group consisting of a763insFQEA, a767insASV, S768dupSVD, V769insASV, D770insSVD, D770insNPG, H773insNPH, N771del insGY, N771del insFH, and N771 dupph.

In certain embodiments, the genomic sample from the subject may be from saliva, blood, urine, normal tissue, or tumor tissue. In particular aspects, the presence of EGFR exon 20 mutants is determined by nucleic acid sequencing (e.g., DNA sequencing of tumor tissue or plasma cycle free DNA) or Polymerase Chain Reaction (PCR) analysis.

Other embodiments provide methods of treating cancer in a patient comprising administering to the subject an effective amount of bosutinib and a secondary cytotoxic agent, wherein the subject has been determined to have one or more exon 19 or exon 20 mutants.

As used herein, "overexpression" means that the protein is expressed at a higher level than in normal cells. Expression levels can be measured using immunohistochemistry, Fluorescence In Situ Hybridization (FISH), or Chromogenic In Situ Hybridization (CISH). The tumor may be lung cancer (including non-small cell lung cancer), breast cancer, stomach cancer, colon cancer, pancreatic cancer, prostate cancer, myeloma, head and neck cancer, ovarian cancer, esophageal cancer, or metastatic cell carcinoma (metastic cell carcinoma).

The subject may be a mammal. The mammal may be a human.

Administration of a therapeutically effective amount of a composition of the present invention is preferred over administration of the individual component compounds because the composition will provide one or more of the following improved properties when compared to the administration of a therapeutically effective amount of the component compounds alone: i) greater anti-cancer effect than the single agent with the highest activity, ii) synergistic or highly synergistic anti-cancer activity, iii) a dosing regimen that provides enhanced anti-cancer activity with low side effects, iv) reduced toxicity, v) increased therapeutic window, vi) increased bioavailability of one or both component compounds, or vii) increased apoptosis relative to the single component compound.

The compounds of the invention may contain one or more chiral atoms or may exist in the form of two enantiomers. Thus, the compounds of the present invention include mixtures of enantiomers as well as purified enantiomers or enantiomerically enriched mixtures. Furthermore, it is to be understood herein that each of the terms "polazitinib", "cytotoxic agent" and "molecular targeting agent" includes all closely related forms of the compound that may be suitable for the situation. For example, all tautomers and mixtures of tautomers, and pharmaceutically acceptable solvates and/or salts thereof, are included within this range by each of the terms "polazinib", "cytotoxic agent" and "molecular targeting agent" herein. The principles of this coverage apply not only to the terms "cytotoxic agent" and "molecular targeting agent," but also to any particular drug member belonging to the group defined by these two terms. For example, the term "cisplatin" as used herein includes all tautomers and mixtures of tautomers thereof, as well as pharmaceutically acceptable solvates and/or salts thereof, as is the case with the term "cytotoxic agent" when combined with bosutinib.

The compounds of the invention may form solvates, which are understood to be complexes of variable stoichiometry formed by a solute (in the present invention, the solute may be poqitinib or a salt thereof and a cytotoxic agent or a salt thereof and/or a molecular targeting agent or a salt thereof) and a solvent. Such solvents for the purposes of the present invention do not interfere with the biological activity of the solute. Examples of suitable solvents include, but are not limited to, water, methanol, dimethyl sulfoxide, ethanol, and acetic acid. Suitably, the solvent used is a pharmaceutically acceptable solvent. Suitably, the solvent used is water.

Pharmaceutically acceptable salts of the compounds of the present invention are readily prepared by those skilled in the art.

In one embodiment, the at least one cytotoxic agent is selected from the group consisting of taxanes, base analogs, platinum-based antineoplastic agents and vinca alkaloid antineoplastic agents, and aromatase inhibitors. In at least one embodiment, the aromatase inhibitor comprises letrozole and anastrozole (anastrozole).

In another embodiment, the at least one molecular targeting agent is selected from the group consisting of Epidermal Growth Factor Receptor (EGFR) family inhibitors and mammalian target of rapamycin (mTOR) inhibitors (e.g., everolimus, temsirolimus, and sirolimus).

In a specific embodiment, the composition comprises bosutinib and an anti-EGFR family antibody. In another specific embodiment, the composition comprises bosutinib and a taxane. In another specific embodiment, the composition comprises bosutinib and a base analog. In another embodiment, the composition comprises bosutinib and a platinum-based anti-tumor drug. In another specific embodiment, the composition comprises bosutinib and a vinca alkaloid.

In one embodiment, the composition may comprise bosutinib and an anti-EGFR family antibody. The anti-EGFR family antibody can be trastuzumab, cetuximab, margetuximab, matuzumab, panitumumab, cetuximab, or pertuzumab. One example of the composition may be bosutinib and trastuzumab; or bordetella and cetuximab. The brigatinib may be in the form of a hydrochloride salt. The composition may further comprise a cytotoxic agent. The cytotoxic agent may be a mitotic inhibitor. The mitotic inhibitor may be a taxane, vinca alkaloid, epothilone, or a combination thereof. The vinca alkaloid can be at least one of vinblastine, vincristine, vindesine and vinorelbine. Examples of the composition include bosutinib, trastuzumab, and vinorelbine. Vinorelbine may be in the form of an injection. The taxane may be paclitaxel or docetaxel. Examples of the composition may include bosutinib, cetuximab, and paclitaxel. Paclitaxel may be in the form of an injection.

The bovatinib may be administered in an amount of 0.1mg to 50 mg. Trastuzumab can be administered in an amount of 0.5mg to 10mg per kg body weight. Cetuximab may be present at 100mg/m²Body surface area to 500mg/m²The amount of body surface area is administered.

The vinorelbine can be in the range of 0.5mg/m²Body surface area to 50mg/m²The amount of body surface area is administered. In addition, paclitaxel may be present at 100mg/m²Body surface area to 300mg/m²The amount of body surface area is administered.

Herceptin among other drugs^TMTrastuzumab is sold under the trademark monoclonal antibody for the treatment of breast cancer. In particular, it is used for HER2 receptor positive breast cancer. Trastuzumab is administered by intravenous slow injection and subcutaneous injection.

Cetuximab is an Epidermal Growth Factor Receptor (EGFR) inhibitor for treatmentMetastatic colorectal cancer, metastatic non-small cell lung cancer, and head and neck cancer. Cetuximab is a chimeric (mouse/human) monoclonal antibody produced by intravenous infusion under the trade name Erbitux by the pharmaceutical company Bristol-Myers Squibb^TMSupplied in the united states and canada, by the pharmaceutical company Merck KGaA, outside of the united states and canada. In Japan, Merck KGaA, Bristol-Myers Squibb and Eli Lilly are supplied together.

In other drugs as Taxol^TMPaclitaxel (PTX), sold under the trademark paclitaxel, is a chemotherapeutic drug used to treat a variety of cancers. This includes ovarian cancer, breast cancer, lung cancer, Kaposi's sarcoma (Kaposi sarcoma), cervical cancer, and pancreatic cancer. Paclitaxel is administered by intravenous injection.

In one embodiment, the composition may comprise bosutinib and a mitotic inhibitor. The mitotic inhibitor is selected from BT-062, HMN-214, eribulin mesylate, vindesine, EC-1069, EC-1456, EC-531, vintafolide, 2-methoxyestradiol, GTx-230, trastuzumab-maytansine conjugate (T-DM1), crolibulin, D1302A-maytansine conjugate, IMGN-529, moxon-lovoruzumab, SAR-3419, SAR-566658, IMP-03138, topotecan/vincristine combination, BPH-8, fosbretylin tromethamine, estramustine sodium phosphate, vincristine, vinflunine, vinorelbine, RX-21101, cabazitaxel, STA-9584, vinblastine, epothilone A, paclitaxel, ixabepilone, epothilone D, paclitaxel, docetaxel, DJ-927, discodermolide, alcladhol, pharmaceutically acceptable salts thereof or combinations thereof. An example of the composition may comprise bosutinib and a taxane, a vinca alkaloid, or a combination thereof. The vinca alkaloid may be at least one drug selected from vinblastine, vincristine, vindesine and vinorelbine. The taxane may be paclitaxel or docetaxel. One example of the composition may comprise bosutinib and paclitaxel; or bosutinib and vinorelbine. The tumor may be breast cancer that is over-expressed by Her 2.

In another embodiment, the composition may comprise bosutinib and a mitotic inhibitor (e.g., paclitaxel). In another embodiment, the cytotoxic agent may be an EGFR inhibitor, e.g., a Raf inhibitor, a RAS inhibitor, a MEK inhibitor, a MAPK inhibitor, or an ERK inhibitor, in combination with pozzinib alone or in combination with pozzinib and a second cytotoxic agent.

The bovatinib may be administered in an amount of 0.1mg to 50 mg. In addition, vinorelbine may be present at 0.5mg/m²Body surface area to 50mg/m²The amount of body surface area is administered. In addition, paclitaxel may be present at 100mg/m²Body surface area to 300mg/m²The amount of body surface area is administered.

Navelbine is used as the main component of other medicines^TMVinorelbine (NVB) is sold under the trademark vinorelbine is a chemotherapeutic drug used in the treatment of various cancers. This includes breast cancer and non-small cell lung cancer. Vinorelbine is administered intravenously or orally. Vinorelbine belongs to the alkaloid drugs of vinca. It is believed that the effect of vinorelbine is to disrupt the normal function of cellular microtubules, thereby preventing cell division.

In one embodiment, the composition may comprise bosutinib and an mTOR inhibitor, and an AKT and/or P13k system inhibitor. The mTOR inhibitor can be selected from zotarolimus (zotarolimus), umirolimus (umirolimus), temsirolimus, sirolimus, and sirolimus nanocrytal^TMSirolimus TransDerm^TMsirolimus-PNP, everolimus, biolimus A9, ridaforolimus, rapamycin (rapamycin), TCD-10023, DE-109, MS-MS-R003, Perceiva, XL-765, quinacrine (quinacrine), PKI-587, PF-04691502, GDC-0980, daculisib, CC-223, PWT-33597, P-7170, LY-3023414, INK-128, GDC-0084, DS-7423, DS-3078, CC-115, CBLC-137, AZD-2014, X-480, X-414, EC-0371, VS-5584, VS-401, PQR-316, PQR-311, PQR-309, PF-06465603, NV-128, nPT-MTOR, WABC-210, Y-600, LOYE-354, HMYER-518, HMYER-220, HMYER-518, GNE-317, EC-0565, CC-214, ABTL-0812 and pharmaceutically acceptable salts thereof or compositions thereof. One example of a composition may comprise bosutinib and rapamycin. Rapamycin may be in the form of an injection. Rapamycin, also known as sirolimusLimus, a compound produced by streptomyces hygroscopicus.

The bovatinib may be administered in an amount of 0.1mg to 50 mg. Further, rapamycin may be present at 0.5mg/m²Body surface area to 10mg/m²The amount of body surface area is administered.

In one embodiment, the composition may comprise bosutinib and an antimetabolite. The antimetabolite may be selected from the group consisting of capecitabine, 5-fluorouracil, gemcitabine, pemetrexed, methotrexate, 6-mercaptopurine, cladribine, cytarabine, doxifluridine, floxuridine, fludarabine, hydroxyurea, dacarbazine, hydroxyurea, and asparaginase. An example of the composition may comprise bosutinib and 5-fluorouracil. The 5-fluorouracil may be in the form of an injection.

The bovatinib may be administered in an amount of 0.1mg to 50 mg. 5-Fluorouracil in an amount of 100mg/m²Body surface area to 3000mg/m²The amount of body surface area is administered.

Adrucil as an additional drug^TMFluorouracil (5-FU) sold under the trademark Fluorouracil is a drug used in the treatment of cancer. It can be used for colon cancer, esophageal cancer, gastric cancer, pancreatic cancer, breast cancer and cervical cancer by intravenous injection. As a cream, it is used for basal cell carcinoma. Fluorouracil belongs to the family of antimetabolites and pyrimidine analogue drugs. The mechanism of operation is not completely understood, but is believed to involve blocking the action of thymidylate synthase, thereby preventing DNA production.

In one embodiment, the composition may comprise bosutinib and a platinum-based anti-neoplastic agent. The platinum antineoplastic agent may be selected from cisplatin, carboplatin, dicycloplatin, eptaplatin, lobaplatin, miboplatin, nedaplatin, oxaliplatin, picoplatin and satraplatin. An example of the composition may comprise bosutinib and cisplatin. Cisplatin may be in the form of an injection.

The bovatinib may be administered in an amount of 0.1mg to 50 mg. Cisplatin can be used at 1mg/m²Body surface area to 100mg/m²The amount of body surface area is administered.

Cisplatin is a chemotherapeutic drug used to treat a variety of cancers. This includes testicular, ovarian, cervical, breast, bladder, head and neck, esophageal, lung, mesothelioma, brain and neuroblastoma cancer. Cisplatin is administered intravenously. Cisplatin belongs to a platinum antineoplastic medicine family. It functions in part by binding to DNA and inhibiting DNA replication.

The composition may further comprise at least one pharmaceutically acceptable carrier. The carrier must be acceptable, i.e., compatible with the other ingredients of the formulation, capable of being manufactured into a pharmaceutical formulation, and not deleterious to the recipient thereof. According to another aspect of the present invention, there is also provided a method of preparing a pharmaceutical formulation comprising admixing a pharmaceutical composition of a combination of poecitinib, or a pharmaceutically acceptable solvate or salt thereof, and a cytotoxic agent and/or a molecular-targeting agent, wherein said cytotoxic agent and/or molecular-targeting agent comprises a pharmaceutically acceptable salt or solvate thereof, with one or more pharmaceutically acceptable carriers. As mentioned above, the ingredients of the pharmaceutical composition used may be present in separate pharmaceutical compositions or formulated together into one pharmaceutical preparation.

The pharmaceutical preparations may be presented in unit dosage forms containing a predetermined amount of the active ingredient. As known to those skilled in the art, the amount of active ingredient per dose will depend on the condition being treated, the route of administration, and the age, weight and condition of the patient. Preferred unit dosage formulations are those containing a daily dose or sub-dose, or an appropriate fraction thereof, of the active ingredient. In addition, such pharmaceutical formulations may be prepared by any method known in the pharmaceutical art.

The composition of bosutinib with a cytotoxic agent and/or a molecular targeting agent may be administered by any suitable route. Suitable routes of administration include oral, rectal, nasal, topical (including buccal and sublingual), vaginal and parenteral (including subcutaneous, intramuscular, intravenous, intradermal, intrathecal and epidural). It will be appreciated that the preferred route of administration may vary with, for example, the condition of the recipient of the composition and the cancer to be treated. It is also understood that each administered agent may be administered by the same or different routes, and that all component compounds may be mixed together in one pharmaceutical composition/formulation. Suitably, all component compounds are administered in separate pharmaceutical compositions.

The compounds or compositions of the present invention are incorporated into convenient dosage forms such as capsules, tablets or injectable formulations. Solid or liquid pharmaceutical carriers are used. Solid carriers include starch, lactose, calcium sulfate dihydrate, terra alba, sucrose, talc, gelatin, agar, pectin, acacia, magnesium stearate, and stearic acid. Liquid carriers include syrup, peanut oil, olive oil, saline and water. Similarly, the carrier may include a slow release material, such as glyceryl monostearate or glyceryl distearate, alone or with a wax. The amount of solid carrier can vary widely, but suitably can be from about 25mg to about 1g per dosage unit. When a liquid carrier is used, the preparation will suitably be in the form of a syrup, elixir, emulsion, soft gelatin capsule, sterile injectable solution (e.g., ampoule) or aqueous or nonaqueous liquid suspension.

For example, for oral administration in the form of a tablet or capsule, the active pharmaceutical ingredient may be combined with an oral, non-toxic, pharmaceutically acceptable inert carrier (e.g., ethanol, glycerol, water, etc.). Powders are prepared by comminuting the compound into suitable fine particles and mixing with a similarly comminuted pharmaceutical carrier such as an edible carbohydrate, for example starch or mannitol. One or more of flavoring agents, preservatives, dispersing agents and coloring agents may also be included.

It will be appreciated that, in respect of the type of formulation in question, the formulation may include, in addition to the above ingredients, other agents conventional in the art, for example those suitable for oral administration may include flavouring agents.

In another aspect, a therapeutically effective amount of a composition of the invention (a combination of brigatinib, or a pharmaceutically acceptable solvate or salt thereof, and a cytotoxic agent, or a pharmaceutically acceptable salt or solvate thereof, and/or a molecular targeting agent, or a pharmaceutically acceptable salt or solvate thereof) is administered to a human in a simultaneous or sequential manner. In general, a therapeutically effective amount of an administered agent of the invention will depend on a number of factors including, for example, the age and weight of the subject, the exact condition in need of treatment, the severity of the condition, the nature of the formulation, and the route of administration. Ultimately, the therapeutically effective amount will be determined by the attending physician.

Suitably, the invention relates to the treatment of a tumour associated with overexpression or amplification of at least one of the genes HER1, HER2 and HER4, either wild type or mutant. This includes the following patients: one of HER1, HER2 and HER4 is a wild type patient, two of HER1, HER2 and HER4 are mutant patients, two of HER1, HER2 and HER4 are wild type patients, one of HER1, HER2 and HER4 is a mutant patient, and all of HER1, HER2 and HER4 are mutant patients.

As used herein, the term "wild-type" refers in the art to a polypeptide or polynucleotide sequence that is present in the natural population without genetic modification. It is also understood in the art that a "mutant" includes a polypeptide or polynucleotide sequence having at least one modification to an amino acid or nucleic acid as compared to the corresponding amino acid or nucleic acid found in the wild-type polypeptide or polynucleotide, respectively. Included within the term mutant are Single Nucleotide Polymorphisms (SNPs) in which there is a single base pair difference in the sequence of a nucleic acid strand as compared to the most commonly found (wild-type) nucleic acid strand. Tumors comprising wild-type or mutant forms of HER1, HER2 or HER4 or cancers with amplification of the HER1, HER2 or HER4 gene or overexpression of the HER1, HER2 or HER4 protein can be identified by known methods.

For example, wild-type or mutant HER1, HER2, and HER4 tumor cells can be identified by DNA amplification and sequencing techniques, DNA and RNA detection techniques (including but not limited to Northern and Southern blots, respectively), and/or various biochip array techniques or in situ hybridization. Wild-type and mutant polypeptides may be detected by a variety of techniques including, but not limited to, immunodiagnostic techniques such as ELISA, Western blot (Western blot), or immunocytochemistry.

According to another embodiment, there is provided the use of the combination medicament for the manufacture of a medicament for the treatment of one or more diseases associated with the overexpression or amplification of at least one gene of HER1, HER2 and HER4 or a mutant of HER1, HER2 or HER4 in a subject.

According to another embodiment, there is provided a method of treating a tumor associated with overexpression or amplification of HER1, HER2 or HER4 or a mutant of HER1, HER2 or HER4 in a subject, the method comprising administering a composition of active ingredients comprising a therapeutically effective amount of bosutinib and at least one agent selected from cytotoxic agents and molecular targeting agents. The description of the compositions and tumors relating to the overexpression or amplification of HER1, HER2 or HER4 or mutants of HER1, HER2 or HER4 is the same as defined in the above description.

In a specific embodiment, the tumor is selected from the group consisting of:

(i) non-small cell lung cancer that carries one or more EGFR mutants selected from the group consisting of L858R substitutions, T790M substitutions and/or deletions in exon 19;

(ii) estrogen receptor negative breast cancer with HER1 and/or HER2 overexpression;

(iii) estrogen receptor and progestin receptor double positive breast cancers that are expressed with HER2 but not over-expressed;

(iv) trastuzumab-resistant breast cancer with overexpression of HER 2;

(v) HER2, progesterone receptor and estrogen receptor triple negative HER1 overexpressing breast cancer;

(vi) esophageal cancer with HER2 overexpression;

(vii) gastric cancer with HER2 overexpression;

(viii) one or more EGFR mutants carrying a point mutant, insertion and/or deletion comprising 3-18 nucleotides between amino acids 763-778, or a non-small cell lung cancer having 2, 3 or 4 EGFR exon 20 mutants at one or more residues selected from a763, a767, S768, V769, D770, N771, P772 and H773, and/or no EGFR mutant at residue C797.

(ix) Non-small cell lung cancer carrying one or more exon 20 mutants selected from the group consisting of A763insFQEA, A767insASV, S768dupSVD, V769insASV, D770insSVD, D770insNPG, H773insNPH, N771del insGY, N771del insFH and N771 dupNPH.

In another specific embodiment, the composition and the tumor to be treated are selected from the group consisting of:

(1) boqitinib and paclitaxel for treatment

(a) Non-small cell lung cancer of EGFR mutants carrying L858R substitutions, T790M substitutions and/or deletions in exon 19,

(b) estrogen receptor negative breast cancer accompanied by overexpression of HER2, or

(c) Trastuzumab-resistant breast cancer with overexpression of HER 2;

(2) bozitinib and cisplatin for treatment

(c) Trastuzumab-resistant breast cancer with overexpression of HER 2;

(3) boqitinib and 5-fluorouracil for the treatment of

(a) HER2 is associated with an estrogen receptor negative breast cancer that is overexpressed,

(b) trastuzumab-resistant breast cancer with overexpression of HER2, or

(c) Esophageal cancer with HER2 overexpression;

(4) bosutinib and cetuximab for the treatment of non-small cell lung cancer carrying EGFR substitutions L858R and T790M accompanied by HER1 overexpression;

(5) boqitinib and trastuzumab for treatment

(a) Trastuzumab-resistant breast cancer with overexpression of HER2, or

(b) Gastric cancer with HER2 overexpression; and

(6) bozitinib and vinorelbine for the treatment of cancer

(a) Estrogen receptor negative breast cancer accompanied by overexpression of HER2,

(b) (ii) a trastuzumab-resistant breast cancer that is doubly positive for estrogen receptor and progestin receptor with HER2 overexpression,

(c) HER1 and HER2 overexpress double negative estrogen receptor and progestin receptor double positive breast cancer, or

(d) HER2, estrogen receptor and progestin receptor triple negative HER1 overexpresses breast cancer.

Furthermore, the present disclosure relates to methods of treating cancer using the compositions of the present invention, wherein the component drugs of the pharmaceutical composition are administered in the form of prodrugs. Pharmaceutically acceptable prodrugs of the compounds of the present invention are readily prepared by those skilled in the art.

The terms "day", "daily", and the like, when referring to a dosing regimen, refer to the time of a calendar day, i.e., the time beginning at midnight and ending at a subsequent midnight.

The term "treatment" and derivatives thereof, as used herein, refers to a therapeutically effective regimen for a patient in need thereof. For certain conditions, treatment refers to: (1) a disorder that ameliorates or prevents one or more biological manifestations of the disorder, (2) interfering with (a) one or more points in a biological cascade that leads to or causes the disorder, or (b) one or more biological manifestations of the disorder; (3) alleviating one or more symptoms, effects or side effects associated with the disorder or treatment thereof; or (4) slowing the progression of the disorder or one or more biological manifestations of the disorder. Thus also relates to prophylactic treatment. The skilled person will appreciate that "prevention" is not an absolute term. Medically, "preventing" is understood to mean prophylactically administering a drug to substantially reduce the likelihood or severity of a condition or biological manifestation thereof, or to delay the onset of the condition or biological manifestation thereof. Prophylactic treatment is suitable, for example, when the subject is considered to be at high risk of developing cancer, for example when the subject has a strong family history of cancer or when the subject has been exposed to a carcinogen.

As used herein, the term "effective amount" refers to an amount of a drug or agent that will elicit the biological or medical response of a tissue, system, animal or human that is being sought, for example, by a researcher or clinician. Furthermore, the term "therapeutically effective amount" refers to any amount that results in improved treatment, cure, prevention, or amelioration of a disease, disorder, or side effect, or a decrease in the rate of progression of a disease or disorder, as compared to a corresponding subject not receiving that amount. The term also includes within its scope an amount effective to enhance normal physiological function.

The term "composition" as used herein refers to a combination of the component drugs bosutinib and a cytotoxic agent and/or a molecular targeting agent or a pharmaceutically acceptable salt or solvate thereof, administered simultaneously or sequentially in any manner, respectively. Preferably, the compounds are administered within close proximity to each other if the administration is not simultaneous. Furthermore, it is also possible if the compounds are administered in the same dosage form, for example one compound may be administered topically and the other compound may be administered orally. Suitably, both compounds are administered orally.

Suitably, the compositions of the present invention are administered within a "specified time (specific period)".

The term "specific time" and derivatives thereof, as used herein, refers to the time interval between administration of one component drug and the other component drug of the compositions of the present invention. Unless otherwise specified, a particular time may include simultaneous administration. In an embodiment of the dual component pharmaceutical composition, when the two compounds of the invention are administered once daily, the specific time refers to the time of administration of the bosutinib and the other compound in the relevant order during the day. When one or both compounds of the invention are administered more than once per day, the specified time is calculated based on the first administration of each compound at the specified time. All administrations of the compounds of the invention after the first administration at a particular time are not considered when calculating the particular time.

Suitably, if the compounds are administered within a "specified time" and not simultaneously, they are all administered within about 24 hours, 12 hours, 11 hours, 10 hours, 9 hours, 8 hours, 7 hours, 6 hours, 5 hours, 4 hours, 3 hours, 2 hours, or 1 hour of each other — in this case, the specified time is about 24 hours, 12 hours, 11 hours, 10 hours, 9 hours, 8 hours, 7 hours, 6 hours, 5 hours, 4 hours, 3 hours, 2 hours, or 1 hour. As used herein, for embodiments of the dual component pharmaceutical composition, administration of the bosutinib and the other component drug within less than about 45 minutes of each other is considered to be simultaneous administration.

Suitably, when a composition of the invention is administered for a "specific time", the compounds will be co-administered for a "duration of time".

The term "duration of time" and derivatives thereof as used herein means that two compounds of the invention are administered within a "specified time" on the indicated consecutive days, optionally followed by administration of only one of the constituent compounds on the following consecutive days.

With respect to administration at a "particular time", in at least one embodiment, during the course of treatment, both compounds will be administered within a particular time period for at least 1 day, 2 days, 3 days, 5 days, 7 days, 14 days, or 30 days — in this case, the duration of time will be at least 1 day, 2 days, 3 days, 5 days, 7 days, 14 days, or 30 days. During the course of treatment, when both compounds are administered over a period of 30 days within a specified time, the treatment is considered chronic and will continue until an event (event) such as a reevaluation of the cancer state or a change in the patient's condition warrants a modification of the regimen.

Further with respect to administration at a "specific time", in another embodiment, during treatment with the two-component pharmaceutical composition, the two-component drug will be administered within the specific time for at least 1 day, followed by at least 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, or 7 days of the bozitinib alone — in this case, for a duration of at least 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, or 8 days; suitably, during the course of treatment, both compounds will be administered continuously for at least 2 days over a specified period of time, followed by at least 1,2, 3,4, 5, 6 or 7 days of bosutinib alone-in this case, for a duration of at least 3,4, 5, 6, 7, 8 or 9 days; suitably, during the course of treatment, both compounds will be administered continuously for at least 3 days over a specified period of time, followed by at least 1,2, 3,4, 5, 6 or 7 days of bosutinib alone-in this case, for a duration of at least 4, 5, 6, 7, 8, 9 or 10 days; suitably, during the course of treatment, both compounds will be administered continuously for at least 4 days over a specified period of time, followed by separate administrations for at least 1,2, 3,4 or 7 days — in this case, for a duration of at least 5, 6, 7, 8 or 11 days; suitably, during the course of treatment, both compounds will be administered continuously for at least 5 days over a specified period of time, followed by separate administrations for at least 1,2, 3,4 or 5 days — in this case, for a duration of at least 6, 7, 8, 9 or 10 days. In another embodiment, during the course of treatment, both compounds will be administered for 1 to 3 consecutive days within a specified time period, followed by 3 to 7 consecutive days of administration of bosutinib alone.

In another embodiment, during the course of treatment, both compounds will be administered for 3 to 6 consecutive days within a specified time period, followed by 1 to 4 consecutive days of administration of bosutinib alone. Suitably, during the course of treatment, both compounds will be administered for 2 consecutive days over a specified period of time, followed by 3 to 7 consecutive days of administration of bosutinib alone. Suitably, during the course of treatment, both compounds will be administered within a specified time period of 1 to 3 days over a7 day period, and throughout the other of the 7 day periods, bosutinib will be administered separately. In another embodiment, during the course of treatment, both compounds will be administered within a specified time of 2 days over a7 day period and within the other days of the 7 day period, bosutinib will be administered separately.

Further with respect to administration at a "specified time", during the course of treatment, for one embodiment of a two component pharmaceutical composition, the two compounds will be administered within the specified time for at least 1 day, followed by administration of the other component drug alone for at least 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, or 7 days — in this case, the duration of time will be at least 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, or 8 days; suitably, during the course of treatment, both compounds will be administered continuously for at least 2 days over a specified period of time, followed by administration of the other component drug alone for at least 1,2, 3,4, 5, 6 or 7 days — in this case, for a duration of at least 3,4, 5, 6, 7, 8 or 9 days; suitably, during the course of treatment, both compounds will be administered within a specified time period for at least 3 consecutive days, followed by separate consecutive administration of the other component drug for at least 1,2, 3,4, 5, 6 or 7 days — in this case, for a duration of at least 4, 5, 6, 7, 8, 9 or 10 days; suitably, during the course of treatment, both compounds will be administered within a specified period of time for at least 4 consecutive days, followed by separate consecutive administration of the other component drug for at least 1,2, 3,4 or 7 days — in this case, for a duration of at least 5, 6, 7, 8 or 11 days; suitably, during the course of treatment, both compounds will be administered within a specified period of time for at least 5 consecutive days, followed by separate consecutive administration of the other component drug for at least 1,2, 3,4 or 5 days — in this case, for a duration of at least 6, 7, 8, 9 or 10 days. Suitably, during the course of treatment, both compounds will be administered for a specified period of time for 1 to 3 consecutive days, followed by separate consecutive administration of the other component drugs for 3 to 7 days. Suitably, during the course of treatment, both compounds will be administered for a specified period of time for a period of 3 to 6 consecutive days, followed by separate consecutive administration of the other component drugs for a period of 1 to 4 days. Suitably, during the course of treatment, both compounds will be administered for 2 consecutive days over a specified period of time, followed by 3 to 7 consecutive days of administration of the other component drug alone. Suitably, during the course of treatment, both compounds will be administered within a specified time period of 1 to 3 days over 7 days, and the other component drugs will be administered separately over the other days of the 7 day period. Suitably, during the course of treatment, both compounds will be administered within a specified time period of 2 days over a7 day period, and the other component drugs will be administered separately over the other days of the 7 day period.

Furthermore, with respect to "specific time" administration, during the course of treatment, for one embodiment of the two component pharmaceutical composition, the bosutinib and the other component drug will be administered within a specific time of 1 to 3 days of a7 day period, and the bosutinib will be administered separately within the other days of the 7 day period. Suitably, the 7-day protocol repeats for 2 cycles or 14 days; suitably, 4 cycles or 28 days; suitably, the administration is continuous.

Suitably, during the course of treatment, for one embodiment of the two-component pharmaceutical composition, the bosutinib and the other component drug are administered within a specified time period of 1 to 3 days during 7 days, and the other component drug is administered separately during the other days during 7 days. Suitably, the 7-day protocol repeats for 2 cycles or 14 days; suitably, 4 cycles or 28 days; suitably, the administration is continuous.

Suitably, during the course of treatment, for one embodiment of the two-component pharmaceutical composition, the bosutinib and the other component drug will be administered within a specified time period of 1 to 5 days during 14 days, and the bosutinib will be administered separately within the other days during 14 days. Suitably, the 14-day protocol repeats for 2 cycles or 28 days; suitably, the administration is continuous.

Suitably, during the course of treatment, for one embodiment of the dual component pharmaceutical composition, the bosutinib and the other component drug are administered within a specified time period of 1 to 5 days during 14 days, and the other component drug is administered separately during the other days during 14 days. Suitably, the 14-day protocol repeats for 2 cycles or 28 days; suitably, the administration is continuous.

Suitably, if the compounds are not administered during a "particular time", they are administered sequentially. The term "sequential administration" as used herein refers to, for example, an embodiment of a two-component pharmaceutical composition wherein the administration of the one of the wave-zitinib and the other of the component drugs is continued for 1 day or more, followed by the administration of the other of the wave-zitinib and the other of the component drugs for 1 day or more. Unless otherwise specified, in the case of a two-component pharmaceutical composition, "sequential administration" and all dosing regimens described herein need not be performed from the beginning of treatment to the end of treatment, but need only be administered at some point during the course of treatment, one component drug of the other component drug of the group of bosutinib, followed by the other component drug, or the specified dosing regimen. Further, contemplated herein are drug holidays (drug holidays) used between sequential administration of one and the other of the bosutinib and the other component drug. As used herein, a drug holiday is a period of several days after the continuous administration of one of the wave-zitinib and the other component drug and before the administration of the other, wherein neither the wave-zitinib nor the other component drug is administered. Suitably, the drug holiday will be a time selected from the following days: 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, and 14 days.

With respect to sequential administration:

suitably, for example for one embodiment of a dual component pharmaceutical composition, one of the bosutinib and the other component drug is administered continuously for 1 to 30 days, followed by an optional drug holiday, followed by the other being administered continuously for 1 to 30 days.

Suitably, for example for one embodiment of a dual component pharmaceutical composition, the other component drugs will be administered first, followed by an optional drug holiday, followed by the administration of bosutinib, in that order.

Suitably, the bosutinib will be administered first, followed by an optional drug holiday, and then the other component drugs in the sequence.

It is understood that "specific time" administration and "sequential" administration may be followed by repeated administrations of one or more cycles, or may be an alternative dosing regimen, and that a drug holiday may precede the repeated administrations or the alternative dosing regimen.

Suitably, the amount of boresinib administered as part of a composition according to the present invention will be selected from about 0.1mg to about 50 mg; suitably, the amount will be selected from about 0.5mg to about 50 mg; suitably, the amount will be selected from about 1mg to about 50 mg; suitably, the amount will be selected from about 5mg to about 50 mg; suitably, the amount will be selected from about 1mg to about 30 mg; suitably, the amount will be selected from about 5mg to about 20 mg; suitably, the amount will be selected from about 1mg to about 10 mg; suitably, the amount will be selected from about 0.1mg to about 5 mg; suitably, the amount is 1mg, suitably, the amount is 5mg, suitably, the amount is 10mg, suitably, the amount is 20mg, suitably, the amount is 30 mg; suitably, the amount is 50 mg. Accordingly, the amount of boresinib administered as part of a composition according to the present invention will be selected from about 0.1mg to about 50 mg. For example, the amount of borutinib administered as part of a composition according to the invention is suitably selected from 1mg, 5mg, 10mg, 20mg, 30mg and 50 mg. Suitably, the selected amount of bosutinib is administered from 1 to 4 times per day in the form of one or more tablets. Suitably, the selected amount of bosutinib is administered twice daily in one or more tablets. Suitably, the selected amount of bosutinib is administered once daily in one or more tablets. Suitably, administration of the bovatinib will begin with a loading dose. Suitably, the loading dose will be 2 to 100 times the maintenance dose; suitably 2 to 10 times; suitably 2 to 5 times; suitably 2 times; suitably 3 times; suitably 4 times; suitably 5 times. Suitably, the loading dose is administered for 1 to 7 days; suitably 1 to 5 days; suitably 1 to 3 days; suitably 1 day; suitably 2 days; suitably 3 days, followed by a maintenance dosage regimen.

Suitably, the amount of the second component drug (which is not bosutinib) administered as part of a composition according to the invention will be selected from about 0.1mg/m²To about 3500mg/m²About 0.5mg/m²To about 3500mg/m²(ii) a Suitably, the amount will be selected from about 1.0mg/m²To about 3500mg/m²(ii) a Suitably, the amount will be selected from about 10.0mg/m²To about 3500mg/m²(ii) a Suitably, the amount will be selected from about 50.0mg/m²To about 3500mg/m²(ii) a Suitably, the amount will be selected from about 100.0mg/m²To about 3500mg/m²(ii) a Suitably, the amount will be selected from about 100.0mg/m²To about 3000mg/m²(ii) a Suitably, the amount will be selected from about 0.1mg/m²To about 20mg/m²(ii) a Suitably, the amount will be selected from about 0.5mg/m²To about 10mg/m²(ii) a Suitably, the amount will be selected from about 0.5mg/m²To about 10mg/m²(ii) a Suitably, the amount will be selected from about 0.5mg/m²To about 50mg/m²(ii) a Suitably, the amount will be selected from about 50mg/m²To about 1000mg/m²(ii) a Suitably, the amount will be selected from about 100mg/m²To about 500mg/m²(ii) a Suitably, the amount will be selected from about 100mg/m²To about 300mg/m². Suitably, the selected amount of the second component medicament which is not bortinib is administered from 1 to 4 times a day. SuitablyThe selected amount of the second component medicament is administered from 1 to 4 times a day.

As used herein, the specific amounts of all of the bosutinib and other component drugs are expressed as the amount of free or unsalted and undissolved compound administered per dose.

Accordingly, the present invention further provides a pharmaceutical composition comprising bosutinib and other component drugs together with one or more pharmaceutically acceptable carriers.

According to another embodiment, there is provided a kit for the treatment of a tumor associated with overexpression or amplification of HER1, HER2 or HER4 or a mutant of HER1, HER2 or HER4 in a subject, the kit comprising a first part and a second part, wherein the first part comprises bosutinib and the second part comprises at least one active ingredient selected from a cytotoxic agent and a molecular targeting agent. The kit may further comprise a package insert comprising instructions for treating a tumor associated with overexpression or amplification of HER1, HER2, or HER4, or a mutant thereof, in a subject.

The descriptions of bosutinib, cytotoxic agents and molecular targeting agents and of the expressions "HER 1, HER2 or HER4 or mutants of HER1, HER2 or HER 4" and "overexpression" are the same as defined in the above description.

The term "composition kit" as used herein refers to one or more pharmaceutical compositions for administration. For example, the brigatinib, or a pharmaceutically acceptable solvate and/or salt thereof, is conjugated to at least one member selected from the group consisting of cytotoxic agents and molecular targeting agents, which also include pharmaceutically acceptable salts or solvates of cytotoxic agents and molecular targeting agents. When the compounds can be administered simultaneously, at least two compounds can be in the same pharmaceutical composition, e.g., a tablet. When the compounds are not administered simultaneously, the combination kit will contain the bosutinib and the other component drugs or pharmaceutically acceptable salts or solvates thereof in different pharmaceutical compositions. The composition kit may comprise the bosutinib and the other component drugs or pharmaceutically acceptable salts or solvates thereof in different pharmaceutical compositions in the same package or in different pharmaceutical compositions in different packages.

In the composition kit, the components may be provided in a suitable sequential, separate and/or simultaneous administration.

The "composition kit" may also be provided with instructions, such as dosages and instructions for administration. Such dosages and administration instructions may be of the kind provided to the physician, for example by a pharmaceutical label, or they may be of the kind provided by the physician, for example instructions to the patient.

According to another embodiment, a method for treating a tumor associated with overexpression or amplification of HER1, HER2 or HER4 or a mutant of HER1, HER2 or HER4 in a subject in need thereof is described, wherein the genotype and/or phenotypic status of EGFR, such as HER1, HER2, HER4, is determined in the subject prior to the appropriate combination therapy described herein. In another embodiment, such status of HER1, HER2 and HER4 in a subject may be determined by suitable immunohistochemistry or in situ rehybridization methods. In another embodiment, the invention also provides a method of treating a human patient at risk of an overexpression or amplification-related disorder of HER1, HER2 or HER4 or a mutant of HER1, HER2 or HER4 by first determining the susceptibility of the individual human patient to HER1, HER2 or HER4 mutant and then administering a therapeutically effective composition as described herein.

One or more embodiments will now be described in more detail with reference to the following examples. However, these examples are not intended to limit the scope of one or more embodiments.

Example 1: cell growth inhibition assay and composition data analysis

The effect of the compositions of the present invention on the inhibition of cancer cell growth was tested in EGFR-overexpressing or mutated cancer cell lines.

1. Cell lines and cell growth conditions and IC50 determination of Individual Compounds and compositions thereof

The information on cancer cells and cell growth conditions is shown in table 1 below.

TABLE 1

Abbreviations

ER: an estrogen receptor; PR: a progestogen receptor; HER 2: human epidermal growth factor receptor 2; HER 1: human epidermal growth factor receptor 1; PIK3 CA: phosphatidylinositol 3-kinase (phosphoinositide-3-kinase), catalyst, alpha polypeptide; and (4) PTEN: a homologous phosphatase; +: positive (over-expression); -: negative; m: a mutant; ATCC: american Type Culture Collection (American Type Culture Collection); FBS: fetal bovine serum.

All cells were cultured for 72 hours prior to cell plating, except for H1975. The cells are in a size of 4-15X 10³Individual cells/well were seeded in 96-well plates. Approximately 24 hours after plating, cells were exposed to 10-fold, 2-fold, or 3-fold serial dilutions of either the individual test compounds or the two test compounds in a composition at a constant or non-constant molar ratio of the two agents. The cells were cultured for 3 days in the presence of the compound except for H1975, which was 3X 10³Cells/well were cultured for 2 days.

2. Measurement of cell growth inhibition (CI)

Cell growth inhibition rate based on cellular protein content measurements according to "Nat protoc.2006; 1, (3) and 1112-6' are measured by a sulforhodamine B colorimetric method. Absorbance at 540nm in SpectraMax^TMRead on a plate reader. For H1975 cells, according to the manufacturer's protocol, by adding CellTiter^TM96 Single Solution Cell Proliferation Assay reagent (Aqueous One Solution Cell Proliferation Assay reagent) (Promega) was used to determine the Cell growth rate (%).

After treatment of the incubation time with test agent or test agent composition (2-3 days) and comparison of the signal to cells treated with vehicle (DMSO), the rate of cell growth inhibition was estimated. The cell growth rate (%) was calculated by the following equation.

The percentage of growth inhibition was calculated as:

[ (Ti-Tz)/(C-Tz) ] × 100, concentration for Ti-Tz

[ (Ti-Tz)/Tz ] x 100 for concentrations of Ti < Tz

Each T value represents absorbance. Tz: time zero (pre-treatment cell density panel for test panel), C: growth was controlled with 100% test reagent, Ti: growth was tested in the presence of concentration levels of drug. GI (GI tract)₅₀The concentration at which 50% inhibition occurs is selected from the group consisting of [ (Ti-Tz)/(C-Tz)]X 100 is calculated as 50.

The effect of the combination effect was evaluated by the combination index using Calcusyn Software. Table 2 describes the Combination Index (CI) range for the two compounds.

TABLE 2

CI (combination index) range	Description of the invention
		<0.1	Strong synergy
0.1-0.3	Strong synergy
		0.3-0.7	Collaboration
0.7-0.85	Moderate synergy
		0.85-0.9	Slight synergy
0.9-1.1	Approximate addition
		1.1-1.2	Slight antagonism
1.2-1.45	Moderate antagonism
		1.45-3.3	Antagonism of
3.3-10	Strong antagonism
		>10	Strong antagonism

3. Results

Tables 3a-3f summarize the growth inhibitory effect of each single agent and Combination (CI) on cell growth in vitro.

TABLE 3a

TABLE 3b

TABLE 3c

TABLE 3d

TABLE 3e

TABLE 3f

Using GraphPad Prism^TMv6 calculated the 50% growth inhibition value (GI) of the combination of Bo-zitinib and partner drugs (partner drugs)₅₀) And calculates the combined GI₅₀Single dose growth inhibition (%) at dose. Using Calcusyn^TM(Biosoft) evaluation of Combined GI₅₀Combination Index (CI) at dose. Table 4 summarizes the combined effect of cell growth in vitro.

Table 4 summarizes

Example 2: synergistic study of cell proliferation on selected cell lines

In this example, the direct cell killing effect of bocetinib alone or in combination with other drugs for the treatment of selected cancer cell lines Calu-3, BT-474 and SK-BR-3 was evaluated.

1. Cell lines and cell growth conditions as well as IC50 assays for individual compounds and combinations thereof.

All cells were incubated at the listed temperature of 37 ℃ with 5% CO₂And 95% humidity. The cells are in a size of 4-15X 10³Individual cells/well were seeded in 96-well plates. Approximately 24 hours after plating, cells were exposed to 10-fold, 2-fold, or 3-fold serial dilutions of either the individual test compounds or the two test compounds in a composition with either a constant or non-constant molar ratio of the two agents. Cells were cultured in the presence of the compound for 3 days.

The growth inhibitory effect of each of the individual and combined regimens is provided below.

Watch 6(a)

Watch 6(b)

Watch 6(c)

The cytostatic activity of a single test compound representing multiple mechanisms of action was examined on Calu-3, BT-474 and SK-BR-3 cell lines. Bozitinib showed greater inhibition potential of cell proliferation in all three cell lines, with IC50 for Calu-3 being 0.001M to 0.009M, IC50 for BT-474 being less than 0.0015M and 0.001M, and IC50 for SK-BR-3 being 0.001M. IC50 for sorafenib, temsirolimus, and trametinib were 4.475, 0.167, and 0.042M, respectively, for Calu-3 cell lines, while BT-474 showed weak or no response to letrozole, palbociclib, and trametinib treatment. Pemetrexed was not effective on these two cells, whereas T-DM1 was most responsive, with the IC50 for Calu-3 being 90.449g/mL and the IC50 for SK-BR-3 being less than 0.030 g/mL.

2. Measurement of cell growth inhibition Rate (CI)

After treatment of the incubation time (2-3 days) with the test agent or combination of test agents and comparison of the signal to cells treated with vehicle (DMSO), the rate of inhibition of cell growth was estimated. The cell growth rate (%) was calculated by the following equation.

Percentage growth inhibition was also calculated using a Bliss independence model and a Lowewe adaptability model. A score above 5 indicates synergy and a score below-5 indicates antagonism.

TABLE 5

3. Results

Calu-3, BT-474 or SK-BR-3 cell lines with the following Bo-zitinib in combination with a second compound in a 6X 6 combination matrix at 37 ℃ with 5% CO, respectively₂And testing at 95% humidity for 3 days:

watch 6(d)

TABLE 6(e)

The table below shows the CI values (or cell viability table) for the two compound compositions for selected cancer cell lines.

TABLE 6(f) Calu-3 Bo-zitinib + Sorafenib

TABLE 6(g) Calu-3 Bevacizinib + Tacrolimus

TABLE 6(h) Calu-3 Bo-zitinib + Trametinib

Fig. 8(a) and (B) show the effect of the combination of bosutinib and pemetrexed or letrozole on the Calu3 cell line.

TABLE 6(i) Calu-3 Bo-zitinib + T-DM1

TABLE 6(j) BT474 Boqitinib + Pabociclib

TABLE 6(k) BT474 Bozitinib + Trametinib

TABLE 6(l) BT474 Bozitinib + Pemetrexed

TABLE 6(m) SK-BR-3 Bo zitinib + T-DM1

The study was aimed at further evaluating the possible synergistic effects of the above compositions on Calu-3, BT-474 and SK-BR-3 cell lines. From the analysis using the Combination Index (CI), most compositions showed synergy, particularly when the two test compounds were treated in the mid concentration range separately.

4. Discussion of the related Art

As shown in the above table, according to one embodiment, the combination of bosutinib and another drug exhibits a synergistic effect in a growth inhibition assay on breast, gastric, lung or esophageal cancer cells, wherein the HER1 or HER2 gene is overexpressed or mutated.

In the case of non-small cell lung cancer (NSCLC), bosutinib exhibited excellent synergy when administered in combination with paclitaxel or cisplatin in HCC827 cells with an exon 19del mutation, a Her1 activating mutation. When bosutinib is combined with 5-FU, no synergy is observed under the same test conditions. In H1975 cells (i.e., one)Cell lines that have resistance to first generation EGFR TKis due to the presence of T790M mutant) when bosutinib is administered at or below GI of bosutinib alone₅₀Shows excellent synergistic effect when combined with the EGFR antibody cetuximab. In H1975 cells, synergy was shown when bosutinib was combined with paclitaxel or 5-FU at certain concentrations, but no synergy was observed when bosutinib was combined with cisplatin under the same test conditions.

In the case of breast cancer, bosutinib has a high synergistic effect when used in combination with paclitaxel, cisplatin or 5-FU in BT-474 cells, which are Her2 overexpressed and negative for Estrogen Receptors (ER). When bosutinib is combined with trastuzumab, at or below GI when bosutinib is used alone₅₀At certain concentrations of (a), a weak synergistic effect is exhibited. (ii) at or below GI of Boletinib alone when combined with vinorelbine₅₀No synergistic effect was observed at the concentrations of (a). Similarly, when vinorelbine was used in combination with poecitinib in Her2 over-expressed and ER-negative SK-BR-3 cells, a significant effect was observed, indicating a synergistic effect. However, the combination of bosutinib with vinorelbine showed synergy at certain concentrations in MDA-MB-361 cells of HERr2 overexpressed, ER positive and trastuzumab resistant cell line, such combination showed synergy at each concentration in MCF cells that were not HER1 and HER2 overexpressed and not ER positive.

Furthermore, a synergistic effect was observed by administering certain concentrations of a combination of bosutinib and vinorelbine in MDA-MB-468 cells, which are HER2 negative, ER negative, and HER1 overexpressed triple negative breast cancer cells. In MBA-MB-453 cells (which are trastuzumab-resistant breast cancer cells in which HER2 is overexpressed), when paclitaxel, 5-FU, cisplatin, or trastuzumab is combined with paucitinib, the GI at or below that of paucitinib alone₅₀An excellent synergistic effect was observed at the concentration of (a).

In TE cells, an esophageal cancer cell line with overexpression of HER2, the synergistic effect of the combination of bosutinib with other drugs is also very good when bosutinib is combined with 5-FU. Furthermore, in N-87 cells, a gastric cancer cell line with overexpression of HER2, synergy was observed at certain concentrations when bosutinib was administered in combination with trastuzumab.

These results indicate that, according to one embodiment, the combination of bosutinib and other targeted or cytotoxic anticancer agents is very effective against cancers such as breast cancer, gastric cancer, lung cancer and esophageal cancer, and can effectively inhibit cancers resistant to conventional therapeutic agents. The composition according to one embodiment has been shown to be effective even on cancer cells that do not have overexpression of HER1 or HER2 or mutants thereof.

As described above, according to one or more embodiments, a combination drug for treating a tumor associated with overexpression or amplification of at least one gene of HER1, HER2 and HER4 or a mutant of HER1, HER2 or HER4 in a subject is provided, and the combination drug can be used for effectively treating a tumor.

According to another embodiment, a method is provided for treating a tumor associated with overexpression or amplification of at least one gene of HER1, HER2 and HER4 or a mutant of HER1, HER2 or HER4 in a subject; when the method is used in a patient in need thereof, the tumor can be effectively treated.

According to another embodiment, a composition kit and a pharmaceutical composition for treating a tumor associated with overexpression or amplification of at least one gene of HER1, HER2 and HER4 or a mutant of HER1, HER2 or HER4 in a subject may be used for effective treatment of the tumor.

Other aspects of the invention include novel compositions and pharmaceutical regimens of a composition of bosutinib as an active ingredient with at least one cytotoxic agent and/or at least one molecular targeting agent, wherein the at least one cytotoxic agent is selected from the group consisting of a taxane, a base analog, a platinum-based antineoplastic agent, and a vinca alkaloid; and wherein the at least one molecular targeting agent is selected from at least one Epidermal Growth Factor Receptor (EGFR) family inhibitor. The EGFR family inhibitor may be an anti-EGFR family antibody. The composition may comprise bosutinib and an anti-EGFR family antibody, wherein the anti-EGFR family antibody may be trastuzumab, margetuximab cetuximab, matuzumab, panitumumab, nixituzumab, or pertuzumab. The Epidermal Growth Factor Receptor (EGFR) family inhibitor may be an mTOR inhibitor. The composition may comprise bosutinib and a taxane. The taxane may be selected from paclitaxel, docetaxel and cabazitaxel. The composition may comprise bosutinib and a base analog. The base analog can be selected from 5-fluorouracil, 6-mercaptopurine, capecitabine, gemcitabine, pemetrexed, methotrexate, cladribine, cytarabine, doxifluridine, floxuridine, fludarabine, and dacarbazine. The composition may comprise bosutinib and a platinum antineoplastic agent. The platinum antineoplastic agent is selected from cisplatin, carboplatin, dicycloplatin, eptaplatin, lobaplatin, miboplatin, nedaplatin, oxaliplatin, picoplatin and satraplatin. The composition may comprise bosutinib and a vinca alkaloid. The vinca alkaloid is selected from vinblastine, vincristine, vinflunine, vinorelbine, vinamidol, vinblastine, vindoline and vindesine. The mTOR inhibitor is selected from zotarolimus, pimecrolimus, temsirolimus, sirolimus Nanocrystal, sirolimus TransDerm, sirolimus-PNP, everolimus, biolimus A9, ridaforolimus, rapamycin, TCD-10023, DE-109, MS-R001, MS-R002, MS-R003, Perceiva, XL-765, quinacrine, PKI-587, PF-04691502, GDC-0980, dacylisib, CC-223, PWT-33597, P-7170, LY-3023414, INK-128, GDC-0084, DS-7423, DS-3078, CC-115, CBLC-137, AZD-2014, X-480, X-414, EC-0371, VS-5584, PQR-316, PT-309, MTR-06465603, MTNR-35311, NV-nOR, NV-51, NV-R-35311, and PQNnR, BC-210, WAY-600, WYE-354, WYE-687, LOR-220, HMPL-518, GNE-317, EC-0565, CC-214, ABTL-0812, and pharmaceutically acceptable salts thereof or combinations thereof. The at least one cytotoxic agent may be selected from the group consisting of paclitaxel, cisplatin, 5-fluorouracil, vinorelbine, and any combination thereof. The at least one molecular targeting agent may be selected from the group consisting of cetuximab, trastuzumab, and any combination thereof. The combination may be selected from the following: (A) bosutinib and paclitaxel; (B) bosutinib and cisplatin; (C) bosutinib and 5-fluorouracil; (D) bosutinib and paclitaxel; (E) bosutinib and trastuzumab; and (F) bosutinib and vinorelbine.

For a combination drug regimen, the tumor can be selected from non-small cell lung cancer, breast cancer, gastric cancer, colon cancer, pancreatic cancer, prostate cancer, myeloma, head and neck cancer, ovarian cancer, esophageal cancer, and metastatic cell cancer. The tumor can be selected from non-small cell lung cancer, breast cancer, gastric cancer and esophageal cancer. The tumor may be selected from: (i) non-small cell lung cancer carrying one or more EGFR mutants selected from the group consisting of L858R substitutions, T790M substitutions and/or deletions in exon 19, and/or one or more EGFR mutants selected from the group consisting of a763, a767, S768, V769, D770, N771, P772 and H773 substitutions and/or deletions in exon 20; (ii) estrogen receptor negative breast cancer with HER1 and/or HER2 overexpression; (iii) estrogen receptor and progestin receptor double positive breast cancers that are expressed with HER2 but not over-expressed; (iv) trastuzumab-resistant breast cancer with overexpression of HER 2; (v) HER1 overexpressing breast cancer that is triple negative for HER2, progesterone receptor, and estrogen receptor; (vi) esophageal cancer with HER2 overexpression; and (vii) gastric cancer with HER2 overexpression.

The combination drug and the tumour to be treated may be selected from: (1) bosutinib and paclitaxel for use in the treatment of (a) a non-small cell lung cancer that carries an EGFR mutant with L858R substitution, T790M substitution and/or deletion in exon 19 and/or one or more EGFR mutants selected from a763, a767, S768, V769, D770, N771, P772 and H773 substitution and/or deletion in exon 20, (b) an estrogen receptor negative breast cancer that is accompanied by overexpression of HER2, or (c) trastuzumab-resistant breast cancer that is accompanied by overexpression of HER 2; (2) bosutinib and cisplatin for use in the treatment of non-small cell lung cancer carrying an EGFR mutant with L858R substitution, T790M substitution and/or deletion in exon 19 and/or one or more EGFR mutants selected from a763, a767, S768, V769, D770, N771, P772 and H773 substitution and/or deletion in exon 20, (b) estrogen receptor negative breast cancer with HER2 overexpression, or (c) trastuzumab-resistant breast cancer with HER2 overexpression; (3) bosutinib and 5-fluorouracil for the treatment of (a) estrogen receptor negative breast cancer with overexpression of HER2, (b) trastuzumab-resistant breast cancer with overexpression of HER2, or (c) esophageal cancer with overexpression of HER 2; (4) bosutinib and cetuximab for the treatment of non-small cell lung cancer carrying EGFR substitutions of L858R and T790M accompanied by overexpression of HER1 and/or carrying one or more EGFR mutants selected from substitutions and/or deletions of a763, a767, S768, V769, D770, N771, P772 and H773in exon 20; (5) bosutinib and trastuzumab for the treatment of (a) trastuzumab-resistant breast cancer with overexpression of HER2 or (b) gastric cancer with overexpression of HER 2; and (6) bosutinib and vinorelbine for the treatment of (a) estrogen receptor negative breast cancer that is overexpressed with HER2, (b) estrogen receptor and progestin receptor double positive and trastuzumab-resistant breast cancer that is overexpressed with HER2, (c) estrogen receptor and progestin receptor double positive breast cancer that is overexpressed with HER1 and HER2, or (d) HER1 overexpressed that is triple negative with HER2, estrogen receptor and progestin receptor.

Another aspect of the invention relates to a method of treating a tumor in a subject in need or at risk of developing HER1, HER2, an overexpressing cancer, wherein said method comprises administering to said subject a therapeutically effective amount of pazitinib as an active ingredient and at least one cytotoxic agent and/or at least one molecular targeting agent, alone or in combination, wherein said at least one cytotoxic agent is selected from the group consisting of a taxane, a base analog, a platinum antineoplastic agent and a vinca alkaloid; and wherein the at least one molecular targeting agent is selected from the group consisting of an Epidermal Growth Factor Receptor (EGFR) family inhibitor and a mammalian target of rapamycin (mTOR) inhibitor. The at least one cytotoxic agent of the method may be selected from the group consisting of paclitaxel, cisplatin, 5-fluorouracil, vinorelbine, and any combination thereof. The at least one molecular targeting agent of the method may be selected from the group consisting of cetuximab, trastuzumab, and any combination thereof. Administration alone or in combination may be selected from the following: (a) bosutinib and paclitaxel; (b) bosutinib and cisplatin; (c) bosutinib and 5-fluorouracil; (d) bosutinib and cetuximab; (e) bosutinib and trastuzumab; (f) bosutinib and vinorelbine. The tumor of the method may be non-small cell lung cancer, breast cancer, gastric cancer, colon cancer, pancreatic cancer, prostate cancer, myeloma, head and neck cancer, ovarian cancer, esophageal cancer, or metastatic cell cancer. The tumor can be selected from non-small cell lung cancer, breast cancer, gastric cancer and esophageal cancer. The tumor may be selected from: (i) non-small cell lung cancer carrying one or more EGFR mutants selected from the group consisting of L858R substitutions, T790M substitutions and/or deletions in exon 19, and/or one or more EGFR mutants selected from the group consisting of a763, a767, S768, V769, D770, N771, P772 and H773 substitutions and/or deletions in exon 20; (ii) estrogen receptor negative breast cancer with HER1 and/or HER2 overexpression; (iii) estrogen receptor and progestin receptor double positive breast cancers that are expressed with HER2 but not over-expressed; (iv) trastuzumab-resistant breast cancer with overexpression of HER 2; (v) HER1, HER2, and HER1 that are triple negative for estrogen receptor overexpress breast cancer; (vi) esophageal cancer with HER2 overexpression; and (vii) gastric cancer with HER2 overexpression. The tumor may be selected from: (i) non-small cell lung cancer carrying one or more EGFR mutants selected from the group consisting of L858R substitutions, T790M substitutions and/or deletions in exon 19, and/or one or more EGFR mutants selected from the group consisting of a763, a767, S768, V769, D770, N771, P772 and H773 substitutions and/or deletions in exon 20; (ii) breast cancer that is estrogen receptor negative with HER1 and/or HER2 overexpression; (iii) estrogen receptor and progestin receptor double positive breast cancers that are expressed with HER2 but not over-expressed; (iv) trastuzumab-resistant breast cancer with overexpression of HER 2; (v) HER1, HER2, and HER1 that are triple negative for estrogen receptor overexpress breast cancer; (vi) esophageal cancer with HER2 overexpression; and (vii) gastric cancer with HER2 overexpression.

The administration and the tumor to be treated of the method are selected from: (1) pazitinib and paclitaxel for use in the treatment of (a) a non-small cell lung cancer that carries an EGFR mutant with a L858R substitution, a T790M substitution and/or deletion in exon 19, and/or one or more EGFR mutants selected from the group consisting of a763, a767, S768, V769, D770, N771, P772 and H773 mutations and/or deletions in exon 20, (b) an estrogen receptor negative breast cancer that is accompanied by overexpression of HER2, or (c) a trastuzumab-resistant breast cancer that is accompanied by overexpression of HER 2; (2) bosutinib and cisplatin for use in the treatment of (a) a non-small cell lung cancer that carries an EGFR mutant with L858R substitution, T790M substitution and/or deletion in exon 19, and/or one or more EGFR mutants selected from a763, a767, S768, V769, D770, N771, P772, and H773 substitution and/or deletion in exon 20, (b) an estrogen receptor negative breast cancer that accompanies HER2 overexpression, or (c) trastuzumab-resistant breast cancer that accompanies HER2 overexpression; (3) bosutinib and 5-fluorouracil for the treatment of (a) estrogen receptor negative breast cancer with overexpression of HER2, (b) trastuzumab-resistant breast cancer with overexpression of HER2, or (c) esophageal cancer with overexpression of HER 2; (4) bosutinib and cetuximab for the treatment of non-small cell lung cancer carrying EGFR substitutions of L858R and T790M accompanied by HER1 overexpression and/or carrying one or more EGFR mutants selected from substitutions and/or deletions of a763, a767, S768, V769, D770, N771, P772 and H773in exon 20; (5) bosutinib and trastuzumab for the treatment of (a) trastuzumab-resistant breast cancer with overexpression of HER2, or (b) gastric cancer with overexpression of HER 2; (6) bosutinib and vinorelbine for use in the treatment of (a) estrogen receptor negative breast cancer which is accompanied by overexpression of HER2, (b) estrogen receptor and progestin receptor double positive and trastuzumab-resistant breast cancer which is accompanied by overexpression of HER2, (c) estrogen receptor and progestin receptor double positive breast cancer which is accompanied by overexpression of HER1 and HER2, or (d) HER1 overexpressed breast cancer which is triple negative for HER2, estrogen receptor and progestin receptor.

Another aspect of the invention relates to a kit for treating a tumor in a subject, the kit comprising a first part and a second part, wherein the first part comprises bosutinib and the second part comprises at least one active ingredient selected from the group consisting of a cytotoxic agent and a molecular targeting agent, wherein the at least one cytotoxic agent is selected from the group consisting of a taxane, a base analog, a platinum antineoplastic agent and a vinca alkaloid; wherein at least one molecular targeting agent is selected from the group consisting of inhibitors of the Epidermal Growth Factor Receptor (EGFR) family and inhibitors of the mammalian target of rapamycin (mTOR). The kit may further comprise a package insert comprising instructions for treating a tumor associated with overexpression or amplification of HER1, HER2, or HER4, or a mutant thereof, in a subject.

It should be understood that the embodiments described herein should be considered only as illustrative and not limiting the scope of the invention. Descriptions of features or aspects in each embodiment should generally be considered as available for other similar features or aspects in other embodiments. Although one or more embodiments have been described with reference to the accompanying drawings, it will be understood by those of ordinary skill in the art that various changes in form and details may be made therein without departing from the spirit and scope of the present invention as defined by the following claims.

Claims

1. A combination for the treatment of a tumour in a subject, said combination comprising as active ingredients a combination of bosutinib and at least one cytotoxic agent and/or at least one molecular targeting agent,

wherein the at least one cytotoxic agent is selected from the group consisting of a taxane, a base analog, a platinum antineoplastic agent, and a vinca alkaloid; and

wherein the at least one molecular targeting agent is selected from at least one Epidermal Growth Factor Receptor (EGFR) family inhibitor.

2. The combination as defined in claim 1, wherein the EGFR family inhibitor is an anti-EGFR family antibody.

3. The combination as described in claim 1, wherein the composition comprises bosutinib and an anti-EGFR family antibody.

4. The combination as described in claim 3, wherein the anti-EGFR family antibody is trastuzumab, T-DM1, margetuximab cetuximab, matuzumab, panitumumab, nixituzumab, or pertuzumab.

5. The combination as described in claim 1, wherein the Epidermal Growth Factor Receptor (EGFR) family inhibitor is an mTOR inhibitor.

6. The combination as described in claim 1, wherein the composition comprises bosutinib and a taxane.

7. The combination as claimed in claim 6, wherein the taxane is selected from paclitaxel, docetaxel and cabazitaxel.

8. The combination as described in claim 1, wherein the composition comprises bosutinib and a base analog.

9. The combination as described in claim 8, wherein the base analog is selected from the group consisting of 5-fluorouracil, 6-mercaptopurine, capecitabine, gemcitabine, pemetrexed, methotrexate, cladribine, cytarabine, doxifluridine, floxuridine, fludarabine, and dacarbazine.

10. The combination as described in claim 1, wherein the composition comprises bosutinib and a platinum antineoplastic agent.

11. The combination as claimed in claim 10, wherein the platinum antineoplastic agent is selected from cisplatin, carboplatin, dicycloplatin, eptaplatin, lobaplatin, miboplatin, nedaplatin, oxaliplatin, picoplatin and satraplatin.

12. The combination as recited in claim 1, wherein the composition comprises bosutinib and a vinca alkaloid.

13. The combination as claimed in claim 12, wherein the vinca alkaloid is selected from vinblastine, vincristine, vinflunine, vinorelbine, vincamine, vinblastine, vindoline and vindesine.

14. The combination as described in claim 5, wherein the mTOR inhibitor is selected from zotarolimus, pimecrolimus, temsirolimus, sirolimus NanoCrystal, sirolimus TransDerm, sirolimus-PNP, everolimus, biolimus A9, ridaforolimus, rapamycin, TCD-10023, DE-109, MS-R001, MS-R002, MS-R003, Perceiva, XL-765, quinacrine, PKI-587, PF-04691502, GDC-0980, dacolisib, CC-223, PWT-33597, P-7170, LY-3023414, INK-128, GDC-0084, DS-7423, DS-3078, CC-115, CBLC-137, AZD-480, X-414, EC-550371, VS-84, PQR-309, PQR-401, PQR-2014R-2014, PQR-309, PQR-2014, and/or combinations thereof, PF-06465603, NV-128, nPT-MTOR, BC-210, WAY-600, WYE-354, WYE-687, LOR-220, HMPL-518, GNE-317, EC-0565, CC-214, ABTL-0812, and pharmaceutically acceptable salts or compositions thereof.

15. The combination according to claim 1, wherein the at least one cytotoxic agent is selected from the group consisting of paclitaxel, cisplatin, 5-fluorouracil, vinorelbine and any combination thereof.

16. The combination as described in claim 1, wherein the at least one molecular targeting agent is selected from the group consisting of cetuximab, trastuzumab, T-DM1, and any combination thereof.

17. The combination as described in claim 1, wherein the composition is selected from the following:

(A) bosutinib and paclitaxel;

(B) bosutinib and cisplatin;

(C) bosutinib and 5-fluorouracil;

(D) bosutinib and cetuximab;

(E) bosutinib and trastuzumab;

(F) bosutinib and T-DM 1; and

(G) bosutinib and vinorelbine.

18. The combination as described in claim 1, wherein the tumor is selected from the group consisting of non-small cell lung cancer, breast cancer, gastric cancer, colon cancer, pancreatic cancer, prostate cancer, myeloma, head and neck cancer, ovarian cancer, esophageal cancer, and metastatic cell cancer.

19. The combination as described in claim 18, wherein the tumor is selected from the group consisting of non-small cell lung cancer, breast cancer, gastric cancer, and esophageal cancer.

20. The combination as described in claim 19, wherein the tumor is selected from the group consisting of:

(i) non-small cell lung cancer carrying one or more EGFR mutants selected from the group consisting of L858R substitutions, T790M substitutions and/or deletions in exon 19, and/or one or more EGFR mutants selected from the group consisting of a763, a767, S768, V769, D770, N771, P772 and H773 substitutions and/or deletions in exon 20;

(iv) trastuzumab-resistant breast cancer with overexpression of HER 2;

(v) HER1 overexpressing breast cancer that is triple negative for HER2, progesterone receptor, and estrogen receptor;

(vi) esophageal cancer with HER2 overexpression; and

(vii) gastric cancer accompanied by overexpression of HER 2.

21. The combination as described in claim 1, wherein the composition and the tumor to be treated are selected from the group consisting of:

(1) boqitinib and paclitaxel for treatment

(a) A non-small cell lung cancer which carries an EGFR mutant which carries a substitution L858R, a substitution T790M and/or a deletion in exon 19, and/or one or more EGFR mutants which are selected from a substitution and/or a deletion A763, A767, S768, V769, D770, N771, P772 and H773in exon 20,

(c) Trastuzumab-resistant breast cancer with overexpression of HER 2;

(2) bozitinib and cisplatin for treatment

(c) Trastuzumab-resistant breast cancer with overexpression of HER 2;

(3) boqitinib and 5-fluorouracil for the treatment of

(b) trastuzumab-resistant breast cancer with overexpression of HER2, or

(c) Esophageal cancer with HER2 overexpression;

(4) bosutinib and cetuximab for the treatment of non-small cell lung cancer carrying EGFR mutants with L858R and T790M substitutions and concomitant HER1 overexpression, and/or carrying one or more EGFR mutants selected from a763, a767, S768, V769, D770, N771, P772 and H773 substitutions and/or deletions in exon 20;

(5) boqitinib and trastuzumab for treatment

(a) Trastuzumab-resistant breast cancer with overexpression of HER2, or

(b) Gastric cancer with HER2 overexpression; and

(6) bozitinib and vinorelbine for the treatment of cancer

22. A method of treating a tumor in a subject, the method comprising administering to the subject a therapeutically effective amount of bosutinib as an active ingredient and at least one cytotoxic agent and/or at least one molecular targeting agent, alone or in combination,

wherein the at least one cytotoxic agent is selected from the group consisting of a taxane, a base analog, a platinum antineoplastic agent and a vinca alkaloid; and

wherein the at least one molecular targeting agent is selected from the group consisting of an Epidermal Growth Factor (EGFR) family inhibitor and a mammalian target of rapamycin (mTOR) inhibitor.

23. The method of claim 22, wherein the at least one cytotoxic agent is selected from the group consisting of paclitaxel, cisplatin, 5-fluorouracil, vinorelbine, and any combination thereof.

24. The method of claim 22, wherein the at least one molecular targeting agent is selected from the group consisting of cetuximab, trastuzumab, T-DM1, and any combination thereof.

25. The method of claim 22, wherein administration is selected from the group consisting of:

(a) bosutinib and paclitaxel;

(b) bosutinib and cisplatin;

(c) bosutinib and 5-fluorouracil;

(d) bosutinib and cetuximab;

(e) bosutinib and trastuzumab;

(f) bosutinib and T-DM 1; and

(g) bosutinib and vinorelbine.

26. The method of claim 22, wherein the tumor is non-small cell lung cancer, breast cancer, gastric cancer, colon cancer, pancreatic cancer, prostate cancer, myeloma, head and neck cancer, ovarian cancer, esophageal cancer, or metastatic cell cancer.

27. The method of claim 26, wherein the tumor is selected from the group consisting of non-small cell lung cancer, breast cancer, gastric cancer, and esophageal cancer.

28. The method of claim 27, wherein the tumor is selected from the group consisting of:

(i) non-small cell lung cancer carrying one or more EGFR mutants selected from the group consisting of substitutions L858R, substitutions T790M and/or deletions in exon 19, and/or one or more EGFR mutants selected from the group consisting of substitutions A763, A767, S768, V769, D770, N771, P772 and H773 and/or deletions in exon 20,

(iv) trastuzumab-resistant breast cancer with overexpression of HER 2;

(v) HER1, HER2, and HER1 that are triple negative for estrogen receptor overexpress breast cancer;

(vi) esophageal cancer with HER2 overexpression; and

(vii) gastric cancer accompanied by overexpression of HER 2.

29. The method of claim 22, wherein the administration and the tumor to be treated are selected from the group consisting of:

(1) boqitinib and paclitaxel for treatment

(c) Trastuzumab-resistant breast cancer with overexpression of HER 2;

(2) bozitinib and cisplatin for treatment

(c) Trastuzumab-resistant breast cancer with overexpression of HER 2;

(3) boqitinib and 5-fluorouracil for the treatment of

(b) trastuzumab-resistant breast cancer with overexpression of HER2, or

(c) Esophageal cancer with HER2 overexpression;

(5) boqitinib and trastuzumab for treatment

(a) Trastuzumab-resistant breast cancer with overexpression of HER2, or

(b) Gastric cancer with HER2 overexpression; and

(6) bozitinib and vinorelbine for the treatment of cancer

30. A kit for treating a tumor in a subject, the kit comprising a first part and a second part, wherein the first part comprises bosutinib and the second part comprises at least one active ingredient selected from a cytotoxic agent and a molecular targeting agent, and

31. The kit of claim 30, further comprising a package insert comprising instructions for treating a tumor associated with overexpression or amplification of HER1, HER2, or HER4, or a mutant thereof, in a subject.