U.S. patent application number 14/578804 was filed with the patent office on 2015-04-23 for pharmaceutical combination of mek inhibitor and b-raf inhibitors.
The applicant listed for this patent is GlaxoSmithKline LLC. Invention is credited to Melissa Dumble, Rakesh Kumar, Sylvie Laquerre, Peter Lebowitz.
Application Number | 20150111904 14/578804 |
Document ID | / |
Family ID | 43876568 |
Filed Date | 2015-04-23 |
United States Patent
Application |
20150111904 |
Kind Code |
A1 |
Dumble; Melissa ; et
al. |
April 23, 2015 |
PHARMACEUTICAL COMBINATION OF MEK INHIBITOR AND B-RAF
INHIBITORS
Abstract
A novel combination comprising the MEK inhibitor
N-{3-[3-cyclopropyl-5-(2-fluoro-4-iodo-phenylamino)6,8-dimethyl;-2,4,7-tr-
ioxo-3,4,6,7-tetrahydro-2H-pyrido[4,3-d]pyrimidin-1-yl]phenyl}acetamide,
or solvate thereof, with a B-Raf inhibitor, particularly
N-{3-[5-(2-Amino-4-pyrimidinyl)-2-(1,1-dimethylethyl)-1,3-thiazol-4-yl]-2-
-fluorophenyl}-2,6-difluorobenzenesulfonamide or a pharmaceutically
acceptable salt thereof, pharmaceutical compositions comprising the
same and methods of using such combinations and compositions in the
treatment of conditions in which the inhibition of MEK and/or B-Raf
is beneficial, eg. melanoma.
Inventors: |
Dumble; Melissa;
(Collegeville, PA) ; Kumar; Rakesh; (Collegeville,
PA) ; Laquerre; Sylvie; (Collegeville, PA) ;
Lebowitz; Peter; (Collegeville, PA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
GlaxoSmithKline LLC |
Wilmington |
DE |
US |
|
|
Family ID: |
43876568 |
Appl. No.: |
14/578804 |
Filed: |
December 22, 2014 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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14197421 |
Mar 5, 2014 |
8952018 |
|
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14578804 |
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13499779 |
Apr 2, 2012 |
8703781 |
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PCT/US2010/052808 |
Oct 15, 2010 |
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14197421 |
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61252213 |
Oct 16, 2009 |
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Current U.S.
Class: |
514/264.1 |
Current CPC
Class: |
A61K 45/06 20130101;
A61K 31/506 20130101; A61K 31/519 20130101; A61K 31/506 20130101;
A61K 31/519 20130101; A61K 2300/00 20130101; A61P 35/00 20180101;
A61K 2300/00 20130101 |
Class at
Publication: |
514/264.1 |
International
Class: |
A61K 31/519 20060101
A61K031/519; A61K 31/506 20060101 A61K031/506 |
Claims
1. A combination comprising: (i) a compound of formula (I)
##STR00026## or solvate thereof; and (ii) a compound of formula
(II) ##STR00027## or a pharmaceutically acceptable salt
thereof.
2. A combination according to claim 1 wherein compound (i) is in
the form of the dimethylsulfoxide solvate and the compound (ii) is
in the form of the methanesulfonate salt.
3. A combination kit comprising a combination according claim 1
together with a pharmaceutically acceptable carrier or
carriers.
4. A pharmaceutical composition comprising a combination according
to claim 1 together with a pharmaceutically acceptable diluent or
carrier.
5. A method of treating melanoma in a human in need thereof which
comprises the administration of a therapeutically effective amount
of (i) a compound of formula (I) ##STR00028## or solvate thereof;
and (ii) a compound of formula (II) ##STR00029## or a
pharmaceutically acceptable salt thereof.
6. The method of claim 5, wherein compound (i) is in the form of
the dimethylsulfoxide solvate.
7. The method of claim 5, wherein the compound (ii) is in the form
of the methanesulfonate salt.
8. The method of claim 5, wherein the melanoma is metastatic
melanoma.
9. The method of claim 5, wherein the melanoma is BRAFV600E mutant
melanoma.
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application is filed as a continuation application of
U.S. Ser. No. 14/197,421 filed Mar. 5, 2014, which is a
continuation application of U.S. Ser. No. 13/499,779, filed Apr. 2,
2012, which is a National Phase Application of International Patent
Application Serial No. PCT/US2010/052808 filed Oct. 15, 2010, which
claims priority from 61,252,213 filed on Oct. 16, 2009 in the
United States.
FIELD OF THE INVENTION
[0002] The present invention relates to a method of treating cancer
in a mammal and to combinations useful in such treatment. In
particular, the method relates to a novel combination comprising
the MEK inhibitor
N-{3-[3-cyclopropyl-5-(2-fluoro-4-iodo-phenylamino)6,8-dimethyl;-2,4,7-tr-
ioxo-3,4,6,7-tetrahydro-2H-pyrido[4,3-d]pyrimidin-1-yl]phenyl}acetamide,
or a pharmaceutically acceptable salt or solvate thereof, with a
B-Raf inhibitor, particularly
N-{3-[5-(2-Amino-4-pyrimidinyl)-2-(1,1-dimethylethyl)-1,3-thiazol-4-yl]-2-
-fluorophenyl}-2,6-difluorobenzenesulfonamide or a pharmaceutically
acceptable salt thereof, pharmaceutical compositions comprising the
same and methods of using such combinations and compositions in the
treatment of conditions in which the inhibition of MEK and/or B-Raf
is beneficial, eg. cancer.
BACKGROUND OF THE INVENTION
[0003] Effective treatment of hyperproliferative disorders
including cancer is a continuing goal in the oncology field.
Generally, cancer results from the deregulation of the normal
processes that control cell division, differentiation and apoptotic
cell death and is characterized by the proliferation of malignant
cells which have the potential for unlimited growth, local
expansion and systemic metastasis. Deregulation of normal processes
include abnormalities in signal transduction pathways and response
to factors which differ from those found in normal cells.
[0004] An important large family of enzymes is the protein kinase
enzyme family. Currently, there are about 500 different known
protein kinases. Protein kinases serve to catalyze the
phosphorylation of an amino acid side chain in various proteins by
the transfer of the .gamma.-phosphate of the ATP-Mg.sup.2+ complex
to said amino acid side chain. These enzymes control the majority
of the signaling processes inside cells, thereby governing cell
function, growth, differentiation and destruction (apoptosis)
through reversible phosphorylation of the hydroxyl groups of
serine, threonine and tyrosine residues in proteins.
[0005] Studies have shown that protein kinases are key regulators
of many cell functions, including signal transduction,
transcriptional regulation, cell motility, and cell division.
Several oncogenes have also been shown to encode protein kinases,
suggesting that kinases play a role in oncogenesis. These processes
are highly regulated, often by complex intermeshed pathways where
each kinase will itself be regulated by one or more kinases.
Consequently, aberrant or inappropriate protein kinase activity can
contribute to the rise of disease states associated with such
aberrant kinase activity including benign and malignant
proliferative disorders as well as diseases resulting from
inappropriate activation of the immune and nervous systems. Due to
their physiological relevance, variety and ubiquitousness, protein
kinases have become one of the most important and widely studied
family of enzymes in biochemical and medical research.
[0006] The protein kinase family of enzymes is typically classified
into two main subfamilies: Protein Tyrosine Kinases and Protein
Serine/Threonine Kinases, based on the amino acid residue they
phosphorylate. The protein serine/threonine kinases (PSTK),
includes cyclic AMP- and cyclic GMP-dependent protein kinases,
calcium and phospholipid dependent protein kinase, calcium- and
calmodulin-dependent protein kinases, casein kinases, cell division
cycle protein kinases and others. These kinases are usually
cytoplasmic or associated with the particulate fractions of cells,
possibly by anchoring proteins. Aberrant protein serine/threonine
kinase activity has been implicated or is suspected in a number of
pathologies such as rheumatoid arthritis, psoriasis, septic shock,
bone loss, many cancers and other proliferative diseases.
Accordingly, serine/threonine kinases and the signal transduction
pathways which they are part of are important targets for drug
design. The tyrosine kinases phosphorylate tyrosine residues.
Tyrosine kinases play an equally important role in cell regulation.
These kinases include several receptors for molecules such as
growth factors and hormones, including epidermal growth factor
receptor, insulin receptor, platelet derived growth factor receptor
and others. Studies have indicated that many tyrosine kinases are
transmembrane proteins with their receptor domains located on the
outside of the cell and their kinase domains on the inside. Much
work is also in progress to identify modulators of tyrosine kinases
as well.
[0007] Receptor tyrosine kinases (RTKs) catalyze phosphorylation of
certain tyrosyl amino acid residues in various proteins, including
themselves, which govern cell growth, proliferation and
differentiation.
[0008] Downstream of the several RTKs lie several signaling
pathways, among them is the Ras-Raf-MEK-ERK kinase pathway. It is
currently understood that activation of Ras GTPase proteins in
response to growth factors, hormones, cytokines, etc. stimulates
phosphorylation and activation of Raf kinases. These kinases then
phosphorylate and activate the intracellular protein kinases MEK1
and MEK2, which in turn phosphorylate and activate other protein
kinases, ERK1 and 2. This signaling pathway, also known as the
mitogen-activated protein kinase (MAPK) pathway or cytoplasmic
cascade, mediates cellular responses to growth signals. The
ultimate function of this is to link receptor activity at the cell
membrane with modification of cytoplasmic or nuclear targets that
govern cell proliferation, differentiation, and survival.
[0009] The constitutive activation of this pathway is sufficient to
induce cellular transformation. Disregulated activation of the MAP
kinase pathway due to aberrant receptor tyrosine kinase activation,
Ras mutations or Raf mutations has frequently been found in human
cancers, and represents a major factor determining abnormal growth
control. In human malignances, Ras mutations are common, having
been identified in about 30% of cancers. The Ras family of GTPase
proteins (proteins which convert guanosine triphosphate to
guanosine diphosphate) relay signals from activated growth factor
receptors to downstream intracellular partners. Prominent among the
targets recruited by active membrane-bound Ras are th Raf family of
serine/threonine protein kinases. The Raf family is composed of
three relarted kinases (A-, B- and C-Raf) that act as downstream
effectors of Ras. Ras-medicated Raf activation in turn triggers
activation of MEK1 and MEK2 (MAP/ERK kinases 1 and 2) which in turn
phosphorylate ERK1 and ERK2 (extracellular signal-regulated kinases
1 and 2) on th tyrosine-185 and threonine-183. Activated ERK1 and
ERK2 translocate and accumulate in the nucleus, where they can
phosphorylate a variety of substrates, including transcription
factors that control cellular growth and survival. Given the
importance of the Ras/Raf/MEK/ERK pathway in the development of
human cancers, the kinase components of the signaling cascade are
merging as potentially important targets for the modulation of
disease progression in cancer and other proliferative diseases.
[0010] MEK1 and MEK2 are members of a larger family of
dual-specificity kinases (MEK1-7) that phosphorylate threonine and
tyrosine residues of various MAP kinases. MEK1 and MEK2 are encoded
by distinct genes, but they share high homology (80%) both within
the C-terminal catalytic kinase domains and the most of the
N-terminal regulatory region. Oncogenis forms of MEK1 and MEK2 have
not been found in human cancers, but constitutive activation of MEK
has been shown to result in cellular transformation. In addition to
Raf, MEK can also be activated by other oncognese as well. So far,
the only known substrates of MEK1 and MEK2 are ERK1 and ERK2. This
unusual substrate specificity in addition to the unique ability to
phosphorylate both tyrosine and threonine residues places MEK1 and
MEK2 at a critical point in the signal transduction cascade which
allows it to integrate many extracellular signals into the MAPK
pathway.
[0011] Accordingly, it has been recognized that an inhibitor of a
protein of the MAPK kinase pathway (eg. MEK) should be of value
both as an anti-proliferative, pro-apoptotic and anti-invasive
agent for use in the containment and/or treatment of proliferative
or invasive disease.
[0012] Moreover, it is also known that a compound having MEK
inhibitory activity effectively induces inhibition of ERK1/2
activity and suppression of cell proliferation (The Journal of
Biological Chemistry, vol. 276, No. 4 pp. 2686-2692, 2001), and the
compound is expected to show effects on diseases caused by
undesirable cell proliferation, such as tumor genesis and/or
cancer.
[0013] Mutations in various Ras GTPases and the B-Raf kinase have
been identified that can lead to sustained and constitutive
activation of the MAPK pathway, ultimately resulting in increased
cell division and survival. As a consequence of this, these
mutations have been strongly linked with the establishment,
development, and progression of a wide range of human cancers. The
biological role of the Raf kinases, and specifically that of B-Raf,
in signal transduction is described in Davies, H., et al., Nature
(2002) 9:1-6; Garnett, M. J. & Marais, R., Cancer Cell (2004)
6:313-319; Zebisch, A. & Troppmair, J., Cell. Mol. Life Sci.
(2006) 63:1314-1330; Midgley, R. S. & Kerr, D. J., Crit. Rev.
Onc/Hematol. (2002) 44:109-120; Smith, R. A., et al., Curr. Top.
Med. Chem. (2006) 6:1071-1089; and Downward, J., Nat. Rev. Cancer
(2003) 3:11-22.
[0014] Naturally occurring mutations of the B-Raf kinase that
activate MAPK pathway signaling have been found in a large
percentage of human melanomas (Davies (2002) supra) and thyroid
cancers (Cohen et al J. Nat. Cancer Inst. (2003) 95(8) 625-627 and
Kimura et al Cancer Res. (2003) 63(7) 1454-1457), as well as at
lower, but still significant, frequencies in the following:
Barret's adenocarcinoma (Garnett et al., Cancer Cell (2004) 6
313-319 and Sommerer et al Oncogene (2004) 23(2) 554-558), billiary
tract carcinomas (Zebisch et al., Cell. Mol. Life Sci. (2006) 63
1314-1330), breast cancer (Davies (2002) supra), cervical cancer
(Moreno-Bueno et al Clin. Cancer Res. (2006) 12(12) 3865-3866),
cholangiocarcinoma (Tannapfel et al Gut (2003) 52(5) 706-712),
central nervous system tumors including primary CNS tumors such as
glioblastomas, astrocytomas and ependymomas (Knobbe et al Acta
Neuropathol. (Berl.) (2004) 108(6) 467-470, Davies (2002) supra,
and Garnett et al., Cancer Cell (2004) supra) and secondary CNS
tumors (i.e., metastases to the central nervous system of tumors
originating outside of the central nervous system), colorectal
cancer, including large intestinal colon carcinoma (Yuen et al
Cancer Res. (2002) 62(22) 6451-6455, Davies (2002) supra and
Zebisch et al., Cell. Mol. Life Sci. (2006), gastric cancer (Lee et
al Oncogene (2003) 22(44) 6942-6945), carcinoma of the head and
neck including squamous cell carcinoma of the head and neck (Cohen
et al J. Nat. Cancer Inst. (2003) 95(8) 625-627 and Weber et al
Oncogene (2003) 22(30) 4757-4759), hematologic cancers including
leukemias (Garnett et al., Cancer Cell (2004) supra, particularly
acute lymphoblastic leukemia (Garnett et al., Cancer Cell (2004)
supra and Gustafsson et al Leukemia (2005) 19(2) 310-312), acute
myelogenous leukemia (AML) (Lee et al Leukemia (2004) 18(1)
170-172, and Christiansen et al Leukemia (2005) 19(12) 2232-2240),
myelodysplastic syndromes (Christiansen et al Leukemia (2005)
supra) and chronic myelogenous leukemia (Mizuchi et al Biochem.
Biophys. Res. Commun. (2005) 326(3) 645-651); Hodgkin's lymphoma
(Figl et al Arch. Dermatol. (2007) 143(4) 495-499), non-Hodgkin's
lymphoma (Lee et al Br. J. Cancer (2003) 89(10) 1958-1960),
megakaryoblastic leukemia (Eychene et al Oncogene (1995) 10(6)
1159-1165) and multiple myeloma (Ng et al Br. J. Haematol. (2003)
123(4) 637-645), hepatocellular carcinoma (Garnett et al., Cancer
Cell (2004), lung cancer (Brose et al Cancer Res. (2002) 62(23)
6997-7000, Cohen et al J. Nat. Cancer Inst. (2003) supra and Davies
(2002) supra), including small cell lung cancer (Pardo et al EMBO
J. (2006) 25(13) 3078-3088) and non-small cell lung cancer (Davies
(2002) supra), ovarian cancer (Russell & McCluggage J. Pathol.
(2004) 203(2) 617-619 and Davies (2002) supr), endometrial cancer
(Garnett et al., Cancer Cell (2004) supra, and Moreno-Bueno et al
Clin. Cancer Res. (2006) supra), pancreatic cancer (Ishimura et al
Cancer Lett. (2003) 199(2) 169-173), pituitary adenoma (De Martino
et al J. Endocrinol. Invest. (2007) 30(1) RC1-3), prostate cancer
(Cho et al Int. J. Cancer (2006) 119(8) 1858-1862), renal cancer
(Nagy et al Int. J. Cancer (2003) 106(6) 980-981), sarcoma (Davies
(2002) supra), and skin cancers (Rodriguez-Viciana et al Science
(2006) 311(5765) 1287-1290 and Davies (2002) supra). Overexpression
of c-Raf has been linked to AML (Zebisch et al., Cancer Res. (2006)
66(7) 3401-3408, and Zebisch (Cell. Mol. Life Sci. (2006)) and
erythroleukemia (Zebisch et al., Cell. Mol. Life Sci. (2006).
[0015] By virtue of the role played by the Raf family kinases in
these cancers and exploratory studies with a range of preclinical
and therapeutic agents, including one selectively targeted to
inhibition of B-Raf kinase activity (King A. J., et al., (2006)
Cancer Res. 66:11100-11105), it is generally accepted that
inhibitors of one or more Raf family kinases will be useful for the
treatment of such cancers or other condition associated with Raf
kinase.
[0016] Mutation of B-Raf has also been implicated in other
conditions, including cardio-facio cutaneous syndrome
(Rodriguez-Viciana et al Science (2006) 311(5765) 1287-1290) and
polycystic kidney disease (Nagao et al Kidney Int. (2003) 63(2)
427-437).
[0017] Though there have been many recent advances in the treatment
of cancer with compounds such as the MEK and B-Raf inhibitors,
there remains a need for more effective and/or enhanced treatment
of an individual suffering the effects of cancer.
SUMMARY OF THE INVENTION
[0018] The present inventors have identified a combination of
chemotherapeutic agents that provides increased activity over
monotherapy. In particular, the drug combination that includes the
MEK inhibitor
N-{3-[3-cyclopropyl-5-(2-fluoro-4-iodo-phenylamino)6,8-dimethyl-
-2,4,7-trioxo-3,4,6,7-tetrahydro-2H-pyrido[4,3-d]pyrimidin-1-yl]phenyl}ace-
tamide, or a pharmaceutically acceptable salt or solvate thereof,
in combination with the B-Raf inhibitor
N-{3-[5-(2-Amino-4-pyrimidinyl)-2-(1,1-dimethylethyl)-1,3-thiazol-4-yl]-2-
-fluorophenyl}-2,6-difluorobenzenesulfonamide or a pharmaceutically
acceptable salt thereof is described.
[0019] The MEK inhibitor of the invention is represented by the
structure of formula (I):
##STR00001##
or a pharmaceutically acceptable salt or solvate thereof
(collectively referred to herein as "compound A"),
[0020] The B-Raf inhibitor of the invention is represented by the
structure of formula (II):
##STR00002##
or a pharmaceutically acceptable salt thereof (collectively
referred to herein as "compound B").
[0021] In a first aspect of the present invention, there is
provided a combination comprising:
(i) a compound of formula (I)):
##STR00003##
or a pharmaceutically acceptable salt or solvate thereof; and (ii)
a compound of formula (II)
##STR00004##
or a pharmaceutically acceptable salt thereof.
[0022] In another aspect of the invention, there is provided a
combination comprising
N-{3-[3-cyclopropyl-5-(2-fluoro-4-iodo-phenylamino)6,8-dimethyl-2,4,7-tri-
oxo-3,4,6,7-tetrahydro-2H-pyrido[4,3-d]pyrimidin-1-yl]phenyl}acetamide
dimethyl sulfoxide (solvate) and
N-{3-[5-(2-Amino-4-pyrimidinyl)-2-(1,1-dimethylethyl)-1,3-thiazol-4-yl]-2-
-fluorophenyl}-2,6-difluorobenzenesulfonamide methanesulfonate.
[0023] In another aspect of the present invention, there is
provided a combination, comprising:
(i) a compound of formula (I):
##STR00005##
or a pharmaceutically acceptable salt or solvate thereof; and (ii)
a compound of formula (II):
##STR00006##
or a pharmaceutically acceptable salt thereof for use in
therapy.
[0024] In another aspect of the present invention, there is
provided a combination, comprising:
(i) a compound of formula (I):
##STR00007##
or a pharmaceutically acceptable salt or solvate thereof; and (ii)
a compound of formula (II):
##STR00008##
or a pharmaceutically acceptable salt thereof for use in the
treatment of cancer.
[0025] In another aspect of the present invention, there is
provided a pharmaceutical composition, comprising:
(i) a compound of formula (I):
##STR00009##
or a pharmaceutically acceptable salt or solvate thereof; and (ii)
a compound of formula (II):
##STR00010##
or a pharmaceutically acceptable salt thereof together with a
pharmaceutically acceptable diluent or carrier.
[0026] In a another aspect there is provided the use of a
combination comprising
i) a compound of formula (I)
##STR00011##
or a pharmaceutically acceptable salt or solvate thereof; and (ii)
a compound of formula (II):
##STR00012##
or a pharmaceutically acceptable salt thereof in the manufacture of
a medicament for the treatment of cancer.
[0027] In another aspect there is provided a method of treatment of
cancer in a mammal comprising administering to said mammal:
(i) a therapeutically effective amount of a compound of formula
(I)
##STR00013##
or a pharmaceutically acceptable salt or solvate thereof: and (ii)
a compound of formula (II):
##STR00014##
or a pharmaceutically acceptable salt thereof.
[0028] In another aspect, there is provided a method of treating
cancer in a human in need thereof comprising the administration of
a therapeutically effective amount of a combination of
N-{3-[3-cyclopropyl-5-(2-fluoro-4-iodo-phenylamino)6,8-dimethyl-2,4,7-tri-
oxo-3,4,6,7-tetrahydro-2H-pyrido[4,3-d]pyrimidin-1-yl]phenyl}acetamide,
or a pharmaceutically acceptable salt or solvate thereof, and
N-{3-[5-(2-Amino-4-pyrimidinyl)-2-(1,1-dimethylethyl)-1,3-thiazol-4-yl]-2-
-fluorophenyl}-2,6-difluorobenzenesulfonamide or a pharmaceutically
acceptable salt thereof.
[0029] In another aspect, there is provided a method of treating
cancer in a human in need thereof comprising the administration of
a therapeutically effective amount of a combination of
N-{3-[3-cyclopropyl-5-(2-fluoro-4-iodo-phenylamino)6,8-dimethyl-2,4,7-tri-
oxo-3,4,6,7-tetrahydro-2H-pyrido[4,3-d]pyrimidin-1-yl]phenyl}acetamide
dimethyl sulfoxide solvate and
N-{3-[5-(2-Amino-4-pyrimidinyl)-2-(1,1-dimethylethyl)-1,3-thiazol-4-yl]-2-
-fluorophenyl}-2,6-difluorobenzenesulfonamide methanesulfonate.
[0030] In a further aspect of this invention is provided a method
of treating cancer in a mammal in need thereof which comprises
administering a therapeutically effective amount of a combination
of the invention wherein the combination is administered within a
specific period and for a duration of time.
BRIEF DESCRIPTION OF THE FIGURE
[0031] FIG. 1 is a graph showing the tumor growth inhibition due to
administration of the MEK inhibitor of Compound A, the B-Raf
inhibitor of Compound B, and the combination thereof.
[0032] FIG. 2 is a graph showing the tumor growth inhibition due to
administration of the MEK inhibitor of Compound A, the B-Raf
inhibitor of Compound B, and the combination thereof.
DETAILED DESCRIPTION OF THE INVENTION
[0033] As used herein, the MEK inhibitor
N-{3-[3-cyclopropyl-5-(2-fluoro-4-iodo-phenylamino)6,8-dimethyl-2,4,7-tri-
oxo-3,4,6,7-tetrahydro-2H-pyrido[4,3-d]pyrimidin-1-yl]phenyl}acetamide,
or a pharmaceutically acceptable salt or solvate thereof, is
represented by a compound of formula (I):
##STR00015##
or pharmaceutically acceptable salt or solvate thereof. For
convenience, the group of possible compound and salts or solvates
is collectively referred to as Compound A, meaning that reference
to Compound A will refer to any of the compound or pharmaceutically
acceptable salt or solvate thereof in the alternative.
[0034] Depending on naming convention, the compound of formula (I)
may also properly be referred to as
N-{3-[3-cyclopropyl-5-[(2-fluoro-4-iodophenyl)amino]-6,8-dimethyl-2,4,7-t-
rioxo-3,4,6,7-tetrahydropyrido[4,3-d]pyrimidin-1(2H)-yl]phenyl}acetamide.
[0035] As used herein, the BRaf inhibitor
N-{3-[5-(2-Amino-4-pyrimidinyl)-2-(1,1-dimethylethyl)-1,3-thiazol-4-yl]-2-
-fluorophenyl}-2,6-difluorobenzenesulfonamide or pharmaceutically
acceptable salt thereof, is represented by a compound formula
(II):
##STR00016##
or a pharmaceutically acceptable salt thereof, For convenience, the
group of possible compound and salts is collectively referred to as
Compound B, meaning that reference to Compound B will refer to any
of the compound or pharmaceutically acceptable salt thereof in the
alternative.
[0036] As used herein the term "combination of the invention"
refers to a combination comprising Compound A and Compound B.
[0037] As used herein the term "neoplasm" refers to an abnormal
growth of cells or tissue and is understood to include benign,
i.e., non-cancerous growths, and malignant, i.e., cancerous
growths. The term "neoplastic" means of or related to a
neoplasm.
[0038] As used herein the term "agent" is understood to mean a
substance that produces a desired effect in a tissue, system,
animal, mammal, human, or other subject. Accordingly, the term
"anti-neoplastic agent" is understood to mean a substance producing
an anti-neoplastic effect in a tissue, system, animal, mammal,
human, or other subject. It is also to be understood that an
"agent" may be a single compound or a combination or composition of
two or more compounds.
[0039] By the term "treating" and derivatives thereof as used
herein, is meant therapeutic therapy. In reference to a particular
condition, treating means: (1) to ameliorate the condition or one
or more of the biological manifestations of the condition, (2) to
interfere with (a) one or more points in the biological cascade
that leads to or is responsible for the condition or (b) one or
more of the biological manifestations of the condition (3) to
alleviate one or more of the symptoms, effects or side effects
associated with the condition or one or more of the symptoms,
effects or side effects associated with the condition or treatment
thereof, or (4) to slow the progression of the condition or one or
more of the biological manifestations of the condition.
[0040] As used herein, "prevention" is understood to refer to the
prophylactic administration of a drug to substantially diminish the
likelihood or severity of a condition or biological manifestation
thereof, or to delay the onset of such condition or biological
manifestation thereof. The skilled artisan will appreciate that
"prevention" is not an absolute term. Prophylactic therapy is
appropriate, for example, when a subject is considered at high risk
for developing cancer, such as when a subject has a strong family
history of cancer or when a subject has been exposed to a
carcinogen.
[0041] As used herein, the term "effective amount" means that
amount of a drug or pharmaceutical agent that will elicit the
biological or medical response of a tissue, system, animal or human
that is being sought, for instance, by a researcher or clinician.
Furthermore, the term "therapeutically effective amount" means any
amount which, as compared to a corresponding subject who has not
received such amount, results in improved treatment, healing,
prevention, or amelioration of a disease, disorder, or side effect,
or a decrease in the rate of advancement of a disease or disorder.
The term also includes within its scope amounts effective to
enhance normal physiological function.
[0042] Compounds A and/or B may contain one or more chiral atoms,
or may otherwise be capable of existing as enantiomers.
Accordingly, the compounds of this invention include mixtures of
enantiomers as well as purified enantiomers or enantiomerically
enriched mixtures. Also, it is understood that all tautomers and
mixtures of tautomers are included within the scope of Compound A
and Compound B.
[0043] Also, it is understood that compounds A and B may be
presented, separately or both, as solvates. As used herein, the
term "solvate" refers to a complex of variable stoichiometry formed
by a solute (in this invention, compounds of formula (I) or (II) or
a salt thereof and a solvent. Such solvents for the purpose of the
invention may not interfere with the biological activity of the
solute. Examples of suitable solvents include, but are not limited
to, water, methanol, dimethylsulforide. ethanol and acetic acid. In
one embodiment, the solvent used is a pharmaceutically acceptable
solvent. Examples of suitable pharmaceutically acceptable solvents
include, without limitation, water, ethanol and acetic acid. In
another embodiment, the solvent used is water.
[0044] Compounds A and B may have the ability to crystallize in
more than one form, a characteristic, which is known polymorphism,
and it is understood that such polymorphic forms ("polymorphs") are
within the scope of Compounds A and B. Polymorphism generally can
occur as a response to changes in temperature or pressure or both
and can also result from variations in the crystallization process.
Polymorphs can be distinguished by various physical characteristics
known in the art such as x-ray diffraction patterns, solubility,
and melting point.
[0045] Compound A is disclosed and claimed, along with
pharmaceutically acceptable salts thereof, and also as solvates
thereof, as being useful as an inhibitor of MEK activity,
particularly in treatment of cancer, in WO 2005/121142. Compound A
is the compound of Example 4-1. Compound A can be prepared as
described in WO 2005/121142.
[0046] Suitably, Compound A is in the form of a dimethyl sulfoxide
solvate. Suitably, Compound A is in the form of a sodium salt.
Suitably, Compound A is in the form of a solvate selected from:
hydrate, acetic acid, ethanol, nitromethane, chlorobenzene,
1-pentancol, isopropyl alcohol, ethylene glycol and
3-methyl-1-butanol. These solvates and salt forms can be prepared
by one of skill in the art from the description in WO
2005/121142.
[0047] Compound B is disclosed and claimed, along with
pharmaceutically acceptable salts thereof, as being useful as an
inhibitor of BRaf activity, particularly in the treatment of
cancer, in PCT patent application PCT/US09/42682. Compound B is
embodied by Examples 58a through 58e of the application. The PCT
application was published on 12 Nov. 2009 as publication
WO2009/137391, and is hereby incorporated by reference.
[0048] More particularly, Compound B may be prepared according to
the methods below:
[0049] Method 1: Compound B (first crystal
form)--N-{3-[5-(2-Amino-4-pyrimidinyl)-2-(1,1-dimethylethyl)-1,3-thiazol--
4-yl]-2-fluorophenyl}-2,6-difluorobenzenesulfonamide
##STR00017##
[0050] A suspension of
N-{3-[5-(2-chloro-4-pyrimidinyl)-2-(1,1-dimethylethyl)-1,3-thiazol-4-yl]--
2-fluorophenyl}-2,6-difluorobenzenesulfonamide (196 mg, 0.364 mmol)
and ammonia in methanol 7M (8 ml, 56.0 mmol) was heated in a sealed
tube to 90.degree. C. for 24 h. The reaction was diluted with DCM
and added silica gel and concentrated. The crude product was
chromatographed on silica gel eluting with 100% DCM to 1:1
[DCM:(9:1 EtOAc:MeOH)]. The clean fractions were concentrated to
yield the crude product. The crude product was repurified by
reverse phase HPLC (a gradient of acetonitrile:water with 0.1% TFA
in both). The combined clean fractions were concentrated then
partitioned between DCM and saturated NaHCO.sub.3. The DCM layer
was separated and dried over Na.sub.2SO.sub.4. The title compound,
N-{3-[5-(2-amino-4-pyrimidinyl)-2-(1,1-dimethylethyl)-1,3-thiazol-4-yl]-2-
-fluorophenyl}-2,6-difluorobenzenesulfonamide was obtained (94 mg,
47% yield). .sup.1H NMR (400 MHz, DMSO-d6) .delta. ppm 10.83 (s,
1H), 7.93 (d, J=5.2 Hz, 1H), 7.55-7.70 (m, 1H), 7.35-7.43 (m, 1H),
7.31 (t, J=6.3 Hz, 1H), 7.14-7.27 (m, 3H), 6.70 (s, 2H), 5.79 (d,
J=5.13 Hz, 1H), 1.35 (s, 9H). MS (ESI): 519.9 [M+H].sup.+.
[0051] Method 2: Compound B (alternative crystal
form)--N-{3-[5-(2-Amino-4-pyrimidinyl)-2-(1,1-dimethylethyl)-1,3-thiazol--
4-yl]-2-fluorophenyl}-2,6-difluorobenzenesulfonamide 19.6 mg of
N-{3-[5-(2-Amino-4-pyrimidinyl)-2-(1,1-dimethylethyl)-1,3-thiazol-4-yl]-2-
-fluorophenyl}-2,6-difluorobenzenesulfonamide (may be prepared in
accordance with example 58a) was combined with 500 L of ethyl
acetate in a 2-mL vial at room temperature. The slurry was
temperature-cycled between 0-40.degree. C. for 48 hrs. The
resulting slurry was allowed to cool to room temperature and the
solids were collected by vacuum filtration. The solids were
analyzed by Raman, PXRD, DSC/TGA analyses, which indicated a
crystal form different from the crystal form resulting from Example
58a, above.
[0052] Method 3: Compound B (alternative crystal form, large
batch)--N-{3-[5-(2-amino-4-pyrimidinyl)-2-(1,1-dimethylethyl)-1,3-thiazol-
-4-yl]-2-fluorophenyl}-2,6-difluorobenzenesulfonamide
##STR00018##
Step A: methyl
3-{[(2,6-difluorophenyl)sulfonyl]amino}-2-fluorobenzoate
##STR00019##
[0054] Methyl 3-amino-2-fluorobenzoate (50 g, 1 eq) was charged to
reactor followed by dichloromethane (250 mL, 5 vol). The contents
were stirred and cooled to .about.15.degree. C. and pyridine (26.2
mL, 1.1 eq) was added. After addition of the pyridine, the reactor
contents were adjusted to .about.15.degree. C. and the addition of
2,6-diflurorobenzenesulfonyl chloride (39.7 mL, 1.0 eq) was started
via addition funnel. The temperature during addition was kept
<25.degree. C. After complete addition, the reactor contents
were warmed to 20-25.degree. C. and held overnight. Ethyl acetate
(150 mL) was added and dichloromethane was removed by distillation.
Once distillation was complete, the reaction mixture was then
diluted once more with ethyl acetate (5 vol) and concentrated. The
reaction mixture was diluted with ethyl acetate (10 vol) and water
(4 vol) and the contents heated to 50-55.degree. C. with stirring
until all solids dissolve. The layers were settled and separated.
The organic layer was diluted with water (4 vol) and the contents
heated to 50-55.degree. for 20-30 min. The layers were settled and
then separated and the ethyl acetate layer was evaporated under
reduced pressure to .about.3 volumes. Ethyl Acetate (5 vol.) was
added and again evaporated under reduced pressure to .about.3
volumes. Cyclohexane (9 vol) was then added to the reactor and the
contents were heated to reflux for 30 min then cooled to 0.degree.
C. The solids were filtered and rinsed with cyclohexane
(2.times.100 mL). The solids were air dried overnight to obtain
methyl 3-{[(2,6-difluorophenyl)sulfonyl]amino}-2-fluorobenzoate
(94.1 g, 91%).
Step B:
N-{3-[(2-chloro-4-pyrimidinyl)acetyl]-2-fluorophenyl}-2,6-difluoro-
benzenesulfonamide
##STR00020##
[0056] Methyl
3-{[(2,6-difluorophenyl)sulfonyl]amino}-2-fluorobenzoate (490 g, 1
equiv.), prepared generally in accordance with Step A, above, was
dissolved in THF (2.45 L, 5 vols) and stirred and cooled to
0-3.degree. C. 1M lithium bis(trimethylsilyl)amide in THF (5.25 L,
3.7 equiv.) solution was charged to the reaction mixture followed
addition of 2-chloro-4-methylpyrimidine (238 g, 1.3 equiv.) in THF
(2.45 L, 5 vols). The reaction was then stirred for 1 hr. The
reaction was quenched with 4.5M HCl (3.92 L, 8 vols). The aqueous
layer (bootom layer) was removed and discarded. The organic layer
was concentrated under reduced pressure to .about.2 L. IPAC
(isopropyl acetate) (2.45 L) was added to the reaction mixture
which was then concentrated to .about.2 L. IPAC (0.5 L) and MTBE
(2.45 L) was added and stirred overnight under N.sub.2. The solids
were filtered. The solids and mother filtrate added back together
and stirred for several hours. The solids were filtered and washed
with MTBE (.about.5 vol). The solids were placed in vacuum oven at
50.degree. C. overnight. The solids were dried in vacuum oven at
30.degree. C. over weekend to obtain
N-{3-[(2-chloro-4-pyrimidinyl)acetyl]-2-fluorophenyl}-2,6-difluorobenzene-
sulfonamide (479 g, 72%).
Step C:
N-{3-[5-(2-chloro-4-pyrimidinyl)-2-(1,1-dimethylethyl)-1,3-thiazol-
-4-yl]-2-fluorophenyl}-2,6-difluorobenzenesulfonamide
##STR00021##
[0058] To a reactor vessel was charged
N-{3-[(2-chloro-4-pyrimidinyl)acetyl]-2-fluorophenyl}-2,6-difluorobenzene-
sulfonamide (30 g, 1 eq) followed by dichloromethane (300 mL). The
reaction slurry was cooled to .about.10.degree. C. and
N-bromosuccinimide ("NBS") (12.09 g, 1 eq) was added in 3
approximately equal portions, stirring for 10-15 minutes between
each addition. After the final addition of NBS, the reaction
mixture was warmed to .about.20.degree. C. and stirred for 45 min.
Water (5 vol) was then added to the reaction vessel and the mixture
was stirred and then the layers separated. Water (5 vol) was again
added to the dichloromethane layer and the mixture was stirred and
the layers separated. The dichloromethane layers were concentrated
to .about.120 mL. Ethyl acetate (7 vol) was added to the reaction
mixture and concentrated to .about.120 mL. Dimethylacetamide (270
mL) was then added to the reaction mixture and cooled to
.about.10.degree. C. 2,2-Dimethylpropanethioamide (1.3 g, 0.5 eq)
in 2 equal portions was added to the reactor contents with stirring
for .about.5 minutes between additions. The reaction was warmed to
20-25.degree. C. After 45 min, the vessel contents were heated to
75.degree. C. and held for 1.75 hours. The reaction mixture was
then cooled to 5.degree. C. and water (270 ml) was slowly charged
keeping the temperature below 30.degree. C. Ethyl acetate (4 vol)
was then charged and the mixture was stirred and layers separated.
Ethyl acetate (7 vol) was again charged to the aqueous layer and
the contents were stirred and separated. Ethyl acetate (7 vol) was
charged again to the aqueous layer and the contents were stirred
and separated. The organic layers were combined and washed with
water (4 vol) 4 times and stirred overnight at 20-25.degree. C. The
organic layers were then concentrated under heat and vacuum to 120
mL. The vessel contents were then heated to 50.degree. C. and
heptanes (120 mL) were added slowly. After addition of heptanes,
the vessel contents were heated to reflux then cooled to 0.degree.
C. and held for .about.2 hrs. The solids were filtered and rinsed
with heptanes (2.times.2 vol). The solid product was then dried
under vacuum at 30.degree. C. to obtain
N-{3-[5-(2-chloro-4-pyrimidinyl)-2-(1,1-dimethylethyl)-1,3-thiazol-4-yl]--
2-fluorophenyl}-2,6-difluorobenzenesulfonamide (28.8 g, 80%).
Step D:
N-{3-[5-(2-amino-4-pyrimidinyl)-2-(1,1-dimethylethyl)-1,3-thiazol--
4-yl]-2-fluorophenyl}-2,6-difluorobenzenesulfonamide
[0059] In 1 gal pressure reactor, a mixture of
N-{3-[5-(2-chloro-4-pyrimidinyl)-2-(1,1-dimethylethyl)-1,3-thiazol-4-yl]--
2-fluorophenyl}-2,6-difluorobenzenesulfonamide (120 g) prepared in
accordance with Step C, above, and ammonium hydroxide (28-30%, 2.4
L, 20 vol) was heated in the sealed pressure reactor to
98-103.degree. C. and stirred at this temperature for 2 hours. The
reaction was cooled slowly to room temperature (20.degree. C.) and
stirred overnight. The solids were filtered and washed with minimum
amount of the mother liquor and dried under vacuum. The solids were
added to a mixture of EtOAc (15 vol)/water (2 vol) and heated to
complete dissolution at 60-70.degree. C. and the aqueous layer was
removed and discarded. The EtOAC layer was charged with water (1
vol) and neutralized with aq. HCl to .about.pH 5.4-5.5. and added
water (1 vol). The aqueous layer was removed and discarded at
60-70.degree. C. The organic layer was washed with water (1 vol) at
60-70.degree. C. and the aqueous layer was removed and discarded.
The organic layer was filtered at 60.degree. C. and concentrated to
3 volumes. EtOAc (6 vol) was charged into the mixture and heated
and stirred at 72.degree. C. for 10 min, then cooled to 20.degree.
C. and stirred overnight. EtOAc was removed via vacuum distillation
to concentrate the reaction mixture to .about.3 volumes. The
reaction mixture was maintained at .about.65-70.degree. C. for
.about.30 mins. Product crystals having the same crystal form as
those prepared in Example 58b (and preparable by the procedure of
Example 58b), above, in heptanes slurry were charged. Heptane (9
vol) was slowly added at 65-70.degree. C. The slurry was stirred at
65-70.degree. C. for 2-3 hours and then cooled slowly to
0-5.degree. C. The product was filtered, washed with EtOAc/heptane
(3/1 v/v, 4 vol) and dried at 45.degree. C. under vacuum to obtain
N-{3-[5-(2-amino-4-pyrimidinyl)-2-(1,1-dimethylethyl)-1,3-thiazol-4-yl]-2-
-fluorophenyl}-2,6-difluorobenzenesulfonamide (102.3 g, 88%).
[0060] Method 4: Compound B (mesylate
salt)--N-{3-[5-(2-amino-4-pyrimidinyl)-2-(1,1-dimethylethyl)-1,3-thiazol--
4-yl]-2-fluorophenyl}-2,6-difluorobenzenesulfonamide
methanesulfonate
##STR00022##
[0061] To a solution of
N-{3-[5-(2-amino-4-pyrimidinyl)-2-(1,1-dimethylethyl)-1,3-thiazol-4-yl]-2-
-fluorophenyl}-2,6-difluorobenzenesulfonamide (204 mg, 0.393 mmol)
in isopropanol (2 mL), methanesulfonic acid (0.131 mL, 0.393 mmol)
was added and the solution was allowed to stir at room temperature
for 3 hours. A white precipitate formed and the slurry was filtered
and rinsed with diethyl ether to give the title product as a white
crystalline solid (210 mg, 83% yield). .sup.1H NMR (400 MHz,
DMSO-d6) .delta. ppm 10.85 (s, 1H) 7.92-8.05 (m, 1H) 7.56-7.72 (m,
1H) 6.91-7.50 (m, 7H) 5.83-5.98 (m, 1H) 2.18-2.32 (m, 3H) 1.36 (s,
9H). MS (ESI): 520.0 [M+H].sup.+.
[0062] Method 5: Compound B (alternative mesylate salt
embodiment)--N-{3-[5-(2-amino-4-pyrimidinyl)-2-(1,1-dimethylethyl)-1,3-th-
iazol-4-yl]-2-fluorophenyl}-2,6-difluorobenzenesulfonamide
methanesulfonate
[0063]
N-{3-[5-(2-amino-4-pyrimidinyl)-2-(1,1-dimethylethyl)-1,3-thiazol-4-
-yl]-2-fluorophenyl}-2,6-difluorobenzenesulfonamide (as may be
prepared according to example 58a) (2.37 g, 4.56 mmol) was combined
with pre-filtered acetonitrile (5.25 vol, 12.4 mL). A pre-filtered
solution of mesic acid (1.1 eq., 5.02 mmol, 0.48 g) in H.sub.2O
(0.75 eq., 1.78 mL) was added at 20.degree. C. The temperature of
the resulting mixture was raised to 50-60.degree. C. while
maintaining a low agitation speed. Once the mixture temperature
reached to 50-60.degree. C., a seed slurry of
N-{3-[5-(2-amino-4-pyrimidinyl)-2-(1,1-dimethylethyl)-1,3-thiazol-4-yl]-2-
-fluorophenyl}-2,6-difluorobenzenesulfonamide methanesulfonate
(1.0% w/w slurried in 0.2 vol of pre-filtered acetonitrile) was
added, and the mixture was aged while agitating at a speed fast
enough to keep solids from settling at 50-60.degree. C. for 2 hr.
The mixture was then cooled to 0-5.degree. C. at 0.25.degree.
C./min and held at 0-5.degree. C. for at 6 hr. The mixture was
filtered and the wet cake was washed twice with pre-filtered
acetonitrile. The first wash consisted of 14.2 ml (6 vol)
pre-filtered acetonitrile and the second wash consisted of 9.5 ml
(4 vol) pre-filtered acetonitrile. The wet solid was dried at
50.degree. C. under vacuum, yielding 2.39 g (85.1% yield) of
product.
[0064] Typically, the salts of the present invention are
pharmaceutically acceptable salts. Salts encompassed within the
term "pharmaceutically acceptable salts" refer to non-toxic salts
of the compounds of this invention. Salts of the compounds of the
present invention may comprise acid addition salts derived from a
nitrogen on a substituent in a compound of the present invention.
Representative salts include the following salts: acetate,
benzenesulfonate, benzoate, bicarbonate, bisulfate, bitartrate,
borate, bromide, calcium edetate, camsylate, carbonate, chloride,
clavulanate, citrate, dihydrochloride, edetate, edisylate,
estolate, esylate, fumarate, gluceptate, gluconate, glutamate,
glycollylarsanilate, hexylresorcinate, hydrabamine, hydrobromide,
hydrochloride, hydroxynaphthoate, iodide, isethionate, lactate,
lactobionate, laurate, malate, maleate, mandelate, mesylate,
methylbromide, methylnitrate, methylsulfate, monopotassium maleate,
mucate, napsylate, nitrate, N-methylglucamine, oxalate, pamoate
(embonate), palmitate, pantothenate, phosphate/diphosphate,
polygalacturonate, potassium, salicylate, sodium, stearate,
subacetate, succinate, tannate, tartrate, teoclate, tosylate,
triethiodide, trimethylammonium and valerate. Other salts, which
are not pharmaceutically acceptable, may be useful in the
preparation of compounds of this invention and these form a further
aspect of the invention. Salts may be readily prepared by a person
skilled in the art.
[0065] While it is possible that, for use in therapy, compounds A
and B, may be administered as the raw chemical, it is possible to
present the active ingredient as a pharmaceutical composition.
Accordingly, the invention further provides pharmaceutical
compositions, which include a compound A and/or a compound B, and
one or more pharmaceutically acceptable carriers, diluents, or
excipients. The compounds A and B are as described above. The
carrier(s), diluent(s) or excipient(s) must be acceptable in the
sense of being compatible with the other ingredients of the
formulation, capable of pharmaceutical formulation, and not
deleterious to the recipient thereof. In accordance with another
aspect of the invention there is also provided a process for the
preparation of a pharmaceutical composition including admixing a
Compound A and/or Compound B, with one or more pharmaceutically
acceptable carriers, diluents or excipients. Such elements of the
pharmaceutical compositions utilized may be presented in separate
pharmaceutical combinations or formulated together in one
pharmaceutical composition. Accordingly, the invention further
provides a combination of pharmaceutical compositions one of which
includes Compound A and one or more pharmaceutically acceptable
carriers, diluents, or excipients and a pharmaceutical composition
containing Compound B and one or more pharmaceutically acceptable
carriers, diluents, or excipients.
[0066] Compound A and Compound B are as described above and may be
utilized in any of the compositions described above.
[0067] Pharmaceutical compositions may be presented in unit dose
forms containing a predetermined amount of active ingredient per
unit dose. As is known to those skilled in the art, the amount of
active ingredient per dose will depend on the condition being
treated, the route of administration and the age, weight and
condition of the patient. Preferred unit dosage compositions are
those containing a daily dose or sub-dose, or an appropriate
fraction thereof, of an active ingredient. Furthermore, such
pharmaceutical compositions may be prepared by any of the methods
well known in the pharmacy art.
[0068] Compounds A and B may be administered by any appropriate
route. Suitable routes include oral, rectal, nasal, topical
(including buccal and sublingual), vaginal, and parenteral
(including subcutaneous, intramuscular, intraveneous, intradermal,
intrathecal, and epidural). It will be appreciated that the
preferred route may vary with, for example, the condition of the
recipient of the combination and the cancer to be treated. It will
also be appreciated that each of the agents administered may be
administered by the same or different routes and that the Compounds
A and B may be compounded together in a pharmaceutical
composition.
[0069] Pharmaceutical compositions adapted for oral administration
may be presented as discrete units such as capsules or tablets;
powders or granules; solutions or suspensions in aqueous or
non-aqueous liquids; edible foams or whips; or oil-in-water liquid
emulsions or water-in-oil liquid emulsions.
[0070] For instance, for oral administration in the form of a
tablet or capsule, the active drug component can be combined with
an oral, non-toxic pharmaceutically acceptable inert carrier such
as ethanol, glycerol, water and the like. Powders are prepared by
comminuting the compound to a suitable fine size and mixing with a
similarly comminuted pharmaceutical carrier such as an edible
carbohydrate, as, for example, starch or mannitol. Flavoring,
preservative, dispersing and coloring agent can also be
present.
[0071] Capsules are made by preparing a powder mixture as described
above, and filling formed gelatin sheaths. Glidants and lubricants
such as colloidal silica, talc, magnesium stearate, calcium
stearate or solid polyethylene glycol can be added to the powder
mixture before the filling operation. A disintegrating or
solubilizing agent such as agar-agar, calcium carbonate or sodium
carbonate can also be added to improve the availability of the
medicament when the capsule is ingested.
[0072] Moreover, when desired or necessary, suitable binders,
lubricants, disintegrating agents and coloring agents can also to
granulating, the powder mixture can be run through the tablet
machine and the result is imperfectly formed slugs broken into
granules. The granules can be lubricated be incorporated into the
mixture. Suitable binders include starch, gelatin, natural sugars
such as glucose or beta-lactose, corn sweeteners, natural and
synthetic gums such as acacia, tragacanth or sodium alginate,
carboxymethylcellulose, polyethylene glycol, waxes and the like.
Lubricants used in these dosage forms include sodium oleate, sodium
stearate, magnesium stearate, sodium benzoate, sodium acetate,
sodium chloride and the like. Disintegrators include, without
limitation, starch, methyl cellulose, agar, bentonite, xanthan gum
and the like. Tablets are formulated, for example, by preparing a
powder mixture, granulating or slugging, adding a lubricant and
disintegrant and pressing into tablets. A powder mixture is
prepared by mixing the compound, suitably comminuted, with a
diluent or base as described above, and optionally, with a binder
such as carboxymethylcellulose, an aliginate, gelatin, or polyvinyl
pyrrolidone, a solution retardant such as paraffin, a resorption
accelerator such as a quaternary salt and/or an absorption agent
such as bentonite, kaolin or dicalcium phosphate. The powder
mixture can be granulated by wetting with a binder such as syrup,
starch paste, acadia mucilage or solutions of cellulosic or
polymeric materials and forcing through a screen. As an alternative
to prevent sticking to the tablet forming dies by means of the
addition of stearic acid, a stearate salt, talc or mineral oil. The
lubricated mixture is then compressed into tablets. The compounds
of the present invention can also be combined with free flowing
inert carrier and compressed into tablets directly without going
through the granulating or slugging steps. A clear or opaque
protective coating consisting of a sealing coat of shellac, a
coating of sugar or polymeric material and a polish coating of wax
can be provided. Dyestuffs can be added to these coatings to
distinguish different unit dosages.
[0073] Oral fluids such as solution, syrups and elixirs can be
prepared in dosage unit form so that a given quantity contains a
predetermined amount of the compound. Syrups can be prepared by
dissolving the compound in a suitably flavored aqueous solution,
while elixirs are prepared through the use of a non-toxic alcoholic
vehicle. Suspensions can be formulated by dispersing the compound
in a non-toxic vehicle. Solubilizers and emulsifiers such as
ethoxylated isostearyl alcohols and polyoxy ethylene sorbitol
ethers, preservatives, flavor additive such as peppermint oil or
natural sweeteners or saccharin or other artificial sweeteners, and
the like can also be added.
[0074] Where appropriate, compositions for oral administration can
be microencapsulated. The composition can also be prepared to
prolong or sustain the release as for example by coating or
embedding particulate material in polymers, wax or the like.
[0075] The agents for use according to the present invention can
also be administered in the form of liposome delivery systems, such
as small unilamellar vesicles, large unilamellar vesicles and
multilamellar vesicles. Liposomes can be formed from a variety of
phospholipids, such as cholesterol, stearylamine or
phosphatidylcholines.
[0076] Agents for use according to the present invention may also
be delivered by the use of monoclonal antibodies as individual
carriers to which the compound molecules are coupled. The compounds
may also be coupled with soluble polymers as targetable drug
carriers. Such polymers can include polyvinylpyrrolidone, pyran
copolymer, polyhydroxypropylmethacrylamide-phenol,
polyhydroxyethylaspartamidephenol, or polyethyleneoxidepolylysine
substituted with palmitoyl residues. Furthermore, the compounds may
be coupled to a class of biodegradable polymers useful in achieving
controlled release of a drug, for example, polylactic acid,
polepsilon caprolactone, polyhydroxy butyric acid, polyorthoesters,
polyacetals, polydihydropyrans, polycyanoacrylates and cross-linked
or amphipathic block copolymers of hydrogels.
[0077] Pharmaceutical compositions adapted for transdermal
administration may be presented as discrete patches intended to
remain in intimate contact with the epidermis of the recipient for
a prolonged period of time. For example, the active ingredient may
be delivered from the patch by iontophoresis as generally described
in Pharmaceutical Research, 3(6), 318 (1986).
[0078] Pharmaceutical compositions adapted for topical
administration may be formulated as ointments, creams, suspensions,
lotions, powders, solutions, pastes, gels, sprays, aerosols or
oils.
[0079] For treatments of the eye or other external tissues, for
example mouth and skin, the compositions are preferably applied as
a topical ointment or cream. When formulated in an ointment, the
active ingredient may be employed with either a paraffinic or a
water-miscible ointment base. Alternatively, the active ingredient
may be formulated in a cream with an oil-in-water cream base or a
water-in-oil base.
[0080] Pharmaceutical compositions adapted for topical
administrations to the eye include eye drops wherein the active
ingredient is dissolved or suspended in a suitable carrier,
especially an aqueous solvent.
[0081] Pharmaceutical compositions adapted for topical
administration in the mouth include lozenges, pastilles and mouth
washes.
[0082] Pharmaceutical compositions adapted for rectal
administration may be presented as suppositories or as enemas.
[0083] Pharmaceutical compositions adapted for nasal administration
wherein the carrier is a solid include a coarse powder having a
particle size for example in the range 20 to 500 microns which is
administered in the manner in which snuff is taken, i.e. by rapid
inhalation through the nasal passage from a container of the powder
held close up to the nose. Suitable compositions wherein the
carrier is a liquid, for administration as a nasal spray or as
nasal drops, include aqueous or oil solutions of the active
ingredient.
[0084] Pharmaceutical compositions adapted for administration by
inhalation include fine particle dusts or mists that may be
generated by means of various types of metered dose pressurised
aerosols, nebulizers or insufflators.
[0085] Pharmaceutical compositions adapted for vaginal
administration may be presented as pessaries, tampons, creams,
gels, pastes, foams or spray compositions.
[0086] Pharmaceutical compositions adapted for parenteral
administration include aqueous and non-aqueous sterile injection
solutions which may contain anti-oxidants, buffers, bacteriostats
and solutes which render the formulation isotonic with the blood of
the intended recipient; and aqueous and non-aqueous sterile
suspensions which may include suspending agents and thickening
agents. The compositions may be presented in unit-dose or
multi-dose containers, for example sealed ampoules and vials, and
may be stored in a freeze-dried (lyophilized) condition requiring
only the addition of the sterile liquid carrier, for example water
for injections, immediately prior to use. Extemporaneous injection
solutions and suspensions may be prepared from sterile powders,
granules and tablets.
[0087] It should be understood that in addition to the ingredients
particularly mentioned above, the compositions may include other
agents conventional in the art having regard to the type of
formulation in question, for example those suitable for oral
administration may include flavoring agents. Unless otherwise
defined, in all dosing protocols described herein, the regimen of
compounds administered does not have to commence with the start of
treatment and terminate with the end of treatment, it is only
required that the number of consecutive days in which both
compounds are administered and the optional number of consecutive
days in which only one of the component compounds is administered,
or the indicated dosing protocol--including the amount of compound
administered, occur at some point during the course of
treatment.
[0088] Compounds A and B may be employed in combination in
accordance with the invention by administration simultaneously in a
unitary pharmaceutical composition including both compounds.
Alternatively, the combination may be administered separately in
separate pharmaceutical compositions, each including one of the
compounds A and B in a sequential manner wherein, for example,
Compound A or Compound B is administered first and the other
second. Such sequential administration may be close in time (eg.
simultaneously) or remote in time. Furthermore, it does not matter
if the compounds are administered in the same dosage form, e.g. one
compound may be administered topically and the other compound may
be administered orally. Suitably, both compounds are administered
orally.
[0089] Thus in one embodiment, one or more doses of Compound A are
administered simultaneously or separately with one or more doses of
Compound B.
[0090] Unless otherwise defined, in all dosing protocols described
herein, the regimen of compounds administered does not have to
commence with the start of treatment and terminate with the end of
treatment, it is only required that the number of consecutive days
in which both compounds are administered and the optional number of
consecutive days in which only one of the component compounds is
administered, or the indicated dosing protocol--including the
amount of compound administered, occur at some point during the
course of treatment.
[0091] In one embodiment, multiple doses of Compound A are
administered simultaneously or separately with multiple doses of
Compound B.
[0092] In one embodiment, multiple doses of Compound A are
administered simultaneously or separately with one dose of Compound
B.
[0093] In one embodiment, one dose of Compound A is administered
simultaneously or separately with multiple doses of Compound B.
[0094] In one embodiment one dose of Compound A is administered
simultaneously or separately with one dose of Compound B.
[0095] In all the above embodiments Compound A may be administered
first or Compound B may be administered first.
[0096] The combinations may be presented as a combination kit. By
the term "combination kit" "or kit of parts" as used herein is
meant the pharmaceutical composition or compositions that are used
to administer Compound A and Compound B according to the invention.
When both compounds are administered simultaneously, the
combination kit can contain Compound A and Compound B in a single
pharmaceutical composition, such as a tablet, or in separate
pharmaceutical compositions. When Compounds A and B are not
administered simultaneously, the combination kit will contain
Compound A and Compound B in separate pharmaceutical compositions
either in a single package or Compound A and Compound B in separate
pharmaceutical compositions in separate packages.
[0097] In one aspect there is provided a kit of parts comprising
components: [0098] Compound A in association with a
pharmaceutically acceptable excipients, diluents or carrier; and
[0099] Compound B in association with a pharmaceutically acceptable
excipients, diluents or carrier.
[0100] In one embodiment of the invention the kit of parts
comprising the following components: [0101] Compound A in
association with a pharmaceutically acceptable excipients, diluents
or carrier; and [0102] Compound B in association with a
pharmaceutically acceptable excipients, diluents or carrier,
wherein the components are provided in a form which is suitable for
sequential, separate and/or simultaneous administration.
[0103] In one embodiment the kit of parts comprises: [0104] a first
container comprising Compound A in association with a
pharmaceutically acceptable excipient, diluent or carrier; and
[0105] a second container comprising Compound B in association with
a pharmaceutically acceptable excipient, diluent or carrier, and a
container means for containing said first and second
containers.
[0106] The combination kit can also be provided by instruction,
such as dosage and administration instructions. Such dosage and
administration instructions can be of the kind that are provided to
a doctor, for example by a drug product label, or they can be of
the kind that are provided by a doctor, such as instructions to a
patient.
[0107] The term "loading dose" as used herein will be understood to
mean a single dose or short duration regimen of Compound A or
Compound B having a dosage higher than the maintenance dose
administered to the subject to, for example, rapidly increase the
blood concentration level of the drug. Suitably, a short duration
regimen for use herein will be from: 1 to 14 days; suitably from 1
to 7 days; suitably from 1 to 3 days; suitably for three days;
suitably for two days; suitably for one day. In some embodiments,
the "loading dose" can increase the blood concentration of the drug
to a therapeutically effective level. In some embodiments, the
"loading dose" can increase the blood concentration of the drug to
a therapeutically effective level in conjunction with a maintenance
dose of the drug. The "loading dose" can be administered once per
day, or more than once per day (e.g., up to 4 times per day).
Suitably the "loading dose" will be administered once a day.
Suitably, the loading dose will be an amount from 2 to 100 times
the maintenance dose; suitably from 2 to 10 times; suitably from 2
to 5 times; suitably 2 times; suitably 3 times; suitably 4 times;
suitably 5 times. Suitably, the loading dose will be administered
for from 1 to 7 days; suitably from 1 to 5 days; suitably from 1 to
3 days; suitably for 1 day; suitably for 2 days; suitably for 3
days, followed by a maintenance dosing protocol.
[0108] The term "maintenance dose" as used herein will be
understood to mean a dose that is serially administered (for
example; at least twice), and which is intended to either slowly
raise blood concentration levels of the compound to a
therapeutically effective level, or to maintain such a
therapeutically effective level. The maintenance dose is generally
administered once per day and the daily dose of the maintenance
dose is lower than the total daily dose of the loading dose.
[0109] Suitably the combinations of this invention are administered
within a "specified period".
[0110] By the term "specified period" and derivatives thereof, as
used herein is meant the interval of time between the
administration of one of Compound A and Compound B and the other of
Compound A and Compound B. Unless otherwise defined, the specified
period can include simultaneous administration. When both compounds
of the invention are administered once a day the specified period
refers to administration of Compound A and Compound B during a
single day. When one or both compounds of the invention are
administered more than once a day, the specified period is
calculated based on the first administration of each compound on a
specific day. All administrations of a compound of the invention
that are subsequent to the first during a specific day are not
considered when calculating the specific period.
[0111] Suitably, if the compounds are administered within a
"specified period" and not administered simultaneously, they are
both administered within about 24 hours of each other--in this
case, the specified period will be about 24 hours; suitably they
will both be administered within about 12 hours of each other--in
this case, the specified period will be about 12 hours; suitably
they will both be administered within about 11 hours of each
other--in this case, the specified period will be about 11 hours;
suitably they will both be administered within about 10 hours of
each other--in this case, the specified period will be about 10
hours; suitably they will both be administered within about 9 hours
of each other--in this case, the specified period will be about 9
hours; suitably they will both be administered within about 8 hours
of each other--in this case, the specified period will be about 8
hours; suitably they will both be administered within about 7 hours
of each other--in this case, the specified period will be about 7
hours; suitably they will both be administered within about 6 hours
of each other--in this case, the specified period will be about 6
hours; suitably they will both be administered within about 5 hours
of each other--in this case, the specified period will be about 5
hours; suitably they will both be administered within about 4 hours
of each other--in this case, the specified period will be about 4
hours; suitably they will both be administered within about 3 hours
of each other--in this case, the specified period will be about 3
hours; suitably they will be administered within about 2 hours of
each other--in this case, the specified period will be about 2
hours; suitably they will both be administered within about 1 hour
of each other--in this case, the specified period will be about 1
hour. As used herein, the administration of Compound A and Compound
B in less than about 45 minutes apart is considered simultaneous
administration.
[0112] Suitably, when the combination of the invention is
administered for a "specified period", the compounds will be
co-administered for a "duration of time".
[0113] By the term "duration of time" and derivatives thereof, as
used herein is meant that both compounds of the invention are
administered for an indicated number of consecutive days.
[0114] Regarding "specified period" administration:
[0115] Suitably, both compounds will be administered within a
specified period for at least one day--in this case, the duration
of time will be at least one day;
suitably, during the course to treatment, both compounds will be
administered within a specified period for at least 3 consecutive
days--in this case, the duration of time will be at least 3 days;
suitably, during the course to treatment, both compounds will be
administered within a specified period for at least 5 consecutive
days--in this case, the duration of time will be at least 5 days;
suitably, during the course to treatment, both compounds will be
administered within a specified period for at least 7 consecutive
days--in this case, the duration of time will be at least 7 days;
suitably, during the course to treatment, both compounds will be
administered within a specified period for at least 14 consecutive
days--in this case, the duration of time will be at least 14 days;
suitably, during the course to treatment, both compounds will be
administered within a specified period for at least 30 consecutive
days--in this case, the duration of time will be at least 30
days.
[0116] Further regarding "specified period" administration:
[0117] Suitably, during the course of treatment, Compound A and
Compound B will be administered within a specified period for from
1 to 4 days over a 7 day period, and during the other days of the 7
day period Compound A will be administered alone. Suitably, this 7
day protocol is repeated for 2 cycles or for 14 days; suitably for
4 cycles or 28 days; suitably for continuous administration.
[0118] Suitably, during the course of treatment, Compound A and
Compound B will be administered within a specified period for from
1 to 4 days over a 7 day period, and during the other days of the 7
day period Compound B will be administered alone. Suitably, this 7
day protocol is repeated for 2 cycles or for 14 days; suitably for
4 cycles or 28 days; suitably for continuous administration.
Suitably, Compound B is administered for consecutive days during
the 7 day period. Suitably, Compound B is administered in a pattern
of every other day during each 7 day period.
[0119] Suitably, during the course of treatment, Compound A and
Compound B will be administered within a specified period for 3
days over a 7 day period, and during the other days of the 7 day
period Compound B will be administered alone. Suitably, this 7 day
protocol is repeated for 2 cycles or for 14 days; suitably for 4
cycles or 28 days; suitably for continuous administration.
Suitably, Compound A will be administered 3 consecutive days during
the 7 day period.
[0120] Suitably, during the course of treatment, Compound A and
Compound B will be administered within a specified period for 2
days over a 7 day period, and during the other days of the 7 day
period Compound B will be administered alone. Suitably, this 7 day
protocol is repeated for 2 cycles or for 14 days; suitably for 4
cycles or 28 days; suitably for continuous administration.
Suitably, Compound A will be administered 2 consecutive days during
the 7 day period.
[0121] Suitably, during the course of treatment, Compound A and
Compound B will be administered within a specified period for 1 day
during a 7 day period, and during the other days of the 7 day
period Compound B will be administered alone. Suitably, this 7 day
protocol is repeated for 2 cycles or for 14 days; suitably for 4
cycles or 28 days; suitably for continuous administration.
[0122] Suitably, if the compounds are not administered during a
"specified period", they are administered sequentially. By the term
"sequential administration", and derivates thereof, as used herein
is meant that one of Compound A and Compound B is administered for
two or more consecutive days and the other of Compound A and
Compound B is subsequently administered for two or more consecutive
days. Also, contemplated herein is a drug holiday utilized between
the sequential administration of one of Compound A and Compound B
and the other of Compound A and Compound B. As used herein, a drug
holiday is a period of days after the sequential administration of
one of Compound A and Compound B and before the administration of
the other of Compound A and Compound B where neither Compound A nor
Compound B is administered. Suitably the drug holiday will be a
period of days selected from: 1 day, 2 days, 3 days, 4 days, 5
days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13
days and 14 days.
[0123] Regarding sequential administration:
[0124] Suitably, one of Compound A and Compound B is administered
for from 1 to 30 consecutive days, followed by an optional drug
holiday, followed by administration of the other of Compound A and
Compound B for from 1 to 30 consecutive days. Suitably, one of
Compound A and Compound B is administered for from 2 to 21
consecutive days, followed by an optional drug holiday, followed by
administration of the other of Compound A and Compound B for from 2
to 21 consecutive days. Suitably, one of Compound A and Compound B
is administered for from 2 to 14 consecutive days, followed by a
drug holiday of from 1 to 14 days, followed by administration of
the other of Compound A and Compound B for from 2 to 14 consecutive
days. Suitably, one of Compound A and Compound B is administered
for from 3 to 7 consecutive days, followed by a drug holiday of
from 3 to 10 days, followed by administration of the other of
Compound A and Compound B for from 3 to 7 consecutive days.
[0125] Suitably, Compound B will be administered first in the
sequence, followed by an optional drug holiday, followed by
administration of Compound A. Suitably, Compound B is administered
for from 1 to 21 consecutive days, followed by an optional drug
holiday, followed by administration of Compound A for from 1 to 21
consecutive days. Suitably, Compound B is administered for from 3
to 21 consecutive days, followed by a drug holiday of from 1 to 14
days, followed by administration of Compound A for from 3 to 21
consecutive days. Suitably, Compound B is administered for from 3
to 21 consecutive days, followed by a drug holiday of from 3 to 14
days, followed by administration of Compound A for from 3 to 21
consecutive days. Suitably, Compound B is administered for 21
consecutive days, followed by an optional drug holiday, followed by
administration of Compound A for 14 consecutive days. Suitably,
Compound B is administered for 14 consecutive days, followed by a
drug holiday of from 1 to 14 days, followed by administration of
Compound A for 14 consecutive days. Suitably, Compound B is
administered for 7 consecutive days, followed by a drug holiday of
from 3 to 10 days, followed by administration of Compound A for 7
consecutive days. Suitably, Compound B is administered for 3
consecutive days, followed by a drug holiday of from 3 to 14 days,
followed by administration of Compound A for 7 consecutive days.
Suitably, Compound B is administered for 3 consecutive days,
followed by a drug holiday of from 3 to 10 days, followed by
administration of Compound A for 3 consecutive days.
[0126] Suitably, Compound A will be administered first in the
sequence, followed by an optional drug holiday, followed by
administration of Compound B. Suitably, Compound A is administered
for from 1 to 21 consecutive days, followed by an optional drug
holiday, followed by administration of Compound B for from 1 to 21
consecutive days. Suitably, Compound A is administered for from 3
to 21 consecutive days, followed by a drug holiday of from 1 to 14
days, followed by administration of Compound B for from 3 to 21
consecutive days. Suitably, Compound A is administered for from 3
to 21 consecutive days, followed by a drug holiday of from 3 to 14
days, followed by administration of Compound B for from 3 to 21
consecutive days. Suitably, Compound A is administered for 21
consecutive days, followed by an optional drug holiday, followed by
administration of Compound B for 14 consecutive days. Suitably,
Compound A is administered for 14 consecutive days, followed by a
drug holiday of from 1 to 14 days, followed by administration of
Compound B for 14 consecutive days. Suitably, Compound A is
administered for 7 consecutive days, followed by a drug holiday of
from 3 to 10 days, followed by administration of Compound B for 7
consecutive days. Suitably, Compound A is administered for 3
consecutive days, followed by a drug holiday of from 3 to 14 days,
followed by administration of Compound B for 7 consecutive days.
Suitably, Compound A is administered for 3 consecutive days,
followed by a drug holiday of from 3 to 10 days, followed by
administration of Compound B for 3 consecutive days.
[0127] It is understood that a "specified period" administration
and a "sequential" administration can be followed by repeat dosing
or can be followed by an alternate dosing protocol, and a drug
holiday may precede the repeat dosing or alternate dosing
protocol.
[0128] Suitably, the amount of Compound A (based on weight of
unsalted/unsolvated amount) administered as part of the combination
according to the present invention will be an amount selected from
about 0.125 mg to about 10 mg; suitably, the amount will be
selected from about 0.25 mg to about 9 mg; suitably, the amount
will be selected from about 0.25 mg to about 8 mg; suitably, the
amount will be selected from about 0.5 mg to about 8 mg; suitably,
the amount will be selected from about 0.5 mg to about 7 mg;
suitably, the amount will be selected from about 1 mg to about 7
mg; suitably, the amount will be about 5 mg. Accordingly, the
amount of Compound A administered as part of the combination
according to the present invention will be an amount selected from
about 0.125 mg to about 10 mg. For example, the amount of Compound
A administered as part of the combination according to the present
invention can be 0.125 mg, 0.25 mg, 0.5 mg, 0.75 mg, 1 mg, 1.5 mg,
2 mg, 2.5 mg, 3 mg, 3.5 mg, 4 mg, 4.5 mg, 5 mg, 5.5 mg, 6 mg, 6.5
mg, 7 mg, 7.5 mg, 8 mg, 8.5 mg, 9 mg, 9.5 mg, 10 mg.
[0129] Suitably, the amount of Compound B (based on weight of
unsalted/unsolvated amount) administered as part of the combination
according to the present invention will be an amount selected from
about 10 mg to about 600 mg. Suitably, the amount will be selected
from about 30 mg to about 300 mg; suitably, the amount will be
selected from about 30 mg to about 280 mg; suitably, the amount
will be selected from about 40 mg to about 260 mg; suitably, the
amount will be selected from about 60 mg to about 240 mg; suitably,
the amount will be selected from about 80 mg to about 220 mg;
suitably, the amount will be selected from about 90 mg to about 210
mg; suitably, the amount will be selected from about 100 mg to
about 200 mg, suitably, the amount will be selected from about 110
mg to about 190 mg, suitably, the amount will be selected from
about 120 mg to about 180 mg, suitably, the amount will be selected
from about 130 mg to about 170 mg, suitably, the amount will be
selected from about 140 mg to about 160 mg, suitably, the amount
will be 150 mg. Accordingly, the amount of Compound B administered
as part of the combination according to the present invention will
be an amount selected from about 10 mg to about 300 mg. For
example, the amount of Compound B administered as part of the
combination according to the present invention is suitably selected
from 10 mg, 20 mg, 30 mg, 40 mg, 50 mg, 60 mg, 70 mg, 80 mg, 85 mg,
90 mg, 95 mg, 100 mg, 105 mg, 110 mg, 115 mg, 120 mg, 125 mg, 130
mg, 135 mg, 140 mg, 145 mg, 150 mg, 155 mg, 160 mg, 165 mg, 170 mg,
175 mg, 180 mg, 185 mg, 190 mg, 195 mg, 200 mg, 205 mg, 210 mg, 215
mg, 220 mg, 225 mg, 230 mg, 235 mg, 240 mg, 245 mg, 250 mg, 255 mg,
260 mg, 265 mg, 270 mg, 275 mg, 280 mg, 285 mg, 290 mg, 295 mg and
300 mg. Suitably, the selected amount of Compound B is administered
from 1 to 4 times a day. Suitably, the selected amount of Compound
B is administered twice a day. Suitably, Compound B is administered
at an amount of 150 mg twice a day. Suitably, the selected amount
of Compound B is administered once a day.
[0130] As used herein, all amounts specified for Compound A and
Compound B are indicated as the amount of free or unsalted
compound.
Method of Treatment
[0131] The combinations of the invention, are believed to have
utility in disorders wherein the inhibition of MEK and/or B-Raf is
beneficial.
[0132] The present invention thus also provides a combination of
the invention, for use in therapy, particularly in the treatment of
disorders wherein the inhibition of MEK and/or B-Raf activity is
beneficial, particularly cancer.
[0133] A further aspect of the invention provides a method of
treatment of a disorder wherein to inhibition of MEK and/or B-Raf
is beneficial, comprising administering a combination of the
invention.
[0134] A further aspect of the present invention provides the use
of a combination of the invention in the manufacture of a
medicament for the treatment of a disorder wherein the inhibition
of MEK and/or B-Raf is beneficial.
[0135] Typically, the disorder is a cancer such that inhibition of
MEK and/or B-Raf has a beneficial effect. Examples of cancers that
are suitable for treatment with combination of the invention
include, but are limited to, both primary and metastatic forms of
head and neck, breast, lung, colon, ovary, and prostate cancers.
Suitably the cancer is selected from: brain (gliomas),
glioblastomas, astrocytomas, glioblastoma multiforme,
Bannayan-Zonana syndrome, Cowden disease, Lhermitte-Duclos disease,
breast, inflammatory breast cancer, Wilm's tumor, Ewing's sarcoma,
Rhabdomyosarcoma, ependymoma, medulloblastoma, colon, head and
neck, kidney, lung, liver, melanoma, ovarian, pancreatic, prostate,
sarcoma, osteosarcoma, giant cell tumor of bone, thyroid cancer,
lymphoblastic T cell leukemia, Chronic myelogenous leukemia,
Chronic lymphocytic leukemia, Hairy-cell leukemia, acute
lymphoblastic leukemia, acute myelogenous leukemia, AML, Chronic
neutrophilic leukemia, Acute lymphoblastic T cell leukemia,
plasmacytoma, Immunoblastic large cell leukemia, Mantle cell
leukemia, Multiple myeloma Megakaryoblastic leukemia, multiple
myeloma, acute megakaryocytic leukemia, promyelocytic leukemia,
Erythroleukemia, malignant lymphoma, hodgkins lymphoma,
non-hodgkins lymphoma, lymphoblastic T cell lymphoma, Burkitt's
lymphoma, follicular lymphoma, neuroblastoma, bladder cancer,
urothelial cancer, lung cancer, vulval cancer, cervical cancer,
endometrial cancer, renal cancer, mesothelioma, esophageal cancer,
salivary gland cancer, hepatocellular cancer, gastric cancer,
nasopharangeal cancer, buccal cancer, cancer of the mouth, GIST
(gastrointestinal stromal tumor) and testicular cancer.
[0136] Additionally, examples of a cancer to be treated include
Barret's adenocarcinoma; billiary tract carcinomas; breast cancer;
cervical cancer; cholangiocarcinoma; central nervous system tumors
including primary CNS tumors such as glioblastomas, astrocytomas
(e.g., glioblastoma multiforme) and ependymomas, and secondary CNS
tumors (i.e., metastases to the central nervous system of tumors
originating outside of the central nervous system); colorectal
cancer including large intestinal colon carcinoma; gastric cancer;
carcinoma of the head and neck including squamous cell carcinoma of
the head and neck; hematologic cancers including leukemias and
lymphomas such as acute lymphoblastic leukemia, acute myelogenous
leukemia (AML), myelodysplastic syndromes, chronic myelogenous
leukemia, Hodgkin's lymphoma, non-Hodgkin's lymphoma,
megakaryoblastic leukemia, multiple myeloma and erythroleukemia;
hepatocellular carcinoma; lung cancer including small cell lung
cancer and non-small cell lung cancer; ovarian cancer; endometrial
cancer; pancreatic cancer; pituitary adenoma; prostate cancer;
renal cancer; sarcoma; skin cancers including melanomas; and
thyroid cancers.
[0137] Suitably, the present invention relates to a method for
treating or lessening the severity of a cancer selected from: brain
(gliomas), glioblastomas, Bannayan-Zonana syndrome, Cowden disease,
Lhermitte-Duclos disease, breast, colon, head and neck, kidney,
lung, liver, melanoma, ovarian, pancreatic, prostate, sarcoma and
thyroid.
[0138] Suitably, the present invention relates to a method for
treating or lessening the severity of a cancer selected from
ovarian, breast, pancreatic and prostate.
[0139] The combination of the invention may be used alone or in
combination with one or more other therapeutic agents. The
invention thus provides in a further aspect a further combination
comprising a combination of the invention with a further
therapeutic agent or agents, compositions and medicaments
comprising the combination and use of the further combination,
compositions and medicaments in therapy, in particular in the
treatment of diseases susceptible to inhibition of MEK and/or
kinase B.
[0140] In the embodiment, the combination of the invention may be
employed with other therapeutic methods of cancer treatment. In
particular, in anti-neoplastic therapy, combination therapy with
other chemotherapeutic, hormonal, antibody agents as well as
surgical and/or radiation treatments other than those mentioned
above are envisaged. Combination therapies according to the present
invention thus include the administration of Compound A and
Compound B as well as optional use of other therapeutic agents
including other anti-neoplastic agents. Such combination of agents
may be administered together or separately and, when administered
separately this may occur simultaneously or sequentially in any
order, both close and remote in time. In one embodiment, the
pharmaceutical combination includes Compound A and Compound B, and
optionally at least one additional anti-neoplastic agent.
[0141] As indicated, therapeutically effective amounts of Compound
A and Compound B are discussed above. The therapeutically effective
amount of the further therapeutic agents of the present invention
will depend upon a number of factors including, for example, the
age and weight of the mammal, the precise condition requiring
treatment, the severity of the condition, the nature of the
formulation, and the route of administration. Ultimately, the
therapeutically effective amount will be at the discretion of the
attendant physician or veterinarian. The relative timings of
administration will be selected in order to achieve the desired
combined therapeutic effect.
[0142] In one embodiment, the further anti-cancer therapy is
surgical and/or radiotherapy.
[0143] In one embodiment, the further anti-cancer therapy is at
least one additional anti-neoplastic agent.
[0144] Any anti-neoplastic agent that has activity versus a
susceptible tumor being treated may be utilized in the combination.
Typical anti-neoplastic agents useful include, but are not limited
to, anti-microtubule agents such as diterpenoids and vinca
alkaloids; platinum coordination complexes; alkylating agents such
as nitrogen mustards, oxazaphosphorines, alkylsulfonates,
nitrosoureas, and triazenes; antibiotic agents such as
anthracyclins, actinomycins and bleomycins; topoisomerase II
inhibitors such as epipodophyllotoxins; antimetabolites such as
purine and pyrimidine analogues and anti-folate compounds;
topoisomerase I inhibitors such as camptothecins; hormones and
hormonal analogues; signal transduction pathway inhibitors;
non-receptor tyrosine angiogenesis inhibitors; immunotherapeutic
agents; proapoptotic agents; and cell cycle signaling
inhibitors.
[0145] Anti-microtubule or anti-mitotic agents: Anti-microtubule or
anti-mitotic agents are phase specific agents active against the
microtubules of tumor cells during M or the mitosis phase of the
cell cycle. Examples of anti-microtubule agents include, but are
not limited to, diterpenoids and vinca alkaloids.
[0146] Diterpenoids, which are derived from natural sources, are
phase specific anti-cancer agents that operate at the G.sub.2/M
phases of the cell cycle. It is believed that the diterpenoids
stabilize the .beta.-tubulin subunit of the microtubules, by
binding with this protein. Disassembly of the protein appears then
to be inhibited with mitosis being arrested and cell death
following. Examples of diterpenoids include, but are not limited
to, paclitaxel and its analog docetaxel.
[0147] Paclitaxel,
5.beta.,20-epoxy-1,2.alpha.,4,7.beta.,10.beta.,13.alpha.-hexa-hydroxytax--
11-en-9-one 4,10-diacetate 2-benzoate 13-ester with
(2R,3S)--N-benzoyl-3-phenylisoserine; is a natural diterpene
product isolated from the Pacific yew tree Taxus brevifolia and is
commercially available as an injectable solution TAXOL.RTM.. It is
a member of the taxane family of terpenes. Paclitaxel has been
approved for clinical use in the treatment of refractory ovarian
cancer in the United States (Markman et al., Yale Journal of
Biology and Medicine, 64:583, 1991; McGuire et al., Ann. Intern,
Med., 111:273, 1989) and for the treatment of breast cancer (Holmes
et al., J. Nat. Cancer Inst., 83:1797, 1991.) It is a potential
candidate for treatment of neoplasms in the skin (Einzig et. al.,
Proc. Am. Soc. Clin. Oncol., 20:46) and head and neck carcinomas
(Forastire et. al., Sem. Oncol., 20:56, 1990). The compound also
shows potential for the treatment of polycystic kidney disease (Woo
et. al., Nature, 368:750. 1994), lung cancer and malaria. Treatment
of patients with paclitaxel results in bone marrow suppression
(multiple cell lineages, Ignoff, R. J. et. al, Cancer Chemotherapy
Pocket Guide, 1998) related to the duration of dosing above a
threshold concentration (50 nM) (Kearns, C. M. et. al., Seminars in
Oncology, 3(6) p. 16-23, 1995).
[0148] Docetaxel, (2R,3S)-- N-carboxy-3-phenylisoserine,
N-tert-butyl ester, 13-ester with
5.beta.-20-epoxy-1,2.alpha.,4,7.beta.,10.beta.,13.alpha.-hexahydroxytax-1-
1-en-9-one 4-acetate 2-benzoate, trihydrate; is commercially
available as an injectable solution as TAXOTERE.RTM.. Docetaxel is
indicated for the treatment of breast cancer. Docetaxel is a
semisynthetic derivative of paclitaxel q.v., prepared using a
natural precursor, 10-deacetyl-baccatin III, extracted from the
needle of the European Yew tree.
[0149] Vinca alkaloids are phase specific anti-neoplastic agents
derived from the periwinkle plant. Vinca alkaloids act at the M
phase (mitosis) of the cell cycle by binding specifically to
tubulin. Consequently, the bound tubulin molecule is unable to
polymerize into microtubules. Mitosis is believed to be arrested in
metaphase with cell death following. Examples of vinca alkaloids
include, but are not limited to, vinblastine, vincristine, and
vinorelbine.
[0150] Vinblastine, vincaleukoblastine sulfate, is commercially
available as VELBAN.RTM. as an injectable solution. Although, it
has possible indication as a second line therapy of various solid
tumors, it is primarily indicated in the treatment of testicular
cancer and various lymphomas including Hodgkin's Disease; and
lymphocytic and histiocytic lymphomas. Myelosuppression is the dose
limiting side effect of vinblastine.
[0151] Vincristine, vincaleukoblastine, 22-oxo-, sulfate, is
commercially available as ONCOVIN.RTM. as an injectable solution.
Vincristine is indicated for the treatment of acute leukemias and
has also found use in treatment regimens for Hodgkin's and
non-Hodgkin's malignant lymphomas. Alopecia and neurologic effects
are the most common side effect of vincristine and to a lesser
extent myelosupression and gastrointestinal mucositis effects
occur.
[0152] Vinorelbine,
3',4'-didehydro-4'-deoxy-C'-norvincaleukoblastine[R--(R*,R*)-2,3-dihydrox-
ybutanedioate (1:2)(salt)], commercially available as an injectable
solution of vinorelbine tartrate (NAVELBINE.RTM.), is a
semisynthetic vinca alkaloid. Vinorelbine is indicated as a single
agent or in combination with other chemotherapeutic agents, such as
cisplatin, in the treatment of various solid tumors, particularly
non-small cell lung, advanced breast, and hormone refractory
prostate cancers. Myelosuppression is the most common dose limiting
side effect of vinorelbine.
[0153] Platinum coordination complexes: Platinum coordination
complexes are non-phase specific anti-cancer agents, which are
interactive with DNA. The platinum complexes enter tumor cells,
undergo, aquation and form intra- and interstrand crosslinks with
DNA causing adverse biological effects to the tumor. Examples of
platinum coordination complexes include, but are not limited to,
oxaliplatin, cisplatin and carboplatin.
[0154] Cisplatin, cis-diamminedichloroplatinum, is commercially
available as PLATINOL.RTM. as an injectable solution. Cisplatin is
primarily indicated in the treatment of metastatic testicular and
ovarian cancer and advanced bladder cancer.
[0155] Carboplatin, platinum,
diammine[1,1-cyclobutane-dicarboxylate(2-)-O,O'], is commercially
available as PARAPLATIN.RTM. as an injectable solution. Carboplatin
is primarily indicated in the first and second line treatment of
advanced ovarian carcinoma.
[0156] Alkylating agents: Alkylating agents are non-phase
anti-cancer specific agents and strong electrophiles. Typically,
alkylating agents form covalent linkages, by alkylation, to DNA
through nucleophilic moieties of the DNA molecule such as
phosphate, amino, sulfhydryl, hydroxyl, carboxyl, and imidazole
groups. Such alkylation disrupts nucleic acid function leading to
cell death. Examples of alkylating agents include, but are not
limited to, nitrogen mustards such as cyclophosphamide, melphalan,
and chlorambucil; alkyl sulfonates such as busulfan; nitrosoureas
such as carmustine; and triazenes such as dacarbazine.
[0157] Cyclophosphamide,
2-[bis(2-chloroethyl)amino]tetrahydro-2H-1,3,2-oxazaphosphorine
2-oxide monohydrate, is commercially available as an injectable
solution or tablets as CYTOXAN.RTM.. Cyclophosphamide is indicated
as a single agent or in combination with other chemotherapeutic
agents, in the treatment of malignant lymphomas, multiple myeloma,
and leukemias.
[0158] Melphalan, 4-[bis(2-chloroethyl)amino]-L-phenylalanine, is
commercially available as an injectable solution or tablets as
ALKERAN.RTM.. Melphalan is indicated for the palliative treatment
of multiple myeloma and non-resectable epithelial carcinoma of the
ovary. Bone marrow suppression is the most common dose limiting
side effect of melphalan.
[0159] Chlorambucil, 4-[bis(2-chloroethyl)amino]benzenebutanoic
acid, is commercially available as LEUKERAN.RTM. tablets.
Chlorambucil is indicated for the palliative treatment of chronic
lymphatic leukemia, and malignant lymphomas such as lymphosarcoma,
giant follicular lymphoma, and Hodgkin's disease.
[0160] Busulfan, 1,4-butanediol dimethanesulfonate, is commercially
available as MYLERAN.RTM. TABLETS. Busulfan is indicated for the
palliative treatment of chronic myelogenous leukemia.
[0161] Carmustine, 1,3-[bis(2-chloroethyl)-1-nitrosourea, is
commercially available as single vials of lyophilized material as
BiCNU.RTM.. Carmustine is indicated for the palliative treatment as
a single agent or in combination with other agents for brain
tumors, multiple myeloma, Hodgkin's disease, and non-Hodgkin's
lymphomas.
[0162] Dacarbazine,
5-(3,3-dimethyl-1-triazeno)-imidazole-4-carboxamide, is
commercially available as single vials of material as
DTIC-Dome.RTM.. Dacarbazine is indicated for the treatment of
metastatic malignant melanoma and in combination with other agents
for the second line treatment of Hodgkin's Disease.
[0163] Antibiotic anti-neoplastics: Antibiotic anti-neoplastics are
non-phase specific agents, which bind or intercalate with DNA.
Typically, such action results in stable DNA complexes or strand
breakage, which disrupts ordinary function of the nucleic acids
leading to cell death. Examples of antibiotic anti-neoplastic
agents include, but are not limited to, actinomycins such as
dactinomycin, anthrocyclins such as daunorubicin and doxorubicin;
and bleomycins.
[0164] Dactinomycin, also know as Actinomycin D, is commercially
available in injectable form as COSMEGEN.RTM.. Dactinomycin is
indicated for the treatment of Wilm's tumor and
rhabdomyosarcoma.
[0165] Daunorubicin,
(8S-cis-)-8-acetyl-10-[(3-amino-2,3,6-trideoxy-.alpha.-L-lyxo-hexopyranos-
yl)oxy]-7,8,9,10-tetrahydro-6,8,11-trihydroxy-1-methoxy-5,12
naphthacenedione hydrochloride, is commercially available as a
liposomal injectable form as DAUNOXOME.RTM. or as an injectable as
CERUBIDINE.RTM.. Daunorubicin is indicated for remission induction
in the treatment of acute nonlymphocytic leukemia and advanced HIV
associated Kaposi's sarcoma.
[0166] Doxorubicin,
(8S,10S)-10-[(3-amino-2,3,6-trideoxy-.alpha.-L-lyxo-hexopyranosyl)oxy]-8--
glycoloyl, 7,8,9,10-tetrahydro-6,8,11-trihydroxy-1-methoxy-5,12
naphthacenedione hydrochloride, is commercially available as an
injectable form as RUBEX.RTM. or ADRIAMYCIN RDF.RTM.. Doxorubicin
is primarily indicated for the treatment of acute lymphoblastic
leukemia and acute myeloblastic leukemia, but is also a useful
component in the treatment of some solid tumors and lymphomas.
[0167] Bleomycin, a mixture of cytotoxic glycopeptide antibiotics
isolated from a strain of Streptomyces verticillus, is commercially
available as BLENOXANE.RTM.. Bleomycin is indicated as a palliative
treatment, as a single agent or in combination with other agents,
of squamous cell carcinoma, lymphomas, and testicular
carcinomas.
[0168] Topoisomerase II inhibitors: Topoisomerase II inhibitors
include, but are not limited to, epipodophyllotoxins.
[0169] Epipodophyllotoxins are phase specific anti-neoplastic
agents derived from the mandrake plant. Epipodophyllotoxins
typically affect cells in the S and G.sub.2 phases of the cell
cycle by forming a ternary complex with topoisomerase II and DNA
causing DNA strand breaks. The strand breaks accumulate and cell
death follows. Examples of epipodophyllotoxins include, but are not
limited to, etoposide and teniposide.
[0170] Etoposide, 4'-demethyl-epipodophyllotoxin
9[4,6-0-(R)-ethylidene-.beta.-D-glucopyranoside], is commercially
available as an injectable solution or capsules as VePESID.RTM. and
is commonly known as VP-16. Etoposide is indicated as a single
agent or in combination with other chemotherapy agents in the
treatment of testicular and non-small cell lung cancers.
[0171] Teniposide, 4'-demethyl-epipodophyllotoxin
9[4,6-0-(R)-thenylidene-.beta.-D-glucopyranoside], is commercially
available as an injectable solution as VUMON.RTM. and is commonly
known as VM-26. Teniposide is indicated as a single agent or in
combination with other chemotherapy agents in the treatment of
acute leukemia in children.
[0172] Antimetabolite neoplastic agents: Antimetabolite neoplastic
agents are phase specific anti-neoplastic agents that act at S
phase (DNA synthesis) of the cell cycle by inhibiting DNA synthesis
or by inhibiting purine or pyrimidine base synthesis and thereby
limiting DNA synthesis. Consequently, S phase does not proceed and
cell death follows. Examples of antimetabolite anti-neoplastic
agents include, but are not limited to, fluorouracil, methotrexate,
cytarabine, mecaptopurine, thioguanine, and gemcitabine.
[0173] 5-fluorouracil, 5-fluoro-2,4-(1H,3H) pyrimidinedione, is
commercially available as fluorouracil. Administration of
5-fluorouracil leads to inhibition of thymidylate synthesis and is
also incorporated into both RNA and DNA. The result typically is
cell death. 5-fluorouracil is indicated as a single agent or in
combination with other chemotherapy agents in the treatment of
carcinomas of the breast, colon, rectum, stomach and pancreas.
Other fluoropyrimidine analogs include 5-fluoro deoxyuridine
(floxuridine) and 5-fluorodeoxyuridine monophosphate.
[0174] Cytarabine,
4-amino-1-.beta.-D-arabinofuranosyl-2(1H)-pyrimidinone, is
commercially available as CYTOSAR-U.RTM. and is commonly known as
Ara-C. It is believed that cytarabine exhibits cell phase
specificity at S-phase by inhibiting DNA chain elongation by
terminal incorporation of cytarabine into the growing DNA chain.
Cytarabine is indicated as a single agent or in combination with
other chemotherapy agents in the treatment of acute leukemia. Other
cytidine analogs include 5-azacytidine and
2',2'-difluorodeoxycytidine (gemcitabine).
[0175] Mercaptopurine, 1,7-dihydro-6H-purine-6-thione monohydrate,
is commercially available as PURINETHOL.RTM.. Mercaptopurine
exhibits cell phase specificity at S-phase by inhibiting DNA
synthesis by an as of yet unspecified mechanism. Mercaptopurine is
indicated as a single agent or in combination with other
chemotherapy agents in the treatment of acute leukemia. A useful
mercaptopurine analog is azathioprine.
[0176] Thioguanine, 2-amino-1,7-dihydro-6H-purine-6-thione, is
commercially available as TABLOID.RTM.. Thioguanine exhibits cell
phase specificity at S-phase by inhibiting DNA synthesis by an as
of yet unspecified mechanism. Thioguanine is indicated as a single
agent or in combination with other chemotherapy agents in the
treatment of acute leukemia. Other purine analogs include
pentostatin, erythrohydroxynonyladenine, fludarabine phosphate, and
cladribine.
[0177] Gemcitabine, 2'-deoxy-2',2'-difluorocytidine
monohydrochloride (0-isomer), is commercially available as
GEMZAR.RTM.. Gemcitabine exhibits cell phase specificity at S-phase
and by blocking progression of cells through the G1/S boundary.
Gemcitabine is indicated in combination with cisplatin in the
treatment of locally advanced non-small cell lung cancer and alone
in the treatment of locally advanced pancreatic cancer.
[0178] Methotrexate,
N-[4[(2,4-diamino-6-pteridinyl)methyl]methylamino]benzoyl]-L-glutamic
acid, is commercially available as methotrexate sodium.
Methotrexate exhibits cell phase effects specifically at S-phase by
inhibiting DNA synthesis, repair and/or replication through the
inhibition of dyhydrofolic acid reductase which is required for
synthesis of purine nucleotides and thymidylate. Methotrexate is
indicated as a single agent or in combination with other
chemotherapy agents in the treatment of choriocarcinoma, meningeal
leukemia, non-Hodgkin's lymphoma, and carcinomas of the breast,
head, neck, ovary and bladder.
[0179] Topoisomerase I inhibitors: Camptothecins, including,
camptothecin and camptothecin derivatives are available or under
development as Topoisomerase I inhibitors. Camptothecins cytotoxic
activity is believed to be related to its Topoisomerase I
inhibitory activity. Examples of camptothecins include, but are not
limited to irinotecan, topotecan, and the various optical forms of
7-(4-methylpiperazino-methylene)-10,11-ethylenedioxy-20-camptothecin
described below.
[0180] Irinotecan HCl,
(4S)-4,11-diethyl-4-hydroxy-9-[(4-piperidinopiperidino)
carbonyloxy]-1H-pyrano[3',4',6,7]indolizino[1,2-b]quinoline-3,14(4H,12H)--
dione hydrochloride, is commercially available as the injectable
solution CAMPTOSAR.RTM.. Irinotecan is a derivative of camptothecin
which binds, along with its active metabolite SN-38, to the
topoisomerase I-DNA complex. It is believed that cytotoxicity
occurs as a result of irreparable double strand breaks caused by
interaction of the topoisomerase I:DNA:irintecan or SN-38 ternary
complex with replication enzymes. Irinotecan is indicated for
treatment of metastatic cancer of the colon or rectum.
[0181] Topotecan HCl,
(S)-10-[(dimethylamino)methyl]-4-ethyl-4,9-dihydroxy-1H-pyrano[3',4',6,7]-
indolizino[1,2-b]quinoline-3,14-(4H,12H)-dione monohydrochloride,
is commercially available as the injectable solution HYCAMTIN.RTM..
Topotecan is a derivative of camptothecin which binds to the
topoisomerase I-DNA complex and prevents religation of singles
strand breaks caused by Topoisomerase I in response to torsional
strain of the DNA molecule. Topotecan is indicated for second line
treatment of metastatic carcinoma of the ovary and small cell lung
cancer.
[0182] Hormones and hormonal analogues: Hormones and hormonal
analogues are useful compounds for treating cancers in which there
is a relationship between the hormone(s) and growth and/or lack of
growth of the cancer. Examples of hormones and hormonal analogues
useful in cancer treatment include, but are not limited to,
adrenocorticosteroids such as prednisone and prednisolone which are
useful in the treatment of malignant lymphoma and acute leukemia in
children; aminoglutethimide and other aromatase inhibitors such as
anastrozole, letrazole, vorazole, and exemestane useful in the
treatment of adrenocortical carcinoma and hormone dependent breast
carcinoma containing estrogen receptors; progestrins such as
megestrol acetate useful in the treatment of hormone dependent
breast cancer and endometrial carcinoma; estrogens, androgens, and
anti-androgens such as flutamide, nilutamide, bicalutamide,
cyproterone acetate and 5.alpha.-reductases such as finasteride and
dutasteride, useful in the treatment of prostatic carcinoma and
benign prostatic hypertrophy; anti-estrogens such as tamoxifen,
toremifene, raloxifene, droloxifene, iodoxyfene, as well as
selective estrogen receptor modulators (SERMS) such those described
in U.S. Pat. Nos. 5,681,835, 5,877,219, and 6,207,716, useful in
the treatment of hormone dependent breast carcinoma and other
susceptible cancers; and gonadotropin-releasing hormone (GnRH) and
analogues thereof which stimulate the release of leutinizing
hormone (LH) and/or follicle stimulating hormone (FSH) for the
treatment prostatic carcinoma, for instance, LHRH agonists and
antagagonists such as goserelin acetate and luprolide.
[0183] Signal transduction pathway inhibitors: Signal transduction
pathway inhibitors are those inhibitors, which block or inhibit a
chemical process which evokes an intracellular change. As used
herein this change is cell proliferation or differentiation. Signal
tranduction inhibitors useful in the present invention include
inhibitors of receptor tyrosine kinases, non-receptor tyrosine
kinases, SH2/SH3domain blockers, serine/threonine kinases,
phosphotidyl inositol-3 kinases, myo-inositol signaling, and Ras
oncogenes.
[0184] Several protein tyrosine kinases catalyse the
phosphorylation of specific tyrosyl residues in various proteins
involved in the regulation of cell growth. Such protein tyrosine
kinases can be broadly classified as receptor or non-receptor
kinases.
[0185] Receptor tyrosine kinases are transmembrane proteins having
an extracellular ligand binding domain, a transmembrane domain, and
a tyrosine kinase domain. Receptor tyrosine kinases are involved in
the regulation of cell growth and are generally termed growth
factor receptors. Inappropriate or uncontrolled activation of many
of these kinases, i.e. aberrant kinase growth factor receptor
activity, for example by over-expression or mutation, has been
shown to result in uncontrolled cell growth. Accordingly, the
aberrant activity of such kinases has been linked to malignant
tissue growth. Consequently, inhibitors of such kinases could
provide cancer treatment methods. Growth factor receptors include,
for example, epidermal growth factor receptor (EGFr), platelet
derived growth factor receptor (PDGFr), erbB2, erbB4, ret, vascular
endothelial growth factor receptor (VEGFr), tyrosine kinase with
immunoglobulin-like and epidermal growth factor homology domains
(TIE-2), insulin growth factor--I (IGFI) receptor, macrophage
colony stimulating factor (cfms), BTK, ckit, cmet, fibroblast
growth factor (FGF) receptors, Trk receptors (TrkA, TrkB, and
TrkC), ephrin (eph) receptors, and the RET protooncogene. Several
inhibitors of growth receptors are under development and include
ligand antagonists, antibodies, tyrosine kinase inhibitors and
anti-sense oligonucleotides. Growth factor receptors and agents
that inhibit growth factor receptor function are described, for
instance, in Kath, John C., Exp. Opin. Ther. Patents (2000)
10(6):803-818; Shawver et al DDT Vol 2, No. 2 Feb. 1997; and Lofts,
F. J. et al, "Growth factor receptors as targets", New Molecular
Targets for Cancer Chemotherapy, ed. Workman, Paul and Kerr, David,
CRC press 1994, London.
[0186] Tyrosine kinases, which are not growth factor receptor
kinases are termed non-receptor tyrosine kinases. Non-receptor
tyrosine kinases useful in the present invention, which are targets
or potential targets of anti-cancer drugs, include cSrc, Lck, Fyn,
Yes, Jak, cAbl, FAK (Focal adhesion kinase), Brutons tyrosine
kinase, and Bcr-Abl. Such non-receptor kinases and agents which
inhibit non-receptor tyrosine kinase function are described in
Sinh, S. and Corey, S. J., (1999) Journal of Hematotherapy and Stem
Cell Research 8 (5): 465-80; and Bolen, J. B., Brugge, J. S.,
(1997) Annual review of Immunology. 15: 371-404.
[0187] SH2/SH3 domain blockers are agents that disrupt SH2 or SH3
domain binding in a variety of enzymes or adaptor proteins
including, PI3-K p85 subunit, Src family kinases, adaptor molecules
(Shc, Crk, Nck, Grb2) and Ras-GAP. SH2/SH3 domains as targets for
anti-cancer drugs are discussed in Smithgall, T. E. (1995), Journal
of Pharmacological and Toxicological Methods. 34(3) 125-32.
[0188] Inhibitors of Serine/Threonine Kinases including MAP kinase
cascade blockers which include blockers of Raf kinases (rafk),
Mitogen or Extracellular Regulated Kinase (MEKs), and Extracellular
Regulated Kinases (ERKs); and Protein kinase C family member
blockers including blockers of PKCs (alpha, beta, gamma, epsilon,
mu, lambda, iota, zeta). IkB kinase family (IKKa, IKKb), PKB family
kinases, akt kinase family members, and TGF beta receptor kinases.
Such Serine/Threonine kinases and inhibitors thereof are described
in Yamamoto, T., Taya, S., Kaibuchi, K., (1999), Journal of
Biochemistry. 126 (5) 799-803; Brodt, P, Samani, A., and Navab, R.
(2000), Biochemical Pharmacology, 60. 1101-1107; Massague, J.,
Weis-Garcia, F. (1996) Cancer Surveys. 27:41-64; Philip, P. A., and
Harris, A. L. (1995), Cancer Treatment and Research. 78: 3-27,
Lackey, K. et al Bioorganic and Medicinal Chemistry Letters, (10),
2000, 223-226; U.S. Pat. No. 6,268,391; and Martinez-lacaci, L., et
al, Int. J. Cancer (2000), 88(1), 44-52.
[0189] Inhibitors of Phosphotidyl inositol-3 Kinase family members
including blockers of PI3-kinase, ATM, DNA-PK, and Ku are also
useful in the present invention. Such kinases are discussed in
Abraham, R. T. (1996), Current Opinion in Immunology. 8 (3) 412-8;
Canman, C. E., Lim, D. S. (1998), Oncogene 17 (25) 3301-3308;
Jackson, S. P. (1997), International Journal of Biochemistry and
Cell Biology. 29 (7):935-8; and Zhong, H. et al, Cancer res, (2000)
60(6), 1541-1545.
[0190] Also useful in the present invention are Myo-inositol
signaling inhibitors such as phospholipase C blockers and
Myoinositol analogues. Such signal inhibitors are described in
Powis, G., and Kozikowski A., (1994) New Molecular Targets for
Cancer Chemotherapy ed., Paul Workman and David Kerr, CRC press
1994, London.
[0191] Another group of signal transduction pathway inhibitors are
inhibitors of Ras Oncogene. Such inhibitors include inhibitors of
farnesyltransferase, geranyl-geranyl transferase, and CAAX
proteases as well as anti-sense oligonucleotides, ribozymes and
immunotherapy. Such inhibitors have been shown to block ras
activation in cells containing wild type mutant ras, thereby acting
as antiproliferation agents. Ras oncogene inhibition is discussed
in Scharovsky, O. G., Rozados, V. R., Gervasoni, S. I. Matar, P.
(2000), Journal of Biomedical Science. 7(4) 292-8; Ashby, M. N.
(1998), Current Opinion in Lipidology. 9 (2) 99-102; and BioChim.
Biophys. Acta, (19899) 1423(3):19-30.
[0192] As mentioned above, antibody antagonists to receptor kinase
ligand binding may also serve as signal transduction inhibitors.
This group of signal transduction pathway inhibitors includes the
use of humanized antibodies to the extracellular ligand binding
domain of receptor tyrosine kinases. For example Imclone C225 EGFR
specific antibody (see Green, M. C. et al, Monoclonal Antibody
Therapy for Solid Tumors, Cancer Treat. Rev., (2000), 26(4),
269-286); Herceptin.RTM. erbB2 antibody (see Tyrosine Kinase
Signalling in Breast cancer:erbB Family Receptor Tyrosine Kinases,
Breast cancer Res., 2000, 2(3), 176-183); and 2CB VEGFR2 specific
antibody (see Brekken, R. A. et al, Selective Inhibition of VEGFR2
Activity by a monoclonal Anti-VEGF antibody blocks tumor growth in
mice, Cancer Res. (2000) 60, 5117-5124).
[0193] Anti-angiogenic agents: Anti-angiogenic agents including
non-receptorMEKngiogenesis inhibitors may also be useful.
Anti-angiogenic agents such as those which inhibit the effects of
vascular edothelial growth factor, (for example the anti-vascular
endothelial cell growth factor antibody bevacizumab [Avastin.TM.],
and compounds that work by other mechanisms (for example linomide,
inhibitors of integrin .alpha.v.beta.3 function, endostatin and
angiostatin);
[0194] Immunotherapeutic agents: Agents used in immunotherapeutic
regimens may also be useful in combination with the compounds of
formula (I). Immunotherapy approaches, including for example
ex-vivo and in-vivo approaches to increase the immunogenecity of
patient tumour cells, such as transfection with cytokines such as
interleukin 2, interleukin 4 or granulocyte-macrophage colony
stimulating factor, approaches to decrease T-cell anergy,
approaches using transfected immune cells such as
cytokine-transfected dendritic cells, approaches using
cytokine-transfected tumour cell lines and approaches using
anti-idiotypic antibodies
[0195] Proapoptotoc agents: Agents used in proapoptotic regimens
(e.g., bcl-2 antisense oligonucleotides) may also be used in the
combination of the present invention.
[0196] Cell cycle signalling inhibitors: Cell cycle signalling
inhibitors inhibit molecules involved in the control of the cell
cycle. A family of protein kinases called cyclin dependent kinases
(CDKs) and their interaction with a family of proteins termed
cyclins controls progression through the eukaryotic cell cycle. The
coordinate activation and inactivation of different cyclin/CDK
complexes is necessary for normal progression through the cell
cycle. Several inhibitors of cell cycle signalling are under
development. For instance, examples of cyclin dependent kinases,
including CDK2, CDK4, and CDK6 and inhibitors for the same are
described in, for instance, Rosania et al, Exp. Opin. Ther. Patents
(2000) 10(2):215-230.
[0197] In one embodiment, the combination of the present invention
comprises a compound of formula I or a salt or solvate thereof and
at least one anti-neoplastic agent selected from anti-microtubule
agents, platinum coordination complexes, alkylating agents,
antibiotic agents, topoisomerase II inhibitors, antimetabolites,
topoisomerase I inhibitors, hormones and hormonal analogues, signal
transduction pathway inhibitors, non-receptor tyrosine
MEKngiogenesis inhibitors, immunotherapeutic agents, proapoptotic
agents, and cell cycle signaling inhibitors.
[0198] In one embodiment, the combination of the present invention
comprises a compound of formula I or a salt or solvate thereof and
at least one anti-neoplastic agent which is an anti-microtubule
agent selected from diterpenoids and vinca alkaloids.
[0199] In a further embodiment, the at least one anti-neoplastic
agent agent is a diterpenoid.
[0200] In a further embodiment, the at least one anti-neoplastic
agent is a vinca alkaloid.
[0201] In one embodiment, the combination of the present invention
comprises a compound of formula I or a salt or solvate thereof and
at least one anti-neoplastic agent, which is a platinum
coordination complex.
[0202] In a further embodiment, the at least one anti-neoplastic
agent is paclitaxel, carboplatin, or vinorelbine.
[0203] In a further embodiment, the at least one anti-neoplastic
agent is carboplatin.
[0204] In a further embodiment, the at least one anti-neoplastic
agent is vinorelbine.
[0205] In a further embodiment, the at least one anti-neoplastic
agent is paclitaxel.
[0206] In one embodiment, the combination of the present invention
comprises a compound of formula I and salts or solvates thereof and
at least one anti-neoplastic agent which is a signal transduction
pathway inhibitor.
[0207] In a further embodiment the signal transduction pathway
inhibitor is an inhibitor of a growth factor receptor kinase
VEGFR2, TIE2, PDGFR, BTK, erbB2, EGFr, IGFR-1, TrkA, TrkB, TrkC, or
c-fms.
[0208] In a further embodiment the signal transduction pathway
inhibitor is an inhibitor of a serine/threonine kinase rafk, akt,
or PKC-zeta.
[0209] In a further embodiment the signal transduction pathway
inhibitor is an inhibitor of a non-receptor tyrosine kinase
selected from the src family of kinases.
[0210] In a further embodiment the signal transduction pathway
inhibitor is an inhibitor of c-src.
[0211] In a further embodiment the signal transduction pathway
inhibitor is an inhibitor of Ras oncogene selected from inhibitors
of farnesyl transferase and geranylgeranyl transferase.
[0212] In a further embodiment the signal transduction pathway
inhibitor is an inhibitor of a serine/threonine kinase selected
from the group consisting of PI3K.
[0213] In a further embodiment the signal transduction pathway
inhibitor is a dual EGFr/erbB2 inhibitor, for example
N-{3-Chloro-4-[(3-fluorobenzyl)oxy]phenyl}-6-[5-({[2-(methanesulphonyl)et-
hyl]amino}methyl)-2-furyl]-4-quinazolinamine (structure below):
##STR00023##
In one embodiment, the combination of the present invention
comprises a compound of formula I or a salt or solvate thereof and
at least one anti-neoplastic agent which is a cell cycle signaling
inhibitor.
[0214] In further embodiment, cell cycle signaling inhibitor is an
inhibitor of CDK2, CDK4 or CDK6.
[0215] In one embodiment the mammal in the methods and uses of the
present invention is a human.
[0216] Suitably, the present invention relates to a method of
treating or lessening the severity of a cancer that is either wild
type or mutant for each of Raf, Ras, MEK, and PI3K/Pten. This
includes but is not limited to patients having cancers that are
mutant for RAF, wild type for RAS, wild type for MEK, and wild type
for PI3K/PTEN; mutant for RAF, mutant for RAS, wild type for MEK,
and wild type for PI3K/PTEN; mutant for RAF, mutant for RAS, mutant
for MEK, and wild type for PI3K/PTEN; and mutant for RAF, wild type
for RAS, mutant for MEK, and wild type PI3K/PTEN.
[0217] The term "wild type" as is understood in the art refers to a
polypeptide or polynucleotide sequence that occurs in a native
population without genetic modification. As is also understood in
the art, a "mutant" includes a polypeptide or polynucleotide
sequence having at least one modification to an amino acid or
nucleic acid compared to the corresponding amino acid or nucleic
acid found in a wild type polypeptide or polynucleotide,
respectively. Included in the term mutant is Single Nucleotide
Polymorphism (SNP) where a single base pair distinction exists in
the sequence of a nucleic acid strand compared to the most
prevalently found (wild type) nucleic acid strand.
[0218] Cancers that are either wild type or mutant for Raf, Ras,
MEK, or mutant for PI3K/Pten are identified by known methods. For
example, wild type or mutant tumor cells can be identified by DNA
amplification and sequencing techniques, DNA and RNA detection
techniques, including, but not limited to Northern and Southern
blot, respectively, and/or various biochip and array technologies.
Wild type and mutant polypeptides can be detected by a variety of
techniques including, but not limited to immunodiagnostic
techniques such as ELISA, Western blot or immunocyto chemistry.
Suitably, Pyrophosphorolysis-activated polymerization (PAP) and/or
PCR methods may be used. Liu, Q et al; Human Mutation 23:426-436
(2004).
[0219] The following examples are intended for illustration only
and are not intended to limit the scope of the invention in any
way.
EXAMPLES
Example 1
Capsule Composition
[0220] An oral dosage form for administering a combination of the
present invention is produced by filing a standard two piece hard
gelatin capsule with the ingredients in the proportions shown in
Table A, below.
TABLE-US-00001 TABLE A INGREDIENTS AMOUNTS
N-{3-[3-cyclopropyl-5-(2-fluoro-4-iodo- 5 mg
phenylamino)6,8-dimethyl-2,4,7-trioxo-3,4,6,7-
tetrahydro-2H-pyrido[4,3-d]pyrimidin-1- yl]phenyl}acetamide
hydrochloride (Compound A)
N-{3-[5-(2-Amino-4-pyrimidinyl)-2-(1,1-dimethylethyl)- 100 mg
1,3-thiazol-4-yl]-2-fluorophenyl}-2,6- difluorobenzenesulfonamide
methanesulfonate (the methanesulfonate salt of Compound B) Mannitol
250 mg Talc 125 mg Magnesium Stearate 8 mg
[0221] While the preferred embodiments of the invention are
illustrated by the above, it is to be understood that the invention
is not limited to the precise instructions herein disclosed and
that the right to all modifications coming within the scope of the
following claims is reserved.
Assays
[0222] In Vitro Combination Studies of BRAF and MEK Inhibitors on
Cancer Cells Lines from Multiple Origins Encoding Different
Mutations
A. Concentration Ranges A
[0223] Drug combinations experiments were carried out in 384-well
plates. Cell were plated in 384-well plates at 500 cells/well in
culture media appropriate for each cell type, supplemented with 10%
FBS and 1% penicillin/streptomycin, and incubated overnight at
37.degree. C., 5% CO.sub.2. Sixteen concentrations of 2 folds
dilution of each drug were tested in matrix for cell growth
inhibition. Concentrations tested for compound A (free form). were
1 .mu.M-0.03 nM and for compound B (DMSO solvate) were 10 .mu.M-0.3
nM. Cells were treated with compound combination and incubated at
37.degree. C. for 72 hours. Cell growth was measured using
CellTiter-Glo.RTM. reagent according to the manufacturer's protocol
and signals were read on a Perkin Elmer EnVision.TM. reader set for
luminescence mode with a 0.5-second read. Results are expressed as
a percentage inhibition compared to DMSO treated cells and
background correction was made by subtraction of values from wells
containing no cells.
[0224] The response (percent inhibition compared to untreated
samples and normalized to media alone) of compound "A" at "a"
concentration (Ra) and that of compound "B" at "b" concentration
(Rb) is compared to response of the mixture of compounds "A" and
"B" at concentrations "a" and "b" respectively (Rab). Using these
values, Excess Over Highest Single Agent (EOHSA) was calculated for
each concentration of each of the tested cell lines:
Rab>10% of the higher value among Ra and Rb=more than
additive
Rab<-10% of the higher value among Ra and Rb=antagonism
[0225] Using this formula, if Rab is greater by 10% or more than
the highest value between Ra and Rb the drug combination is
considered more than additive. If Rab is smaller by 10% or more
than the highest value between Ra and Rb the drug combination is
antagonistic.
[0226] For each of the cell lines tested, the number of
combinations in the 16.times.16 matrix with more than additive
response (those where the Rab is greater than 10% higher than the
higher value among Ra and Rb) were enumerated. The number of more
than additive combinations (out of the 264 tested) are summarized
in Table 1. On this table, concentration combinations on a given
cell line were found to be particularly beneficial (gray square) if
more than 20% (51 combination out of 256 tested) of combinations
tested showed >10% EOHSA.
TABLE-US-00002 TABLE 1 Combination effect of MEK and BRAF
inhibitors on multiple cancer cell lines. ##STR00024##
These data demonstrate that the combination of Compound A and
Compound B is favourable on multiple cancer cell lines from
multiple origins independent of the mutational status of key
oncogenes within the MAPK or the AKT/PI3K/PTEN pathways.
B. Concentration Ranges B
[0227] Evaluation similar to Section A above of combination of
Compound A and Compound B were performed using the data generated
in Section A, but only for drug concentrations which were deemed to
be clinically relevant (100 nM-3 nM). These concentrations were
chosen as those which tended to be efficacious but non-toxic in
pre-clinical mice xenograft models. Using these concentrations, a
total of 25 drug combinations were evaluated for each cell line,
and results are summarized in Table 2.
[0228] The number of combinations having Rab>10% of the higher
value among Ra and Rb of the 25 clinically relevant combinations
tested were calculated and expressed as a percentage in Table
2.
TABLE-US-00003 TABLE 2 in vitro combination of MEK and BRAF
inhibitors using clinically relevant drug concentrations
##STR00025##
The data demonstrate that the combination of Compound A and
Compound B is favourable in most cell lines tested at relevant
clinical drug concentrations and highly favourable on all
BRAF.sup.V600E and KRAS mutant cell lines tested, independent of
the PI3K/PTEN pathway mutational status.
In Vitro Cell Growth Inhibition in Tumor Cell Lines
Methods:
[0229] Cell lines and growth conditions--Human colon tumor lines,
Colo-205, DLD-1, HCT-8, HT-29, LS-1034, NCI-HSO8, RKO, SW1417,
SW1463, SW480 and SW837, and human melanoma line A375 were from
ATCC. A375PF11 was derived from A375. 12R5-1, 12R5-3, 12R8-1,
12R8-3, 16R5-2, 16R6-3 and 16R6-4 are single cell clones derived
from mixed populations of A375PF11 cells that were selected to grow
in Compound A to concentrations of 1200 and 1600 nM, thereby
exhibiting acquired resistance to Compound B. All lines were
cultured in RPMI 1640 medium containing 10% fetal bovine serum
(FBS).
[0230] Cell growth inhibition assay and combination data
analysis--All cells were cultured for a minimum of 72 hours prior
to cell plating. Cells were assayed in a 96-well tissue culture
plate (NUNC 136102) of RPMI medium containing 10% FBS for all cells
at 1,000 cells per well. Approximately 24 hours after plating,
cells were exposed to ten, three-fold serial dilutions of compound
or the combination of the two agents at a constant molar to molar
ratio of 1:10 Compound A (DMSO solvate) to Compound B (free form)
in RPMI media containing 10% FBS. Concentrations tested for
Compound A were 1 .mu.M-0.05 nM and for Compound B were 10
.mu.M-0.5 nM. Cells were incubated in the presence of compounds for
3 days. ATP levels were determined by adding Cell Titer Glo.RTM.
(Promega) according to the manufacturer's protocol. Briefly, Cell
Titer Glo.RTM. was added each plate, incubated for 30 minutes then
luminescent signal was read on the SpectraMax L plate reader with a
0.5 sec integration time.
[0231] Inhibition of cell growth was estimated after treatment with
compound or combination of compounds for three days and comparing
the signal to cells treated with vehicle (DMSO). Cell growth was
calculated relative to vehicle (DMSO) treated control wells.
Concentration of compound that inhibits 50% of control cell growth
(IC.sub.50) was interpolated when y=50% of the vehicle control
using nonlinear regression with the equation, y=(A+(B-A)/(1+(C/x)
D))), where A is the minimum response (y.sub.min), B is the maximum
response (y.sub.max), C is the inflection point of the curve
(EC.sub.50) and D is the Hill coefficient.
[0232] Combination effects on potency were evaluated using
Combination Index (CI) which was calculated with the
back-interpolated IC.sub.50 values and the mutually non-exclusive
equation derived by Chou and Talalay (1):
CI=Da/IC.sub.50(a)+Db/IC.sub.50(b)+(Da.times.Db)/(IC.sub.50(a).times.IC.-
sub.50(b))
where IC.sub.50(a) is the IC.sub.50 of Compound A; IC.sub.50(b) is
the IC.sub.50 for Compound B;
[0233] Da is the concentration of Compound A in combination with
Compound B that inhibited 50% of cell growth; and Db is the
concentration of Compound B in combination with Compound A that
inhibited 50% of cell growth. In general, a CI value <0.9,
between 0.9 and 1.1, or >1.1 indicates synergy, additivity and
antagonism, respectively. In general, the smaller the CI number,
the greater is the strength of synergy.
[0234] The combination effects on the response scale were
quantified by Excess Over Highest Single Agent (EOHSA) based on the
concept of nonlinear blending as described in detail by Peterson
and Novick (2007) and Peterson (2010) [(2; 3) [Peterson and Novick,
2007; Peterson, 2010]. EOHSA values are defined as increases in
improvement (here, in `percentage points` (ppts) difference)
produced by the combination over the best single agent at its
component dose level for the combination. For single agent and
combination treatments, cells were exposed to compounds at a
fixed-dose-ratio, and dose response curves were fit to the
experimental data and analyzed using regression models. At
specified total dose levels of IC.sub.50 along the dose response
curve, the dose combination (corresponding to IC.sub.50) was
determined for making EOHSA statistical inferences. More
specifically, for a combination drug experiment involving drug 1 at
dose d1 and drug 2 at dose d2, (i.e., total dose equals d1+d2) is
said to have a positive EOHSA if the mean response at the
combination is better than the mean response to drug 1 at dose d1
or drug 2 at dose d2.
Results:
[0235] The effect of cell growth inhibition by a MEK inhibitor
Compound A, a BRAF inhibitor Compound B and their combination was
determined in a panel of human tumor cell lines. The mean
IC.sub.50s (from at least two independent experiments) and the
combination effects at IC.sub.50s are summarized in Table 3 with
BRAF and KRAS mutation status.
[0236] Referring to Table 3, the four colon cell lines with BRAF
V600E mutation displayed sensitivity to Compound A with IC.sub.50
values between 0.001 .mu.M and 0.025 .mu.M, and to Compound B with
IC.sub.50 values between 0.018 .mu.M and 5.654 .mu.M. The
combination of Compound A and Compound B was synergistic with CI
values between 0.25 and 0.73 and/or enhanced cell growth inhibition
with EOHSA values between 7 and 26 ppts in these four lines with
BRAF V600E mutation. The seven colon lines without BRAF V600E
mutation (either with BRAF G596R or KRAS mutations) were
insensitive to Compound B (IC.sub.50>10 .mu.M), however highly
sensitive to cell growth inhibition by Compound A with IC.sub.50s
ranging from 0.001 to 0.093 .mu.M in six out of the seven lines.
The combination of Compound A and Compound B showed enhanced cell
growth inhibition in DLD1 colon tumor line, and minimal to no added
benefit over Compound A single compound treatment in the other six
colon cell lines.
[0237] For the melanoma lines listed in Table 3, A375PF11 cells
with BRAF V600E mutation were highly sensitive to either Compound A
(IC.sub.50=0.001 .mu.M) or Compound B (IC.sub.50=0.012 .mu.M)
single agent. The combination of Compound A and Compound B were
synergistic with CI value of 0.3 in A375PF11 cells. The melanoma
lines 12R8-3, 12R8-1, 12R5-3, and 16R6-3 were resistant to Compound
B (IC.sub.50>10 .mu.M), moderately sensitive to Compound A with
IC.sub.50s ranging from 0.058 .mu.M to 0.109, and responded to the
combination of Compound A and Compound B with IC.sub.50s ranging
from 0.018 to 0.023 .mu.M for Compound A and 0.178-0.234 .mu.M for
Compound B. The melanoma lines 16R5-2, 16R6-4 and 12R5-1 were
resistant or insensitive to either Compound A or Compound B alone,
however became sensitive to the combination of Compound A and
Compound B with IC.sub.50s ranging from 0.018 to 0.039 .mu.M for
Compound A and 0.177-0.386 .mu.M for Compound B. The combination of
Compound A and Compound B also showed enhancement of cell growth
inhibition in all these melanoma lines. Note, CI values could not
be calculated therefore not applicable where the single agent
values were outside of the range tested.
[0238] Of interest, the combined administration of Compound A and
Compound B in the BRAF-V600E mutant colon and melanoma cell lines
showed synergistic effect demonstrated by the CI values <0.9, or
resulted in a reduced IC50 value comparing to that of Compound A or
Compound B, administered alone, where at least one of the single
agents did not result in 50% inhibition within the tested
range.
TABLE-US-00004 TABLE 3 Cell growth inhibition by Compound A,
Compound B and their combination in human tumor cell lines.
IC.sub.50 values in micromolar (mean .+-. std) Compound A or B =
1:10 molar Combination Effects at Tumor Cell Mutation Status Single
Agent ratio combination IC.sub.50 Lines KRAS/BRAF Compound A
Compound B Compound A Compound B CI EOHSA (ppt) Colon Colo-205
BRAF_V600E 0.001 .+-. 0.000 0.018 .+-. 0.013 0.0005 .+-. 0.0002
0.005 .+-. 0.002 0.65 .+-. 0.10 26 .+-. 10.6 HT-29 BRAF_V600E 0.001
.+-. 0.001 0.022 .+-. 0.022 0.0004 .+-. 0.0003 0.004 .+-. 0.003
0.73 .+-. 0.11 15 .+-. 4.3 SW1417 BRAF_V600E 0.003 .+-. 0.002 0.203
.+-. 0.015 0.001 .+-. 0.0003 0.014 .+-. 0.003 0.69 .+-. 0.44 7 .+-.
8.1 RKO BRAF_V600E 0.025 .+-. 0.007 5.654 .+-. 3.900 0.006 .+-.
0.001 0.057 .+-. 0.015 0.25 .+-. 0.01 18 .+-. 2.7 DLD1 KRAS_G13D
0.557 .+-. 0.393 >10 0.044 .+-. 0.003 0.443 .+-. 0.030 N/A 22
.+-. 2.3 HCT-8 KRAS_G13D 0.050 .+-. 0.011 >10 0.035 .+-. 0.013
0.348 .+-. 0.132 N/A 7 .+-. 2.9 SW480 KRAS_G12C 0.042 .+-. 0.003
>10 0.024 .+-. 0.003 0.235 .+-. 0.034 N/A 5 .+-. 0.7 NCI-H508
BRAF_G596R 0.010 .+-. 0.005 >10 0.008 .+-. 0.005 0.081 .+-.
0.054 N/A 4 .+-. 4.1 SW837 KRAS_G12C 0.093 .+-. 0.063 >10 0.087
.+-. 0.037 0.874 .+-. 0.368 N/A 0 .+-. 2.8 LS-1034 KRAS_A146T 0.011
.+-. 0.007 >10 0.029 .+-. 0.022 0.295 .+-. 0.221 N/A -21 .+-.
2.7 SW1463 KRAS_G12C 0.005 .+-. 0.004 >10 0.021 .+-. 0.013 0.210
.+-. 0.133 N/A -26 .+-. 4.2 Melanoma A375PF11 BRAF_V600E 0.001 .+-.
0.001 0.012 .+-. 0.012 0.0002 .+-. 0.000 0.002 .+-. 0.003 0.30 .+-.
0.32 18 .+-. 3.0 12R8-1 BRAF_V600E 0.058 .+-. 0.030 >10 0.019
.+-. 0.017 0.186 .+-. 0.173 N/A 22 .+-. 0.2 12R8-3 BRAF_V600E 0.059
.+-. 0.037 >10 0.018 .+-. 0.018 0.178 .+-. 0.182 N/A 27 .+-. 0.7
12R5-3 BRAF_V600E 0.092 .+-. 0.052 >10 0.021 .+-. 0.015 0.210
.+-. 0.153 N/A 30 .+-. 5.8 16R6-3 BRAF_V600E 0.109 .+-. 0.022
>10 0.023 .+-. 0.021 0.234 .+-. 0.210 N/A 25 .+-. 6.8 16R5-2
BRAF_V600E >1 >10 0.018 .+-. 0.013 0.177 .+-. 0.128 N/A 35
.+-. 21 16R6-4 BRAF_V600E >1 >10 0.021 .+-. 0.013 0.215 .+-.
0.135 N/A 44 .+-. 12.4 12R5-1 BRAF_V600E >1 >10 0.039 .+-.
0.021 0.386 .+-. 0.211 N/A 61 .+-. 14.9 Table 3 Key: IC.sub.50: the
concentration of Compound as single agent, or the concentration of
Compound A or B in combination when Compound A and Compound B =
1:10 molar ratio that reduces cell growth by 50%; CI; Combination
Index; N/A = not applicable EOHSA: Excess over Highest Single
Agent, measured as a percentage.
REFERENCE LIST
[0239] (1) Chou T C, Talalay P. Quantitative analysis of
dose-effect relationships: the combined effects of multiple drugs
or enzyme inhibitors. Adv Enzyme Regul 1984; 22:27-55. [0240] (2)
Peterson J J, Novick S J. Nonlinear blending: a useful general
concept for the assessment of combination drug synergy. J Recept
Signal Transduct Res 2007; 27(2-3):125-46. [0241] (3) Peterson J. A
Review of Synergy Concepts of Nonlinear Blending and Dose-Reduction
Profiles. Frontiers of Bioscience S2, 483-503. 2010.
Mouse Xenograft Model A
[0242] A xenograft models using A375P F11 (human melanoma cell line
encoding BRaf.sup.V600E mutation) cells were established from cells
grown in tissue culture and harvested aseptically using a trypsin
digest. The tumor cells were injected subcutaneously into female
athymic mice (strain nu/nu) with between 5.times.10.sup.6 and
10.sup.7 cells in 50% martigel. Tumors were allowed to establish.
Dosing began on day 24 after implantation, corresponding to a mean
tumor volume of .about.200 mm.sup.3.
[0243] This human xenograft tumor model utilized 4 groups of mice,
with 8 mice per group. The animals were identified via a
subcutaneous (sc) microchip or tattoos.
[0244] A first group of untreated or placebo-treated tumor-bearing
control animals acted as controls. A second group was dosed once
daily orally with
N-{3-[5-(2-Amino-4-pyrimidinyl)-2-(1,1-dimethylethyl)-1,3-thiazol-4--
yl]-2-fluorophenyl}-2,6-difluorobenzenesulfonamide (crystaline free
base form) (Compound B). A third group was dosed once daily orally
with
N-{3-[3-cyclopropyl-5-(2-fluoro-4-iodo-phenylamino)6,8-dimethyl-2,4,7-tri-
oxo-3,4,6,7-tetrahydro-2H-pyrido[4,3-d]pyrimidin-1-yl]phenyl}acetamide
dimethyl sulfoxide solvate (Compound A). A fourth group was dosed
once daily orally with a combination of
N-{3-[5-(2-Amino-4-pyrimidinyl)-2-(1,1-dimethylethyl)-1,3-thiazol-4-yl]-2-
-fluorophenyl}-2,6-difluorobenzenesulfonamide (crystaline free base
form) and
N-{3-[3-cyclopropyl-5-(2-fluoro-4-iodo-phenylamino)6,8-dimethyl-2,4,7-
-trioxo-3,4,6,7-tetrahydro-2H-pyrido[4,3-d]pyrimidin-1-yl]phenyl}acetamide
dimethyl sulfoxide solvate. Each drug was provided in a suspension
of 0.5% HPMC/0.2% TWEEN 80.
[0245] The tumor sizes were measured twice weekly using Vernier
callipers. Tumor volume was calculated from two-dimensional
measurements using an equation approximating the volume of a
prolate ellipsoid:
Tumor volume in cubic mm=(length.times.width.sup.2).times.0.5
The measurements are reported in FIG. 1 following 36 day treatment.
The data showed that combination of the MEK and B-Raf inhibitors is
advantageous compared to each agent administrated individually.
Mouse Xenograft Model B
[0246] A375P cells were harvested from culture flasks by exposure
to 0.25% trypsin/EDTA for 5 min at 37.degree. C. Detached cells
were collected, centrifuged (1500 rpm, 5 min, 4.degree. C.) and
rinsed to remove the trypsin solution. Cells were resuspended in
PBS without magnesium or calcium and counted. Cells were spun as
previously to remove PBS and a single cell suspension was created
either in 50% Matrigel: 50% PBS (v:v) or 100% PBS so that a 100
.mu.L subcutaneous injection would deliver the required number of
cells per mouse. The A375P melanoma line was injected with Matrigel
at 1.75 million cells per mouse subcutaneously into 8-10 week old,
female CD-1 nu/nu mice. Tumors were established (.about.150-300
mm.sup.3) for all cell lines within 2-4 weeks post-injection.
[0247] Compound A (DMSO solvate) and Compound B (free form) were
administered orally to mice at the indicated doses in 0.2 ml/20
gram body weight in 0.5% HPMC (hydroxypropylmethylcellulose, Sigma
cat # H7509) and 0.2% Tween 80 (Sigma cat # P1754) in distilled
water, pH 7.0-8.0.
[0248] Mice with similar sized tumors (150-200 mm.sup.3) were
identified. The length and width of tumors were measured by
handheld calipers and body weights of mice were measured using a
bench top weighing scale. Mice were placed in groups of eight or
seven accordingly and dosed orally with either vehicle, individual
compound or compound combination. Mice were weighed and tumors
measured twice weekly for the duration of the study. Data presented
in FIG. 2 demonstrates that combination of Compound A (0.1 mg/kg)
and Compound B (30 mg/kg) daily for 33 days (days 24 to 56 after
implantation) is more efficacious than each agent alone.
* * * * *