BR112015002384B1 - PHARMACEUTICAL COMBINATIONS COMPRISING A B-RAF INHIBITOR AND AN EGFR INHIBITOR, AND THEIR USES - Google Patents

Abstract

PHARMACEUTICAL COMBINATIONS COMPRISING A B-RAF INHIBITOR, AN EGFR INHIBITOR AND, OPTIONALLY, A PI3K-ALPHA INHIBITOR, AND THEIR USES. The present invention relates to a pharmaceutical combination comprising (a) a B-Raf inhibitor, (b) an EGFR inhibitor and, optionally, (c) a PI3K inhibitor; uses of such a combination in the treatment of proliferative diseases; and methods of treating a subject suffering from a proliferative disease comprise administering a therapeutically effective amount of such a combination.

Description

FIELD OF INVENTION

[0001] The present invention relates to a combination of a B-Raf kinase inhibitor and an epidermal growth factor receptor (EGFR also known as ErbB-1 or HER-1) inhibitor and, optionally, an inhibitor of phosphatidylinositol 3-kinase (PI or PI3K3-kinases) which is used to treat proliferative diseases. This invention also relates to the uses of such a combination in the treatment of proliferative diseases; to pharmaceutical compositions of combining agents and methods for treating an individual suffering from a proliferative disease comprising administering a therapeutically effective amount of such a combination to the individual.

BACKGROUND OF THE INVENTION

[0002] Protein kinases represent a large family of proteins, which play a central role in regulating a wide variety of cellular processes and maintaining control over cellular function. Aberrant kinase activity has been observed in many disease states including benign and malignant proliferative disorders as well as diseases resulting from inappropriate activation of the immune and nervous systems.

[0003] The Raf family of serine/threonine kinases includes three members: C-Raf (or Raf-1), B-Raf and A-Raf. B-Raf activating alleles have been identified in ~70% of melanomas, 40% of papillary thyroid carcinoma, 30% of low-grade ovarian carcinoma, and 10% of colorectal cancers. The majority of B-Raf mutations are found within the kinase domain, with a single substitution (V600E) accounting for 80%. Mutated B-Raf proteins activate the Raf-MEK-ERK pathway either through increased kinase activity for MEK or through activation of C-Raf. The B-Raf inhibitor in the present combination therapy inhibits cellular processes involving B-Raf kinase, blocking the signal cascade in these cancer cells and ultimately inducing stasis and/or cell death. B-Raf inhibitors useful in the present combinations are generally and specifically described in published PCT patent application WO2011/025927, which is hereby incorporated by reference.

[0004] There are three classes of PI3-kinases (PI3K). Class I enzymes consist of heterodimers having a regulatory domain (p85) and a catalytic subunit (p110), of which there are four isoforms: p110α, p110β, p110δ and p110Y. The α and β isoforms are ubiquitously expressed; α is linked upstream mainly to receptor tyrosine kinases, while β can mediate signals from G protein-coupled receptors and receptor tyrosine kinases. The δ and Y isoforms are primarily expressed in lymphocytes and play important roles in the regulation of immune responses.

[0005] A gain of function in PI3K signaling is common in many types of human cancer and includes inactivation of the PTEN tumor suppressor gene, amplification/upexpression or activation of mutations of some receptor tyrosine kinases (e.g. erbB3, erbB2, EGFR), amplification of genomic regions containing AKT, amplification of PIK3CA (the gene encoding p110α) and mutations in p110α. More than 30% of various types of solid tumors have recently been discovered to contain PIK3CA mutations. Of these mutation frequencies, PIK3CA is one of the most commonly mutated genes identified in human cancers. PI3K inhibitors useful in the present method, which have inhibitory activity towards the α isoform of PI3-kinases, are described in WO2010/029082 which is hereby incorporated by reference.

[0006] EGFRs are transmembrane receptors present on cell membranes. They have an extracellular binding component, a transmembrane component, and an intracellular tyrosine kinase component. EGFRs play an important role in controlling normal cell growth, apoptosis and other cellular functions. Dysregulation of EGFR activity can lead to uninterrupted or abnormal activation of receptors that cause unregulated cell division.

[0007] Epidermal growth factor receptor inhibitors are known in the art. Typically, they are small molecule tyrosine kinase inhibitors, such as erlotinib and gefitinib, or monoclonal antibodies. Anti-EGFR monoclonal antibodies, such as cetuximab and panitumumab, are especially useful EGFR inhibitors for use in the present invention. Cetuximab, its preparation and use to treat proliferative diseases, is disclosed in U.S. patent 6,217,866, which is incorporated herein by reference. Panitumumab, its preparation and use to treat proliferative diseases, is disclosed in U.S. patent 6,235,883, which is incorporated herein by reference.

SUMMARY OF THE INVENTION

[0008] The present invention relates to a therapeutic combination comprising: (a) a B-Raf inhibitor, (b) an EGFR inhibitor and, optionally, (c) a PI3K inhibitor, useful for separate simultaneous or sequential administration to an individual in need of it to treat or prevent a proliferative disease.

[0009] The present invention especially relates to a therapeutic combination comprising: (a) a B-Raf inhibitor of the Formula: or a pharmaceutically acceptable salt thereof (hereinafter referred to as Compound A), (b) an EGFR inhibitor, and, optionally, (c) a PI3K inhibitor.

DETAILED DESCRIPTION OF THE INVENTION

[0010] The present invention relates to a therapeutic combination comprising: (a) a B-Raf inhibitor, (b) an EGFR inhibitor and, optionally, (c) a PI3K inhibitor, useful for separate simultaneous administration or sequential to an individual in need of it to treat or prevent a proliferative disease.

[0011] The present invention especially relates to a therapeutic combination comprising: (a) a B-Raf inhibitor of the Formula: or a pharmaceutically acceptable salt thereof (hereinafter referred to as “Compound A”), (b) an EGFR inhibitor, and, optionally, (c) a PI3K inhibitor, especially a selective PI3Kα inhibitor.

[0012] The present invention especially relates to a therapeutic combination in which the EGFR inhibitor is a tyrosine kinase inhibitor, such as erlotinib or gefitinib, especially erolitinib, and especially in which the EGFR inhibitor is a monoclonal antibody, e.g. example, cetuximab or panitumumab, especially cetuximab.

[0013] PI3K inhibitors are known in the art. The optional PI3K inhibitor is especially a selective PI3K-α inhibitor which is a cycloamine urea 2-carboxamide derivative described in WO2010/029082, particularly the compounds of Formula (I): in which: A represents a heteroaryl selected from the group consisting of: R1 represents one of the following substituents: (1) unsubstituted or substituted C1-C7-alkyl, preferably substituted, wherein said substituents are independently selected from one or more, preferably one to nine of the following moieties: deuterium, fluorine, or one to two of the following C3-C5-cycloalkyl moieties; (2) Optionally substituted C3-C5-cycloalkyl wherein said substituents are independently selected from one or more, preferably one to four of the following moieties: deuterium, C1-C4-alkyl (preferably methyl), fluorine, cyano, aminocarbonyl; (3) optionally substituted phenyl wherein said substituents are independently selected from one or more, preferably one to two of the following moieties: deuterium, halo, cyano, C1-C7-alkyl, C1-C7-alkylamino, di(C1- C7-alkyl)amino, C1-C7-alkylaminocarbonyl, di(C1-C7-alkyl)aminocarbonyl, C1-C7-alkoxy; (4) optionally mono- or disubstituted amine; wherein said substituents are independently selected from the following moieties: deuterium, C1-C7-alkyl (which is unsubstituted or substituted by one or more substituents selected from the group of deuterium, fluorine, chlorine, hydroxy), phenylsulfonyl (which is unsubstituted or substituted by one or more, preferably one, C1-C7-alkyl, C1-C7-alkoxy, di(C1-C7-alkyl)amino-C1-C7-alkoxy); (5) substituted sulfonyl; wherein said substituent is selected from the following moieties: C1-C7-alkyl (which is unsubstituted or substituted by one or more substituents selected from the group of deuterium, fluorine), pyrrolidine, (which is unsubstituted or substituted by one or more substituents selected from the group of deuterium, hydroxy, oxo; particularly an oxo); (6) fluorine, chlorine; R2 represents hydrogen; R3 represents (1) hydrogen, (2) fluorine, chlorine, (3) optionally substituted methyl, wherein said substituents are independently selected from one or more, preferably one to three of the following moieties: deuterium, fluorine, chlorine, dimethylamino .

[0014] Radicals and symbols as used in defining a compound of Formula I have the meanings as described in WO2010/029082.

[0015] A preferred PI3K inhibitor is a compound that is specifically described in WO2010/029082. A most preferred selective PI3K-α inhibitor of the present invention is 1-({4-methyl-5-[2-(2,2,2-trifluoro-1,1-dimethyl-ethyl)-pyridin-4-yl] -thiazol-2-yl}-amide) of (S)-pyrrolidine-1,2-dicarboxylic acid 2-amide or a pharmaceutically acceptable salt thereof, of the Formula:

[0016] Hereinafter, dual combinations of Compound A and an EGFR inhibitor, triple combinations of Compound A, an EGFR inhibitor and a PI3K inhibitor of Formula I and more specifically dual combinations of Compound A and cetuximab and the triple combination of Compound A, cetuximab and Compound B will be referred to as a COMBINATION OF THE INVENTION.

[0017] The present invention particularly relates to a COMBINATION OF THE INVENTION useful for separate simultaneous or sequential administration to an individual in need thereof to treat or prevent a proliferative disease.

[0018] The present invention also relates to a COMBINATION OF THE INVENTION for use in preparing a pharmaceutical composition or medicine for treating or preventing a proliferative disease in an individual in need thereof.

[0019] The present invention further relates to the use of a COMBINATION OF THE INVENTION for the preparation of a pharmaceutical composition or medicine for the treatment or prevention of a proliferative disease.

[0020] The present invention relates to a method for treating an individual suffering from a proliferative disease comprising administering to said individual a COMBINATION OF THE INVENTION in an amount that is jointly therapeutically effective against a proliferative disease.

[0021] The present invention further provides a commercial packaging comprising as therapeutic agents a COMBINATION OF THE INVENTION, together with instructions for simultaneous, separate or sequential administration thereof for use in delaying the progression or treatment of a proliferative disease.

[0022] The general terms used here are defined with the following meanings, unless explicitly stated otherwise:

[0023] The terms “comprising” and “including” are used herein in their open and non-limiting sense unless otherwise noted.

[0024] The terms “a” and “the” and similar references in the context of describing the invention (especially in the context of the following claims) are to be interpreted to encompass both the singular and the plural, unless otherwise indicated here or clearly contradicted by the context. Where the plural form is used for compounds, salts, and the like, it is considered to also mean a compound, salt, or the like alone.

[0025] The terms “combination”, “therapeutic combination” or “pharmaceutical combination”, as used herein, either define a fixed combination in a dosage unit form or a kit of parts for combined administration wherein Compound A and Compound B can be administered independently at the same time or separately within time intervals that allow the combination pairs to show a cooperative, e.g., synergistic, effect.

[0026] The term “pharmaceutical composition” is defined herein to refer to a mixture or solution containing at least one therapeutic agent to be administered to an individual, for example, a mammal or human in order to prevent or treat a disease or particular condition affecting the mammal.

[0027] The term “pharmaceutically acceptable” is defined herein to refer to those compounds, materials, compositions and/or dosage forms, which are, within the scope of legal medical judgment, suitable for contact with the tissues of an individual , for example, a mammal or human, without excessive toxicity, irritant allergic response, and other complications of commensurate problem with a reasonable benefit/risk ratio.

[0028] The term “a combined preparation” is defined herein especially to refer to a “kit of parts” in the sense that combination pairs (a) and (b) as defined above can be dosed independently or by use of different fixed combinations with distinct quantities of combination pairs (a) and (b), i.e. simultaneously or at different points of time. The parts of the kit of parts can be administered later, for example, simultaneously or chronologically in stages, that is, at different time points and with the same or different time intervals for any part of the kit of parts. The ratio of total amounts of combination pair (a) to combination pair (b) to be administered in the combined preparation may be varied, for example, to meet the needs of a subpopulation of patients to be treated or the needs of the isolated patient.

[0029] The terms “coadministration” or “combined administration” as used herein are defined to encompass the administration of the selected therapeutic agents to a single patient, and are intended to include treatment regimens in which the agents are not necessarily administered by the same route of administration. or at the same time.

[0030] The terms “treatment” or “treatment” as used herein comprise a treatment that slows down, reduces or alleviates at least one symptom in an individual or delays the progression of a disease. For example, treatment may be the lessening of one or several symptoms of a disorder or complete eradication of a disorder, such as cancer. Within the meaning of the present invention, the term “treat” also denotes delaying, delaying the onset (that is, the period before the clinical manifestation of a disease) and/or reducing the risk of developing or worsening a disease. The term “protect” is used herein to mean preventing, delaying or treating, or all, as appropriate, the development or continuation or worsening of a disease in an individual.

[0031] The terms “therapeutically active together” or “therapeutically active effect together” mean that the therapeutic agents can be given separately (in a chronologically staged manner, especially a sequence-specific manner) at such time intervals as preferred, in the warm-blooded animal, especially human, being treated, it still shows a (preferably synergistic) interaction (joint therapeutic effect). Whether this is the case can, inter alia, be determined by following blood levels, showing that both compounds are present in the blood of the human being treated at least during certain time intervals.

[0032] The terms “pharmaceutically effective amount” or “clinically effective amount” or “therapeutically effective amount” of a combination of therapeutic agents is an amount sufficient to provide a clinically observable improvement in the baseline observable signs and symptoms of the disorder treated with the combination.

[0033] The terms “individual” or “patient” as used herein include animals that are capable of suffering or being afflicted with cancer or any disorder involving, directly or indirectly, cancer. Examples of individuals include mammals, for example, humans, dogs, cows, horses, pigs, sheep, goats, cats, mice, rabbits, rats and transgenic non-human animals. In the preferred embodiment, the subject is a human, for example, a human suffering from, at risk of suffering from, or potentially capable of suffering from cancers.

[0034] The terms “about” or “approximately” will have the meaning of within 10%, more preferably within 5%, of a certain value or range.

[0035] Compound A and/or Compound B can be administered in free form or in pharmaceutically acceptable salt form.

[0036] A “pharmaceutically acceptable salt”, as used herein, unless otherwise indicated, includes salts of acidic and basic groups that may be present in the compounds of the present invention. The compounds of the present invention which are basic in nature are capable of forming a wide variety of salts with various inorganic and organic acids. Acids that can be used to prepare pharmaceutically acceptable acid addition salts of such basic compounds of the present invention are those that form non-toxic acid addition salts, that is, salts containing pharmaceutically acceptable anions, such as acetate salts. , benzoate, bromide, chloride, citrate, fumarate, hydrobromide, hydrochloride, iodide, lactate, maleate, mandelate, nitrate, oxalate, salicylate, succinate, and tartrate.

[0037] Unless otherwise specified, or clearly indicated by the text, or not applicable, reference to therapeutic agents useful in the COMBINATION OF THE INVENTION includes both the free base of the compounds, and all pharmaceutically acceptable salts of the compounds.

[0038] The present invention particularly relates to a COMBINATION OF THE INVENTION useful for treating or preventing a proliferative disease in an individual in need thereof. In this embodiment of the present invention, the COMBINATION OF THE INVENTION is used for treating or preventing a proliferative disease comprising administering to the subject a combination therapy, comprising an effective amount of Compound A and an effective amount of an EGFR inhibitor, such as an EGFR monoclonal antibody, especially cetuximab or panitumumab, especially cetuximab. Preferably, these agents are administered in therapeutically effective dosages that, when combined, provide a beneficial effect. Administration can be separate, simultaneous or sequential.

[0039] The present invention further relates to a COMBINATION OF THE INVENTION useful for treating or preventing a proliferative disease in an individual in need thereof. In this embodiment of the present invention, the COMBINATION OF THE INVENTION is used for treating or preventing a proliferative disease comprising administering to the subject a triple combination therapy, comprising an effective amount of Compound A, an effective amount of an EGFR inhibitor, such as an of monoclonal antibody EGFR, especially cetuximab or panitumumab, especially cetuximab, and an effective amount of a selective PI3K-α inhibitor, especially a compound of Formula I, preferably Compound B. Preferably, these agents are administered in typical dosages. rapeutically effective which, when combined, provide a beneficial effect. Administration can be separate, simultaneous and/or sequential.

[0040] Therefore, the present invention more particularly relates to a combination of: (a) a B-Raf inhibitor of the Formula: or a pharmaceutically acceptable salt thereof, and (b) an EGFR inhibitor which is cetuximab.

[0041] Furthermore, the present invention particularly relates to a triple combination of: (a) a B-Raf inhibitor of the Formula: or a pharmaceutically acceptable salt thereof, (b) an EGFR inhibitor which is cetuximab, and (c) a PI3K inhibitor of Formula I, especially a PI3K inhibitor of Formula:

[0042] In one embodiment, the proliferative disease is cancer. The term “cancer” is used herein to mean a broad spectrum of tumors, including all solid tumors and hematological malignancies. Examples of such tumors include, but are not limited to, benign or malignant tumors of the brain, lung (in particular small cell lung cancer and non-small cell lung cancer), squamous cell, bladder, gastric, pancreatic, breast, head and neck , renal, kidney, ureter, ovarian, prostate, colorectal, esophageal, testicular, gynecological (e.g., uterine sarcomas, carcinoma of the fallopian tubes, endometrial, cervix, vagina, or vulva), thyroid, pancreatic, bone, skin, melanoma, uterine, ovarian, rectal, anal, colon, testicular, Hodgkin's disease, esophageal, small intestine, endocrine system (e.g., thyroid, parathyroid, or adrenal gland), soft tissue sarcomas, urethra, penis, leukemia , lymphomas, neoplasms of the central nervous system, sarcomas, myeloma, biliary, liver, neurofibromatosis, acute myeloid leukemia (AML), myelodysplastic syndromes (MDS), and Kaposi's sarcoma.

[0043] In a further embodiment of the present invention, the proliferative disease is melanoma, lung cancer (including non-small cell lung cancer (NSCLC), colorectal cancer (CRC), breast cancer, kidney cancer such as, e.g. renal cell carcinoma (RCC), liver cancer, endometrial cancer, acute myeloid leukemia (AML), myelodysplastic syndromes (MDS), thyroid cancer, particularly papillary thyroid cancer, pancreatic cancer, neurofibromatosis or hepatocellular carcinoma.

[0044] In an additional embodiment of the present invention, the proliferative disease is a solid tumor. The term “solid tumor” means especially melanoma, breast cancer, ovarian cancer, colorectal cancer, and generally gastrointestinal tract cancer, cervical cancer, lung cancer (including small cell lung cancer and non-small cell lung cancer), head and neck, bladder cancer, prostate cancer or Kaposi's sarcoma. The present combination inhibits the growth of solid tumors as well as liquid tumors. Furthermore, depending on the type of tumor and particular combination used, a decrease in tumor volume may be achieved. THE COMBINATION OF THE INVENTION disclosed herein is also adapted to prevent the metastatic expansion of tumors and the growth or development of micrometastases. THE COMBINATION OF THE INVENTION disclosed herein is suitable for treating patients with poor prognoses, especially such patients with poor prognoses having metastatic melanoma, colorectal or pancreatic cancer.

[0045] In an additional embodiment, the proliferative disease is melanoma or colorectal cancer, particularly colorectal cancer.

[0046] THE COMBINATION OF THE INVENTION is particularly useful for treating cancers having a genetic alteration in the RAS / RAF / MEK signal transduction pathway such as, for example, a B-Raf mutation or gene amplification.

[0047] In an important embodiment, the cancer to be treated is characterized by a B-Raf mutation, for example, B-Raf mutated colorectal cancer. In particular, the B-Raf mutation is a V600 mutation, for example, a V600E, V600K or V600G mutation.

[0048] Thus, the present invention particularly relates to a method for treating colorectal cancer characterized by a B-Raf mutation comprising administering a therapeutically effective amount of a COMBINATION OF THE INVENTION to a patient in need thereof.

[0049] More particularly, the present invention relates to a method for treating colorectal cancer characterized by a B-Raf mutation comprising administering a therapeutically effective amount of a combination of: (a) a B-Raf inhibitor of the Formula: or a pharmaceutically acceptable salt thereof, and (b) an EGFR inhibitor which is cetuximab.

[0050] Furthermore, the present invention particularly relates to a method for treating colorectal cancer characterized by a B-Raf mutation comprising administering a therapeutically effective amount of a triple combination of: (a) a B-Raf inhibitor of the Formula: or a pharmaceutically acceptable salt thereof, (b) an EGFR inhibitor which is cetuximab, and (c) a PI3K inhibitor of Formula I, especially a PI3K inhibitor of Formula:

[0051] In an important embodiment of each of these methods, the B-Raf mutation is a V600 mutation, for example, a V600E, V600K or V600G mutation.

[0052] The nature of proliferative diseases is multifactorial. Under certain circumstances, drugs can be combined with different mechanisms of action. However, just considering any combination of therapeutic agents having different mode of action does not necessarily lead to combinations with advantageous effects.

[0053] The administration of a pharmaceutical combination of the invention can not only result in a beneficial effect, for example, a synergistic therapeutic effect, for example with respect to relief, delay of progression or inhibition of symptoms, but also in other beneficial effects surprising, for example fewer side effects, more durable response, an improved quality of life or decreased morbidity, compared to a monotherapy that applies only one of the pharmaceutical therapeutic agents used in the combination of the invention.

[0054] An additional benefit is that lower doses of the therapeutic agents of the COMBINATION OF THE INVENTION can be used, for example, that dosages not only need to be frequently lower, but are also applied less frequently, or can be used to decrease the incidence of side effects seen with either combination pair alone. This is in accordance with the wishes and requirements of the patients to be treated.

[0055] It can be shown by established test models that a COMBINATION OF THE INVENTION results in the beneficial effects described hereinbefore. The person skilled in the art is fully permitted to select a relevant test model to prove such beneficial effects. The pharmacological activity of a COMBINATION OF THE INVENTION can be, for example, demonstrated in a clinical study or in an animal model as essentially described hereinafter.

[0056] By determining a synergistic interaction between one or more components, the optimal range for effect and the absolute dose ranges of each component for effect can be definitively measured by administering the components in different w/w ratio ranges and doses to patients in need of treatment. For humans, the complexity and cost of conducting clinical studies on patients may make it impractical to use this form of testing as a primary model for synergy. However, observation of synergy in one species can be predictive of the effect in other species and existing animal models, as described here, to measure a synergistic effect, and the results of such studies can also be used to predict ratio ranges. effective dose and absolute doses and plasma concentrations required in other species by applying pharmacokinetic/pharmacodynamic methods. Correlations established between tumor models and effects seen in man suggest that synergy in animals can be demonstrated, for example, through xenograft models or in appropriate cell lines.

[0057] Compound A is generally administered orally at a dose in the range of 10 mg to 1000 mg per day, for example 50 mg to 450 mg per day, or 100 mg to 400 mg per day. The daily dose can be administered on a qd or bid schedule.

[0058] The prescribing information for the ERBUTUX® brand of cetuximab advises that it is initially administered at a dose of 400 mg/m2 as a 120-min intravenous infusion followed by weekly doses at 250 mg/m2 infused over 60 minutes. Cetuximab is administered according to the prescribing information when used in the present combinations. However, dose reduction is also a possibility. Therefore, in accordance with the present invention, cetuximab is initially administered at a dose of 200 to 400 mg/m2 followed by weekly doses of 125 to 250 mg/m2.

[0059] Compound B is generally administered orally at a dose in the range of 30 mg to 450 mg per day, for example 100 to 400 mg per day. The daily dose can be administered on a qd or bid schedule.

[0060] It is an object of this invention to provide a pharmaceutical composition, comprising the COMBINATION OF THE INVENTION that is jointly therapeutically effective against a proliferative disease. In this composition, the Compound A and/or Compound B combination pairs may be administered in a single formulation or unit dosage form, may be administered simultaneously but separately, or may be administered sequentially by any suitable route. Preferably, the oral dosage form of Compound A and Compound B are administered simultaneously, but separately.

[0061] The monoclonal antibody EGFR inhibitor is typically administered separately as an intravenous infusion, preferably on a once-a-week schedule when the EGFR inhibitor is cetuximab.

[0062] In one embodiment, the present invention also relates to a COMBINATION OF THE INVENTION for use in preparing a pharmaceutical composition or medicament for treating or preventing a proliferative disease in an individual in need thereof.

[0063] The individual combination pairs of the COMBINATION OF THE INVENTION can be administered separately at different times during the course of therapy or simultaneously in split or single combination forms. The invention is to be understood, therefore, as covering all such simultaneous or alternating treatment regimens and the term “administer” is to be interpreted accordingly.

[0064] The effective dosage of each of the combination pairs employed in the COMBINATION OF THE INVENTION may vary, depending on the particular compound or pharmaceutical composition employed, the mode of administration, the condition being treated, and the severity of the condition being treated. Therefore, the dosage regimen of the COMBINATION OF THE INVENTION is selected according to a variety of factors including the route of administration and the patient's renal and hepatic function. The clinician or physician of ordinary skill can easily determine and prescribe the effective amount of simple therapeutic agents required to alleviate, neutralize or arrest the progress of the condition.

[0065] The optimal ratios, individual and combined dosages, and concentrations of combination pairs (a) and (b) of the COMBINATION OF THE INVENTION that yield efficacy without toxicity are based on the kinetics of availability of the therapeutic agents to the target sites, and are determined using methods known to those of skill in the art.

[0066] Effective dosing of each of the combination pairs may require more frequent administration of one of the compound(s) when compared to the other compound(s) in the combination. Therefore, to allow for proper dosing, packaged pharmaceutical products may contain one or more dosage forms containing the combination of compounds, and one or more dosage forms containing one of the combination of compounds, but not the other. s) compound(s) of the combination.

[0067] When the combination pairs that are used in the COMBINATION OF THE INVENTION are applied in the form thus marketed as for simple drugs, their dosage and mode of administration may be in accordance with the information provided in the leaflet of the respective marketed drug, if here from otherwise not mentioned.

[0068] The optimal dosage of each pair of combinations for treating a proliferative disease can be determined empirically for each individual using known methods and will depend on a variety of factors, including, but not limited to, the degree of advancement of the disease; the individual's age, body weight, general health, gender, and diet; the time and route of administration; and other medications that the individual is taking. Optimal dosages can be established using routine testing and procedures that are well known in the art.

[0069] The amount of each pair of combinations that can be combined with the carrier materials to produce a simple dosage form will vary, depending on the individual treated and the particular mode of administration. In some embodiments, the unit dosage form containing the combination of agents as described herein will contain the amounts of each agent in the combination typically being administered when the agents are administered alone.

[0070] The dosage frequency may vary, depending on the compound used and the particular condition to be treated or prevented. Patients can generally be monitored for therapeutic effectiveness using assays appropriate to the condition being treated or prevented that will be familiar to those of ordinary skill in the art.

[0071] The present invention relates to a method for treating an individual suffering from a proliferative disease comprising administering to said individual a COMBINATION OF THE INVENTION in an amount that is jointly therapeutically effective against a proliferative disease. In particular, the proliferative disease to be treated with a COMBINATION OF THE INVENTION is colorectal cancer, particularly a B-Raf mutated colorectal cancer, for example, a B-Raf mutated V600 colorectal cancer. In addition, treatment may include surgery or radiotherapy.

[0072] The present invention further relates to the COMBINATION OF THE INVENTION for use in the treatment of a proliferative disease, particularly cancer, in particular mutated B-Raf colorectal cancer, such as a V600 mutated B-Raf colorectal cancer.

[0073] The present invention further provides a commercial package comprising as therapeutic agents a COMBINATION OF THE INVENTION, together with instructions for simultaneous, separate or sequential administration thereof for use in delaying the progression or treatment of a proliferative disease in an individual in need of the same.

[0074] The following Examples illustrate the invention described above; however, they are not intended to limit the scope of the invention in any way. The beneficial effects of the pharmaceutical combination of the present invention can also be determined by other test models known as such to the person skilled in the relevant art.

EXAMPLE 1

[0075] This is a phase Ib, open-label, multicenter, randomized phase II dose escalation study that will enroll approximately 124 patients with B-Raf mutant metastatic colorectal cancer (mCRC).

[0076] The phase Ib (n~24) goal is to determine the maximum tolerated dose (MTD) and/or the recommended phase two dose (RP2D) of Compound A in combination with cetuximab (dual combination) and the MTD and /or RP2D of Compound A in combination with Compound B and cetuximab (triple combination). In the first stage of dose escalation, patient cohorts will be treated with the dual combination until the MTD/RP2D of the dual combination is identified. Then, patient cohorts will be treated with the triple combination during the second stage of dose escalation until the MTD/RP2D of the triple combination is identified.

[0077] Phase II (n~100) will evaluate the clinical efficacy of the dual combination and triple combination and will also characterize the safety of the drug combinations. Treatment will be administered in 28-day cycles until disease progression, unacceptable toxicity, withdrawal of informed consent, or death.

[0078] Tumor response will be assessed locally by the investigator according to a guideline based on RECIST version 1.1. Each patient will be evaluated for all potential sites of tumor lesions at Scan/Baseline and every 6 weeks after starting study treatment until disease progression. Scan/Baseline imaging assessments can be performed within 21 days of the start of treatment. Study tumor assessments have a window of ±7 days, with the exception of the first post-baseline tumor assessment. The first post-baseline tumor assessment should be performed 6 weeks (+7 day window allowed) after initiating treatment. There will be a tumor assessment at the end of Treatment (± 3 days) if the patient discontinues for any reason other than disease progression and the last tumor assessment was performed > 21 days prior to this day. Patients enrolled in the phase II portion of the study who discontinue study treatment due to a reason other than disease progression should be followed up monthly via phone call and should have tumor assessments every 6 weeks (± 7 days) until disease progression or initiation of subsequent antineoplastic therapy, or death, whichever occurs first.

Molecular pre-screening

[0079] To enter the screening phase of the study, patients must have written documentation of KRAS wild-type and BRAF V600 mutation status which should be obtained locally from a recent tumor biopsy (preferred) or the Most recent tumor files available. Molecular pre-screening informed consent must be signed prior to any study-related molecular pre-screening procedures (not applicable if mutational status has already been assessed outside of the study).

Treatment period

[0080] The treatment period will begin on Cycle 1 Day 1. During phase II, study treatment should be initiated <1 week following randomization. Study treatments will be administered over 28-day cycles and will continue until disease progression, unacceptable toxicity, withdrawal of informed consent, or death.

End of treatment (EOT)

[0081] The EOT visit occurs within 14 days after the last administration of study treatment (Section 7.1.5). All participating patients must complete this visit even if they had to discontinue prematurely.

Follow-up period

[0082] The follow-up period begins after the end of the Treatment visit and continues until the completion of all follow-up assessments, including survival follow-up.

POPULATION a) PATIENT POPULATION

[0083] Both phases of the study, phase Ib and phase II, will be conducted in adult patients with metastatic colorectal cancer (mCRC) harboring wild-type KRAS and a BRAF V600 mutation whose disease has progressed despite prior antineoplastic therapy or to for whom no effective standard therapy is yet available.

[0084] Patients enrolled in this study are not permitted to participate in parallel investigational drug or device studies. Additionally, patients who have completed the study should not be re-assigned to a second course of treatment.

b) INCLUSION CRITERIA

[0085] Patients eligible for inclusion in this study must meet ALL of the following criteria: 1. Age > 18 years at the start of dosing (phase Ib) or at the time of randomization (phase II) 2. Histological or cytological proof of cancer metastatic colorectal cancer (mCRC) 3. Progression after at least one prior standard of care regimen or being intolerant to irinotecan-based regimens 4. Written documentation of wild-type KRAS and BRAF V600E mutation, or any other mutation of BRAF V600 5. Phase II only: recent tumor biopsy at baseline 6. Evidence of measurable disease as determined by RECIST v1.1. Note: Lesions in areas of prior radiotherapy or other locoregional therapies (e.g., percutaneous separation) should not be considered measurable unless progression of the lesion has been documented since therapy. 7. Life expectancy > 3 months 8. ECOG performance status < 2 9. Seronegative pregnancy test within 72 hours prior to the first dose of study treatment in all women of childbearing potential 10. Able to understand and voluntarily sign the informed consent form, and the ability to comply with study visit schedules and other protocol requirements. Written informed consent must be obtained prior to screening procedures.

c) EXCLUSION CRITERIA

[0086] Patients eligible for this study must not meet ANY of the following criteria: 1. Phase II only: prior treatment with cetuximab, pan-nitumumab, and/or other EGFR inhibitors 2. Phase II only: prior treatment with EGFR inhibitors RAF, PI3K inhibitors and/or MEK inhibitors 3. Symptomatic or untreated leptomeningeal disease 4. Symptomatic brain metastasis. Previously treated or treatment-naïve patients for these conditions who are asymptomatic in the absence of corticosteroid therapy are permitted to enroll. Brain metastasis must be stable with verification through imaging (e.g., MRI or CT of the brain completed at screening to demonstrate no current evidence of progressive brain metastasis). Patients are not permitted to receive enzyme-inducing antiepileptic drugs. 5. Patients with diabetes mellitus requiring insulin treatment and/or with clinical signs or fasting glucose > 140 mg/dL / 7.8 mmol/L, history of clinically significant gestational diabetes mellitus or documented steroid-induced diabetes mellitus. 6. Known acute or chronic pancreatitis 7. Clinically significant cardiac disease including any of the following: • Congestive cardiac arrest requiring treatment (NYHA type > 2), LVEF < 45% as determined by MUGA scan or ECHOE, or uncontrolled hypertension ( refer to SHO-ISH guidelines) • History or presence of clinically significant ventricular arrhythmias or atrial fibrillation • Clinically significant resting bradycardia • Unstable angina pectoris < 3 months before starting study drug • Acute myocardial infarction (AMI) < 3 months before starting study drug • QTcF > 480 msec 8. Patients with any of the following laboratory values at Scan/Baseline: • Absolute neutrophil count (ANC) < 1,500/mm3 [1.5 x 109/ L] • Platelets < 100,000/mm3 [100 x 109/L] • Hemoglobin < 9.0 g/dL • Serum creatinine > 1.5 x ULN or calculated or directly measured CrCl < 50% LLN (lower limit of normal) • Total serum bilirubin > 1.5 x ULN • AST/SGOT and/or ALT/SGPT > 2.5 x ULN, or > 5 x ULN if liver metastasis is present. 9. Impairment of gastrointestinal (GI) function or GI disease that may significantly alter oral Compound A/Compound B absorption (e.g., ulcerative diseases, uncontrolled nausea, vomiting, diarrhea, malabsorption syndrome, bowel resection slender). 10. Previous or concurrent malignancy. Exceptions: adequately treated basal cell or squamous cell skin cancer; in situ carcinoma of the cervix, curatively treated and without evidence of recurrence for at least 3 years before entry into the study; or other curatively treated solid tumor, and without evidence of recurrence for at least 3 years prior to study entry. 11. Pregnant or lactating (lactating) women where pregnancy has been defined as the state of a female after conception and until the end of pregnancy, confirmed by a positive hCG laboratory test (>5 mIU/mL). Women of childbearing potential, defined as all women physiologically capable of becoming pregnant, are not permitted to participate in this study UNLESS they are using highly effective methods of contraception throughout the study and for 3 months after discontinuation of study drug. Highly effective methods of contraception include: • Total abstinence • Male or female sterilization • Combination of any two of the following (a+b or a+c or b+c) a. Use of oral, injected, or implanted hormonal contraception methods b. Placement of an intrauterine device (IUD) or intrauterine system (IUS) c. Barrier methods of contraception: condom or occlusive cap (diaphragm or cervical/valve caps) with spermicidal foam/gel/film/cream/vaginal suppository Postmenopausal women were allowed to participate in this study. Women are considered postmenopausal and not of childbearing potential if they have 12 months of natural (spontaneous) amenorrhea with an appropriate clinical profile (e.g., appropriate age, history of vasomotor symptoms) or six months of spontaneous amenorrhea with serum Follicle Stimulating Hormone (FSH) levels > 40 mIU/mL or have had surgical bilateral ovariectomy (with or without hysterectomy) or tubal ligation for at least six weeks prior to screening. In the case of o-ophorectomy alone, only when the woman's reproductive status has been confirmed by follow-up hormonal level assessment is she considered not to be of childbearing potential. 12. Sexually active males must use condoms during the relationship while taking the medicine and for 3 months after stopping treatment and should not father a child during this period. The use of condoms by vasectomized men is also required to prevent the release of the drug through the seminal fluid. 13. History of thromboembolic or cerebrovascular events within the last 6 months, including transient ischemic attack, cerebrovascular accident, deep vein thrombosis, or pulmonary embolism. 14. Patients who have received radiation therapy (which includes > 30% of bone marrow reserve), chemotherapy, biological therapy (e.g., antibodies) within < 4 weeks (6 weeks for nitrosourea, mitomycin-C), or have been treated with continuous or intermittent small molecule therapeutics or investigational agents within 5 half-lives of the agent (or < 4 weeks when the half-life is unknown) before initiating study drug or who have not recovered from the side effects of such therapy (except alopecia). 15. Patients who have undergone any major surgery within the last 2 weeks before starting study drug or who would not have fully recovered from previous surgery. 16. Known human immunodeficiency virus (HIV) infection. 17. Other severe, acute, or chronic medical or psychiatric condition or laboratory abnormality that may increase the risk associated with study participation or study drug administration or that may interfere with the interpretation of study results and, in the judgment of the investigator, would make the patient unsuitable for the study. 2) TREATMENT a) TREATMENT OF THE STUDY

[0087] The investigational drugs to be used in this study are Compound A and Compound B. The other drug to be used in this study is cetuximab.

[0088] The study treatments are: - Dual combination: Compound A and cetuximab - Triple combination: Compound A, Compound B, and cetuximab

1) DOSING REGIME

[0089] Patients will be assigned (phase Ib) or randomized (phase II) to one of the following regimens. - Dual combination: Compound A (QD or BID) and cetuximab (QW) - Triple combination: Compound A (QD or BID), Compound B (QD or BID), and cetuximab (QW) DOSE AND TREATMENT SCHEME

INSTRUCTIONS FOR ADMINISTRATION OF COMPOUND A OR COMPOUND A + COMPOUND B

[0090] Compound A and Compound B will be administered orally on a daily schedule (QD) as a fixed dose, and not by body weight or body surface area. Should new evidence from ongoing studies indicate which twice daily (BID) regimen(s) may be preferred, a BID regimen of either Compound A and/or Compound B in combination with cetuximab may be explored. opening new cohorts in the phase Ib portion of the study. A simple RP2D and schematic for the phase II portion will be explored for each drug. • QD DOSING: Patients should be instructed to take Compound A capsules (and Compound B tablets, if applicable) daily with a large glass of water (~250 mL) in the morning approximately 1 hour after completing a breakfast. morning light (e.g., non-grapefruit-based juice, toast, and jam, at approximately the same time each day. Patients should continue fasting for 1 h after administration. If the patient forgets to take the morning dose , then he should take the dose within 6 hours of the missed dose. If more than 6 hours passed, then the dose should be continued that day and the patient should continue treatment with the next scheduled dose. If, for any reason, breakfast was not eaten, then the patient should still take the scheduled morning dose with a glass of water. If this occurred on total PK sampling days, it should be documented in the eCRF. • BID DOSING: Compound A doses (and Compound B, if applicable) should be taken 12 ± 2 hours separately. Patients will be instructed to take daily doses with a large glass of water (~250 mL) in the morning approximately 1 hour after finishing a light breakfast and in the evening approximately 1 hour after a light meal or snack, at approximately the same time. of each day. Patients should continue fasting for 1 h after administration. If, for any reason, an evening meal was not consumed, then the patient should still take the scheduled evening dose with a glass of water. If only one of the two oral drugs (Compound A, Compound B) is administered BID, both drugs should be taken together in the morning and only the drug administered BID should be taken in the evening. • Doses should be taken at approximately the same time each day, except on days when blood collection is scheduled at the clinic, on which occasions patients should take their morning doses at the clinic. • Compound A and Compound B will be dosed at the same time for patients who are assigned/randomized to the triple combination. • On days when blood collection is scheduled at the clinic, patients will take oral study medicine at the clinic under the supervision of the investigator or designee. On all other days, patients will take oral study drugs at home. • FASTING PLASMA GLUCOSE MONITORING: On fasting plasma glucose monitoring days, patients must fast overnight for at least 8 hours before blood collection. Fasting plasma glucose should be collected before administering any steroid if given on the same day for cetuximab premedication. A light breakfast can be consumed after fasting plasma glucose has been removed. Compound A (and Compound B, if applicable) can be administered 1 hour after breakfast. Patients should continue fasting for 1 hour after administration of Compound A (and Compound B, if applicable). • PK SAMPLING: On PK sampling days, patients must be fasting overnight for at least 8 hours before the light meal to achieve light eating conditions. Pre-dose PK samples should only be collected prior to input of Compound A (and Compound B, if applicable). • At each visit, on-site staff will ensure that the appropriate dose of each study drug is administered and will provide the patient with the correct amount of study drug(s) for subsequent dosing. Patients will be instructed to return unused study drugs to the site at each visit. • Patients should be taught to swallow capsules/tablets whole and not to chew or crush. • Any dose that is missed should be skipped and should not be replaced or topped up during the next scheduled dose or on a subsequent day, wherever applicable. • Patients must avoid consumption of grapefruit, pomegranates, star fruits, Seville oranges or products containing the juice of each during the entire study and preferably 7 days before the first dose of study medications, due to potential CYP3A4 interaction with the medications of the study. Orange juice is allowed. • If vomiting occurs during the course of treatment, no redosing of the patient is permitted before the next scheduled dose. The occurrence and frequency of any vomiting and/or diarrhea (or increased stool frequency) should be noted in the AEs section of the eCRF. Additionally, on total PK sampling days, the time of onset of any vomiting episode within the first 4 hours post-dosing on that day should be noted in the corresponding PK eCRF Dose Administration Log. • The investigator or responsible local personnel should instruct the patient to take the study medicine as per protocol (promote compliance). All dosages prescribed and dispensed to the patient and all dose changes and all missed doses during the study must be recorded in the eCRF Dosage Administration Log. Drug liability must be performed on a regular basis. Patients will be instructed to return unused study drugs to the site at the end of each cycle. Site personnel will ensure that the appropriate dose of each study drug is administered at each visit and will provide the patient with the correct amount of drugs for subsequent dosing.

ADMINISTRATION OF CETUXIMAB

[0091] Cetuximab will be administered intravenously weekly on Days 1, 8, 15 and 22 (± 3 days) of each cycle at the study site in accordance with institutional standards. Premedication should be administered as described following institutional standards 30 minutes before cetuximab infusion. The initial administered dose of cetuximab (Cycle 1 Day 1) is 400 mg/m2 as a 120-min intravenous infusion followed by a 250 mg/m2 weekly dose infused over 60 minutes. The infusion rate should not exceed 10 mg/min. Close monitoring is required during the infusion and for at least 1 hour after the end of the infusion.

[0092] If an infusion reaction occurs while cetuximab is being administered, the infusion should be stopped immediately and patients should be strictly monitored and treated in line with institutional standards.

SEQUENCE OF DRUG ADMINISTRATION

[0093] Premedication having the potential to alter the pH of the upper gastrointestinal (GI) tract may alter the solubility of Compound A and/or Compound B and consequently their bioavailability. These agents include, but are not limited to, proton pump inhibitors (e.g., omeprazole), H2 antagonists (e.g., ranitidine), and antacids. Therefore, oral dosing of Compound A (and Compound B, if applicable) will be administered prior to cetuximab and its premedication should preferably be based on a combination of an H1 antagonist (e.g. diphenhydramine) and dexamethasone (10 mg IV). A minimum of 1 hour must pass from the time of administration of Compound A (and Compound B, if applicable) to the administration of cetuximab premedication. Cetuximab infusion is recommended to occur 0.5 h post-premedication (i.e., 1.5 h post-Compound A/Compound B intake).

DURATION OF TREATMENT

[0094] Patients may continue treatment with the study drug until they experience unacceptable toxicity, disease progression and/or treatment is discontinued at the discretion of the investigator or withdrawal of consent.

DOSE ESCALATION GUIDELINES INITIAL DOSE RATIO (1) DUAL COMBINATION

[0095] The starting dose for the dual combination study drugs is 100 mg QD for Compound A, and 400 mg/m2 initial dose (Cycle 1 Day 1) and 250 mg/m2 subsequent weekly doses as an intravenous infusion for cetuximab. These starting doses are based on available data from the first continuous in-human study of Compound A and the recommended cetuximab dose for metastatic colorectal cancer according to the cetuximab label. Currently taking into account all available information regarding the dose-DLT relationships of Compound A and cetuximab as single agents and the uncertainty surrounding the toxicity of the combination, the previous distribution of DLT rates indicates that the proposed starting dose combination satisfies the lation with overdose control criteria (EWOC).

(2) TRIPLE COMBINATION

[0096] The starting doses of Compound A, Compound B, and cetuximab during the triple combination are based on all available data for all three drugs. Compound A and cetuximab will be administered at 50% and 100% of the determined MTD/RP2D of the dual combination, respectively. The starting dose of Compound B is expected to be 100 mg QD which is 25% of the single-agent MTD identified during a phase I clinical study of Compound B administered to patients with solid tumors.

[0097] No DDI at the PK level is expected between Compound B and cetuximab. Since Compound A is a BCRP inhibitor and Compound B is a BCRP substrate, there is a potential for increased Compound B exposure when coadministered with Compound A. Given the favorable bioavailability observed preclinically (58% at ADME rat) and clinically, the maximum possible increase in Compound B exposure is expected to be less than 60%. Therefore, the starting dose of Compound B is set at 100 mg QD to provide sufficient safety margin. Furthermore, Compound B is a time-dependent CYP3A4 inhibitor. Compound A is mainly metabolized by CYP3A4. According to the Food and Drug Administration (FDA) recommended static mechanistic model, a 100 mg QD dose of Compound B when administered concomitantly with Compound A can increase the plasma AUC of Compound A by up to 3-fold. To mitigate the potential increase in Compound A exposure when Compound B is added, the starting dose of Compound A in the triple combination (Compound A, Compound B, cetuximab) will be decreased to 50% of its identified MTD/RP2D during the dual combination. , as stated above. Additionally, rapid PK assessment through in-life PK analysis will be implemented in the triple combination dose escalation phase to monitor PK on Day 1 and Day 8 of Compound A and Compound B to inform dose escalation decisions. In the event that a DDI effect is observed and suggests overexposure of Compound A, a dose reduction of Compound A may be implemented.

[0098] Before the first patient is dosed with the triple combination, the Bayesian model will be updated with the latest dual phase combination dose escalation data to confirm that the proposed starting doses for Compound A and Compound B are still are appropriate (i.e., meet EWOC criteria) when administered with the identified cetuximab dose of the dual combination. If the proposed starting dose does not meet the criteria, a lower dose combination will be used that meets the EWOC criteria.

INTERIM DOSE LEVELS

[0099] The tables below describe the starting doses and interim dose levels of the study treatments for the dual (Compound A, cetuximab) and triple (Compound A, Compound B, cetuximab) combinations that may be evaluated during this trial. The dose of cetuximab will not be escalated, but may be reduced. Additional dose levels not currently specified may be added and additional patients may be enrolled at an already tested dose level if such changes are deemed necessary to provide optimal safety and tolerability, pharmacokinetic data, and pharmacodynamics.

[0100] If at any time during the phase Ib portion of the study, emerging data from other clinical trials with Compound A and/or Compound B indicate that a BID dosing regimen of Compound A and/or Compound B regimen should If preferred, cohorts evaluating BID dosing regimen(s) could be explored in the phase Ib portion of the study. If the decision is made to switch to BID, then the initial total daily dose (to be administered as two divided doses for BID) will be a dose that has previously been observed to be tolerated well as a single daily dose, will be below the MTD, and will allow by BLRM.

[0101] Dose levels beyond the MTDs/RP2Ds determined during the previous single-agent studies of Compound A and Compound B will not be evaluated in this study.

EXAMPLE 2

[0102] The effect of combining Compound A with the PI3Kα-specific inhibitor - Compound B, or the EGFR inhibitor - erlotinib, on the proliferation of BRAF mutant CRC-derived cell lines is examined. Both combinations synergistically inhibited proliferation in the majority of cells tested with the pairings of Compound A/Compound B and Compound A/erlotinib active in 7/8 and 6/9 cell lines, respectively. Combinations were active in cells harboring mutant and wild-type alleles of the PI3Kα gene. In all cell lines tested, only Compound A exhibited significant single agent activity, although cell lines with any of the activating mutations in PI3Kα or loss of PTEN were largely refractory to all three compounds. Finally, the synergy between Compound A and Compound B was maintained, but the overall strength of the antiproliferative effect was increased, when the EGFR inhibitory antibody - cetuximab - was added as a third agent. Collectively these data support the combination of Compound A with any of the EGFR or PI3Kα inhibitors. Furthermore, these results suggest that additional benefit may be gained by the simultaneous addition of all three types of inhibitor.

[0103] Single agent and combinatorial effects on the proliferation of RAF (Compound A), PI3Kα (Compound B) and EGFR (erlotinib, cetuximab) inhibitors in nine BRAF mutant CRC-derived cell lines. All cell lines express the BRAFV600E protein, except MDST8 which expresses the BRAFV600K variant. Cells harboring known or putative activating mutations in the PI3Kα gene are marked with a (*) and cells with loss of PTEN marked with a (#). Cell proliferation was measured in 72h cell titration Glo™ assays and all results shown are the result of at least triplicate measurements. Shown are single agent IC50 values for each compound and synergy score measurements for each combination (described in Lehar J, Krueger AS, Avery W, et al (2009). Synergistic drug combinations tend to improve selectivity therapeutically relevant. Nat Biotechnol 27, 659-666.). Interactions were judged synergistic when ratings > 2.0 where observed. For triple combination synergy measurements, the synergy between Compound A and Compound B was measured in a standard dose matrix format in the presence of a fixed concentration of cetuximab (50 nM).

EXAMPLE 3

[0104] Assessment of therapeutic interactions between a B-RAF inhibitor (Compound A), PI3K-α inhibitor (Compound B), and cetuximab is evaluated in the subcutaneous HT-29 human colorectal adenocarcinoma xenograft model. The cells were reported to be heterozygous for V600E mutant B-RAF (1799T>A in BRAF), P449T mutant PI3K-α (1345C>A in PIK3CA), and a point and insertion mutation in APC; and be homozygous for point mutations in SMAD4 and TP53. No additional likely oncogenic mutations were observed in the 59 other genes whose alterations are frequently associated with neoplasia.

[0105] Compound A was stored at room temperature and suspended at 2.0 mg/mL in 0.5% carboxymethyl cellulose (CMC) and 0.5% Tween® 80 in deionized water (Vehicle 1). A fresh suspension was prepared every two weeks and stored at room temperature.

[0106] Compound B was stored at 4° C and suspended at 2.5 mg/mL in 0.5% methylcellulose in deionized water (Vehicle 2). A fresh suspension was prepared weekly and stored at 4°C.

[0107] Cetuximab (ERBITUX®, ImClone/Bristol Myers Squibb, 2 mg/mL, Lot #10COO39SA) was aliquoted at the beginning of the study and stored at 4° C; a fresh aliquot was used on each day of dosing.

[0108] Paclitaxel dosing solutions at 3 mg/mL were prepared fresh on each day of treatment by diluting aliquots of a home-prepared paclitaxel raw material (30 mg/mL paclitaxel in 50% ethanol: 50% of Cremophor® EL) ten times with 5% dextrose in water. Dosing solutions have recently been prepared for a group at a time.

[0109] Each agent was administered at a single dose level, individually and in dual and triple combinations that were initiated on Day 1 (D1) in female athymic mice with established subcutaneous tumors. Body weight (BW) and health were monitored, and tumor volume was measured twice weekly throughout the study. Animals were euthanized on D29 at 4 and 24 h after final dosing with Compound A and tumors were harvested from three animals per group at each time point. Efficacy was determined from mean tumor volume changes between D1 and D29. Prisma summarizes test results as non-significant (ns) at P > 0.05, significant (symbolized by “*”) at 0.01 < P < 0.05, very significant (“**”) at 0.001 < P < 0.01, and extremely significant (“***”) at P < 0.001. The results are reported in the table below.

[0110]

[0111] Treatment efficacy was determined on D29, the day dosing Compound A was completed. For the purpose of statistical analysis, ΔTV, the difference in tumor volume between D1 (the beginning of dosing) and the final day, was determined for each animal. For each treatment group, the response on the final day was calculated by one of the following relationships: ΔT = (treated group mean tumor volume on the final day) - (treated group mean tumor volume on D1), ΔC = (control group mean tumor volume on the final day) - (group mean tumor volume control on D1), and T0 = average tumor volume of the group treated on D1. T/T0 values that are negative represent net tumor reduction for a group. A T/C value of 40% or less suggests potential therapeutic activity.

RESPONSE TO COMBINATION THERAPIES (GROUPS 5-8)

[0112] In Group 5, Compound A in dual combination with Compound B resulted in a ΔT of 366 mm3, corresponding to 39% T/C, and median inhibition of tumor growth was not significant. The combination improved non-significantly under Compound A monotherapy in Group 2 and Compound B monotherapy in Group 3.

[0113] In Group 6, Compound A in dual combination with cetu ximab resulted in a ΔT of 114 mm3, corresponding to 12% T/C, and significant inhibition (P < 0.01). The combination significantly improved Compound A monotherapy in Group 2 and cetuximab monotherapy in Group 4 (P < 0.01).

[0114] In Group 7, Compound B in the dual combination with cetuxi mab resulted in a ΔT of 459 mm3, corresponding to 49% T/C, and non-significant inhibition. The combination non-significantly improved Compound B monotherapy in Group 3 and cetuximab monotherapy in Group 4.

[0115] Group 8, the triple combination of Compound A, Compound B, and cetuximab resulted in a ΔT of -3 mm3, corresponding to -2% of T/T0, and significant activity (P < 0.001). This treatment significantly improved Compound A monotherapy in Group 2 (P < 0.001), Compound B monotherapy in Group 3 (P < 0.01), and cetuximab monotherapy in Group 4 (P < 0.001). Furthermore, it improved significantly in dual combinations with Compound A/Compound B in Group 5 and Compound B/cetuximab in Group 7 (P < 0.01), and not significantly in the dual combination of Compound A/cetuximab in Group 6.

Claims (10)

1. Pharmaceutical combination, characterized by the fact that it comprises: (a) a B-Raf inhibitor of the Formula: or a pharmaceutically acceptable salt thereof, and (b) an EGFR inhibitor, which is cetuximab, for simultaneous administration. 2. Combination, according to claim 1, characterized by the fact that it is for use in the treatment of a proliferative disease in an individual in need thereof. 3. Combination, according to claim 1, characterized by the fact that it is for use in the preparation of a medicine for treating a proliferative disease. 4. Combination, according to claim 3, characterized by the fact that the proliferative disease comprises a B-Raf mutation. 5. Combination, according to claim 4, characterized by the fact that the B-Raf mutation is a V600 mutation. 6. Pharmaceutical combination, according to claim 5, characterized by the fact that the proliferative disease is colorectal cancer. 7. Use of the combination, as defined in claim 1, characterized by the fact that it is for manufacturing a medicine for treating a proliferative disease. 8. Use of a therapeutically effective amount of: (a) a B-Raf inhibitor of the Formula: or a pharmaceutically acceptable salt thereof, and (b) an EGFR inhibitor, which is cetuximab, said use being characterized by the fact that it is for preparing a pharmaceutical combination for treating a proliferative disease in a human patient, being that the proliferative disease comprises a B-Raf mutation, and that said combination is administered simultaneously to said patient. 9. Use according to claim 8, characterized by the fact that the B-Raf mutation is a V600 mutation. 10. Use according to claim 9, characterized by the fact that the proliferative disease is colorectal cancer.