HK1181321A - Composition for combination anti-cancer therapy - Google Patents

Description

Combination anticancer therapy

Technical Field

The present invention relates to a combination therapy for the treatment of a patient suffering from a proliferative disorder, in particular a solid tumor, such as colorectal cancer, melanoma and thyroid cancer, comprising administering to said patient propane-1-sulfonic acid {3- [5- (4-chlorophenyl) -1H-pyrrolo [2,3-b ] pyridine-3-carbonyl ] -2, 4-difluoro-phenyl } -amide and an EGFR inhibitor.

Background

Normally functioning b-Raf is a kinase that participates in signal relaying from the cell membrane to the nucleus and is only active when it is needed to relay the signal. However, mutant b-Raf with the V600E mutation was always active and therefore plays a role in tumor development. The above mutant b-Raf has been implicated in a variety of tumors, such as colorectal cancer, melanoma, and thyroid cancer.

Propane-1-sulfonic acid {3- [5- (4-chlorophenyl) -1H-pyrrolo [2,3-b ] pyridine-3-carbonyl ] -2, 4-difluoro-phenyl } -amide (hereinafter also referred to as "compound I") is a b-Raf kinase inhibitor that specifically targets mutant b-Raf having the V600E mutation. See WO2007/002325 for such compounds. Thus, the above inhibitors are useful for inhibiting tumors, in particular solid tumors, such as colorectal cancer, melanoma and thyroid cancer, comprising b-Raf having the V600 mutation and preferably the V600E mutation.

Protein Tyrosine Kinases (PTK) catalyze the phosphorylation of tyrosine residues in a variety of proteins involved in regulating cell Growth and differentiation (Wilks et al, Progress in Growth Factor Research97(1990) 2; Chan, A.C. and Shaw, A.S., curr, Opin. immunol.8(1996) 394-401). PTKs described above can be classified into receptor tyrosine kinases (e.g., EGFR/HER-1, c-erbB-2/HER-2, c-met, PDGFr, and FGFr) and non-receptor tyrosine kinases (e.g., src and lck).

It is known that receptor tyrosine kinases of the HER family, such as HER-2 and EGFR (HER-1), are often aberrantly expressed in common human cancers such as breast, gastrointestinal (colon, rectal or gastric), thyroid, leukemia, ovarian, bronchial, pancreatic and melanoma. High levels of these receptors are associated with poor prognosis and poor therapeutic response (Wright, C. et al, Br. J. cancer65(1992) 118-121).

PTK inhibitors and in particular EGFR inhibitors have been developed. However, tumors containing b-Raf with the V600E mutation are known to be resilient to treatment with EGFR inhibitors. See Prewett et al, clin. cancer Res. (2002),8: 994-. However, the applicant has unexpectedly found that the combination therapy of compound I with an EGFR inhibitor is not only able to reduce the above rebound, but also to induce an improved antitumor effect which is significantly superior to the results obtained with each compound alone without a significant increase in toxicity.

In addition to EGFR inhibitors, topoisomerase inhibitors are also antiproliferative agents. However, tumors containing the V600E mutation are also known to be resilient to treatment with topoisomerase inhibitors. See Prewett et al, Clin. cancer Res. (2002),8: 994-. However, the applicant has unexpectedly found that the combination of compound I with an EGFR inhibitor and a topoisomerase inhibitor not only reduces the above rebound, but also causes an improved antitumor effect which is significantly superior to the results obtained with each compound alone without a significant increase in toxicity.

Disclosure of Invention

The present invention relates to a pharmaceutical product comprising: (A) a first component comprising as an active agent propane-1-sulfonic acid {3- [5- (4-chlorophenyl) -1H-pyrrolo [2,3-b ] pyridine-3-carbonyl ] -2, 4-difluoro-phenyl } -amide or a pharmaceutically acceptable salt thereof; and (B) a second component comprising an EGFR inhibitor as an active agent; the pharmaceutical product is for simultaneous or sequential use as a combined preparation for the treatment of a proliferative disorder, in particular cancer, more particularly colorectal cancer, melanoma and thyroid cancer, comprising b-Raf having the V600 mutation and in particular the V600E mutation.

The invention also relates to a method of treating a patient suffering from a proliferative disorder, said method comprising administering to said patient a combination as described above.

The present invention also relates to a kit comprising: (A) a first component comprising compound I or a pharmaceutically acceptable salt thereof as an active agent; and (B) a second component comprising an EGFR inhibitor as an active agent.

The invention also relates to a pharmaceutical product comprising: (A) a first component comprising compound I or a pharmaceutically acceptable salt thereof as an active agent; (B) a second component comprising an EGFR inhibitor as an active agent; and optionally (C) a third component comprising a topoisomerase inhibitor or a pharmaceutically acceptable salt thereof; the pharmaceutical product is for simultaneous or sequential use as a combined preparation for the treatment of a proliferative disorder, in particular cancer, more particularly colorectal cancer, melanoma and thyroid cancer, comprising b-Raf having the V600 mutation and in particular the V600E mutation.

In addition, the present invention relates to the use of compound I or a pharmaceutically acceptable salt thereof and an EGFR inhibitor for the treatment of proliferative disorders.

Another aspect of the invention is the use of compound I, or a pharmaceutically acceptable salt thereof, and an EGFR inhibitor in the manufacture of a medicament for the treatment of proliferative disorders.

Drawings

Figure 1 shows the tolerability of compound I monotherapy (75mg/kg bid), erlotinib hydrochloride monotherapy (67mg/kg qd), erlotinib hydrochloride monotherapy (100mg/kg qd) and combination therapy of compound I (75mg/kg bid) and erlotinib hydrochloride (67mg/kg qd), expressed as% weight change.

Figure 2 shows the anti-tumor activity of compound I monotherapy (75mg/kg bid), erlotinib hydrochloride monotherapy (67mg/kg qd), erlotinib hydrochloride monotherapy (100mg/kg qd) and combination therapy of compound I (75mg/kg bid) and erlotinib hydrochloride (67mg/kg qd), as indicated by the change in mean tumor volume over time.

Figure 3 shows the effect of compound I monotherapy (75mg/kg bid), erlotinib hydrochloride monotherapy (67mg/kg qd), erlotinib hydrochloride monotherapy (100mg/kg qd) and compound I (75mg/kg bid) and erlotinib hydrochloride (67mg/kg qd) combination therapy on survival as a percentage of surviving mice over time.

Figure 4 shows the tolerability of compound I monotherapy (75mg/kg bid), compound I monotherapy (25mg/kgbid), cetuximab (cetuximab) monotherapy (40mg/kg2 x/wk), compound I (25mg/kg bid) and cetuximab (40mg/kg2 x/wk) combination therapy, and compound I (75mg/kg bid) and cetuximab (40mg/kg2 x/wk) combination therapy, expressed by% weight change.

Figure 5 shows the anti-tumor activity of compound I monotherapy (75mg/kg bid), compound I monotherapy (25mg/kgbid), cetuximab monotherapy (40mg/kg2 x/wk), compound I (25mg/kg bid) and cetuximab (40mg/kg2 x/wk) combination therapy, and compound I (75mg/kg bid) and cetuximab (40mg/kg2 x/wk) combination therapy, as expressed by the change in mean tumor volume over time.

Figure 6 shows the effect on survival of compound I monotherapy (75mg/kg bid), compound I monotherapy (25mg/kgbid), cetuximab monotherapy (40mg/kg2 x/wk), compound I (25mg/kg bid) and cetuximab (40mg/kg2 x/wk) combination therapy, and compound I (75mg/kg bid) and cetuximab (40mg/kg2 x/wk) combination therapy, expressed as a percentage of surviving mice over time.

FIG. 7 shows combination therapy of Compound I monotherapy (25mg/kg bid), cetuximab monotherapy (40mg/kg2 x/wk), irinotecan hydrochloride monotherapy (40mg/kg q4d x 5), Compound I (25mg/kg bid), and irinotecan hydrochloride (40mg/kg q4d x 5), tolerability of the combination therapy of cetuximab (40mg/kg2 x/wk) and irinotecan hydrochloride (40mg/kg q4d x 5), of the combination therapy of compound I (25mg/kg bid) and cetuximab (40mg/kg2 x/wk) and of the combination therapy of compound I (25mg/kgbid), cetuximab (40mg/kg2 x/wk) and irinotecan hydrochloride (40mg/kg q4d x 5), expressed as% weight change.

FIG. 8 shows combination therapy of Compound I monotherapy (25mg/kg bid), Cetuximab monotherapy (40mg/kg2 x/wk), irinotecan hydrochloride monotherapy (40mg/kg q4d x 5), Compound I (25mg/kgbid), and irinotecan hydrochloride (40mg/kg q4d x 5), the antitumor activity of the combination therapy of cetuximab (40mg/kg2 x/wk) and irinotecan hydrochloride (40mg/kg q4d x 5), of the combination therapy of compound I (25mg/kg bid) and cetuximab (40mg/kg2 x/wk) and of the combination therapy of compound I (25mg/kg bid), cetuximab (40mg/kg2 x/wk) and irinotecan hydrochloride (40mg/kg q4d x 5) is expressed as the change in mean tumor volume over time.

FIG. 9 shows combination therapy of Compound I monotherapy (25mg/kg bid), Cetuximab monotherapy (40mg/kg2 x/wk), irinotecan hydrochloride monotherapy (40mg/kg q4d x 5), Compound I (25mg/kgbid), and irinotecan hydrochloride (40mg/kg q4d x 5), cetuximab (40mg/kg2 x/wk) and irinotecan hydrochloride (40mg/kg q4d x 5) combination therapy, compound I (25mg/kg bid) and cetuximab (40mg/kg2 x/wk) combination therapy and the effect of compound I (25mg/kg bid), cetuximab (40mg/kg2 x/wk) and irinotecan hydrochloride (40mg/kg q4d x 5) combination therapy on survival, expressed as a percentage of surviving mice over time.

Detailed Description

As mentioned above, "compound I" means in this application propane-1-sulfonic acid {3- [5- (4-chlorophenyl) -1H-pyrrolo [2,3-b ] pyridine-3-carbonyl ] -2, 4-difluoro-phenyl } -amide. Which is a compound having the following structure.

Compound I is a b-Raf kinase inhibitor that specifically targets b-Raf with the V600E mutation.

The "V600E" mutation of b-Raf as used herein refers to a mutation in the b-Raf protein wherein the valine residue at residue position 600 of b-Raf is replaced by glutamic acid.

When referring to receptor tyrosine kinases of the HER family, such as HER-2 and EGFR (HER-1), the abbreviation "HER" as used herein refers to the human epidermal receptor and the abbreviation "EGFR" refers to the epidermal growth factor receptor.

The term "combined preparation" as used herein means a simultaneous or sequential combination. The active agents, e.g. compound I or topoisomerase inhibitors such as irinotecan or EGFR inhibitors such as erlotinib or pharmaceutically acceptable salts or cetuximab, for the combinations of the present application can be used in any suitable dosage form, e.g. tablets, capsules, solutions, suspensions, etc., depending on the nature of the active ingredient selected. Compound I or a pharmaceutically acceptable salt thereof may be administered, for example, orally. Erlotinib or a pharmaceutically acceptable salt thereof can be administered, for example, orally. Cetuximab may be administered, for example, intraperitoneally or intravenously. Irinotecan or a pharmaceutically acceptable salt thereof can be administered, for example, intraperitoneally or intravenously. Some combinations disclosed herein may exhibit greater than additive therapeutic effects (synergistic therapeutic effects).

The term "pharmaceutically acceptable carrier" as used herein means that the carrier is not of a nature that would allow a reasonably prudent physician to avoid administration to a patient, taking into account the disease or condition to be treated and the corresponding route of administration.

The term "pharmaceutically acceptable salt" of a compound, as used herein, refers to any conventional salt or base addition salt that retains the biological effectiveness and properties of the compound and is formed from a suitable non-toxic organic or inorganic acid or organic or inorganic base. The term "therapeutically effective" as used herein refers to an amount of a drug or combination or composition that is effective to produce a desired therapeutic effect, such as arresting the growth of or shrinking a cancerous tumor or extending the life of a patient, upon administration to the patient.

The terms "cell proliferative disorder" and "proliferative disorder" refer to disorders associated with a degree of abnormal cell proliferation. In one embodiment, the proliferative disorder is cancer.

The terms "cancer" and "cancerous" refer to or describe the physiological state typically characterized by unregulated cell growth/proliferation in a mammal. Examples of cancer include, but are not limited to, colorectal cancer, melanoma, and thyroid cancer.

The term "colorectal tumor" or "colorectal cancer" refers to any tumor or cancer of the large intestine, including the colon (the large intestine from the cecum to the rectum) and rectum, including, for example, adenocarcinoma and less common forms such as lymphoma and squamous cell carcinoma.

By "inhibiting cell growth or proliferation" is meant reducing cell growth or proliferation by at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, or 100% and includes inducing cell death.

The phrase "substantially reduced" or "substantially different," as used herein, refers to a difference between two values (typically one value associated with one molecule and the other value associated with a reference/control molecule) that is sufficiently high that one of skill in the art can consider the difference between the two values to be statistically significant with respect to the biological characteristic represented by the value.

The term "tumor" refers to all neoplastic cell growth and proliferation (whether malignant or benign) and all precancerous and cancerous cells and tissues. The terms "cancer," "cancerous," "cell proliferative disorder," "proliferative disorder," and "tumor" when referred to in this application are not mutually exclusive.

After treatment, "regression" of the tumor is considered to occur when the volume of the tumor is reduced. If the tumor is still present (tumor volume)>0mm³) But its volume and the time of treatment initiationThe comparison was reduced, and it was considered that "Partial Regression (PR)" had occurred. If the tumor is apparently absent after treatment, "Complete Regression (CR)" is considered to have occurred.

The term "small molecule" as used herein refers to a compound having a molecular weight of less than 1000g/mol and preferably less than 700 g/mol. The small molecules of the invention may be obtained by chemical reactions known to those skilled in the art of organic and/or pharmaceutical chemistry. Examples of small molecules may be, but are not limited to, compound I or a compound known as erlotinib, preferably erlotinib hydrochloride.

The term "macromolecule" as used herein refers to a compound having a molecular weight greater than 1000 g/mol. Preferably, "macromolecule" refers to a compound that is obtainable by biotechnological production methods. More preferably, the term "macromolecule" refers to a polypeptide, such as an antibody, more particularly a monoclonal antibody. An example of a macromolecule of the invention is cetuximab.

The present invention relates to a pharmaceutical product comprising: (A) a first component comprising compound I or a pharmaceutically acceptable salt thereof as an active agent; and (B) a second component comprising an EGFR inhibitor as an active agent; the amounts of the active agents are such that the combination thereof is therapeutically effective in treating the proliferative disorder. The invention also relates to the above mentioned product as a combined preparation for simultaneous or sequential use in the treatment of a proliferative disorder, in particular a cancer, more particularly a solid tumor, in particular colorectal cancer, melanoma and/or thyroid cancer, comprising b-Raf having the V600E mutation.

The invention also relates to a method of treating a patient suffering from a proliferative disorder, said method comprising administering to said patient a combination or pharmaceutical formulation as described above.

"treatment of a proliferative disorder" should be understood to include maintaining or reducing tumor size, inducing tumor regression (in part or in whole), inhibiting tumor growth, and/or extending the lifespan of a patient suffering from the disorder.

The present invention also relates to a kit or composition comprising: (A) a first component comprising compound I or a pharmaceutically acceptable salt thereof as an active agent; and (B) a second component comprising an EGFR inhibitor as an active agent. The kits or compositions are useful, for example, in the treatment of proliferative disorders.

In one embodiment of the invention, the proliferative disorder is a solid tumor, in particular colorectal cancer, melanoma and/or thyroid cancer.

In another embodiment of the invention, the proliferative disorder is a tumor comprising b-Raf having the V600 mutation, and preferably the V600E mutation.

In another embodiment of the invention, the proliferative disorder is selected from colorectal cancer, melanoma and thyroid cancer and the cancer relates to a tumor comprising b-Raf having the V600 mutation and preferably the V600E mutation.

In another embodiment of the invention, the proliferative disorder is a solid tumor comprising b-Raf having the V600 mutation, and preferably the V600E mutation.

In another embodiment of the invention, the proliferative disorder is colorectal cancer.

In another embodiment of the invention, the proliferative disorder is colorectal cancer involving a tumor comprising b-Raf having the V600 mutation and preferably the V600E mutation.

In another embodiment of the invention, the EGFR inhibitor is a small molecule EGFR inhibitor. In one such embodiment, the EGFR inhibitor is erlotinib or a pharmaceutically acceptable salt thereof, such as erlotinib hydrochloride (erlotinib HCl). The trade name of erlotinib hydrochloride is also knownAnd is available, for example, from Genentech, Sout in the United statesAnd sold by h San Francisco, U.S.A.

In another embodiment of the invention, the EGFR inhibitor is a macromolecular EGFR inhibitor, e.g., an antibody that targets EGFR. In one such embodiment, the EGFR inhibitor may be a monoclonal antibody targeting EGFR, such as cetuximab. The trade name of cetuximab is also known asAnd is sold, for example, by ImClone Systems, inc.

In some embodiments, the invention relates to a pharmaceutical product for the treatment of colorectal cancer involving a tumor comprising b-Raf having the V600E mutation, comprising: (A) a first component comprising compound I or a pharmaceutically acceptable salt thereof as an active agent; and (B) a second component comprising erlotinib, or a pharmaceutically acceptable salt thereof, as an active agent; the amounts of the active agents are such that the combination thereof is therapeutically effective in treating the cancer.

In another embodiment, the invention relates to a pharmaceutical product for the treatment of colorectal cancer involving a tumor comprising b-Raf having the V600E mutation, comprising: (A) a first component comprising compound I or a pharmaceutically acceptable salt thereof as an active agent; and (B) a second component comprising cetuximab as an active agent; the amounts of the active agents are such that the combination thereof is therapeutically effective in treating the cancer.

The amount of each component administered by the combination or composition of the invention may, but need not, be therapeutically effective per se. That is, the present invention specifically encompasses combinations wherein the amount of compound I or a pharmaceutically acceptable salt thereof and/or the amount of an EGFR inhibitor in said combination can be less than the therapeutically effective amount of each active agent when administered as monotherapy.

The first component (a) and the second component (B) of the present invention are administered in any amount and for any duration, wherein the combined amounts thereof are therapeutically effective in the treatment of a proliferative disorder.

In some embodiments of the invention, compound I or a pharmaceutically acceptable salt thereof is administered in an amount of from about 200 mg/day to about 3000 mg/day, from about 1000 mg/day to about 2500 mg/day, or from about 1700 mg/day to about 2100 mg/day. In another embodiment, the dose is about 1920 mg/day.

In another embodiment of the invention, the above amounts of compound I or a pharmaceutically acceptable salt thereof may be administered daily as a single dose or divided, e.g. into equal doses (although this is not essential) and administered twice daily (bid). For example, compound I or a pharmaceutically acceptable salt thereof may be administered at a dose of about 100mg to about 1500mg bid, about 500mg to about 1250mg bid, about 850mg to about 1050mg bid, or about 960mg bid.

In one embodiment of the invention, compound I or a pharmaceutically acceptable salt thereof is administered until disease progression or unacceptable toxicity.

In some embodiments of the invention, erlotinib, or a pharmaceutically acceptable salt thereof, is administered at a dose of about 20 mg/day to about 500 mg/day, about 100 mg/day to about 400 mg/day, or about 100 mg/day to about 200 mg/day.

In one embodiment of the invention, erlotinib, or a pharmaceutically acceptable salt thereof, is administered until disease progression or unacceptable toxicity.

In another embodiment of the invention, cetuximab is present at about 50mg/m²Weekly to about 700mg/m²Weekly, about 100mg/m²One week to about 600mg/m²Weekly or about 200mg/m²One week to about 500mg/m²Dose per week.

In another embodiment, cetuximab is administered weekly, wherein the first amount administered is about 400mg/m²To about 500mg/m²And then the dosage per administration is about 200mg/m²To about 300mg/m²。

In a further embodiment of the process of the present invention,cetuximab is administered weekly, wherein the first dose is about 450mg/m²And then the dosage per administration is about 250mg/m²。

In another embodiment of the invention, cetuximab is administered until disease progression or unacceptable toxicity.

Accordingly, in another embodiment, the present invention provides a pharmaceutical product comprising: (A) a first component comprising compound I or a pharmaceutically acceptable salt thereof as an active agent; and (B) a second component comprising erlotinib, or a pharmaceutically acceptable salt thereof, as an active agent; the pharmaceutical product is for simultaneous or sequential use as a combined preparation for the treatment of a proliferative disorder, wherein

(A) Is administered in an amount of from about 200 mg/day to about 3000 mg/day, from about 1000 mg/day to about 2500 mg/day, from about 1700 mg/day to about 2100 mg/day, or about 1920 mg/day, and

(B) is administered in an amount of from about 20 mg/day to about 500 mg/day, from about 100 mg/day to about 400 mg/day, or from about 100 mg/day to about 200 mg/day.

In this embodiment, compound I or a pharmaceutically acceptable salt thereof may be administered twice daily. The proliferative disorders to be treated in this way are solid tumors, in particular colorectal, melanoma and thyroid cancers, comprising b-Raf with the V600E mutation. More specifically, the proliferative disorder is colorectal cancer involving a tumor comprising b-Raf having the V600E mutation.

In addition, in this embodiment, the product of the invention may comprise: compound I or a pharmaceutically acceptable salt thereof, administered orally twice daily at a dose of about 850mg to about 1050mg or administered orally twice daily at a dose of about 960 mg; and erlotinib, or a pharmaceutically acceptable salt thereof, that is administered orally at a dose of about 100 mg/day to about 400 mg/day, or about 100 mg/day to about 200 mg/day. Both drugs may be administered, for example, until disease progression or unacceptable toxicity.

The present invention also provides a pharmaceutical product comprising: (A) a first component comprising compound I or a pharmaceutically acceptable salt thereof as an active agent; and (B) a second component comprising cetuximab as an active agent; the pharmaceutical product is for simultaneous or sequential use as a combined preparation for the treatment of a proliferative disorder, wherein

(A) (ii) is administered in an amount of from about 200 mg/day to about 3000 mg/day, from about 1000 mg/day to about 2500 mg/day, from about 1700 mg/day to about 2100 mg/day, or about 1920 mg/day; and is

(B) At about 50mg/m²Weekly to about 700mg/m²Weekly, about 100mg/m²One week to about 600mg/m²Weekly or about 200mg/m²One week to about 500mg/m²The amount per week.

In this embodiment, the proliferative disorder to be treated in this manner is a solid tumor, in particular colorectal cancer, melanoma and thyroid cancer, comprising b-Raf having the V600E mutation. More specifically, the proliferative disorder is colorectal cancer involving a tumor comprising b-Raf having the V600E mutation.

In addition, in this embodiment, the product of the invention may comprise: compound I or a pharmaceutically acceptable salt thereof, administered orally twice daily at a dose of about 850mg to about 1050mg or orally twice daily at a dose of about 960 mg; and cetuximab at about 200mg/m²One week to about 500mg/m²The dose per week was administered intravenously. In one embodiment, cetuximab is initially at 400mg/m²The dose of (A) is administered by intravenous infusion over 120 minutes and at 250mg/m after one week²The doses of (a) were administered by intravenous infusion over 60 minutes per week. Both drugs may be administered, for example, until disease progression or unacceptable toxicity.

The present invention also provides a kit or composition comprising: (A) a first component comprising compound I or a pharmaceutically acceptable salt thereof as an active agent; and (B) a second component comprising erlotinib, or a pharmaceutically acceptable salt or prodrug thereof, as an active agent.

The present invention also provides a kit or composition comprising: (A) a first component comprising compound I or a pharmaceutically acceptable salt thereof as an active agent; and (B) a second component comprising cetuximab as an active agent.

In another aspect of the invention, a pharmaceutical product comprising the above components (a) and (B) is used in combination with radiotherapy and/or in combination with other active agents.

Thus, in some embodiments, the present invention provides a pharmaceutical product comprising the above components (a) and (B) and a third component (C) comprising a topoisomerase inhibitor as an active agent; the pharmaceutical product is for simultaneous or sequential use as a combined preparation in the treatment of a proliferative disorder, such as a solid tumor comprising b-Raf having the V600 mutation, and preferably the V600E mutation. As mentioned above, the amount of each component administered by the combination product of the invention may, but need not, be therapeutically effective per se and the invention specifically includes combinations wherein the amount of each active agent in the combination may be less than the therapeutically effective amount of each active agent when administered as monotherapy.

In one embodiment of the invention, the topoisomerase inhibitor is a type I topoisomerase inhibitor. In one embodiment of the invention, the topoisomerase inhibitor is irinotecan or a pharmaceutically acceptable salt thereof, e.g., irinotecan hydrochloride (irinotecan hydrochloride). Irinotecan hydrochloride manufactured by Pfizer inc, new york, u.s.aAnd (5) selling. Irinotecan or a pharmaceutically acceptable salt thereof can be administered, for example, intraperitoneally or intravenously.

In one embodiment of the invention, irinotecan, or a pharmaceutically acceptable salt thereof, is present at about 1 to about 400mg/m²Weekly or about 1 to about 250mg/m²The amount per week. In another embodiment, irinotecan, or a pharmaceutically acceptable salt thereof, is present at about 50 to about 200mg/m²Dose per week. In another embodiment, irinotecan, or a pharmaceutically acceptable salt thereof, is at about 125mg/m²Dose per week.

In another embodiment, irinotecan, or a pharmaceutically acceptable salt thereof, is administered for a period of six weeks and from about 75 to about 175mg/m per week²E.g. about 125mg/m per week for the first four weeks²E.g., on days 1, 8, 15, and 22. In another embodiment, irinotecan is to be administered for a period of six weeks and from about 130 to about 230mg/m per week²E.g. about 180mg/m per week²Every two weeks, starting on the first week, for example on days 1, 15 and 29. In another embodiment, irinotecan is at a concentration of about 300 to about 400mg/m every three weeks²For example about 350mg/m²The administration is once. In another embodiment, irinotecan is present at about 130 to about 230mg/m every two weeks²For example about 180mg/m²The administration is once. Administration can be by infusion, for example, over a period of about 90 minutes. Treatment may be carried out until disease progression or unacceptable toxicity.

The physician can adjust the dosage level of each of components (a), (B), or (C) below or above the dosage level described herein, depending on the patient's needs and the patient's response to treatment. The dosage may be administered according to any dosing regimen determined by a physician in accordance with the needs of the patient. For example, the dosage of each component may be administered as a single dose or as divided doses over several days or on an alternating daily schedule.

The present invention also provides a pharmaceutical product comprising: (A) a first component comprising compound I or a pharmaceutically acceptable salt thereof as an active agent; (B) a second component comprising cetuximab as an active agent; and (C) a third component comprising irinotecan, or a pharmaceutically acceptable salt thereof, as an active agent; the pharmaceutical product is for simultaneous or sequential use as a combined preparation for the treatment of a proliferative disorder, wherein

(A) (ii) is administered in an amount of from about 200 mg/day to about 3000 mg/day, from about 1000 mg/day to about 2500 mg/day, from about 1700 mg/day to about 2100 mg/day, or about 1920 mg/day;

(B) at about 50mg/m²Weekly to about 700mg/m²Weekly, about 100mg/m²One week to about 600mg/m²Weekly or about 200mg/m²One week to about 500mg/m²(iv) amount per week; and is

(C) At a rate of about 1 to about 250mg/m²Weekly, about 50 to about 200mg/m²Weekly or about 125mg/m²The amount per week.

In this embodiment, the proliferative disorder to be treated in this manner is a solid tumor, in particular colorectal cancer, melanoma and thyroid cancer, comprising b-Raf having the V600E mutation. More specifically, the proliferative disorder is colorectal cancer involving a tumor comprising b-Raf having the V600E mutation.

In addition, in this embodiment, the product of the invention may comprise: compound I or a pharmaceutically acceptable salt thereof, administered orally twice daily at a dose of about 850mg to about 1050mg or orally twice daily at a dose of about 960 mg; cetuximab at about 200mg/m²One week to about 500mg/m²Intravenous administration of the dose per week; and irinotecan at a rate of about 50 to about 200mg/m²Weekly or about 125mg/m²The dose per week was administered intravenously. All drugs can be administered, for example, until disease progression or unacceptable toxicity.

The present invention also provides a kit or composition comprising: (A) a first component comprising compound I or a pharmaceutically acceptable salt thereof as an active agent; (B) a second component comprising cetuximab as an active agent; and (C) a third component comprising irinotecan, or a pharmaceutically acceptable salt thereof, as an active agent.

Compound I exists in its natural state in crystalline form. However, the amorphous form of the compound has greater water solubility than the crystalline form and thus has an improved dissolution rate and thus an improved bioavailability compared to the crystalline form. Thus, amorphous forms of the compounds are preferred. Thus, in preferred embodiments of the methods and kits of the present invention, compound I is in a substantially amorphous form and more preferably in an amorphous form. As used herein, the term "substantially amorphous" material includes materials having a degree of crystallinity of no greater than about 10%; and "amorphous" materials include materials having a degree of crystallinity of no greater than about 2%.

In one embodiment of the invention, compound I is contained in a solid molecular complex with hydroxypropyl methyl cellulose acetate succinate (HPMC-AS). The term "solid molecular complex" AS used herein refers to a composition in which compound I is randomly distributed ("molecularly dispersed") in a matrix formed by HPMC-AS. Preferably, the above-described composition of compound I and HPMC-AS forms a one-phase system, which can be characterized by an X-ray powder diffraction pattern which is substantially free or free of crystallization signals associated with the crystalline form of compound I. In some embodiments, compound I is present in the polymer in a finely divided (subdivisions) final state. In some embodiments, compound I is molecularly dispersed in the HPMC-AS matrix, thereby immobilizing it in its amorphous form. By "immobilized" is meant that the molecules of compound I interact with the molecules of HPMC-AS to keep them in the above-mentioned matrix and prevent nucleation due to lack of mobility. In some embodiments, the polymer may prevent intramolecular hydrogen bonding or weak dispersion forces between two or more molecules of compound I.

In some embodiments, the ratio of the weight of compound I in the solid molecular complex to the weight of HPMC-AS therein is from about 1:9 to about 5: 5. In one embodiment, the ratio is from about 2:8 to about 4: 6. In another embodiment, the ratio is about 3: 7.

In some embodiments of the methods and kits of the present invention, the first component comprises the above-described solid molecular complex of compound I and HPMC-AS, blended with colloidal silica. In some embodiments, the blend comprises at least 0.5 wt.% silica. In one embodiment of the invention, the blend is about 97% composite and about 3% silica.

In another embodiment, the first component comprises a composition comprising a solid molecular complex as described above, with or without silica as described above, and a pharmaceutically acceptable carrier. In some embodiments, the above-described complex or a blend comprising the above-described complex is suspended in a vehicle. An example of a carrier is hydroxypropyl cellulose (HPC). In one embodiment, the vehicle contains about 2 wt.% HPC.

Each component may also contain additional agents such as preservatives, solubilizers, stabilizers, wetting agents, emulsifiers, sweeteners, colorants, flavorants, salts for varying the osmotic pressure, buffers, coating agents and antioxidants.

In some embodiments, the first component may comprise a solid molecular complex of compound I and HPMC-AS blended with colloidal silicon dioxide, hydroxypropyl cellulose, crospovidone (disintegrant), magnesium stearate (lubricant that can be used in tableting and encapsulation operations), and/or croscarmellose sodium (disintegrant).

In one embodiment, the first component is a hard gelatin capsule comprising a solid molecular complex of compound I and HPMC-AS blended with colloidal silicon dioxide, hydroxypropylcellulose, magnesium stearate, and croscarmellose sodium.

In one embodiment, the first component is a tablet comprising compound I or a pharmaceutically acceptable salt thereof. In one embodiment, the tablet comprises a solid molecular complex of compound I or a pharmaceutically acceptable salt thereof and HPMC-AS. For example, the compound may be blended with colloidal silicon dioxide, hydroxypropyl cellulose, magnesium stearate, and croscarmellose sodium. The tablets may be coated, for example, with a film coating. The film coating may, for example, comprise polyvinyl alcohol, titanium dioxide, polyethylene glycol 3350, talc and red iron oxide.

In some embodiments, the second component may comprise cetuximab in solution. In one embodiment, the solution is about 2mg/ml cetuximab.

In some embodiments, the second component may comprise a tablet comprising erlotinib or a pharmaceutically acceptable salt thereof, such as erlotinib hydrochloride.

In some embodiments, the third component may comprise a solution comprising irinotecan, or a pharmaceutically acceptable salt thereof, e.g., irinotecan hydrochloride. In one embodiment, the solution is an about 5% dextrose solution. In one embodiment, the solution contains about 20mg irinotecan hydrochloride, about 45mg sorbitol, and about 0.9mg lactic acid per ml. In one embodiment, the pH of the solution is from about 3.0 to about 3.8, for example about 3.5.

In addition, the present invention provides the use of compound I or a pharmaceutically acceptable salt thereof and an EGFR inhibitor and optionally a topoisomerase inhibitor for the treatment of a proliferative disorder, in particular a solid tumor, more particularly colorectal cancer, melanoma and/or thyroid cancer, all comprising b-Raf having the V600E mutation.

The present invention also provides the use of compound I or a pharmaceutically acceptable salt thereof and an EGFR inhibitor and optionally a topoisomerase inhibitor in the manufacture of a medicament for the treatment of a proliferative disorder, in particular a solid tumor, more particularly colorectal cancer, melanoma and/or thyroid cancer, all comprising b-Raf having the V600E mutation.

The present invention also provides a method of treating a patient suffering from a proliferative disorder, particularly a solid tumor, more particularly colorectal cancer, melanoma and/or thyroid cancer, all comprising B-Raf having the V600E mutation, comprising administering to said patient the combination of (a) and (B) or any of the combinations of (a), (B) and (C) at the dosages and dosing regimens described herein above.

The applicant carried out studies using mice containing human colorectal cancer xenografts.

Applicants found that the combination of compound I (75mg/kg bid) and erlotinib hydrochloride (67mg/kg qd) yielded Tumor Growth Inhibition (TGI) and life extension (ILS) results that were significantly better than the corresponding monotherapy results (p <0.05) and the results obtained with erlotinib hydrochloride monotherapy (100mg/kg qd). In addition, 9 of 10 mice undergoing combination therapy had only partial regression, while no regression (partial or complete) was observed in any of the monotherapy groups.

These studies indicate that treatment of patients with the combination of compound I and erlotinib hydrochloride is superior to treatment with either drug alone. In addition, these studies show that the combination of the two drugs allows at least a reduction of the erlotinib hydrochloride dose with equal or better results.

Applicants have also found that the combination of compound I (25mg/kg bid) and cetuximab (40mg/kg2 ×/wk) gives Tumor Growth Inhibition (TGI) and life extension (ILS) results which are significantly better than the corresponding monotherapy results (p <0.05) and the results obtained with compound I monotherapy (75mg/kg bid). Applicants have also found that the combination of compound I (75mg/kg bid) and cetuximab (40mg/kg2 ×/wk) gives TGI and ILS results which are significantly better than the corresponding monotherapy results (p <0.05) and the results obtained with compound I monotherapy (25mg/kg bid). In addition, 7 of 9 mice undergoing combination therapy of compound I (25mg/kg bid) and cetuximab (40mg/kg2 x/wk) had partial regression and 10 of 10 mice undergoing combination therapy of compound I (75mg/kg bid) and cetuximab (40mg/kg2 x/wk) showed regression, of which 7 were partial regressions and 3 were complete regressions. In contrast, no regression (partial or complete) was observed in any of the monotherapy groups.

In addition, applicants found that the combination of compound I (25mg/kg bid), cetuximab (40mg/kg2 x/wk) and irinotecan hydrochloride (40mg/kg q4d x 5) yielded Tumor Growth Inhibition (TGI) and life extension (ILS) results that were significantly better than the corresponding monotherapy results (p <0.05) and also better than the results achieved by the combination therapy of compound I (25mg/kg bid) and irinotecan hydrochloride (40mg/kg q4d x 5) and the combination therapy of compound I (25mg/kg bid) and cetuximab (40mg/kg2 x/wk). In the study, combination therapy of compound I (25mg/kg bid) and irinotecan hydrochloride (40mg/kg q4d x 5) caused partial but not complete regression in 4 of 10 mice and combination therapy of compound I (25mg/kg bid) and cetuximab (40mg/kg2 x/wk) caused partial but not complete regression in 5 of 10 mice. Combination therapy of cetuximab (40mg/kg2 x/wk) and irinotecan (40mg/kg q4d x 5) and the corresponding compound I monotherapy, cetuximab monotherapy and irinotecan hydrochloride monotherapy did not produce regression. In contrast, treatment with compound I (25mg/kg bid), cetuximab (40mg/kg2 x/wk), and irinotecan hydrochloride (40mg/kg q4d x 5) produced regression in all 10 mice, with 9 partial and 1 complete regressions.

These studies indicate that treatment of patients with a combination of compound I and cetuximab is superior to treatment with either drug alone. In addition, these studies indicate that the combination of the two drugs allows at least a reduction in the dose of compound I with equal or better results. In addition, the study showed that treatment of patients with a combination of compound I, cetuximab and irinotecan hydrochloride yielded even superior results.

Examples

The present invention will be more fully understood by reference to the following examples. However, these examples should not be construed as limiting the scope of the invention.

Abbreviations used herein are as follows:

q.s.: proper amount of

X: number of times

po: is administered orally

ip: intraperitoneal cavity

bid: twice daily

wk: week (week)

qd: once a day

q4d × 5: the preparation is administered once every 4 days for 5 times

BWL: weight loss

In the following examples, weight loss is represented graphically as the percent change in average group weight, using the formula: ((W-W)₀)/W₀) X 100, wherein ' W ' denotes the mean body weight of the treatment group on a particular day and ' W₀' means the average body weight of the same treatment group at the start of treatment. Maximum weight loss is also expressed using the above formula and refers to the maximum percentage of weight loss observed for a particular group at any time throughout the course of the experiment.

Efficacy data are presented graphically as mean tumor volume ± mean Standard Error (SEM). In addition, the tumor volume of the treated group was expressed as a percentage (% T/C) relative to the tumor volume of the control group, using the formula: 100 × ((T-T)₀)/(C-C₀) Where T represents the mean tumor volume of the treatment group on a particular day of the course of the experiment, T₀Represents the mean tumor volume of the same treatment group at the first day of treatment, C represents the mean tumor volume of the control group on that particular day of the experimental procedure and C₀Mean tumor volume at the first day of treatment for the same control group.

Tumor volume (in cubic millimeters) was calculated using the following ellipsoid formula: (D × (D)²)2, wherein "D" represents the long diameter of the tumor and "D" represents the short diameter of the tumor。

In addition, the percent tumor regression and/or tumor volume change was calculated using the following formula: ((T-T)₀)/T₀) X 100, wherein ` T ` denotes mean tumor volume on a particular day in the treatment group and ` T `₀' means the mean tumor volume in the same treatment group at the start of treatment.

Statistical analysis was determined by rank sum test, one-way analysis of variance, and causal Bonferroni t-test (SigmaStat, version 2.0, Jandel Scientific, San Francisco, CA, USA). Differences between groups were considered significant if the probability value (p) was ≦ 0.05.

For survival evaluation, percent increase in survival space (ILS) was calculated as 100 × [ (median survival day for treated group-median survival day for control group)/median survival day for control group ]. Median survival was determined using Kaplan Meier survival analysis. Survival in the treatment groups was statistically compared to survival in the vehicle group and comparison of survival between groups was performed using log-rank test (Graph PadPrism, La Jolla, CA, USA). Differences between groups were considered significant if the probability value (p) was ≦ 0.05.

Example 1

This example describes the formation of a suspension comprising compound I.

A solid molecular complex comprising compound I and hydroxypropylmethylcellulose acetate succinate (HPMC-AS) was first formed.

Compound I and HPMC-AS in a ratio of about 3:7 were dissolved in Dimethylacetamide (DMA) respectively. The resulting solution was then added to very cold dilute hydrochloric acid with stirring, which allowed co-precipitation of compound I and HPMC-AS a solid molecular complex, wherein compound I was present in the nanoparticle size range. The ratio of DMA to acid is from 1:5 to 1: 10.

The co-precipitate was then washed with water to remove DMA, filtered, dried to a moisture content <2% and passed through a # 30 sieve before evaluation. The resulting solid molecular complex was 30 wt% compound I and 70 wt% HPMC.

Then the compound is mixed with colloidal silica (C)200, Evonik Industries AG, Essen, Germany), so that 97g of the compound and 3g of the colloidal silica are present per 100g of the blend.

Then, a composition containing 2% of hydroxypropylcellulose (LF, Aqualon, Wilmington, Delaware, USA) and 1N HCl (q.s. to pH4) for pH adjustment in an aqueous vehicle.

23.2ml of vehicle were equilibrated to room temperature and slowly transferred to 773.2mg of the above blend. The resulting preparation was then slowly mixed until a homogeneous suspension was obtained. The resulting suspension contained 9.375mg/ml of Compound I. The suspension was stored at 2-8 ℃ and protected from light.

Example 2

Mice were implanted with human HT-29 cell xenografts. Mice, cell lines used and implants are described below.

Female athymic Crl: NU-Foxn1NU mice were used for efficacy testing (Charles River, Wilmington, MA, USA). The mice were 10-12 weeks old and 23-25 grams repeated. The health of the mice was evaluated daily as follows: blood samples taken from sentinel animals (sentinel animals on shared shells) on a common grid were observed and analyzed. All animals were acclimatized and recovered from transport-related stress over a week. Autoclaved water and irradiated food (5058-ms Pico Lab mouse chow, Purina Mills, Richmond, IN, USA) were provided ad libitum and animals were kept IN 12-hour light-dark cycles. The cages, bedding and water bottles were autoclaved and replaced weekly prior to use. All animal experiments were conducted in the applicant's AAALAC-approved facility in accordance with the laboratory guidelines for animal care and use, local regulations, and protocols approved by the roche committee for animal care and use.

HT-29 cells (American Type Culture Collection, Rockville, Md.) were grown, expanded, harvested and prepared in McCoy-5 medium supplemented with 10% Fetal Bovine Serum (FBS) and 1%200nM L-glutamine to receive 3X 10 cells per mouse⁶Cells/0.2 ml Phosphate Buffered Saline (PBS) without calcium and magnesium. Cells were implanted subcutaneously in the right flank of each mouse.

Example 3

This example describes the preparation of a suspension of erlotinib hydrochloride.

Add 1 gram tween 80 to about 950ml of water. To the solution was added 3 grams of sodium carboxymethylcellulose under high speed stirring. Mixing was continued until the sodium carboxymethyl cellulose dissolved. Then add the appropriate amount of water until 1 liter. Then 12.5g of erlotinib hydrochloride (A)Genentech) was suspended in solution and passed through a dissolver. The solution was then degassed with nitrogen.

The content of the final suspension is as follows:

the components are as follows: measurement of

Erlotinib hydrochloride: 12.5g

Sodium carboxymethylcellulose: 3g

Tween 80: 1g

Water for injection: proper amount to 1 liter

This gave a solution containing 12.5mg/ml erlotinib hydrochloride. The solution was stored at 2-8 ℃.

Example 4

Suspensions containing compound I were prepared as described in example 1.

A solution containing 12.5mg/ml erlotinib hydrochloride was prepared as described in example 3. A solution containing 8.30mg/ml erlotinib hydrochloride was prepared in a similar manner to example 3, except that 8.30g erlotinib hydrochloride was used instead of 12.5g erlotinib hydrochloride.

Mice containing HT-29 xenografts prepared in the manner described in example 2 were randomly assigned to groups (10 mice per group) according to tumor volume, so that all groups had similar starting mean tumor volumes. The initial mean tumor volume used in this study was about 136mm³。

Treatment of mice began on day 12 post cell implantation and ended on day 29 post cell implantation. Each group underwent the following different treatments:

(1) mice received compound I vehicle bid po and erlotinib hydrochloride vehicle qd po;

(2) mice received compound I75mg/kg bid po;

(3) mice received erlotinib hydrochloride 67mg/kg qd po;

(4) mice received erlotinib hydrochloride 100mg/kg qd po;

(5) mice received compound I75mg/kg bid po and erlotinib hydrochloride 67mg/kg qd po.

The suspension of compound I and the vehicle corresponding thereto were administered twice daily (0.2 ml/animal) using a 1cc sterile syringe and a 18 gauge gavage needle. The solution of erlotinib hydrochloride and the vehicle corresponding thereto were administered once daily (0.2 ml/animal) using a 1cc sterile syringe and 18 gauge needle, starting on day 12 post-implantation and ending on day 29 post-implantation. The 12.5mg/ml solution was used for the erlotinib hydrochloride 100mg/kg group and the 8.30mg/ml solution was used for the erlotinib hydrochloride 67mg/kg qd group. All administrations were based on the average mouse body weight, i.e. 25 g.

Tumors were measured once or twice weekly. All animals were individually tracked throughout the experiment.

Toxicity

Generally, no clear signs of toxicity were observed in any of the dose groups of the study when evaluated by measuring changes in body weight and visual observation of individual animals. Erlotinib hydrochloride 100mg/kg qd in combination was historically not well tolerated (Higgins et al, Anticancer Drugs,15:503-12(2004)), so the use of 67mg/kg qd in combination arms was intended to ensure tolerability. Erlotinib hydrochloride 100mg/kg qd was included in the monotherapy arms for comparison. However, compound I was very well tolerated and was administered at 75mg/kg bid, even when combined with erlotinib hydrochloride. Skin rash associated with EGFR inhibitors is common in mice treated with erlotinib hydrochloride, which is self-limiting even under continuous treatment. See table 1 and fig. 1.

TABLE 1

Tumor Growth Inhibition (TGI)

The group receiving compound I monotherapy (75mg/kg bid) showed 91% TGI, whereas the group receiving erlotinib hydrochloride (100mg/kg qd) showed 51% TGI and the group receiving erlotinib hydrochloride (67mg/kgqd) showed 38% TGI. No tumor regression was observed in all of the above groups. The group receiving the combination therapy of compound I (75mg/kg bid) and erlotinib hydrochloride (67mg/kg qd) showed a TGI of more than 100%, with 9 of 10 mice having Partial Regression (PR). See tables 2 and 3 and fig. 2.

TABLE 2

TABLE 3

Survival evaluation

The group receiving compound I monotherapy (75mg/kg) showed 100% life extension (ILS). The group receiving erlotinib hydrochloride monotherapy (100mg/kg qd) showed 38% ILS. The group receiving erlotinib hydrochloride monotherapy (67mg/kg qd) showed 35% ILS. The group receiving the combination therapy of compound I (75mg/kg bid) and erlotinib hydrochloride (67mg/kg qd) showed 142% ILS. See table 4 and fig. 3.

TABLE 4

Statistical analysis

The% TGI of the combination therapy of Compound I (75mg/kg bid) and erlotinib hydrochloride (67mg/kg qd) was statistically superior to the% TGI of all monotherapies (p < 0.05). The% ILS of the combination therapy of Compound I (75mg/kg bid) and erlotinib hydrochloride (67mg/kg qd) was also statistically superior to the% ILS of all monotherapies tested (p <0.05 for all comparisons). See table 5.

TABLE 5

One-way analysis of variance and causal Bonferroni method

**Breslow-Gehan-Wilcoxon

Example 5

Two suspensions containing compound I were prepared in a similar manner to example 1, except that 20ml of a 9.375mg/ml concentration suspension was prepared using 19.4ml of the vehicle and 644mg of the blend and 20ml of a 3.125mg/ml concentration suspension was prepared using 19.8ml of the vehicle and 214.8mg of the blend.

Cetuximab is available from ImClone Systems, Inc.It is a solution with a concentration of 2 mg/ml.

Mice containing HT-29 xenografts prepared in the manner described in example 2 were randomly assigned to groups (10 mice per group) according to tumor volume, so that all groups had similar starting mean tumor volumes. The initial mean tumor volume used in this study was about 135mm³。

Treatment began on day 12 post-cell implantation and ended on day 34 post-cell implantation. Each group underwent the following different treatments:

(1) mice received compound I vehicle bid po and cetuximab vehicle 2 ×/wk ip;

(2) mice received cetuximab 40mg/kg2 x/wk ip;

(3) mice received compound I25mg/kg bid po;

(4) mice received compound I75mg/kg bid po;

(5) mice received compound I25mg/kg bid po and cetuximab 40mg/kg2 ×/wk ip;

(6) mice received compound I75mg/kg bid po and cetuximab 40mg/kg2 ×/wk ip.

The suspension of compound I and the vehicle corresponding thereto were administered twice daily (0.2 ml/animal) using a 1cc sterile syringe and a 18 gauge gavage needle. The 9.375mg/ml suspension was used in Compound I75mg/kgbid group and the 3.125mg/ml suspension was used in Compound I25mg/kg bid group. Cetuximab and the vehicle corresponding thereto were administered intraperitoneally twice a week (0.5 ml/animal) using a 1cc sterile syringe and a 26-gauge needle on a monday/thursday or tuesday/friday schedule. All administrations were based on the average mouse body weight, i.e. 25 g.

Tumors were measured once or twice weekly. All animals were individually tracked throughout the experiment.

Toxicity

Generally, no clear signs of toxicity were observed in any of the dose groups of the study when evaluated by measuring changes in body weight and visual observation of individual animals. See table 6 and fig. 4. Rashes associated with EGFR inhibitors are common in mice treated with cetuximab, which are self-limiting even under continuous treatment. One mouse developed a bacterial infection in which a progressive weight loss of >20% was due to the sequelae of skin rash, thus requiring humane sacrifice. The mice were deleted from the overall tumor growth inhibition and survival assay.

TABLE 6

Tumor Growth Inhibition (TGI)

The group receiving compound I monotherapy (25mg/kg bid) showed 74% TGI and the group receiving compound I monotherapy (75mg/kg bid) showed 93% TGI. The group receiving cetuximab (40mg/kg2 ×/wk) achieved 51% TGI. No tumor regression was observed in all of the above groups. However, both combination therapy groups showed >100% TGI. The group receiving compound I (25mg/kg bid) and cetuximab (40mg/kg2 ×/wk) showed Partial Regression (PR) but not Complete Regression (CR) in 7 out of 10 mice. The group receiving compound I (75mg/kg bid) and cetuximab (40mg/kg2 ×/wk) showed PR in 7 out of 10 mice and CR in 3 out of 10 mice. See tables 7 and 8 and fig. 5.

TABLE 7

TABLE 8

Survival evaluation

The group receiving compound I monotherapy (25mg/kg bid) showed 44% ILS and the group receiving compound I monotherapy (75mg/kg bid) showed 75% ILS. The group receiving cetuximab (40mg/kg2 ×/wk) achieved 16% ILS. The group receiving compound I (25mg/kg bid) and cetuximab (40mg/kg2 ×/wk) showed 97% ILS. The group receiving compound I (75mg/kg bid) and cetuximab (40mg/kg2 ×/wk) showed 122% ILS. See table 9 and fig. 6.

TABLE 9

Statistical analysis

The% TGI of the combination therapy of Compound I (25mg/kg bid) and cetuximab (40mg/kg2 ×/wk) and the% TGI of the combination therapy of Compound I (75mg/kg bid) and cetuximab (40mg/kg2 ×/wk) were statistically superior to the% TGI of all monotherapies (p < 0.05). The% ILS achieved by both combination therapies was also statistically superior to the% ILS of all monotherapies tested (p <0.05 for all comparisons). See table 10.

Watch 10

One-way analysis of variance and causal Bonferroni method

**Breslow-Gehan-Wilcoxon

Example 6

A suspension containing compound I was prepared in a similar manner to example 1 except that 40ml of a 3.125mg/ml suspension was prepared using 39.6ml of the vehicle and 429.6mg of the blend.

Cetuximab is available from ImClone Systems, Inc.It is a solution with a concentration of 2 mg/ml. Irinotecan hydrochloride was purchased from Pfizer Inc.It is a sterile stock solution at a concentration of 20mg/ml and diluted to 2mg/ml with sterile saline as required.

Mice containing HT-29 xenografts prepared in the manner described in example 2 were randomly assigned to groups (10 mice per group) according to tumor volume, so that all groups had similar starting mean tumor volumes. Mean initial edema for this studyTumor volume is about 135mm³。

Treatment started on day 11 post cell implantation and ended on day 32 post cell implantation. Each group underwent the following different treatments:

(1) mice received compound I vehicle bid po, cetuximab vehicle 2 x/wk ip, and irinotecan hydrochloride vehicle q4d x 5 ip;

(2) mice received irinotecan hydrochloride 40mg/kg q4d × 5 ip;

(3) mice received compound I25mg/kg bid po;

(4) mice received cetuximab 40mg/kg2 x/wk ip;

(5) mice received compound I25mg/kg bid po and irinotecan hydrochloride 40mg/kg q4d × 5 ip;

(6) the mice received cetuximab 40mg/kg2 ×/wk ip and irinotecan hydrochloride 40mg/kgq4d × 5 ip;

(7) mice received compound I25mg/kg bid po and cetuximab 40mg/kg2 ×/wk ip;

(8) the mice received compound I25mg/kg bid po, cetuximab 40mg/kg2 ×/wk ip, and irinotecan hydrochloride 40mg/kg q4d × 5 ip.

The suspension of compound I and the vehicle corresponding thereto were administered twice daily (0.2 ml/animal) using a 1cc sterile syringe and a 18 gauge gavage needle. Cetuximab and the vehicle corresponding thereto were administered intraperitoneally twice a week (0.2 ml/animal) using a 1cc sterile syringe and a 26-gauge needle on a monday/thursday or tuesday/friday schedule. Irinotecan hydrochloride and its corresponding vehicle were administered intraperitoneally (0/2 ml/animal) using a 1cc sterile syringe and 26 gauge needle at a q4d x 5 schedule. All administrations were based on the average mouse body weight, i.e. 25 g.

Tumors were measured once or twice weekly. All animals were individually tracked throughout the experiment.

Toxicity

Generally, no clear signs of toxicity were observed in any of the dose groups of the study when evaluated by measuring changes in body weight and visual observation of individual animals. See table 11 and fig. 7. Rashes associated with EGFR inhibitors are common in mice treated with cetuximab, which are self-limiting even under continuous treatment.

TABLE 11

Tumor Growth Inhibition (TGI)

The group receiving compound I monotherapy (25mg/kg bid) showed 76% TGI. The group receiving cetuximab monotherapy (40mg/kg2 ×/wk) showed 58% TGI. The group receiving irinotecan hydrochloride monotherapy (40mg/kg q4d × 5) showed 59% TGI. The group receiving compound I (25mg/kg bid) and irinotecan hydrochloride (40mg/kg q4d X5) showed 98% TGI. The group receiving cetuximab (40mg/kg2 x/wk) and irinotecan hydrochloride (40mg/kg q4d x 5) showed 92% TGI. The group receiving compound I (25mg/kg bid) and cetuximab (40mg/kg2 ×/wk) showed >100% TGI. The group receiving compound I (25mg/kg bid), cetuximab (40mg/kg2 ×/wk) and irinotecan hydrochloride (40mg/kgq4d × 5) showed >100% TGI. No tumor regression was observed in all monotherapy groups. The group receiving compound I (25mg/kg bid) and cetuximab (40mg/kg2 ×/wk) showed Partial Regression (PR) but not Complete Regression (CR) in 5 out of 10. The group receiving compound I (25mg/kg bid), cetuximab (40mg/kg2 ×/wk), and irinotecan hydrochloride (40mg/kg q4d × 5) showed PR in 9 out of 10 and CR in 1 out of 10. See tables 12 and 13 and fig. 8.

TABLE 12

Watch 13

Survival evaluation

The group receiving compound I monotherapy (25mg/kg bid) showed 80% ILS. The group receiving cetuximab monotherapy (40mg/kg2 ×/wk) showed 27% ILS. The group receiving irinotecan hydrochloride monotherapy (40mg/kg q4d × 5) showed 17% ILS. The group receiving compound I (25mg/kg bid) and irinotecan hydrochloride (40mg/kg q4d X5) showed 163% ILS. The group receiving cetuximab (40mg/kg2 x/wk) and irinotecan hydrochloride (40mg/kg q4d x 5) showed 80% ILS. The group receiving compound I (25mg/kg bid) and cetuximab (40mg/kg2 ×/wk) showed 127% ILS. The group receiving compound I (25mg/kg bid), cetuximab (40mg/kg2 ×/wk) and irinotecan hydrochloride (40mg/kgq4d × 5) showed 259% ILS. See table 14 and fig. 9.

TABLE 14

Statistical analysis

The% TGI of the compound I/cetuximab, compound I/irinotecan hydrochloride and compound I/cetuximab/irinotecan hydrochloride combination therapy was statistically superior to the% TGI of all monotherapies (p < 0.05). The% TGI of the compound I/cetuximab/irinotecan hydrochloride combination therapy was also statistically superior to the% TGI of the compound I/irinotecan hydrochloride and cetuximab/irinotecan hydrochloride combination therapy (p < 0.05).

The% ILS for compound I/cetuximab, compound I/irinotecan hydrochloride, and compound I/cetuximab/irinotecan hydrochloride combination therapy was statistically superior to the% ILS for all monotherapies (p <0.05 for all comparisons). The% ILS of the compound I/cetuximab/irinotecan hydrochloride combination therapy was also statistically superior to the% ILS of the compound I/irinotecan hydrochloride and compound I/cetuximab combination therapy.

See table 15.

Watch 15

One-way analysis of variance and causal Bonferroni method

**Breslow-Gehan-Wilcoxon

Claims (21)

1. A pharmaceutical product, comprising: (a) a first component comprising as an active agent propane-1-sulfonic acid {3- [5- (4-chlorophenyl) -1H-pyrrolo [2,3-b ] pyridine-3-carbonyl ] -2, 4-difluoro-phenyl } -amide (compound I) or a pharmaceutically acceptable salt thereof; and (b) a second component comprising an EGFR inhibitor as an active agent; the pharmaceutical product is for simultaneous or sequential use as a combined preparation for the treatment of a proliferative disorder, in particular cancer, more particularly colorectal cancer, melanoma and thyroid cancer, comprising b-Raf having the V600 mutation.

2. A pharmaceutical product according to claim 1 for use in the treatment of colorectal cancer, melanoma and thyroid cancer comprising b-Raf having the V600E mutation.

3. A pharmaceutical product according to claim 1 or 2 wherein the EGFR inhibitor is erlotinib, or a pharmaceutically acceptable salt thereof.

4. The pharmaceutical product of claim 1 or 2, wherein the EGFR inhibitor is cetuximab.

5. The pharmaceutical product according to any one of claims 1 to 4, further comprising a third component (C) comprising as an active agent a topoisomerase inhibitor, in particular a type I topoisomerase inhibitor.

6. A pharmaceutical product according to claim 5 wherein the topoisomerase inhibitor is irinotecan, or a pharmaceutically acceptable salt thereof.

7. The pharmaceutical product according to any one of claims 1 to 6, wherein propane-1-sulfonic acid {3- [5- (4-chlorophenyl) -1H-pyrrolo [2,3-b ] pyridine-3-carbonyl ] -2, 4-difluoro-phenyl } -amide, or a pharmaceutically acceptable salt thereof, is in an amorphous or substantially amorphous form.

8. A kit, comprising: (a) a first component comprising as an active agent propane-1-sulfonic acid {3- [5- (4-chlorophenyl) -1H-pyrrolo [2,3-b ] pyridine-3-carbonyl ] -2, 4-difluoro-phenyl } -amide or a pharmaceutically acceptable salt thereof; and (b) a second component comprising an EGFR inhibitor as an active agent.

9. The kit of claim 8, further comprising a third component comprising as an active agent a topoisomerase inhibitor, more particularly irinotecan.

10. The kit of claim 8 or 9 for use in the treatment of a proliferative disorder, in particular cancer, more particularly colorectal cancer, melanoma and thyroid cancer, comprising b-Raf having the V600E mutation.

11. The pharmaceutical product according to claim 3, wherein the propane-1-sulfonic acid {3- [5- (4-chlorophenyl) -1H-pyrrolo [2,3-b ] pyridine-3-carbonyl ] -2, 4-difluoro-phenyl } -amide, or a pharmaceutically acceptable salt thereof, is administered in an amount of from about 850mg to about 1050mg twice a day; and the erlotinib or a pharmaceutically acceptable salt thereof is administered in an amount from about 100mg to about 200mg per day.

12. The pharmaceutical product according to claim 4, wherein the propane-1-sulfonic acid {3- [5- (4-chlorophenyl) -1H-pyrrolo [2,3-b ™)]Pyridine-3-carbonyl]-2, 4-difluoro-phenyl } -amide or a pharmaceutically acceptable salt thereof is administered in an amount of about 850mg to about 1050mg twice daily; and said cetuximab is administered weekly, wherein the first dose is about 400mg/m²To about 500mg/m²And then the dosage per administration is about 200mg/m²To about 300mg/m²。

13. The pharmaceutical product according to claim 11 or 12, further comprising irinotecan, or a pharmaceutically acceptable salt thereof.

14. The pharmaceutical product according to claim 13, wherein the irinotecan, or pharmaceutically acceptable salt thereof, is at about 50mg/m²To about 200mg/m²The amounts are administered weekly.

15. The pharmaceutical product according to claim 13, wherein irinotecan, or a pharmaceutically acceptable salt thereof, is to be administered for a period of six weeks and for the first four weeks at a dose of about 75 to about 175mg/m per week²。

16. The pharmaceutical product according to claim 13, wherein irinotecan, or a pharmaceutically acceptable salt thereof, is at a concentration of about 300 to about 400mg/m²Once every three weeks; or at about 130 to about 230mg/m²The administration is once every two weeks.

17. A pharmaceutical product, comprising: (A) a first component comprising compound I or a pharmaceutically acceptable salt thereof as an active agent; and (B) a second component comprising erlotinib, or a pharmaceutically acceptable salt thereof, as an active agent; the pharmaceutical product as a combined preparation for simultaneous or sequential use in the treatment of a proliferative disorder, in particular cancer, more particularly colorectal cancer, melanoma and thyroid cancer, comprising b-Raf having the V600E mutation, wherein

(A) Is administered in an amount of from about 200 mg/day to about 3000 mg/day, from about 1000 mg/day to about 2500 mg/day, from about 1700 mg/day to about 2100 mg/day, or about 1920 mg/day, and

(B) is administered in an amount of from about 20 mg/day to about 500 mg/day, from about 100 mg/day to about 400 mg/day, or from about 100 mg/day to about 200 mg/day.

18. A pharmaceutical product, comprising: (A) a first component comprising compound I or a pharmaceutically acceptable salt thereof as an active agent; and (B) a second component comprising cetuximab as an active agent; the pharmaceutical product is for simultaneous or sequential use as a combined preparation for the treatment of a proliferative disorder, wherein

(A) (ii) is administered in an amount of from about 200 mg/day to about 3000 mg/day, from about 1000 mg/day to about 2500 mg/day, from about 1700 mg/day to about 2100 mg/day, or about 1920 mg/day; and is

(B) At about 50mg/m²Weekly to about 700mg/m²Weekly, about 100mg/m²One week to about 600mg/m²Weekly or about 200mg/m²One week to about 500mg/m²The amount per week.

19. A pharmaceutical product, comprising: (A) a first component comprising compound I or a pharmaceutically acceptable salt thereof as an active agent; (B) a second component comprising cetuximab as an active agent; and (C) a third component comprising irinotecan, or a pharmaceutically acceptable salt thereof, as an active agent; the pharmaceutical product is for simultaneous or sequential use as a combined preparation for the treatment of a proliferative disorder, wherein

(A) (ii) is administered in an amount of from about 200 mg/day to about 3000 mg/day, from about 1000 mg/day to about 2500 mg/day, from about 1700 mg/day to about 2100 mg/day, or about 1920 mg/day;

(B) at about 50mg/m²Weekly to about 700mg/m²Weekly, about 100mg/m²One week to about 600mg/m²Weekly or about 200mg/m²One week to about 500mg/m²(iv) amount per week; and is

(C) At a rate of about 1 to about 250mg/m²Weekly, about 50 to about 200mg/m²Weekly or about 125mg/m²The amount per week.

20. Use of propane-1-sulfonic acid {3- [5- (4-chlorophenyl) -1H-pyrrolo [2,3-b ] pyridine-3-carbonyl ] -2, 4-difluoro-phenyl } -amide, or a pharmaceutically acceptable salt thereof, and an EGFR inhibitor in the manufacture of a medicament for the treatment of proliferative disorders, such as cancer, more particularly colorectal cancer, melanoma, and thyroid cancer, all of which comprise b-Raf having the V600 mutation and, in particular, the V600E mutation.

21. Novel formulations, kits and uses substantially as described herein.