WO2015112705A2

WO2015112705A2 - Therapeutic combinations for treating cancer

Info

Publication number: WO2015112705A2
Application number: PCT/US2015/012442
Authority: WO
Inventors: Andrew D. SIMMMONS; Thomas Christian HARDING
Original assignee: Clovis Oncology Inc
Current assignee: Clovis Oncology Inc
Priority date: 2014-01-24
Filing date: 2015-01-22
Publication date: 2015-07-30
Anticipated expiration: 2016-07-24
Also published as: WO2015112705A3

Abstract

The present invention relates to combination treatment methods for treating cancer, particularly non-small cell lung cancer (NSCLC). More specifically, the invention relates to methods of treating or managing cancer, particularly NSCLC, in a subject, comprising administering an irreversible mutant EGFR inhibitor compound in combination with one or more antineoplastic agents, particularly an Aurora kinase inhibitor.

Description

THERAPEUTIC COMBINATIONS FOR TREATING CANCER

FIELD OF THE INVENTION

[0001] The present invention relates to combination treatment methods for treating cancer, particularly non-small cell lung cancer (NSCLC). More specifically, the invention relates to methods of treating or managing cancer, particularly NSCLC, in a subject, comprising administering an irreversible mutant epidermal growth factor receptor (EGFR) inhibitor compound in combination with one or more antineoplastic agents, particularly an Aurora kinase inhibitor.

[0002] The present invention relates to pharmaceutical compositions and methods comprising said irreversible mutant EGFR inhibitor compounds in combination with one or more antineoplastic agents, such as an Aurora kinase inhibitor, administered simultaneously or sequentially.

BACKGROUND

[0003] Despite years of research and prevention strategies, lung cancer remains the most common cancer worldwide with approximately 1.35 million new cases annually and non-small cell lung cancer (NSCLC) accounting for almost 85% of all lung cancers (Herbst RS et al, N Engl J Med. 2008;359: 1367-80). Additionally, lung cancer continues to be the most common cause of cancer-related deaths worldwide with a 5 -year survival rate of less than 10% in patients with advanced disease.

[0004] Activating mutations in the epidermal growth factor receptor (EGFR) are key drivers of NSCLC malignancy in 10-15% of patients of European descent and approximately 30% of patients of East Asian descent (Rosell R et al, N Engl J Med. 2009;361 :958-67). Patients with the most common EGFR mutations (exon 21 L858R and deletions in exon 19) typically have good responses to therapy with first-generation reversible EGFR tyrosine kinase inhibitors (TKIs), such as erlotinib or gefitinib (Mok TS et al, N Engl J Med.

2009;361 :947-57; Fukuoka M et al, J Clin Oncol. 2011;29:2866-74). Toxicity associated with both erlotinib and gefitinib includes skin rash and diarrhea related to inhibition of wild- type EGFR (WT EGFR) in skin and intestine, respectively (Herbst RS et al., Clin Lung Cancer. 2003;4:366-9). [0005] Despite the impressive initial response to treatment, disease progression generally occurs after 9 to 14 months of erlotinib or gefitinib therapy, driven in

approximately 60% of cases by a second site EGFR point mutation that results in the substitution of threonine with methionine at amino acid position 790 (T790M; Sequist LV et al, Sci Transl Med. 201 l;3:75ra26; Pao W et al, PLoS Med. 2005;2:e73; Sharma SV et al, Nat Rev Cancer. 2007;7: 169-81; Yu HA et al, Clin Cancer Res. 2013;19:2240-7). Research suggests that T790M mediates resistance to first-generation EGFR inhibitors by acting as a "gatekeeper" mutation, inducing steric hindrance in the ATP binding pocket and preventing inhibitor binding (Pao W et al, PLoS Med. 2005;2:e73; Kwak EL et al, Proc Natl Acad Sci U S A. 2005;102:7665-70; Kobayashi S et al, N Engl J Med. 2005;352:786-92). Additional work has indicated that T790M increases the affinity of EGFR for ATP, therefore out- competing ATP-competitive TKIs and restoring enzymatic activity in their presence (Yun CH et al, Proc Natl Acad Sci U S A. 2008;105:2070-5).

[0006] Patients with mutant EGFR NSCLC who have failed treatment with first generation EGFR inhibitors and have acquired resistance through the T790M mutation have few treatment options. Currently, there are no targeted therapies for these patients, who are usually treated with cytotoxic chemotherapy that has limited efficacy, but significant toxicity, in the second- or third-line setting. Although second generation irreversible HER-family TKIs, including dacomitinib (PF299804) and afatinib (BIBW2992), are able to inhibit T790M-mutant EGFR in vitro in biochemical and cell-based assays, in clinical trials these agents have not been shown to induce compelling responses in patients that have failed first- generation TKIs (Miller VA et al, Lancet Oncol. 2012;13:528-38). Due to the potent inhibition of WT EGFR and its associated toxicities, these agents cannot reach exposures in the clinic required to inhibit T790M in tumor tissue.

[0007] To circumvent this problem, third generation irreversible mutant EGFR inhibitors were developed. A covalent inhibitor that inhibits mutant EGFR, including T790M, more potently than the WT receptor, termed WZ4002, was described (Zhou W et al, Nature. 2009;462: 1070-4), but did not progress into human trials. CO-1686 is a potent, small-molecule, irreversible tyrosine kinase inhibitor (TKI) that selectively targets the common EGFR mutations (L858R, del 19, T790M) and has minimal inhibitory activity towards WT EGFR (Walter, A. et al, Cancer Discovery. 2013;3: 1404-15). Oral

administration of CO-1686 leads to tumor regressions in cell-based and patient- derived xenograft models as well as a transgenic mouse model expressing mutant forms of EGFR. [0008] CO- 1686 is currently being evaluated in phase I/II clinical trials in EGFR- mutant NSCLC. CO- 1686 is currently in dose-escalation study to evaluate safety,

pharmacokinetics, and preliminary efficacy in previously treated mutant EGFR NSCLC (ClinicalTrials.gov identifier: NCT01526928). The initial area of clinical study for CO-1686 is the treatment of patients with mutant EGFR NSCLC who have received prior EGFR- directed therapy. Expansion of CO-1686 into the front-line setting of EGFR mutant NSCLC patients may be warranted considering the equivalent potency observed between erlotinib and CO-1686 in the transgenic L858R-EGFR mutant transgenic mouse model. Toxicity associated with erlotinib and gefitinib includes skin rash and diarrhea related to inhibition of WT EGFR in skin and intestine, respectively (Herbst RS et al., N Engl J Med.

2008;359: 1367-80). Use of a WT EGFR sparing, mutant selective inhibitor such as CO-1686 has the potential for preventing some of these side-effects observed with first-generation EGFR inhibitors and therefore improving patient quality-of-life.

[0009] Although disease progression in EGFR mutant NSCLC patients after 9 to 14 months of erlotinib or gefitinib therapy is driven in approximately 60% of cases by T790M, other resistance pathways independent of EGFR signaling are also observed in patients treated with first generation EGFR inhibitors. Some of the reported pre-clinical mechanisms of resistance include MET mutation and amplification, ERBB2/HER2 gene amplification, EMT and small cell histological transformation (Herbst RS et al., Clin Lung Cancer.

2003;4:366-9; Yu HA et al., Clin Cancer Res. 2013;19:2240-7; Suda K et al., Clin Cancer Res. 2010;16:5489-98; Byers LA et al., Clin Cancer Res. 2013;19:279-90; Engelman JA et al., Science. 2007;316: 1039-43) and may represent alternative escape routes. As with most antineoplastic agents, innate and acquired resistance to CO-1686 and other third generation irreversible EGFR inhibitors is likely to be observed in patients. So far, acquired resistance to CO-1686 is not mediated by further mutation or amplification of the EGFR gene and resistant cells appear to have a reduced dependence on EGFR signaling than parental cells. Indeed, initial studies of in vitro resistance to CO-1686 demonstrate an enrichment of a RNA-based signature of EMT derived from resistance to EGFR and PI3K/Akt inhibitors (Walter, A. et al., Cancer Discovery. 2013;3: 1404-15). Recent papers indicate that resistance to WZ4002, an alternative irreversible third EGFR inhibitor, is mediated by IGF1R pathway activation due to the repression of IGFBP3 expression via promoter methylation (Cortot AB et al., Cancer Res. 2013;73:834-43) or genomic amplification of the MAPK1 (ERK2) gene (Ercan D et al, Cancer Discov. 2012;2:934-47). [0010] It is highly likely that resistance to irreversible inhibitors will be observed in the clinic. Hence, there is an unmet and critical need to explore combination therapies for treatment methods comprising irreversible EGFR inhibitors in combination with other antineoplastic agents such as Aurora kinase inhibitors to delay time to progression and improve patient outcomes.

SUMMARY

[0011] The present invention is directed to combination therapies that provide better therapeutic profiles than current single agent therapies or other combination therapies utilizing EGFR inhibitors. Encompassed by the invention are combination therapies, of an irreversible mutant EGFR inhibitor compound with one or more antineoplastic agents, particularly an Aurora kinase inhibitor, that have at least an additive potency or at least an additive therapeutic effect. Preferably, the invention is directed to combination therapies where the therapeutic efficacy is greater than additive, e.g., a synergistic efficacy exists between an irreversible mutant EGFR inhibitor compound and one or more antineoplastic agents, particularly an Aurora kinase inhibitor. Preferably, such combination therapies also reduce or avoid unwanted or adverse effects.

[0012] In certain embodiments, the combination therapies of the invention provide an improved overall therapy relative to the administration of the therapeutic agents by themselves. In certain embodiments, doses of existing chemotherapeutic agents can be reduced or administered less frequently in using the combination therapies of the invention, thereby increasing patient compliance, improving therapy and reducing unwanted or adverse effects.

[0013] Accordingly, this invention is directed to combination therapies designed to treat or manage cancer, particularly NSCLC, in a subject, wherein the combination therapies comprise administering an irreversible mutant EGFR inhibitor to the subject in need thereof in combination with one or more antineoplastic agents, particularly an Aurora kinase inhibitor. In particular, this invention is directed to methods of treating or managing cancer, particularly NSCLC, in a subject, comprising administering a therapeutically effective amount of an irreversible mutant EGFR inhibitor in combination with the administration of therapeutically effective amount of one or more antineoplastic agents, particularly an Aurora kinase inhibitor. [0014] In one embodiment, the present invention provides methods for treating cancer, particularly NSCLC, in a subject comprising administering an EGFR mutant inhibitor compound that covalently modifies Cysteine 797 in EGFR, wherein said compound exhibits at least 2-fold greater inhibition of a drug resistant EGFR mutant relative to wild type EGFR, and an additional antineoplastic agent, particularly an Aurora kinase inhibitor.

[0015] In one embodiment, the present invention provides methods for treating drug resistant cancer, particularly NSCLC, in a subject comprising administering an EGFR mutant inhibitor compound that covalently modifies Cysteine 797 in EGFR, wherein said compound exhibits at least 2-fold greater inhibition of a drug resistant EGFR mutant relative to wild type EGFR, and an additional antineoplastic agent, particularly an Aurora kinase inhibitor.

[0016] In one embodiment, the present invention provides a method for treating cancer having mutant epidermal growth factor receptor (EGFR) in a subject comprising administering an irreversible mutant EGFR inhibitor and an Aurora kinase inhibitor. In one embodiment, the irreversible mutant EGFR inhibitor covalently binds to cysteine 797 of mutant EGFR and exhibits at least 2-fold greater inhibition of a drug-resistant mutant EGFR relative to wild type EGFR.

[0017] In one embodiment, the cancer is non-small cell lung cancer (NSCLC).

[0018] In one embodiment, the irreversible mutant EGFR inhibitor is selected from

CO- 1686, WZ4002, AZD9291, CNX2006, ASP8273, EGF816, HM61713, TAS-2913, and AZ5104. In one embodiment, the irreversible mutant EGFR inhibitor is CO- 1686.

[0019] In one embodiment, the Aurora kinase inhibitor is selected from TAK-901,

AZD1152, PF-03814735, VX-680, MLN-8237, PHA-739358, CYC116, SNS-314, VX-680, AT9283, R763, PF-03814735, GSK1070916, AMG-900, MLN-8237, ENMD-2076, MK- 5108, AZD1152, BI 811283, PHA-680632, VE-465, JNJ-7706621, CCT129202, AKI-001, CHR-3520, Hesperadin, ZM447439, and Jadomycin-B

[0020] In one embodiment, the drug-resistant mutant is selected from L858R/T790M

EGFR and Exon-19 Deletion/T790M. In one embodiment, the drug resistant mutant is selected from T790M, L858R, G719S, G719C, G719A, L861Q, a small in-frame deletion in exon 19, or an insertion in exon 20.

[0021] In one embodiment, the present invention provides a method for treating cancer having mutant EGFR in a subject wherein said subject has relapsed NSCLC. In one embodiment, the subject was previously treated with a first EGFR inhibitor. In one embodiment, the first EGFR inhibitor was selected from erlotinib, gefitinib, afatinib, dacomitinib, cetuximab, panitumumab, CO- 1686, WZ4002, AZD9291, CNX2006, ASP8273, EGF816, HM61713, TAS-2913, and AZ5104.

[0022] In one embodiment, the present invention provides methods for treating cancer, particularly NSCLC, comprising irreversible mutant EGFR inhibitor compounds in combination with one or more antineoplastic agents, particularly an Aurora kinase inhibitor, administered simultaneously or sequentially.

[0023] In one embodiment, the present invention provides pharmaceutical compositions of compounds or pharmaceutically acceptable salts of one or more compounds described herein and a pharmaceutically acceptable carrier or excipient.

BRIEF DESCRIPTION OF THE FIGURES

[0024] FIGURE 1 shows cell viability curves for CO- 1686 in the PC9 parental and

CO- 1686 Resistant (COR) pools. The table indicates 50% growth inhibition (GI₅₀, nM) values for CO- 1686 in each of the cell lines. GI₅₀ values greater than 1000 have been listed as >1000. The parental cell lines are sensitive to CO-1686 whereas the PC9 COR pools are resistant.

[0025] FIGURES 2A-2E show cell viability curves for 5 different Aurora A, B, or

A/B inhibitors in the PC9 parental and PC9 COR pools. The PC9 COR pools are maintained and tested in ΙμΜ CO-1686, and the concentration of each Aurora inhibitor in indicated on the x-axis. The Aurora inhibitors tested all generally provide limited activity (reduction in cellular viability) in the PC9 cell line, whereas reduced cell viability was observed in the PC9 COR cell lines with the combination of CO-1686 and each of the Aurora inhibitors.

[0026] FIGURE 3 shows a three dimensional bar chart illustrating the GI₅₀ values

(nM) for CO-1686 and five different Aurora inhibitors in the PC9 parental and PC9 COR pools. The PC9 COR pools were maintained and tested in ΙμΜ CO-1686. GI₅₀ values greater than ΙΟΟΟηΜ have been graphed at ΙΟΟΟηΜ for comparative purposes. The actual values are indicated above the relevant bars. The PC9 parental cell line is sensitive to single agent CO-1686 whereas the PC9 COR pools are resistant. Single agent Aurora inhibitors provide limited potency in the PC9 cell line, whereas reduced cell viability was observed in the PC9 COR cell lines with the combination of CO-1686 and each of the Aurora inhibitors tested.

[0027] FIGURE 4A shows cell viability curves for various aurora kinase inhibitors in the CO-1686 resistant PC9 2A10 cell line. The PC9 2A10 cell line is maintained and was tested in ΙμΜ CO- 1686, and the concentration of each Aurora inhibitor is indicated on the x- axis. Figure 4B indicates 50% growth inhibition (GI₅₀, nM) values for each Aurora inhibitor evaluated in this experiment. All of the Aurora inhibitors demonstrate increased cytotoxicity when combined with CO- 1686 in the PC9 2A10 resistance cell line.

[0028] FIGURES 5A and 5B show cell viability curves for CO-1686, MLN8237, and the combination of CO-1686 and MLN8237 in the PC9 and CO-1686 resistant PC9 2A10 cell lines, respectively. The PC9 2A10 cell line is maintained and was tested in ΙμΜ CO- 1686. These data demonstrate that the combination of CO-1686 and the Aurora inhibitor MLN8237 is cytotoxic in both the PC9 parental and CO-1686 resistant PC9 2A10 cell lines. PC9 and CO-1686 resistant PC9 2A10 cells exhibit increased sensitivity to the combination of CO-1686 and the Aurora kinase inhibitor MLN8237

[0029] FIGURES 6A and 6B show caspase 3/7 levels for CO-1686, MLN8237, and the combination of CO-1686 and MLN8237 in the PC9 and CO-1686 resistant PC9 2A10 cell lines, respectively, after 24 hr of treatment. The PC9 2A10 cell line is maintained and was tested in ΙμΜ CO-1686. The combination of CO-1686 and MLN8237 exhibits pronounced cell death activation in both PC9 and PC9 2A10 cell lines. CO-1686 and MLN8237 combination increases caspase 3/7 levels in PC9 parental and CO-1686 resistant PC9 2A10 tumor cells.

[0030] FIGURES 7A and 7B illustrate the cell cycle profiles of PC9 cells under single agent or combination treatment of CO-1686 and MLN8237 for 48 hr. FIGURE 7A shows representative fluorescence-activated cell sorting (FACS) plots for each treatment. FIGURE 7B shows a summary of cell cycle analysis for each treatment group. CO-1686 treatment alone causes an accumulations of cells in the Gl phase of the cell cycle, while treatment of MLN8237 causes cells to arrest in the G2 phase and an accumulation of cells in an diploid state (>4N). The combination of both compounds increased the number of cells in GO Phase. CO-1686 and MLN8237 combination reduces polyploidy and drives more cells into apoptosis in PC9 cells.

[0031] FIGURES 8A and 8B illustrate the cell cycle profiles of CO-1686 resistant

PC9 2A10 cells under single agent or combination treatment of CO-1686 and MLN8237 for 48 hr. FIGURE 8A shows representative fluorescence-activated cell sorting (FACS) plots for each treatment. FIGURE 8B shows a summary of cell cycle analysis for each treatment group. CO-1686 treatment resulted in an accumulation of cells in the GO (apoptosis) and Gl phases of the cell cycle, while treatment of MLN8237 predominantly resulted in G2 arrest. The combination of both compounds increased the number of cells in GO arrest. CO-1686 and MLN8237 combination reduces polyploidy and drives more cells into apoptosis in CO- 1686 resistant PC9 2A10 cells

[0032] FIGURE 9 shows tumor volumes from PC9 2A10 tumor bearing mice treated with CO-1686, MLN8237, or the combination of CO-1686 and MLN8237 at the doses and schedules indicated. CO-1686 administration at 100 mg/kg BID resulted in significant tumor growth inhibition, whereas monotherapy treatment with MLN8237 at 20 mg/kg QD demonstrated limited activity. The combination of CO-1686 and MLN8237 demonstrated the greatest tumor growth inhibition of all of the groups evaluated in this study.

DETAILED DESCRIPTION

[0033] The following description is merely exemplary in nature and is not intended to limit the present disclosure, application, or uses.

[0034] Various embodiments of the disclosure could also include permutations of the various elements recited in the claims as if each dependent claim was a multiple dependent claim incorporating the limitations of each of the preceding dependent claims as well as the independent claims. Such permutations are expressly within the scope of this disclosure.

[0035] Unless defined otherwise, technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art(s) to which this invention belongs. Although any methods, devices, and materials similar or equivalent to those described herein can be used in the practice or testing of the invention, the preferred methods, devices and materials are now described.

DEFINITIONS:

[0036] As used herein, "irreversible mutant EGFR inhibitor" or "covalent mutant

EGFR inhibitor" refers to a third generation EGFR inhibitor that covalently binds to a mutant form of epidermal growth factor receptor (EGFR). In some embodiments, an irreversible mutant EGFR inhibitor covalently binds to cysteine 797 of mutant EGFR and exhibits at least 2-fold greater inhibition of a drug-resistant mutant EGFR relative to wild type EGFR.

Irreversible mutant EGFR inhibitors include, but are not limited to, CO-1686, WZ4002, AZD9291, CNX2006, ASP8273, EGF816, HM61713, TAS-2913, AZ5104 and others.

Structures of a few exemplary irreversible mutant EGFR inhibitors are shown in Table 1. [0037] As used herein, "Aurora kinase inhibitor" or "Aurora inhibitor" refers to any compound that inhibits Aurora kinase activity. An Aurora kinase inhibitor can inhibit one or more of Aurora A kinase, Aurora B kinase, or Aurora C kinase. Aurora kinase inhibitors include, but are not limited to, TAK-901, AZD1152, PF-03814735, VX-680, MLN-8237, PHA-739358, CYC116, SNS-314, AT9283, R763, GSK1070916, AMG-900, ENMD-2076, MK-5108, BI 811283, PHA-680632, VE-465, JNJ-7706621, CCT129202, AKI-001, CHR- 3520, Hesperadin, ZM447439, Jadomycin-B, and others. Structures of a few exemplary Aurora kinase inhibitors are shown in Table 2.

[0038] As used herein, the term "non-small cell lung cancer" or "NSCLC" refers to any type of epithelial lung cancer other than small cell lung cancer (SCLC).

[0039] As used herein, the term "kinase inhibitor" refers to any type of enzyme inhibitor that specifically blocks the action of one or more kinases responsible for cancer, particularly NSCLC. In certain embodiments, a kinase inhibitor of the present invention refers to a small molecule, a protein/peptide or an antibody.

[0040] The terms "reduce," "inhibit," "diminish," "suppress," "decrease," "prevent" and grammatical equivalents (including "lower," "smaller," etc.) when in reference to the expression of any symptom in an untreated subject relative to a treated subject, mean that the quantity and/or magnitude of the symptoms in the treated subject is lower than in the untreated subject by any amount that is recognized as clinically relevant by any medically trained personnel. In one embodiment, the quantity and/or magnitude of the symptoms in the treated subject is at least 10% lower than, at least 25% lower than, at least 50% lower than, at least 75% lower than, and/or at least 90% lower than the quantity and/or magnitude of the symptoms in the untreated subject.

[0041] The term "inhibitory compound" as used herein, refers to any compound capable of interacting with (i.e., for example, attaching, binding etc.) to a binding partner under conditions such that the binding partner becomes unresponsive to its natural ligands. Inhibitory compounds may include, but are not limited to, small organic molecules, antibodies, and proteins/peptides.

[0042] The term "irreversible inhibitor" or "covalent inhibitor" as used herein, refers to an inhibitor that covalently modifies an enzyme, and inhibition therefore cannot be reversed.

[0043] The term "drug" or "compound" as used herein, refers to any

pharmacologically active substance capable of being administered which achieves a desired effect. Drugs or compounds can be synthetic or naturally occurring, non-peptide, proteins or peptides, oligonucleotides or nucleotides, polysaccharides or sugars.

[0044] The term "administered" or "administering", as used herein, refers to any method of providing a composition to a patient such that the composition has its intended effect on the patient. An exemplary method of administering is by a direct mechanism such as, local tissue administration (i.e., for example, extravascular placement), oral ingestion, transdermal patch, topical, inhalation, suppository etc.

[0045] The term "therapeutically effective amount" refers to that amount of therapeutic agent sufficient to destroy, modify, control or remove cancer tissue. A

therapeutically effective amount may refer to the amount of therapeutic agent sufficient to delay or minimize the spread of cancer. A therapeutically effective amount may also refer to the amount of therapeutic agent that provides a therapeutic benefit in the treatment or management of cancer. Further, a therapeutically effective amount with respect to an irreversible mutant EGFR inhibitor of the combination therapies of the invention means that amount of an irreversible mutant EGFR inhibitor in combination with one or more antineoplastic agents, particularly an Aurora kinase inhibitor that provides a therapeutic benefit in the treatment or management of cancer, particularly NSCLC, including

amelioration of symptoms associate with cancer, such as NSCLC. Used in connection with an amount of irreversible mutant EGFR inhibitor, the term can encompass an amount that improves overall therapy, reduces or avoids unwanted effects, or enhances the therapeutic efficacy of or synergizes with one or more antineoplastic agents, such as an Aurora kinase inhibitor, utilized in combination therapies of the invention. The "therapeutically effective amount" may vary depending, for example, on the kinase inhibitors selected, the stage of the cancer, the age, weight and/or health of the patient and the judgment of the prescribing physician. An appropriate amount in any given instance may be readily ascertained by those skilled in the art or capable of determination by routine experimentation.

[0046] As used herein, the terms "manage", "managing", and "management" refer to the beneficial effects that a patient derives from a combination therapy of the invention, which does not result in a cure of cancer, such as NSCLC. In certain embodiments, a combination therapy of the invention "manages" cancer, particularly NSCLC, so as to prevent the progression or worsening of the cancer.

[0047] As used herein, the terms "treat", "treating", and "treatment" refer to the eradication, removal, modification or control of cancer, particularly NSCLC, that results from the combination therapy of the invention. In certain embodiments, such terms refer to minimizing or delaying of the spread of cancer, particularly NSCLC.

[0048] As used herein, the term "combination therapy" or "combination treatment" refers to methods of treating cancer, particularly NSCLC, in a subject comprising an irreversible mutant EGFR inhibitor compound with one or more antineoplastic agents, particularly an Aurora kinase inhibitor. As used herein, these terms encompass administering an irreversible mutant EGFR inhibitor and one or more antineoplastic agents simultaneously or sequentially.

[0049] The term "patient", as used herein, is a human or animal and need not be hospitalized. For example, out-patients, persons in nursing homes are "patients." A patient may comprise any age of a human or non-human animal and therefore includes both adult and juveniles (i.e., children). It is not intended that the term "patient" connote a need for medical treatment, therefore, a patient may voluntarily or involuntarily be part of

experimentation whether clinical or in support of basic science studies.

[0050] The term "subject" as used herein refers to a vertebrate, preferably a mammal, more preferably a primate, still more preferably a human. Mammals include, without limitation, humans, primates, wild animals, feral animals, farm animals, sports animals, and pets.

[0051] The term "pharmaceutically" or "pharmacologically acceptable", as used herein, refer to molecular entities and compositions that do not produce adverse, allergic, or other untoward reactions when administered to an animal or a human.

[0052] The term, "pharmaceutically acceptable carrier", as used herein, includes any and all solvents, or a dispersion medium including, but not limited to, water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyethylene glycol, and the like), suitable mixtures thereof, and vegetable oils, coatings, isotonic and absorption delaying agents, liposome, commercially available cleansers, and the like. Supplementary bioactive ingredients also can be incorporated into such carriers.

[0053] The term "salts", as used herein, refers to any salt that complexes with identified compounds contained herein. Examples of such salts include, but are not limited to, acid addition salts formed with inorganic acids (e.g. hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, nitric acid, and the like), and salts formed with organic acids such as, but not limited to, acetic acid, oxalic acid, tartaric acid, succinic acid, malic acid, fumaric acid, maleic acid, ascorbic acid, benzoic acid, tannic acid, pamoic acid, alginic acid, polyglutamic, acid, naphthalene sulfonic acid, naphthalene disulfonic acid, and polygalacturonic acid. Salt compounds can also be administered as pharmaceutically acceptable quaternary salts known by a person skilled in the art, which specifically include the quaternary ammonium salts of the formula ~NR,R',R"+Z -, wherein R, R, R" is independently hydrogen, alkyl, or benzyl, and Z is a counter ion, including, but not limited to, chloride, bromide, iodide, alkoxide, toluenesulfonate, methylsulfonate, sulfonate, phosphate, or carboxylate (such as benzoate, succinate, acetate, glycolate, maleate, malate, fumarate, citrate, tartrate, ascorbate, cinnamoate, mandeloate, and diphenylacetate). Salt compounds can also be administered as pharmaceutically acceptable pyridine cation salts having a substituted or unsubstituted partial formula: wherein Z is a counter ion, including, but not limited to, chloride, bromide, iodide, alkoxide, toluenesulfonate, methylsulfonate, sulfonate, phosphate, or carboxylate (such as benzoate, succinate, acetate, glycolate, maleate, malate, fumarate, citrate, tartrate, ascorbate, cinnamoate, mandeloate, and diphenylacetate).

[0054] The term "prodrug" refers to a derivative of a compound that can hydrolyze, oxidize, or otherwise react under biological conditions (in vitro or in vivo) to provide a compound of the invention. Prodrugs may only become active upon some reaction under biological conditions, but they may have activity in their unreacted forms. Examples of prodrugs contemplated herein include, without limitation, analogs or derivatives of compounds of the invention, and/or their salts when salt formation is possible, but in particular, derivatives of zinc binding thiol moiety. Examples of prodrug moieties include substituted and unsubstituted, branched or unbranched lower alkyl ester moieties, (e.g., propionic acid esters), lower alkenyl esters, di-lower alkyl-amino lower-alkyl esters (e.g., dimethylaminoethyl ester), acylamino lower alkyl esters (e.g., acetyloxymethyl ester), acyloxy lower alkyl esters (e.g., pivaloyloxymethyl ester), aryl esters (phenyl ester), aryl- lower alkyl esters (e.g., benzyl ester), heteroaryl esters (nicotinate ester), substituted (e.g., with methyl, halo, or methoxy substituents) aryl and aryl-lower alkyl esters, amides, lower- alkyl amides, di-lower alkyl amides, and hydroxy amides. Naturally occurring amino acid esters or their enantiomers, dipeptide esters, phosphate esters, methoxyphosphate esters, disulfides and disulfide dimers. Prodrugs and their uses are well known in the art (see, e.g., Berge et al. 1977). Prodrugs can typically be prepared using well-known methods, such as those described in Burger's Medicinal Chemistry and Drug Discovery (Manfred E. Wolff ed.1995) and (Rautio, 2008).

[0055] As used in this disclosure, including the appended claims, the singular forms

"a," "an," and "the" include plural references, unless the content clearly dictates otherwise, and are used interchangeably with "at least one" and "one or more." [0056] Unless otherwise indicated, all numbers expressing quantities of ingredients, dimensions reaction conditions and so forth used in the specification and claims are to be understood as being modified in all instances by the term "about".

[0057] As used herein, the term "activity" refers to the activation, production, expression, synthesis, intercellular effect, and/or pathological or aberrant effect of the referenced molecule, either inside and/or outside of a cell.

[0058] As used herein, the terms "comprises," "comprising," "includes," "including,"

"contains," "containing," and any variations thereof, are intended to cover a non-exclusive inclusion, such that a process, method, product-by-process, or composition of matter that comprises, includes, or contains an element or list of elements does not include only those elements but may include other elements not expressly listed or inherent to such process, method, product-by-process, or composition of matter.

[0059] In this application and the claims, the use of the singular includes the plural unless specifically stated otherwise. In addition, use of "or" means "and/or" unless stated otherwise. Moreover, the use of the term "including", as well as other forms, such as "includes" and "included", is not limiting. Also, terms such as "element" or "component" encompass both elements and components comprising one unit and elements and components that comprise more than one unit unless specifically stated otherwise.

[0060] Methods described herein can be extended to a variety of cancers, particularly

NSCLC. In some instances, cancer can be a metastatic cancer. Examples of additional cancers related to the methods described herein include, but are not limited to, breast, ovarian, pancreatic, sarcoma, prostate cancer, colon cancer (such as a colon carcinoma, including small intestine cancer), glioma, leukemia, liver cancer, melanoma (e.g., metastatic malignant melanoma), acute myeloid leukemia, kidney cancer, bladder cancer, renal cancer (e.g., renal cell carcinoma), glioblastoma, brain tumors, chronic or acute leukemias including acute lymphocytic leukemia (ALL), adult T-cell leukemia (T-ALL), chronic myeloid leukemia, acute lymphoblastic leukemia, chronic lymphocytic leukemia, lymphomas (e.g., Hodgkin's and non-Hodgkin's lymphoma, lymphocytic lymphoma, primary CNS lymphoma, T-cell lymphoma, Burkitt's lymphoma, anaplastic large-cell lymphomas (ALCL), cutaneous T-cell lymphomas, nodular small cleaved-cell lymphomas, peripheral T-cell lymphomas, Lennert's lymphomas, immunoblastic lymphomas, T-cell leukemia/lymphomas (ATLL), entroblastic/centrocytic (cb/cc) follicular lymphomas cancers, diffuse large cell lymphomas of B lineage, angioimmunoblastic lymphadenopathy (AILD)-like T cell lymphoma and HIV associated body cavity based lymphomas), embryonal carcinomas, undifferentiated carcinomas of the rhino-pharynx (e.g., Schmincke's tumor), Castleman's disease, Kaposi's Sarcoma, multiple myeloma, Waldenstrom's macro globulinemia and other B-cell

lymphomas, nasopharangeal carcinomas, bone cancer, skin cancer, cancer of the head or neck, cutaneous or intraocular malignant melanoma, uterine cancer, rectal cancer, cancer of the anal region, stomach cancer, testicular cancer, carcinoma of the fallopian tubes, carcinoma of the endometrium, carcinoma of the cervix, carcinoma of the vagina, carcinoma of the vulva, cancer of the esophagus, cancer of the small intestine, cancer of the endocrine system, cancer of the thyroid gland, cancer of the parathyroid gland, cancer of the adrenal gland, sarcoma of soft tissue, cancer of the urethra, cancer of the penis, solid tumors of childhood, cancer of the bladder, cancer of the kidney or ureter, carcinoma of the renal pelvis, neoplasm of the central nervous system (CNS), tumor angiogenesis, spinal axis tumor, brain stem glioma, pituitary adenoma, epidermoid cancer, squamous cell cancer, or

environmentally induced cancers including those induced by asbestos, e.g., mesothelioma. In another embodiment, methods described herein can be useful for treating a combination of two or more types of cancer. In some aspects the methods are useful to treat individual patients diagnosed with cancer.

[0061] The present invention provides pharmaceutical compositions comprising at least one pharmaceutically-acceptable carrier, in addition to one or more compounds described herein. The composition can take any suitable form for the desired route of administration. Where the composition is to be administered orally, any suitable orally deliverable dosage form can be used, including without limitation tablets, capsules (solid or liquid filled), powders, granules, syrups and other liquids, elixirs, inhalants, troches, lozenges, and solutions. Injectable compositions or i.v. infusions are also provided in the form of solutions, suspensions, and emulsions.

EMBODIMENTS OF THE INVENTION

[0062] Specific embodiments of the invention are described in the following sections.

Mutant NSCLC cell lines with acquired resistance to CO-1686 can be generated

[0063] To study acquired resistance to CO-1686, a novel third generation mutant- selective EGFR TKI, PC9 (EGFR^de119) cells were continuously exposed to increasing doses of CO-1686 until resistance developed (TABLE 3). In addition to generating COR populations by stepwise increases in CO-1686 exposure, PC9 resistant cell populations (each derived from a well in a 24-well dish) were also generated by immediate exposure and maintenance in a high dose of CO- 1686. This approach yielded five resistant clones, designated PC9 COR1, COR2, COR3, COR6, and COR7. A summary of the CO-1686 resistant populations is shown in TABLE 3. A representative example of CO-1686 cell viability data for the PC9 parental and COR pools are shown in FIGURE 1. Cell viability was determined in the parental cell lines and all COR pools using first, second and third generation EGFR TKIs (erlotinib, afatinib, and CO-1686 respectively), as well as the EGFR targeted monoclonal antibody cetuximab. All of the PC9 CORs were resistance to all 4 EGFR inhibitors (TABLE 3).

The Aurora A inhibitor MLN8237 restores CO-1686 sensitivity in multiple PC9 COR populations

[0064] The parental and resistant clones were screened using a compound library

(n=44) to identify drug combinations that could restore CO-1686 sensitivity. Compounds were selected to assess clinically relevant and diverse mechanisms of action. Parental clones were screened in the absence of CO-1686, whereas the PC9 COR populations were maintained and screened in ΙμΜ CO-1686. The GI₅₀ values for the combination of CO-1686 and MLN8237 in the PC9 COR cells lines were at least 10 fold greater than the GI₅₀ value of MLN8237 in the PC9 cell line.

Multiple Aurora A and B inhibitors restored partial drug sensitivity in multiple PC9 COR cell clones

[0065] The screening data indicates that the combination of CO-1686 and the Aurora

A inhibitor MLN8237 restored drug sensitivity in the PC9 COR populations. To validate and expand these data, multiple Aurora A, B, and A/B inhibitors were evaluated in combination with CO-1686 in the PC9 parental and PC9 COR populations (FIGURES 2A-2E). As expected, CO-1686 monotherapy was active in the parental cell line but inactive in the COR populations. As a single agent, the Aurora inhibitors showed limited activity in the parental cell line. The PC9 COR pools were maintained and tested in ΙμΜ CO-1686, and the combination of CO-1686 and an Aurora A, B, or A/B inhibitor showed potent activity in all of the PC9 COR cell lines (FIGURE 3).

CO-1686 resistant PC9 2A10 cells are sensitive to Aurora kinase inhibitors

[0066] CO-1686 resistant PC9 cells were generated by in vitro incubation of PC9 cells with increasing concentrations of CO-1686 over several months to a maximum concentration of ΙμΜ CO-1686. The resistant pool was then subjected to two rounds of in vivo selection, during which mice were dosed with 50 - 100 mg/kg BID CO-1686. Viable cells were collected from a resistant tumor and dissociated to generate the PC9 2A10 cell line. The PC9 2A10 cell line is maintained in ΙμΜ CO-1686 and is tumorigenic in nude and SCID mice.

PC9 and CO-1686 resistant PC9 2A10 cells are sensitive to the combination of CO-1686 and the Aurora A inhibitor MLN8237

[0067] Multiple Aurora A, B, and A/B inhibitors were evaluated in combination with

CO-1686 in the CO-1686 resistant PC9 2A10 cell line using a cellular viability assay. The PC-9 2A10 cells were maintained and tested in ΙμΜ CO-1686. A representative example of CO-1686 cell viability data is shown in FIGURE 4A, and the GI₅₀ (nM) values are

summarized in FIGURE 4B. All of the Aurora inhibitors evaluated demonstrated reduced cellular viability (GI₅₀ = 14 - 379nM) in combination with ΙμΜ CO-1686 as compared to CO-1686 monotherapy (GI50 = 1944nM)

The combination of CO-1686 and MLN8237 reduces tumor cell viability in PC9 and CO-1686 resistant PC9 2A10 cells

[0068] Cellular viability of CO-1686, MLN8237, and the combination was also assessed in the PC9 (FIGURE 5A) and CO-1686 resistant PC9 2A10 (FIGURE 5B) cell lines. CO-1686 demonstrated potent activity in the PC-9 cell line (GI₅₀ = 204nM) and limited activity in the CO-1686 resistant PC-9 2A10 cell line (GI₅₀ = 1869nM). MLN8237 treatment of the PC9 cell line resulted in modest decrease in cell viability (GI₅₀ = 1499nM), whereas a greater impact in cellular viability was observed in the CO-1686 resistant PC9 2A10 cell line (GI₅₀ = 58nM). The combination of CO-1686 and MLN8237 had the greatest impact on cellular viability in both cell lines (GI₅₀ = 28 - 40nM).

The combination of CO-1686 and MLN8237 increases caspase 3/7 activation in PC9 and CO-1686 resistant PC 9 2A10 cells

[0069] PC9 and CO-1686 resistant PC9 2A10 cell lines were treated with CO-1686,

MLN8237, and the combination of CO-1686 and MLN8237. After 24 hr of incubation, caspase 3/7 activation was evaluated. Caspase 3/7 activation is a well-established marker for apoptosis or programmed cell death. The results were consistent with the cell viability data demonstrating that the combination of CO-1686 and MLN8237 had the greatest increase in caspase 3/7 activity in both the PC-9 (FIGURE 6A) and CO-1686 resistant PC-9 2A10 (FIGURE 6B) cell lines.

The combination of CO-1686 and MLN8237 increases cell cycle arrest in PC9 and CO- 1686 resistant PC9 2A10 cells Aurora-A is a mitotic kinase that regulates mitotic spindle formation and segregation, and it has previously been shown that the Aurora A inhibitor MLN8237 results in G2 arrest and an increase in the number of cells with polyploidy (>4N)( Blood. 2010 Jun 24;115(25):5202-13. doi: 10.1182/blood-2009-12-259523. Epub 2010 Apr 9.A novel Aurora-A kinase inhibitor MLN8237 induces cytotoxicity and cell-cycle arrest in multiple myeloma.

http://mct.aacrjoumals.Org/content/l 1/3/763. full). It has also been demonstrated that treatment of the NSCLC cells with erlotinib resulted increased the number of cells in Gl arrest (http://molpharm.aspetioumals.Org/content/72/2/248.full). Cell cycle analysis was performed in PC9 and PC9 2A10 cells treated with single agent and the combination of CO- 1686 and MLN8237 for 48 hr. In the PC9 cell line CO-1686 treatment caused an

accumulation of cells in the Gl phase of the cell cycle, while treatment of MLN8237 increased the percent of G2 and polyploidy cells (FIGURES 7A and 7B). The combination of both compounds reduced the number of polyploidy cells and increased the number of cells in apoptosis (GO). CO-1686 treatment in PC9 2A10 cells increased the number of cells in GO and Gl, whereas MLN8237 increased the percent of G2 and polyploidy cells (FIGURES 8A and 8B). The combination of CO-1686 and MLN8237 in the PC9 2A10 cell line increased the percentage of GO cells. Taken together, these data show that the combination of CO-1686 and MLN8237 results in increased apoptosis (GO arrest) in PC9 and PC9 2A10 cells.

CO-1686 and MLN8237 generate potent in vivo activity in the CO-1686 resistant PC9 2A10 xenograft model

[0070] Mice bearing PC9 2A10 tumors were treated with CO-1686, MLN8237, or the combination of CO-1686 and MLN8237 at the doses and schedules indicated (FIGURE 9). The tumor volumes in mice treated with MLN8237 were comparable to those in vehicle treated animals. CO-1686 administration at 100 mg/kg BID resulted in significant (p < 0.001) tumor growth delay, with a mean tumor volume of 1285 mm³ on Day 71. These results suggest that although the PC9 2A10 cell line is maintained in ΙμΜ CO-1686, it still maintains a degree of dependence on EGFR pathway signaling. The combination of CO- 1686 and MLN8237 resulted in significant (p < 0.001) tumor growth delay, with a mean tumor volume of 365 mm³ on Day 71. On Day 71 the tumor volumes of CO-1686 monotherapy and combination treated animals were 1213 ± 174 (n=8) and 395 ± 65 (n=7), respectively, and significantly (p = 0.0011) different by a two-tailed t test. EXAMPLES

[0071] The following examples are provided for illustrative purposes only and are not intended to limit the scope of the invention.

[0072] Cell culture. PC-9 cells were a kind gift of Dr. F. Koizumi (National Cancer

Center Research Institute & Shien-Lab, Japan). All cell lines were grown in RPMI 1640 (Life Technologies; Carlsbad, CA) supplemented with 10% FBS (HyClone; South Logan, UT), 2mM L-glutamine, and 1% Penicillin-Streptomycin (Mediatech; Corning, NY). All cells were maintained and propagated as monolayer cultures at 37°C in a humidified 5% C0₂ incubator.

[0073] Generation of resistant populations. CO-1686 resistant pools were generated by continuous exposure of PC9 cells to increasing doses of CO-1686 until resistance developed. Additional PC9 resistant pools were generated by plating cells in a 24 well dish, and after 24 hr adding CO-1686 to a final concentration of ΙμΜ. After several months of continuous exposure individual wells were expanded in media containing ΙμΜ CO-1686. This approach yielded five resistant clones, designated PC9 CORl, COR2, COR3, COR6, and COR7. The CO-1686 resistant PC9 COR cells are maintained in the same media with the addition of ΙμΜ CO-1686.

[0074] Cell proliferation assays. Cells were seeded in RPMI- 1640 growth media supplemented with 5% FBS, 2mM L-glutamine, and 1% P/S, allowed to adhere overnight, and treated with a dilution series of test compound for 72 hr. Where indicated, cells were also treated with ΙμΜ CO-1686. Cell viability was determined by CellTiter Glo (Promega; Madison, WI) and results were represented as background-subtracted relative light units normalized to a DMSO-treated control. Growth inhibition (GI₅₀) values were determined by GraphPad Prism 5.04 (GraphPad Software; La Jolla, CA). All compounds were obtained from Selleck Chemical (Houston, TX).

[0075] Generation of CO-1686 resistant PC9 2A10 cell line. CO-1686 resistant

PC9 cells were generated by in vitro incubation of PC9 cells with increasing concentrations of CO-1686 over several months to a maximum of ΙμΜ. The resistant pool was then subjected to two rounds of in vivo selection, during which mice were dosed with 50 - 100 mg/kg BID CO-1686. Viable cells were collected from a resistant tumor and dissociated to generate the PC9 2A10 cell line. The PC9 2A10 cell line is maintained in Ι μΜ CO-1686.

[0076] Combination cell proliferation assays in PC9 and PC9 2A10 cell lines.

PC9 parental cells were seeded in RPMI- 1640 growth media supplemented with 10% FBS, 2mM L-glutamine, and 1% P/S, allowed to adhere overnight, and treated with a dilution series of test compound for 72 hr. The highest concentration of CO- 1686 and MLN8237 evaluated was 20μΜ. The highest dose of the combination was 20μΜ CO- 1686 and 20μΜ of MLN8237, and a 1 : 1 ratio was maintained as compounds were diluted. PC9 2A10 CO- 1686 resistant cells were seeded in media stated above with ΙμΜ CO- 1686 and allowed to adhere overnight. Prior to dosing, media was removed and cells rinsed once with media without drug. Cells were then treated with a dilution series of test compound for 72 hr. The highest dose of the combination was 20μΜ CO- 1686 and 2μΜ MLN8237, respectively, and a 10: 1 ratio was maintained as compounds were diluted. Viability was determined by CellTiter Glo and results were represented as background-subtracted relative light units normalized to a DMSO-treated control.

[0077] Caspase 3/7 assay. Cells were seeded and treated as described for combination cell proliferation assays, and caspase 3 and 7 activity was measured after 24 hours of drug exposure using Caspase-Glo 3/7 assay (Promega; Madison, WI). Results were represented as background-subtracted relative light units normalized to a DMSO-treated control.

[0078] Cell cycle profiling. Cells were seeded in their respective media and allowed to attach overnight. PC9 parental cells were then treated with either DMSO control, 200nM CO- 1686, ΙΟΟηΜ MLN8237 or a combination of 200nM CO- 1686 and ΙΟΟηΜ MLN8237. PC9-2A10 were washed once in RPMI media without drug then placed in media containing either DMSO control, 2000nM CO- 1686, ΙΟΟηΜ MLN8237 or a combination of 2000nM CO- 1686 and ΙΟΟηΜ MLN8237. Cells were drug treated for 48 hr before harvested for PI staining. Briefly, cells were trypsined then washed twice in cold PBS and then fixed for one hour in 70% ethanol, followed by two more PBS washes and staining with a PI/RNAse solution (BD Biosciences; San Jose CA). Flow cytometry was performed on a LSRII (BD Biosciences; San Jose CA) and data analyzed using Flow Jo (Flow Jo; Ashland, OR).

[0079] Xenograft study. Female SCID mice were implanted by sc injection with 5 x

10⁷ PC-9 2A10 tumor cells in 50% Matrigel. Once tumors reached 150 - 200 mm³, animals were sorted into treatment groups (n = 10) and dosed with vehicle, CO- 1686 at 100 mg/kg BID, MLN8237 at 20 mg/kg BID, or a combination of CO- 1686 and MLN8237 at the doses indicated. The formulation for the vehicle and CO- 1686 groups was 5% DMSO, 15% Solutol HS15 in PBS, and the formulation for MLN8237 was 10% Hydroxypropyl Beta Cyclodextrin in water. The study endpoint was a mean tumor volume of 2000 mm³ or Day 71, whichever came first, and efficacy was determined by mean tumor volume at the study endpoint.

[0080] The description of the present invention has been presented for purposes of illustration and description, but is not intended to be exhaustive or limiting of the invention to the form disclosed. The scope of the present invention is limited only by the scope of the following claims. Many modifications and variations will be apparent to those of ordinary skill in the art. The embodiment described and shown in the figures was chosen and described in order to best explain the principles of the invention, the practical application, and to enable others of ordinary skill in the art to understand the invention for various

embodiments with various modifications as are suited to the particular use contemplated.

[0081] While the invention has been particularly shown and described with reference to a number of embodiments, it would be understood by those skilled in the art that changes in the form and details may be made to the various embodiments disclosed herein without departing from the spirit and scope of the invention and that the various embodiments disclosed herein are not intended to act as limitations on the scope of the claims. All references cited herein are incorporated in their entirety by reference.

TABLE 1 - Examples of irreversible EGFR inhibitors

TABLE 2 - Examples of Aurora inhibitors

TABLE 3 - Parental and CO-1686 resistant NSCLC cell lines and pools

Claims

CLAIMS What is claimed is:

1. A method for treating cancer having mutant epidermal growth factor receptor (EGFR) in a subject comprising administering an irreversible mutant EGFR inhibitor and an Aurora kinase inhibitor.

2. The method of claim 1 wherein the irreversible mutant EGFR inhibitor covalently binds to cysteine 797 of mutant EGFR and exhibits at least 2-fold greater inhibition of a drug-resistant mutant EGFR relative to wild type EGFR.

3. The method of claim 1 wherein said cancer is non-small cell lung cancer (NSCLC).

4. The method of claim 1 wherein the irreversible mutant EGFR inhibitor is selected from CO- 1686, WZ4002, AZD9291, CNX2006, ASP8273, EGF816, HM61713, TAS-2913, and AZ5104.

5. The method of claim 1 wherein in the irreversible mutant EGFR inhibitor is CO- 1686.

6. The method of claim 1 where in the Aurora kinase inhibitor is selected from TAK- 901, AZD1152, PF-03814735, VX-680, MLN-8237, PHA-739358, CYC 116, SNS- 314, AT9283, R763, GSK1070916, AMG-900, ENMD-2076, MK-5108, BI 811283, PHA-680632, VE-465, JNJ-7706621, CCT129202, AKI-001, CHR-3520, Hesperadin, ZM447439, and Jadomycin-B

7. The method of claim 2, wherein the drug-resistant mutant is selected from

L858R/T790M EGFR and Exon-19 Deletion/T790M.

8. The method of claim 2, wherein the drug resistant mutant is selected from T790M, L858R, G719S, G719C, G719A, L861Q, a small in-frame deletion in exon 19, or an insertion in exon 20.

9. The method of claim 3 wherein said subject has relapsed NSCLC.

10. The method of claim 9 wherein the subject was previously treated with a first EGFR inhibitor.

11. The method of claim 10 wherein said first EGFR inhibitor was selected from

erlotinib, gefitinib, afatinib, dacomitinib, cetuximab, panitumumab, CO- 1686, WZ4002, AZD9291, CNX2006, ASP8273, EGF816, HM61713, TAS-2913, and AZ5104.