Note : Les descriptions sont présentées dans la langue officielle dans laquelle elles ont été soumises.
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METHODS FOR INHIBITING TIE2 KINASE
USEFUL IN THE TREATMENT OF CANCER
Description of the Text File Submitted Electronically
[0001] The contents of the text file submitted electronically herewith are
incorporated herein
by reference in their entirety: A computer readable format copy of the
Sequence Listing
(filename: DECP 066 00W0 SeqList ST25.txt, date recorded: November 7, 2013,
file size 6
kilobytes).
Field of the Invention
[0002] The present invention relates to methods of inhibiting TIE2 kinase
useful in the
treatment of tumor growth, tumor invasiveness, tumor intravasation, tumor
dissemination, tumor
metastasis, and tumor immunotolerance. Specifically, the invention relates to
methods of using
compositions of Formula I herein described as potent inhibitors of TIE2 for
treating breast cancer
growth, invasiveness, intravasation dissemination, metastasis, and
immunotolerance.
Background of the Invention
[0003] Tunica interna endothelial cell kinase-2 (TIE2) is largely
restricted to expression in
endothelial cells of the vasculature, and in a subset of bone marrow derived
TIE2 expressing
monocytes (TEMs). TIE2 is the receptor for angiopoietin 1 (ANG1), angiopoietin
2 (ANG2),
and angiopoietin 4 (ANG4) and this signaling system plays an important role in
both
angiogenesis (sprouting of new vessels from existing vessels) and
vasculogenesis (de novo new
vessel formation). TEMs are a subset of circulating monocytes and tissue
macrophages that have
proangiogenic and provasculogenic activity in tumor models (De Palma MD et al,
Cancer Cell
2005; 8:211-226). TIE2 inhibition decreases the ability of TEMs to associate
with blood vessels
(Mazzieri R, Cancer Cell 2011; 19:512-526) and markedly decreases the
proangiogenic activity
of this macrophage subset (De Palma M, Clin Cancer Res 2011; 17(16):5226-
5232).
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[0004] Cytotoxic chemotherapy, radiation therapy, and anti-angiogenic
treatments damage
the tumor-associated vasculature thus leading to a hypoxic tumor environment.
The hypoxic
tumor environment leads to rebound tumor vascularization by activating an
angiogenic switch
from the vascular endothelial growth factor (VEGF)NEGFR2 pathway to the
ANG/TIE2
pathway in vascular endothelial cells. The recruitment of pro-vasculogenic
TEMs from the bone
marrow to these hypoxic tumor sites facilitates this revascularization by the
association of TEMs
with endothelial cells within the tumor microenvironment. TEMs and TIE2-
expressing
endothelial cells are thus believed to play an important role in the
revascularization of tumors
after these treatments, leading to progression due to the growth of residual
tumor cells (De Palma
M, et al. Trends Immunol 2007; 28:519-524).
[0005] TIE2 is also a mediator of osteoclast differentiation, and TIE2
inhibition led to
decreased osteolytic bone invasion and decreased tumor growth in the 4T1 mouse
breast cancer
model (Dales JP, et al. Int J Oncol 2003; 22:391-397 ). Beyond the physiologic
expression of
TIE2 on endothelial, monocyte/macrophage, and osteoclast cells of the tumor
microenvironment,
TIE2 has also been demonstrated to be present on breast cancer cells. Tumor
cell expression of
TIE2 was associated with an elevated risk of metastatic disease and an
independent predictor of
prognosis on multivariate analysis (Min Y, et al. Cancer Res 2010; 70:2918-
2828).
[0006] Significantly, a subset of TIE2-expressing tissue macrophages are
located within
specialized vascular structures known as tumor microenvironment for metastases
(TMEMs).
Recent observations have linked TIE2-expressing macrophages within TMEM
structures as
being essential for extravasation of breast cancer cells into the vascular
circulation and
subsequent dissemination to distal metastatic sites (Condeelis J, Pollard JW.
Cell 2006; 124:263-
6; Ginter PS, et al. Cancer Res 2012; 72(24 Suppl):Abstract #P6-02-04). Thus
inhibition of
TIE2, and of the macrophages within TMEM structures, may lead to a decrease in
new
metastases.
[0007] TIE2-expressing tissue macrophages (TEMs) have recently been
demonstrated to
play a role in breast cancer immunotolerance. TEMs from breast tumors are able
to suppress
tumor-specific immune responses. Specifically, suppressive functions of TEMs
are similarly
driven by TIE2 and VEGFR kinase activity. TEMs isolated from breast cancer
tissue can
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function as antigen-presenting cells that elicit only a weak proliferation of
T cells. Blocking
TIE2 and VEGFR kinase activity induced TEMs to change their phenotype into
cells with
features of myeloid dendritic cells with robust antigen-presentation.
Immunosuppressive activity
of TEMs is also associated with high CD86 surface expression and extensive
engagement of T
regulatory cells in breast tumors. TIE2 and VEGFR kinase activities were
required to maintain
high CD86 surface expression levels and to convert T cells into
immunosuppressive regulatory
cells (Ibberson M, et al. Clin Cancer Res 2013; 19:3439-3449).
[0008] The polyoma middle-T antigen (PyMT) syngeneic mouse breast cancer
model utilizes
the mouse mammary tumor virus (MMTV) promoter, a breast specific promoter, to
express
PyMT in mouse breast tissue. In this model, PyMT breast cancer cells are
implanted in the
mouse mammary fat pad, and these cancers metastasize and lead to the death of
the mouse.
Unlike xenograft models, the PyMT model utilizes fully immunocompetent mice.
Metastasis in
this model is known to be modulated by TIE2 expressing macrophages within TMEM
vascular
structures. Thus, there is a need for new treatments for diseases assocaited
with TIE2.
Summary of the Invention
[0009] Methods of the present invention find utility in the inhibition TIE2
kinase. As a result
of this inhibition the present invention is useful in the treatment or
prophylaxis against of tumor
growth, invasiveness, intravasation, dissemination, metastasis, and tumor
immunotolerance. In
particular, the invention relates to methods of using compositions of Formula
I, described below,
as potent inhibitors of TIE2 for treating breast cancer growth, invasiveness,
intravasation,
dissemination, metastasis, and immunotolerance:
0
0
N
N N N
H H F
CN.-
*
H
= nHX
Nk /
3
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Formula I
wherein
n is an integer from 0 to 7;
X is the basic radical of a pharmaceutically acceptable salt;
provided that when n is 0, the composition of Formula I is the parent free
base. In
some embodiments, HX is absent whereby the structure of Formula I is the
parent free
base.
[0010] The compositions of Formula I also find utility in other cancers
wherein TIE2
expression, either in the tumor cell or in the tumor microenvironment, causes
tumor progression
by mechanisms mediating primary tumor growth, primary tumor invasiveness,
tumor
intravasation into the blood stream, tumor cell dissemination, tumor
metastases to distal tissues,
or tumor immunotolerance. Inhibition of TIE2 kinase by the composition of
Formula I therefore
finds utility in the treatment of cancer by inhibiting processes including
primary tumor growth,
primary tumor invasiveness, tumor intravasation into the blood stream, tumor
cell dissemination,
tumor metastases to distal tissues, or tumor immunotolerance.
[0011] TIE2 kinase has been shown to be causative of cancer progression in
gliomas (Liu et
al, Oncotarget (2010) 1: 700-709; Brunckhorst et al, Cancer Research (2010)
70: 7283-7293),
melanomas (Helfrich et al, Clin Cancer Res (2009) 15: 1384-1392 ), ovarian
cancer (Karlan et
al, J. Clinical Oncology (2012) 30: 362-370), colorectal cancer (Ahmad et al,
Cancer (2001) 92:
1138-1143; Hashizume et al, Cancer Research (2010) 70: 2213-2223),
hepatocellular carcinoma
(Matsubara et al, Hepatology (2013) 57: 1416-1425; Mitsuhashi et al,
Hepatology (2003) 37:
1105-1113; Tanaka et al, J. Clin Invest (1999) 103: 341-345), and
hematological cancers (Muller
et al, Leukemia Research (2002) 26: 163-168; Hou et al, Leukemia Research
(2008) 32: 904-
912).
Brief Description of the Drawings
[0012] Figure 1 shows the inhibition of primary PyMT tumor growth using the
composition
of Formula II , paclitaxel, or a combination thereof.
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[0013] Figure 2 shows the inhibition of PyMT tumor macrophage accumulation
using the
composition of Formula II, paclitaxel, or a combination thereof.
[0014] Figure 3 shows the inhibition of PyMT tumor TIE2-expressing cell
accumulation
using the composition of Formula II, paclitaxel, or a combination thereof.
[0015] Figure 4 shows the inhibition of lung metastases in the PyMT breast
cancer model
using the composition of Formula II, paclitaxel, or a combination thereof.
[0016] Figure 5 shows the inhibition of lung metastases in the PyMT breast
cancer model
comparing the activities of paclitaxel and the combination of paclitaxel and
the composition of
Formula II.
[0017] Figure 6 shows the inhibition of lung metastases in the PyMT breast
cancer model
using eribulin as a single agent or in combination with the composition of
Formula II.
[0018] Figure 7 shows enzymatic and in vivo activities of eribulin as a
single agent or in
combination with the composition of Formula II.
Detailed Description of the Invention
Definitions:
[0019] The term "basic radical of a pharmaceutically acceptable salt" in
compositions of
Formula I include, without limitation, water-soluble and water-insoluble
salts, such as substituted
or unsubstituted benzenesulfonate , the acetate, amsonate (4,4-diaminostilbene-
2, 2 -disulfonate),
benzonate, bicarbonate, bisulfate, bitartrate, borate, bromide, butyrate,
calcium, calcium edetate,
camsylate, carbonate, chloride, citrate, clavulariate, dihydrochloride,
edetate, edisylate, estolate,
esylate, fiunarate, gluceptate, gluconate, glutamate, glycollylarsanilate,
hexafluorophosphate,
hexylresorcinate, hydrabamine, hydrobromide, hydrochloride, hydroxynaphthoate,
iodide,
isothionate, lactate, lactobionate, laurate, magnesium, malate, maleate,
mandelate, mesylate,
methylbromide, methylnitrate, methylsulfate, mucate, napsylate, nitrate, N-
methylglucamine
ammonium salt, 3-hydroxy-2-naphthoate, oleate, oxalate, palmitate, pamoate (1
,1 -methene-bis-
2-hydroxy-3-naphthoate, einbonate), pantothenate, phosphate/diphosphate,
picrate,
polygalacturonate, propionate, p-toluenesulfonate, salicylate, stearate,
subacetate, succinate,
sulfate, sulfosalicylate, suramate, tannate, tartrate, teoclate, tosylate,
triethiodide, and valerate
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salts. Specific examples of the basic radicals include para-toluene sulfonate,
triflate, and
methanesulfonate.
[0020] The term "salt' refers to pharmaceutically acceptable salts
[0021] The term "pharmaceutically acceptable salt" also refers to a salt of
the compositions
of the present invention having an acidic functional group, such as a
carboxylic acid functional
group, and a base.
[0022] The term "treating" with regard to a subject, refers to improving at
least one symptom
of the subject's disorder. Treating can be curing, improving, or at least
partially ameliorating the
disorder.
[0023] The term "administer", "administering", or "administration" as used
in this disclosure
refers to either directly administering a composition or pharmaceutically
acceptable salt of the
compound or a composition to a subject, or administering a prodrug derivative
or analog of the
composition or pharmaceutically acceptable salt of the compound or composition
to the subject,
which can form an equivalent amount of active compound within the subject's
body.
[0024] An "effective amount," when used in connection with medical uses is
an amount that
is effective for providing a measurable treatment, prevention, or reduction in
the rate of
pathogenesis of a disease of interest.
[0025] The present invention relates to methods of the treatment (blocking)
or prophylaxis
against tumor growth, invasiveness, intravasation, dissemination, metastasis,
and tumor
immunotolerance. The method comprises administering to a patient in need of
treatment or
reduction of prophylactic effects of these conditions an effective amount of a
composition of
Formula I herein described in a dosing regimen that regulates TIE2 inhibition.
[0026] The amount of composition described herein needed for achieving a
therapeutic effect
may be determined empirically in accordance with conventional procedures for
the particular
purpose. Generally, for administering therapeutic agents (e.g. compositions of
Formulae I or II
(and/or additional agents) described herein) for therapeutic purposes, the
therapeutic agents are
given at a pharmacologically effective dose. A "pharmacologically effective
amount,"
"pharmacologically effective dose," "therapeutically effective amount," or
"effective amount"
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refers to an amount sufficient to produce the desired physiological effect or
amount capable of
achieving the desired result, particularly for treating the disorder or
disease. An effective amount
as used herein would include an amount sufficient to, for example, delay the
development of a
symptom of the disorder or disease, alter the course of a symptom of the
disorder or disease (e.g.,
slow the progression of a symptom of the disease), reduce or eliminate one or
more symptoms or
manifestations of the disorder or disease, and reverse a symptom of a disorder
or disease. For
example, administration of therapeutic agents to a patient suffering from
cancer provides a
therapeutic benefit not only when the underlying condition is eradicated or
ameliorated, but also
when the patient reports a decrease in the severity or duration of the
symptoms associated with
the disease, e.g., a decrease in tumor burden, a decrease in circulating tumor
cells, an increase in
progression free survival. Therapeutic benefit also includes halting or
slowing the progression of
the underlying disease or disorder, regardless of whether improvement is
realized.
[0027] In one embodiment of the invention, the composition of Formula I is
1-(3-tert-butyl-
1 -(quino lin-6 -y1)- 1 H-pyrazol-5-y1)-3-(2-fluoro -4-(2-(methylc arb
amoyl)pyri din-4-
yloxy)phenyOurea para-toluene sulfonic acid salt of Formula II which is a
potent inhibitor of
TIE2, the receptor tyrosine kinase for angiopoietin ligands.
(:)
IN
N N N
H H
410 F ...--
0 N
H
NN/ s SO3H
Formula II
[0028] The compositions of Formula I find utility in cancers wherein TIE2
expression, either
in the tumor cell or in the tumor microenvironment, causes tumor progression
by mechanisms
mediating primary tumor growth, primary tumor invasiveness, tumor
intravasation into the blood
stream, tumor cell dissemination, tumor metastases to distal tissues, or tumor
immunotolerance.
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PCT/US2013/069005
Inhibition of TIE2 kinase by the composition of Formula I therefore finds
utility in the treatment
of cancer by inhibiting processes including primary tumor growth, primary
tumor invasiveness,
tumor intravasation into the blood stream, tumor cell dissemination, tumor
metastases to distal
tissues, or tumor immunotolerance.
[0029] Therapeutic concentrations of the compositions of Formula I block
cells within the
tumor microenvironment known to cause tumor growth, invasion, intravasation,
dissemination,
metastases, of tumor induced immunotolerance. Such cell types within the tumor
microenvironment include TIE2-expressing monocytes, TIE2-expessing
macrophages, and
TIE2-expressing endothelial cells.
[0030] Tumors responsive to angiopoietin/TIE2 signaling include but are not
limited to
breast cancer, ovarian cancer, hepatocellular carcinoma, gliomas, colorectal
cancer, and
hematological malignancies.
[0031] In another embodiment, the composition of Formula I when the HX is
absent is the
free base compound 1-(3-tert-buty1-1-(quinolin-6-y1)-1H-pyrazol-5-y1)-3-(2-
fluoro-4-(2-
(methylcarbamoyOpyridin-4-yloxy)phenyOurea having the structure:
0
Nisi \ 1 N el
N
N N
H H F
410 0 N
H
N N /
[0032] The composition of Formula I may be administered as a single agent
or in
combination with other therapeutic agents known to treat cancers. Such other
therapeutic agents
include radiation therapy, anti-tubulin agents, DNA alkylating agents, DNA
synthesis-inhibiting
agents, DNA intercalating agents, anti-estrogen agents, anti-androgens,
steroids, anti-EGFR
agents, kinase inhibitors, topoisomerase inhibitors, Histone Deacetylase
(HDAC) inhibitors,
DNA methylation inhibitors, anti-HER2 agents, anti-angiogenic agents,
proteasome inhibitors,
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thalidomide, lenalidomide, antibody-drug-conjugates (ADCs), immunomodulating
agents, or
cancer vaccines.
[0033] Effective amounts, toxicity, and therapeutic efficacy can be
determined by standard
pharmaceutical procedures in cell cultures or experimental animals, e.g., for
determining the
LD50 (the dose lethal to about 50% of the population) and the ED50 (the dose
therapeutically
effective in about 50% of the population). The dosage can vary depending upon
the dosage form
employed and the route of administration utilized. The dose ratio between
toxic and therapeutic
effects is the therapeutic index and can be expressed as the ratio LD50/ED50.
In some
embodiments, compositions and methods that exhibit large therapeutic indices
are preferred. A
therapeutically effective dose can be estimated initially from in vitro
assays, including, for
example, cell culture assays. Also, a dose can be formulated in animal models
to achieve a
circulating plasma concentration range that includes the IC50 as determined in
cell culture, or in
an appropriate animal model. Levels of the described compositions in plasma
can be measured,
for example, by high performance liquid chromatography. The effects of any
particular dosage
can be monitored by a suitable bioassay. The dosage can be determined by a
physician and
adjusted, as necessary, to suit observed effects of the treatment.
[0034] In certain embodiments, the prophylactic effect will result in a
quantifiable change of
at least about 10%, at least about 20%, at least about 30%, at least about
50%, at least about
70%, or at least about 90%. In some embodiments, the effect will result in a
quantifiable change
of about 10%, about 20%, about 30%, about 50%, about 70%, or even about 90% or
more.
Therapeutic benefit also includes halting or slowing the progression of the
underlying disease or
disorder, regardless of whether improvement is realized.
[0035] When the compositions of Formulae I or II are used in combination
with other anti-
cancer agents, the other anti-cancer agent may be dosed independently of the
dosing schedule of
the composition of Formulae I or II. The other anti-cancer agent may be dosed
at its previously
established therapeutic dose and dosing schedule, or its dose and dosing
schedule may be
modified to optimize efficacy, safety or tolerability when used in combination
with the
compositions of Formulae I or II.
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[0036] Further, any compositions of Formulae I or II (and/or additional
agents) described
herein can be administered to a subject as a component of a composition that
comprises a
pharmaceutically acceptable carrier or vehicle. Such compositions can
optionally comprise a
suitable amount of a pharmaceutically acceptable excipient so as to provide
the form for proper
administration.
[0037] Pharmaceutical excipients can be liquids, such as water and oils,
including those of
petroleum, animal, vegetable, or synthetic origin, such as peanut oil, soybean
oil, mineral oil,
sesame oil and the like. The pharmaceutical excipients can be, for example,
saline, gum acacia,
gelatin, starch paste, talc, keratin, colloidal silica, urea and the like. In
addition, auxiliary,
stabilizing, thickening, lubricating, and coloring agents can be used. In one
embodiment, the
pharmaceutically acceptable excipients are sterile when administered to a
subject. Water is a
useful excipient when any agent described herein is administered
intravenously. Saline solutions
and aqueous dextrose and glycerol solutions can also be employed as liquid
excipients,
specifically for injectable solutions. Suitable pharmaceutical excipients also
include starch,
glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel,
sodium stearate, glycerol
monostearate, talc, sodium chloride, dried skim milk, glycerol, propylene,
glycol, water, ethanol
and the like. Any agent described herein, if desired, can also comprise minor
amounts of wetting
or emulsifying agents, or pH buffering agents.
[0038] The compositions of Formula I may be used in combination with other
agents
including chemotherapeutic agents, targeted therapeutics, biological agents,
or radiotherapy.
[0039] The compositions of Formula I may be used in combination with
chemotherapeutic
agents including but not limited to anti-tubulin agents (paclitaxel,
paclitaxel protein-bound
particles for injectable suspension, eribulin, docetaxel, ixabepilone,
vincristine), vinorelbine,
DNA-alkylating agents (including cisplatin, carboplatin, oxaliplatin,
cyclophosphamide,
ifosfamide, temozolomide), DNA intercalating agents (including doxorubicin,
pegylated
liposomal doxorubicin, daunorubicin, idarubicin, and epirubicin), 5-
fluorouracil, capecitabine,
cytarabine, decitabine, 5-aza cytadine, gemcitabine and methotrexate.
[0040] The compositions of Formula I may be used in combination with kinase
inhibitors
including but not limited to erlotinib, gefitinib, lapatanib, everolimus,
temsirolimus,
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LY2835219, LEE011, PD 0332991, crizotinib, cabozantinib, sunitinib, pazopanib,
sorafenib,
regorafenib, axitinib, dasatinib, imatinib, nilotinib, vemurafenib,
dabrafenib, trametinib,
idelalisib, and quizartinib.
[0041] The compositions of Formula I may be used in combination with anti-
estrogen agents
including but not limited to tamoxifen, fulvestrant, anastrozole, letrozole,
and exemestane.
[0042] The compositions of Formula I may be used in combination with anti-
androgen
agents including but not limited to abiraterone acetate, enzalutamide,
nilutamide, bicalutamide,
flutamide, cyproterone acetate.
[0043] The compositions of Formula I may be used in combination with
steroid agents
including but not limited to prednisone and dexamethazone.
[0044] The compositions of Formula I may be used in combination with
topoisomerase I
inhibitors including but not limited to irinotecan, camptothecin, and
topotecan.
[0045] The compositions of Formula I may be used in combination with
topoisomerase II
inhibitors including but not limited to etoposide, etoposide phosphate, and
mitoxantrone.
[0046] The compositions of Formula I may be used in combination with
Histone Deacetylase
(HDAC) inhibitors including but not limited to vorinostat, romidepsin,
panobinostat, valproic
acid, and belinostat.
[0047] The compositions of Formula I may be used in combination with DNA
methylation
inhibitors including but not limited to DZNep and 5-aza-2'-deoxycytidine.
[0048] The compositions of Formula I may be used in combination with
proteasome
inhibitors including but not limited to bortezomib and carfilzomib.
[0049] The compositions of Formula I may be used in combination with
thalidomide,
lenalidomide and pomalidomide.
[0050] The compositions of Formula I may be used in combination with
biological agents
including but not limited to trastuzumab, ado-trastuzumab, pertuzumab,
cetuximab,
panitumumab, ipilimumab, anti-PD-1 agents including labrolizumab and
nivolumab, anti-PD-L1
agents including MPDL3280A, anti-angiogenic agents including bevacizumab and
aflibercept,
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and antibody-drug-conjugates (ADCs) including brentuximab vedotin and
trastuzumab
emtansine.
[0051] The compositions of Formula I may be used in combination with
radiotherapy.
[0052] The compositions of Formula I may be used in combination with
therapeutic vaccines
including but not limited to sipuleucel-T.
[0053] In some embodiments, the composition of Formula I or Formula II can
be used in
combination with one or more of the other agents described herein.
Methods for blocking primary breast tumor growth and invasiveness:
[0054] A first aspect of the invention relates to a method of blocking
primary breast tumor
growth and invasiveness which comprises administering to a patient in need
thereof an effective
amount of a composition of Formula I in a dosing regimen sufficient to block
TIE2 kinase in the
tumor microenvironment.
[0055] In one embodiment of this aspect of the invention, the dosing
regimen of the
composition of Formula I is a daily dosing administration.
[0056] In another embodiment of this aspect of the invention the dosing
regimen of the
composition of Formula I is a daily dosing administration. The intermittent
non-daily dosing
regimen may include, without limitation, alternate daily dosing, every third-
day dosing, twice
weekly dosing, or once weekly dosing.
[0057] In another embodiment of this aspect of the invention, a suitable
dosing regimen of
the composition of Formula I includes administration twice weekly, once
weekly, or alternate
weekly.
[0058] In another embodiment of this aspect of the invention, the dosing
regimen of the
composition of Formula I is twice weekly or once weekly.
[0059] In other embodiments of this aspect of the invention, the dosing
regimen of the
composition of Formula I is administration twice weekly.
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[0060] In yet another embodiment of the invention, the method of blocking
primary breast
tumor growth and invasiveness comprises administering a composition of Formula
I in a dosing
regimen sufficient to block TIE2 kinase in the tumor microenvironment in
combination with one
or more agents taken from an anti-tubulin agent, a DNA alkylating agent, a DNA
synthesis-
inhibiting agent, a DNA intercalating agent, an anti-estrogen agent, an anti-
HER2 agent, a kinase
inhibitor or an anti-angiogenic agent.
[0061] In another embodiment of this aspect of the invention, the method of
blocking
primary breast tumor growth and invasiveness comprises administering a
composition of
Formula I in combination with paclitaxel.
[0062] In another embodiment of this aspect of the invention, the method of
blocking
primary breast tumor growth and invasiveness comprises administering a
composition of
Formula I in combination with paclitaxel protein-bound particles for
injectable suspension.
[0063] In another embodiment of this aspect of the invention, the method of
blocking
primary breast tumor growth and invasiveness comprises administering a
composition of
Formula I in combination with docetaxel.
[0064] In another embodiment of this aspect of the invention, a method of
blocking primary
breast tumor growth and invasiveness comprises the administration of a
composition of Formula
I in combination with eribulin.
[0065] In another embodiment of this aspect of the invention, a method of
blocking primary
breast tumor growth and invasiveness comprises the administration of a
composition of Formula
I in combination with ixabepilone.
[0066] In another embodiment of this aspect of the invention, a method of
blocking primary
breast tumor growth and invasiveness comprises the administration of a
composition of Formula
I in combination with vinorelbine.
[0067] In another embodiment of this aspect of the invention, the method of
blocking
primary breast tumor growth and invasiveness comprises the administration of a
composition of
Formula I in combination with capecitabine.
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[0068] In another embodiment of this aspect of the invention, the method of
blocking
primary breast tumor growth and invasiveness comprises the administration of a
composition of
Formula I in combination with gemcitabine.
[0069] In another embodiment of this aspect of the invention, the method of
blocking
primary breast tumor growth and invasiveness comprises the administration of a
compositionof
Formula I in combination with 5-fluorouracil.
[0070] In another embodiment of this aspect of the invention, the method of
blocking
primary breast tumor growth and invasiveness comprises the administration of a
composition of
Formula I in combination with methotrexate.
[0071] In another embodiment of this aspect of the invention, the method of
blocking
primary breast tumor growth and invasiveness comprises the administration of a
composition of
Formula I in combination with cyclophosphamide.
[0072] In another embodiment of this aspect of the invention, the method of
blocking
primary breast tumor growth and invasiveness comprises the administration of a
composition of
Formula I in combination with cisplatin.
[0073] In another embodiment of this aspect of the invention, the method of
blocking
primary breast tumor growth and invasiveness comprises the administration of a
composition of
Formula I in combination with carboplatin.
[0074] In another embodiment of this aspect of the invention, the method of
blocking
primary breast tumor growth and invasiveness comprises administering of a
composition of
Formula I in combination with doxorubicin.
[0075] In another embodiment of this aspect of the invention, the method of
blocking
primary breast tumor growth and invasiveness comprises administering of a
composition of
Formula I in combination with pegylated liposomal doxorubicin.
[0076] In another embodiment of this aspect of the invention, the method of
blocking
primary breast tumor growth and invasiveness comprises administering of a
compositionof
Formula I in combination with epirubicin.
14
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[0077] In yet another embodiment of this aspect of the invention, the
method of blocking
primary breast tumor growth and invasiveness comprises the administration of a
composition of
Formula I in combination with tamoxifen.
[0078] In another embodiment of this aspect of the invention, the method of
blocking
primary breast tumor growth and invasiveness comprises the administration of a
composition of
Formula I in combination with fulvestrant.
[0079] In another embodiment of this aspect of the invention, the method of
blocking
primary breast tumor growth and invasiveness comprises the administration of a
composition of
Formula I in combination with anastrozole.
[0080] In another embodiment of this aspect of the invention, the method of
blocking
primary breast tumor growth and invasiveness comprises the administration of a
composition of
Formula I in combination with letrozole.
[0081] \In another embodiment of this aspect of the invention, the method
of blocking
primary breast tumor growth and invasiveness comprises the administration of a
composition of
Formula I in combination with exemestane.
[0082] In another embodiment of this aspect of the invention, the method of
blocking
primary breast tumor growth and invasiveness comprises the administration of a
composition of
Formula I in combination with trastuzumab.
[0083] In another embodiment of this aspect of the invention, the method of
blocking
primary breast tumor growth and invasiveness comprises the administration of a
composition of
Formula I in combination with ado-trastuzumab emtansine.
[0084] In another embodiment of this aspect of the invention, the method of
blocking
primary breast tumor growth and invasiveness comprises administering a
composition of
Formula I in combination with pertuzumab.
[0085] In another embodiment of this aspect of the invention, the method of
blocking
primary breast tumor growth and invasiveness comprises the administration of a
composition of
Formula I in combination with lapatinib.
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[0086] In another embodiment of this aspect of the invention, the method of
blocking
primary breast tumor growth and invasiveness comprises the administration of a
composition of
Formula I in combination with everolimus.
[0087] In another embodiment of this aspect of the invention, the method of
blocking
primary breast tumor growth and invasiveness comprises the administration of a
composition of
Formula I in combination with temsirolimus.
[0088] In another embodiment of this aspect of the invention, the method of
blocking
primary breast tumor growth and invasiveness comprises the administration of a
composition of
Formula I in combination with the CDK4/6 inhibitor LY2835219.
[0089] In another embodiment of this aspect of the invention, the method of
blocking
primary breast tumor growth and invasiveness comprises the administration of a
composition of
Formula I in combination with the CDK4/6 inhibitor LEE011.
[0090] In another embodiment of this aspect of the invention, the method of
blocking
primary breast tumor growth and invasiveness comprises the administration of a
composition of
Formula I in combination with the CDK4/6 inhibitor PD 0332991.
[0091] In another embodiment of this aspect of the invention, the method of
blocking
primary breast tumor growth and invasiveness comprises the administration of a
composition of
Formula I in combination with bevacizumab.
[0092] Another aspect of the invention relates to a method of blocking
primary breast tumor
growth and invasiveness which comprises the administration of a composition of
Formula I at
doses sufficient to block TIE2 kinase in the tumor microenvironment, wherein
the composition
of Formula I is administered in an intermittent non-daily dosing regimen. In
some embodiments,
the intermittent non-daily dosing regimen, includes alternate daily dosing,
every third daily
dosing, twice weekly dosing, and once weekly dosing.
[0093] In another embodiment of this aspect of the invention, a method of
blocking primary
breast tumor growth and invasiveness comprises the administration of a
composition of Formula
I at doses sufficient to block TIE2 kinase in the tumor microenvironment,
wherein the
composition of Formula I is administered twice weekly, once weekly, or
alternate weekly.
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[0094] In another embodiment of this aspect of the invention, a method of
blocking primary
breast tumor growth and invasiveness comprises the administration of a
composition of Formula
I at doses sufficient to block TIE2 kinase in the tumor microenvironment,
wherein the
composition of Formula I is administered twice weekly or once weekly.
[0095] In another embodiment of this aspect of the invention, a method of
blocking primary
breast tumor growth and invasiveness comprises the administration of a
composition of Formula
I at doses sufficient to block TIE2 kinase in the tumor microenvironment,
wherein the
composition of Formula I is administered twice weekly.
[0096] In another embodiment of this aspect of the invention, a method of
blocking primary
breast tumor growth and invasiveness comprises the administration of a
composition of Formula
I at doses sufficient to block TIE2 kinase in the tumor microenvironment
wherein the
composition of Formula I is administered in combination with one or more
agents taken from an
anti-tubulin agent, a DNA alkylating agent, a DNA synthesis-inhibiting agent,
a DNA
intercalating agent, an anti-estrogen agent, an anti-HER2 agent, a kinase
inhibitor or an anti-
angiogenic agent.
[0097] In another embodiment of this aspect of the invention, the method of
blocking
primary breast tumor growth and invasiveness comprises administering a
composition of
Formula I in combination with paclitaxel.
[0098] In another embodiment of this aspect of the invention, the method of
blocking
primary breast tumor growth and invasiveness comprises administering a
composition of
Formula I in combination with paclitaxel protein-bound particles for
injectable suspension.
[0099] In another embodiment of this aspect of the invention, the method of
blocking
primary breast tumor growth and invasiveness comprises administering a
composition of
Formula I in combination with docetaxel.
[0100] In another embodiment of this aspect of the invention, a method of
blocking primary
breast tumor growth and invasiveness comprises the administration of a
composition of Formula
I in combination with eribulin.
17
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[0101] In another embodiment of this aspect of the invention, a method of
blocking primary
breast tumor growth and invasiveness comprises the administration of a
composition of Formula
I in combination with ixabepilone.
[0102] In another embodiment of this aspect of the invention, a method of
blocking primary
breast tumor growth and invasiveness comprises the administration of a
composition of Formula
I in combination with vinorelbine.
[0103] In another embodiment of this aspect of the invention, the method of
blocking
primary breast tumor growth and invasiveness comprises the administration of a
composition of
Formula I in combination with capecitabine.
[0104] In another embodiment of this aspect of the invention, the method of
blocking
primary breast tumor growth and invasiveness comprises the administration of a
composition of
Formula I in combination with gemcitabine.
[0105] In another embodiment of this aspect of the invention, the method of
blocking
primary breast tumor growth and invasiveness comprises the administration of a
composition of
Formula I in combination with 5-fluorouracil.
[0106] In another embodiment of this aspect of the invention, the method of
blocking
primary breast tumor growth and invasiveness comprises the administration of a
composition of
Formula I in combination with methotrexate.
[0107] In another embodiment of this aspect of the invention, the method of
blocking
primary breast tumor growth and invasiveness comprises the administration of a
composition of
Formula I in combination with cyclophosphamide.
[0108] In another embodiment of this aspect of the invention, the method of
blocking
primary breast tumor growth and invasiveness comprises the administration of a
composition of
Formula I in combination with cisplatin.
[0109] In another embodiment of this aspect of the invention, the method of
blocking
primary breast tumor growth and invasiveness comprises the administration of a
composition of
Formula I in combination with carboplatin.
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[0110] In another embodiment of this aspect of the invention, the method of
blocking
primary breast tumor growth and invasiveness comprises administering of a
composition of
Formula I in combination with doxorubicin.
[0111] In another embodiment of this aspect of the invention, the method of
blocking
primary breast tumor growth and invasiveness comprises administering of a
composition of
Formula I in combination with pegylated liposomal doxorubicin.
[0112] In another embodiment of this aspect of the invention, the method of
blocking
primary breast tumor growth and invasiveness comprises administering of a
composition of
Formula I in combination with epirubicin.
[0113] In yet another embodiment of this aspect of the invention, the
method of blocking
primary breast tumor growth and invasiveness comprises the administration of a
composition of
Formula I in combination with tamoxifen.
[0114] In another embodiment of this aspect of the invention, the method of
blocking
primary breast tumor growth and invasiveness comprises the administration of a
composition of
Formula I in combination with fulvestrant.
[0115] In another embodiment of this aspect of the invention, the method of
blocking
primary breast tumor growth and invasiveness comprises the administration of a
composition of
Formula I in combination with anastrozole.
[0116] In another embodiment of this aspect of the invention, the method of
blocking
primary breast tumor growth and invasiveness comprises the administration of a
composition of
Formula I in combination with letrozole.
[0117] In another embodiment of this aspect of the invention, the method of
blocking
primary breast tumor growth and invasiveness comprises the administration of a
composition of
Formula I in combination with exemestane.
[0118] In another embodiment of this aspect of the invention, the method of
blocking
primary breast tumor growth and invasiveness comprises the administration of a
composition of
Formula I in combination with trastuzumab.
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[0119] In another embodiment of this aspect of the invention, the method of
blocking
primary breast tumor growth and invasiveness comprises the administration of a
composition of
Formula I in combination with ado-trastuzumab emtansine.
[0120] In another embodiment of this aspect of the invention, the method of
blocking
primary breast tumor growth and invasiveness comprises administering a
composition of
Formula I in combination with pertuzumab.
[0121] In another embodiment of this aspect of the invention, the method of
blocking
primary breast tumor growth and invasiveness comprises the administration of a
composition of
Formula I at doses sufficient to block TIE2 kinase in tumor microenvironment
TIE2-expressing
macrophages administered in combination with lapatinib.
[0122] In another embodiment of this aspect of the invention, the method of
blocking
primary breast tumor growth and invasiveness comprises the administration of a
composition of
Formula I at doses sufficient to block TIE2 kinase in tumor microenvironment
TIE2-expressing
macrophages administered in combination with everolimus.
[0123] In another embodiment of this aspect of the invention, the method of
blocking
primary breast tumor growth and invasiveness comprises the administration of a
composition of
Formula I at doses sufficient to block TIE2 kinase in tumor microenvironment
TIE2-expressing
macrophages administered in combination with temsirolimus.
[0124] In another embodiment of this aspect of the invention, the method of
blocking
primary breast tumor growth and invasiveness comprises the administration of a
composition of
Formula I at doses sufficient to block TIE2 kinase in tumor microenvironment
TIE2-expressing
macrophages administered in combination with the CDK4/6 inhibitor LY2835219.
[0125] In another embodiment of this aspect of the invention, the method of
blocking
primary breast tumor growth and invasiveness comprises the administration of a
composition of
Formula I at doses sufficient to block TIE2 kinase in tumor microenvironment
TIE2-expressing
macrophages administered in combination with the CDK4/6 inhibitor LEE011.
[0126] In another embodiment of this aspect of the invention, the method of
blocking
primary breast tumor growth and invasiveness comprises the administration of a
composition of
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Formula I at doses sufficient to block TIE2 kinase in tumor microenvironment
TIE2-expressing
macrophages administered in combination with the CDK4/6 inhibitor PD 0332991.
[0127] In another embodiment of this aspect of the invention, the method of
blocking
primary breast tumor growth and invasiveness comprises the administration of a
composition of
Formula I in combination with bevacizumab.
Methods for blocking breast cancer intravasation, dissemination and
metastasis:
[0128] In yet another aspect of the invention a method of blocking breast
cancer
intravasation, dissemination and metastasis is provided which comprises
administering to patient
in need thereof an effective amount of a composition of Formula I sufficient
to block TIE2
kinase in the tumor microenvironment.
[0129] In one embodiment of this aspect of the invention, the method of
blocking breast
cancer intravasation, dissemination and metastasis comprises administering to
patient in need
thereof an effective amount of a composition of Formula I in a dosing regimen
sufficient to block
TIE2 kinase in the tumor microenvironment.
[0130] In another embodiment of this aspect of the invention, the dosing
regimen sufficient
to block breast cancer intravasation, dissemination and metastasis comprises
the daily
administration of a composition of Formula I.
[0131] In another embodiment of this aspect of the invention, the dosing
regimen of a
composition of Formula I is administered in an intermittent non-daily dosing
manner, including
alternate daily dosing, every third daily dosing, twice weekly dosing, or once
weekly dosing.
[0132] In another embodiment of this aspect of the invention, the dosing
regimen of a
composition of Formula I is administered twice weekly, once weekly, or
alternate weekly.
[0133] In another embodiment of this aspect of the invention, the dosing
regimen of a
composition of Formula I is twice weekly or once weekly administration.
[0134] In another embodiment of this aspect of the invention, the dosing
regimen a
composition of Formula I is administered twice weekly.
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[0135] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer intravasation, dissemination and metastasis comprises administering to
a patient in need
thereof an effective amount of a composition of Formula I in a dosing regimen
sufficient to block
TIE2 kinase in the tumor microenvironment in combination with one or more
agents taken from
an anti-tubulin agent, a DNA alkylating agent, a DNA synthesis-inhibiting
agent, a DNA
intercalating agent, an anti-estrogen agent, an anti-HER2 agent, a kinase
inhibitor or an anti-
angiogenic agent.
[0136] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer intravasation, dissemination and metastasis comprises administering a
composition of
Formula I in combination with paclitaxel.
[0137] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer intravasation, dissemination and metastasis comprises administering a
composition of
Formula I in combination with paclitaxel protein-boundn particles for
injectable suspension.
[0138] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer intravasation, dissemination and metastasis comprises administering a
composition of
Formula I in combination with docetaxel.
[0139] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer intravasation, dissemination and metastasis comprises administering a
composition of
Formula I in combination with eribulin.
[0140] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer intravasation, dissemination and metastasis comprises adminstering a
composition of
Formula I in combination with ixabepilone.
[0141] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer intravasation, dissemination and metastasis comprises adminstering a
composition of
Formula I in combination with vinorelbine.
[0142] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer intravasation, dissemination and metastasis comprises administering a
composition of
Formula I in combination with capecitabine.
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[0143] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer intravasation, dissemination and metastasis comprises administering a
composition of
Formula I in combination with gemcitabine.
[0144] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer intravasation, dissemination and metastasis comprises administering a
composition of
Formula I in combination with 5-fluorouracil.
[0145] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer intravasation, dissemination and metastasis comprises administering a
composition of
Formula I in combination with methotrexate.
[0146] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer intravasation, dissemination and metastasis comprises administering a
composition of
Formula I in combination with cyclophosphamide.
[0147] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer intravasation, dissemination and metastasis comprises administering a
composition of
Formula I in combination with cisplatin.
[0148] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer intravasation, dissemination and metastasis comprises administering a
composition of
Formula I in combination with carboplatin.
[0149] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer intravasation, dissemination and metastasis comprises administering a
composition of
Formula I in combination with doxorubicin.
[0150] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer intravasation, dissemination and metastasis comprises administering a
composition of
Formula I in combination with epirubicin.
[0151] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer intravasation, dissemination and metastasis comprises administering a
composition of
Formula I in combination with tamoxifen.
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[0152] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer intravasation, dissemination and metastasis comprises administering a
composition of
Formula I in combination with fulvestrant.
[0153] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer intravasation, dissemination and metastasis comprises administering a
composition of
Formula I in combination with anastrozole.
[0154] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer intravasation, dissemination and metastasis comprises administering a
composition of
Formula I in combination with letrozole.
[0155] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer intravasation, dissemination and metastasis comprises administering a
composition of
Formula I in combination with exemestane.
[0156] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer intravasation, dissemination and metastasis comprises administering a
composition of
Formula I in combination with trastuzumab.
[0157] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer intravasation, dissemination and metastasis comprises administering a
composition of
Formula I in combination with ado-trastuzumab emtansine.
[0158] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer intravasation, dissemination and metastasis comprises the
administration of a composition
of Formula I in combination with pertuzumab.
[0159] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer intravasation, dissemination and metastasis comprises administering a
composition of
Formula I in combination with lapatinib.
[0160] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer intravasation, dissemination and metastasis comprises administering a
composition of
Formula I in combination with everolimus.
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[0161] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer intravasation, dissemination and metastasis comprises administering a
composition of
Formula I in combination with temsirolimus.
[0162] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer intravasation, dissemination and metastasis comprises administering a
composition of
Formula I in combination with the CDK4/6 inhibitor LY2835219.
[0163] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer intravasation, dissemination and metastasis comprises administering a
composition of
Formula I in combination with the CDK4/6 inhibitor LEE011.
[0164] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer intravasation, dissemination and metastasis comprises administering a
composition of
Formula I in combination with the CDK4/6 inhibitor PD 0332991.
[0165] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer intravasation, dissemination and metastasis comprises administering a
composition of
Formula I in combination with bevacizumab.
[0166] In another aspect of the invention, a method of blocking breast
cancer intravasation,
dissemination and metastasis comprises administration of a composition of
Formula I at doses
sufficient to block TIE2 kinase in the tumor microenvironment, wherein the
dosing regimen of
the composition of Formula I is administered as intermittent non-daily dosing.
In some
embodiments, the alternate daily dosing includes every third daily dosing,
twice weekly dosing,
or once weekly dosing.
[0167] In one embodiment of this aspect of the invention, a method of
blocking breast cancer
intravasation, dissemination and metastasis comprises the administration of a
composition of
Formula I at doses sufficient to block TIE2 kinase in the tumor
microenvironment, with the
dosing regimen of the composition of Formula I being administered twice
weekly, once weekly,
or alternate weekly.
[0168] In another embodiment of this aspect of the invention, a method of
blocking breast
cancer intravasation, dissemination and metastasis comprises the
administration of a composition
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of Formula I at doses sufficient to block TIE2 kinase in the tumor
microenvironment, with the
dosing regimen of the composition of Formula I being administered twice weekly
or once
weekly.
[0169] In another embodiment of this aspect of the invention, a method of
blocking breast
cancer intravasation, dissemination and metastasis comprises the
administration of a composition
of Formula I at doses sufficient to block TIE2 kinase in the tumor
microenvironment, with the
dosing regimen of the composition of Formula I being administered twice
weekly.
[0170] In another embodiment of this aspect of the invention, a method of
blocking breast
cancer intravasation, dissemination and metastasis comprises the
administration of a composition
of Formula I at doses sufficient to block TIE2 kinase in the tumor
microenvironment
administered in combination with one or more agents taken from an anti-tubulin
agent, a DNA
alkylating agent, a DNA synthesis-inhibiting agent, a DNA intercalating agent,
an anti-estrogen
agent, an anti-HER2 agent, a kinase inhibitor or an anti-angiogenic agent.
[0171] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer intravasation, dissemination and metastasis comprises administering a
composition of
Formula I in combination with paclitaxel.
[0172] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer intravasation, dissemination and metastasis comprises administering a
composition of
Formula I in combination with paclitaxel protein-bound particles for
injectable suspension.
[0173] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer intravasation, dissemination and metastasis comprises administering a
composition of
Formula I in combination with docetaxel.
[0174] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer intravasation, dissemination and metastasis comprises the
administration of a composition
of Formula I in combination with eribulin.
[0175] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer intravasation, dissemination and metastasis comprises the
administration of a composition
of Formula I in combination with ixabepilone.
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[0176] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer intravasation, dissemination and metastasis comprises the
administration of a composition
of Formula I in combination with vinorelbine.
[0177] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer intravasation, dissemination and metastasis comprises the
administration of a composition
of Formula I in combination with capecitabine.
[0178] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer intravasation, dissemination and metastasis comprises the
administration of a composition
of Formula I in combination with gemcitabine.
[0179] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer intravasation, dissemination and metastasis comprises the
administration of a composition
of Formula I in combination with 5-fluorouracil.
[0180] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer intravasation, dissemination and metastasis comprises the
administration of a composition
of Formula I in combination with methotrexate.
[0181] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer intravasation, dissemination and metastasis comprises the
administration of a composition
of Formula I in combination with cyclophosphamide.
[0182] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer intravasation, dissemination and metastasis comprises the
administration of a composition
of Formula I in combination with cisplatin.
[0183] In another embodiment of of this aspect of the invention, the method
of blocking
breast cancer intravasation, dissemination and metastasis comprises the
administration of a
composition of Formula I in combination with carboplatin.
[0184] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer intravasation, dissemination and metastasis comprises administering of
a composition of
Formula I in combination with doxorubicin.
27
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[0185] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer intravasation, dissemination and metastasis comprises administering of
a composition of
Formula I in combination with pegylated liposomal doxorubicin.
[0186] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer intravasation, dissemination and metastasis comprises administering of
a composition of
Formula I in combination with epirubicin.
[0187] In yet another embodiment of this aspect of the invention, the
method of blocking
breast cancer intravasation, dissemination and metastasis comprises the
administration of a
composition of Formula I in combination with tamoxifen.
[0188] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer intravasation, dissemination and metastasis comprises the
administration of a
composition of Formula I in combination with fulvestrant.
[0189] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer intravasation, dissemination and metastasis comprises the
administration of a
composition of Formula I in combination with anastrozole.
[0190] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer intravasation, dissemination and metastasis comprises the
administration of a
composition of Formula I in combination with letrozole.
[0191] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer intravasation, dissemination and metastasis comprises the
administration of a
composition of Formula I in combination with exemestane.
[0192] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer intravasation, dissemination and metastasis comprises the
administration of a
composition of Formula I in combination with trastuzumab.
[0193] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer intravasation, dissemination and metastasis comprises the
administration of a
composition of Formula I in combination with ado-trastuzumab emtansine.
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[0194] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer intravasation, dissemination and metastasis comprises administering a
composition of
Formula I in combination with pertuzumab.
[0195] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer intravasation, dissemination and metastasis comprises the
administration of a
composition of Formula I at doses sufficient to block TIE2 kinase in tumor
microenvironment
TIE2-expressing macrophages administered in combination with lapatinib.
[0196] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer intravasation, dissemination and metastasis comprises the
administration of a composition
of Formula I at doses sufficient to block TIE2 kinase in tumor
microenvironment TIE2-
expressing macrophages administered in combination with everolimus.
[0197] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer intravasation, dissemination and metastasis comprises the
administration of a composition
of Formula I at doses sufficient to block TIE2 kinase in tumor
microenvironment TIE2-
expressing macrophages administered in combination with temsirolimus.
[0198] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer intravasation, dissemination and metastasis comprises the
administration of a composition
of Formula I at doses sufficient to block TIE2 kinase in tumor
microenvironment TIE2-
expressing macrophages administered in combination with the CDK4/6 inhibitor
LY2835219.
[0199] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer intravasation, dissemination and metastasis comprises the
administration of a composition
of Formula I at doses sufficient to block TIE2 kinase in tumor
microenvironment TIE2-
expressing macrophages administered in combination with the CDK4/6 inhibitor
LEE011.
[0200] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer intravasation, dissemination and metastasis comprises the
administration of a composition
of Formula I at doses sufficient to block TIE2 kinase in tumor
microenvironment TIE2-
expressing macrophages administered in combination with the CDK4/6 inhibitor
PD 0332991.
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[0201] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer intravasation, dissemination and metastasis comprises the
administration of a
composition of Formula I in combination with bevacizumab.
Methods for blocking breast cancer immunotolerance:
[0202] Another aspect of the invention relates to a method of blocking
breast cancer
immunotolerance. The method comprises administering to a patient in need
thereof an effective
amount of a composition of Formula I. In one embodiment, the dosing regimen of
the salt is
sufficient to block TIE2 kinase in the tumor microenvironment that mediates
immunotolerance.
[0203] In one embodiment of this aspect of the invention, the method of
blocking breast
cancer immunotolerance comprises administering to a patient in need thereof an
effective
amount of a composition of Formula I in a dosing regimen sufficient to block
TIE2 kinase in the
tumor microenvironment that mediates immunotolerance.
[0204] In one embodiment of this aspect of the invention, the method of
blocking breast
cancer immunotolerance comprises the administration of a composition of
Formula I daily.
[0205] In another embodiment of this aspect of the invention,the
composition of Formula I
is administered in an intermittent non-daily manner. In some embodiments, the
intermittent non-
daily manner includes alternate daily dosing, every third daily dosing, twice
weekly dosing, or
once weekly dosing.
[0206] In another embodiment of this aspect of the invention,
administration of the
composition of Formula I is twice weekly, once weekly, or alternate weekly.
[0207] In another embodiment of this aspect of the invention, the
composition of Formula I
is administered twice weekly or once weekly.
[0208] In another embodiment of this aspect of the invention, the
composition of Formula I
is administered twice weekly.
[0209] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer immunotolerance comprises administering to a patient in need thereof an
effective
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amount of a composition of Formula I in a dosing regimen sufficient to block
TIE2 kinase in the
tumor microenvironment in combination with one or more agents taken from an
anti-tubulin
agent, a DNA alkylating agent, a DNA synthesis-inhibiting agent, a DNA
intercalating agent, an
anti-estrogen agent, an anti-HER2 agent, a kinase inhibitor, an anti-
angiogenic agent, or an
immunomodulating agent.
[0210] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer immunotolerance comprises administering a composition of Formula I in
combination
with paclitaxel.
[0211] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer immunotolerance comprises administering a composition of Formula I in
combination
with paclitaxel protein-bound particles for injectable suspension.
[0212] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer immunotolerance comprises administering a composition of Formula I in
combination
with docetaxel.
[0213] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer immunotolerance comprises the administration of a composition of
Formula I in
combination with eribulin.
[0214] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer immunotolerance comprises the administration of a composition of
Formula I in
combination with ixabepilone.
[0215] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer immunotolerance comprises the administration of a composition of
Formula I in
combination with vinorelbine.
[0216] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer immunotolerance comprises the administration of a composition of
Formula I in
combination with capecitabine.
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[0217] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer immunotolerance comprises the administration of a composition of
Formula I in
combination with gemcitabine.
[0218] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer immunotolerance comprises the administration of a composition of
Formula I in
combination with 5-fluorouracil.
[0219] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer immunotolerance comprises the administration of a composition of
Formula I in
combination with 5-methotrexate.
[0220] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer immunotolerance comprises the administration of a composition of
Formula I in
combination with cyclophosphamide.
[0221] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer immunotolerance comprises the administration of a composition of
Formula I in
combination with cisplatin.
[0222] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer immunotolerance comprises the administration of a composition of
Formula I in
combination with carboplatin.
[0223] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer immunotolerance comprises administering of a composition of Formula I
in combination
with doxorubicin.
[0224] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer immunotolerance comprises administering of a composition of Formula I
in combination
with pegylated liposomal doxorubicin.
[0225] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer immunotolerance comprises administering of a composition of Formula I
in combination
with epirubicin.
32
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[0226] In yet another embodiment of this aspect of the invention, the
method of blocking
breast cancer immunotolerance comprises the administration of a composition of
Formula I in
combination with tamoxifen.
[0227] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer immunotolerance comprises the administration of a composition of
Formula I in
combination with fulvestrant.
[0228] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer immunotolerance comprises the administration of a composition of
Formula I in
combination with anastrozole.
[0229] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer immunotolerance comprises the administration of a composition of
Formula I in
combination with letrozole.
[0230] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer immunotolerance comprises the administration of a composition of
Formula I in
combination with exemestane.
[0231] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer immunotolerance comprises the administration of a composition of
Formula I in
combination with trastuzumab.
[0232] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer immunotolerance comprises the administration of a composition of
Formula I in
combination with ado-trastuzumab emtansine.
[0233] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer immunotolerance comprises administering a composition of Formula I in
combination
with pertuzumab.
[0234] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer immunotolerance comprises the administration of a composition of
Formula I at doses
sufficient to block TIE2 kinase in tumor microenvironment TIE2-expressing
macrophages
administered in combination with lapatinib.
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[0235] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer immunotolerance comprises the administration of a composition of
Formula I at doses
sufficient to block TIE2 kinase in tumor microenvironment TIE2-expressing
macrophages
administered in combination with everolimus.
[0236] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer immunotolerance comprises the administration of a composition of
Formula I at doses
sufficient to block TIE2 kinase in tumor microenvironment TIE2-expressing
macrophages
administered in combination with temsirolimus.
[0237] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer immunotolerance comprises the administration of a composition of
Formula I at doses
sufficient to block TIE2 kinase in tumor microenvironment TIE2-expressing
macrophages
administered in combination with the CDK4/6 inhibitor LY2835219.
[0238] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer immunotolerance comprises the administration of a composition of
Formula I at doses
sufficient to block TIE2 kinase in tumor microenvironment TIE2-expressing
macrophages
administered in combination with the CDK4/6 inhibitor LEE011.
[0239] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer immunotolerance comprises the administration of a composition of
Formula I at doses
sufficient to block TIE2 kinase in tumor microenvironment TIE2-expressing
macrophages
administered in combination with the CDK4/6 inhibitor PD 0332991.
[0240] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer immunotolerance comprises the administration of a composition of
Formula I in
combination with bevacizumab.
[0241] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer immunotolerance comprises administering a composition of Formula I in
combination
with an anti-CTLA-4 agent.
34
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[0242] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer immunotolerance comprises administering a composition of Formula I in
combination
with ipilimumab.
[0243] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer immunotolerance comprises administering a composition of Formula I in
combination
with an anti-PD-1 agent.
[0244] In another embodiment of this aspect of the invention, a method of
blocking breast
cancer immunotolerance comprises administering a composition of Formula I in
combination
with lambrolizumab.
[0245] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer immunotolerance comprises administering a composition of Formula I in
combination
with an anti-PD L-1 agent.
[0246] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer immunotolerance comprises administering a composition of Formula I in
combination
with MPDL3280A.
Methods for blocking breast cancer immunotolerance:
[0247] In another aspect of the invention, a method of blocking breast
cancer
immunotolerance comprises the administration of a composition of Formula I at
doses sufficient
to block TIE2 kinase in the tumor microenvironment, wherein the dosing regimen
of the
composition of Formula I is administered in an intermittent non-daily dosing
manner, including
alternate daily dosing, every third daily dosing, twice weekly dosing, or once
weekly dosing.
[0248] In one embodiment of this aspect of the invention, a method of
blocking breast cancer
immunotolerance comprises the administration of a composition of Formula I at
doses sufficient
to block TIE2 kinase in the tumor microenvironment, wherein the composition of
Formula I is
administered twice weekly, once weekly, or alternate weekly.
[0249] In another embodiment of this aspect of the invention, a method of
blocking breast
cancer immunotolerance comprises the administration of a composition of
Formula I at doses
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sufficient to block TIE2 kinase in the tumor microenvironment, wherein the
composition of
Formula I is administered twice weekly or once weekly.
[0250] In another embodiment of this aspect of the invention, a method of
blocking breast
cancer immunotolerance comprises the administration of a composition of
Formula I at doses
sufficient to block TIE2 kinase in the tumor microenvironment, wherein the
composition of
Formula I is administered twice weekly.
[0251] In another embodiment of this aspect of the invention, a method of
blocking breast
cancer immunotolerance comprises the administration of a composition of
Formula I at doses
sufficient to block TIE2 kinase in the tumor microenvironment administered in
combination with
one or more agents taken from an anti-tubulin agent, a DNA alkylating agent, a
DNA synthesis-
inhibiting agent, a DNA intercalating agent, an anti-estrogen agent, an anti-
HER2 agent, a kinase
inhibitor, an anti-angiogenic agent, or an immunomodulating agent.
[0252] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer immunotolerance comprises administering a composition of Formula I in
combination
with paclitaxel.
[0253] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer immunotolerance comprises administering a composition of Formula I in
combination
with paclitaxel protein-bound particles for injectable suspension.
[0254] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer immunotolerance comprises administering a composition of Formula I in
combination
with docetaxel.
[0255] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer immunotolerance comprises the administration of a composition of
Formula I in
combination with eribulin.
[0256] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer immunotolerance comprises the administration of a composition of
Formula I in
combination with ixabepilone.
36
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[0257] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer immunotolerance comprises the administration of a composition of
Formula I in
combination with vinorelbine.
[0258] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer immunotolerance comprises the administration of a composition of
Formula I in
combination with capecitabine.
[0259] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer immunotolerance comprises the administration of a composition of
Formula I in
combination with gemcitabine.
[0260] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer immunotolerance comprises the administration of a composition of
Formula I in
combination with 5-fluorouracil.
[0261] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer immunotolerance comprises the administration of a composition of
Formula I in
combination with methotrexate.
[0262] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer immunotolerance comprises the administration of a composition of
Formula I in
combination with cyclophosphamide.
[0263] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer immunotolerance comprises the administration of a composition of
Formula I in
combination with cisplatin.
[0264] In another embodiment of of this aspect of the invention, the method
of blocking
breast cancer immunotolerance comprises the administration of a composition of
Formula I in
combination with carboplatin.
[0265] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer immunotolerance comprises administering of a composition of Formula I
in combination
with doxorubicin.
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[0266] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer immunotolerance comprises administering of a composition of Formula I
in combination
with pegylated liposomal doxorubicin.
[0267] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer immunotolerance comprises administering of a composition of Formula I
in combination
with epirubicin.
[0268] In yet another embodiment of this aspect of the invention, the
method of blocking
breast cancer immunotolerance comprises the administration of a composition of
Formula I in
combination with tamoxifen.
[0269] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer immunotolerance comprises the administration of a composition of
Formula I in
combination with fulvestrant.
[0270] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer immunotolerance comprises the administration of a composition of
Formula I in
combination with anastrozole.
[0271] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer immunotolerance comprises the administration of a composition of
Formula I in
combination with letrozole.
[0272] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer immunotolerance comprises the administration of a composition of
Formula I in
combination with exemestane.
[0273] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer immunotolerance comprises the administration of a composition of
Formula I in
combination with trastuzumab.
[0274] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer immunotolerance comprises the administration of a a composition of
Formula I in
combination with ado-trastuzumab emtansine.
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[0275] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer immunotolerance comprises administering a composition of Formula I in
combination
with pertuzumab.
[0276] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer immunotolerance comprises the administration of a a composition of
Formula I at doses
sufficient to block TIE2 kinase in tumor microenvironment TIE2-expressing
macrophages
administered in combination with lapatinib.
[0277] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer immunotolerance comprises the administration of a composition of
Formula I at doses
sufficient to block TIE2 kinase in tumor microenvironment TIE2-expressing
macrophages
administered in combination with everolimus.
[0278] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer immunotolerance comprises the administration of a composition of
Formula I at doses
sufficient to block TIE2 kinase in tumor microenvironment TIE2-expressing
macrophages
administered in combination with temsirolimus.
[0279] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer immunotolerance comprises the administration of a composition of
Formula I at doses
sufficient to block TIE2 kinase in tumor microenvironment TIE2-expressing
macrophages
administered in combination with the CDK4/6 inhibitor LY2835219.
[0280] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer immunotolerance comprises the administration of a composition of
Formula I at doses
sufficient to block TIE2 kinase in tumor microenvironment TIE2-expressing
macrophages
administered in combination with the CDK4/6 inhibitor LEE011.
[0281] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer immunotolerance comprises the administration of a composition of
Formula I at doses
sufficient to block TIE2 kinase in tumor microenvironment TIE2-expressing
macrophages
administered in combination with the CDK4/6 inhibitor PD 0332991.
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[0282] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer immunotolerance comprises the administration of a composition of
Formula I in
combination with bevacizumab.
[0283] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer immunotolerance comprises administering a composition of Formula I in
combination
with an anti-CTLA-4 agent.
[0284] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer immunotolerance comprises administering a composition of Formula I in
combination
with ipilimumab.
[0285] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer immunotolerance comprises administering a composition of Formula I in
combination
with an anti-PD-1 agent.
[0286] In another embodiment of this aspect of the invention, a method of
blocking breast
cancer immunotolerance comprises administering a composition of Formula I in
combination
with lambrolizumab.
[0287] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer immunotolerance comprises administering a composition of Formula I in
combination
with an anti-PD L-1 agent.
[0288] In another embodiment of this aspect of the invention, the method of
blocking breast
cancer immunotolerance comprises administering a composition of Formula I in
combination
with MPDL3280A.
Methods for increasing overall survival in breast cancer patients:
[0289] Another aspect of the invention relates to a method of increasing
overall survival in
breast cancer patients compriseing administering to a patient in need thereof
an effective amount
of a composition of Formula I. In one embodiment, the dosing regimen is
sufficient to block
TIE2 kinase in the tumor microenvironment .
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[0290] In another embodiment of this aspect of the invention, the method of
increasing
overall survival in breast cancer patients comprises a dosing regimen wherein
the composition of
Formula I administered daily.
[0291] In another embodiment of this aspect of the invention, the method of
increasing
overall survival in breast cancer patients comprises a composition of Formula
I in a dosing
regimen administered in an intermittent non-daily manner, including alternate
daily dosing,
every third daily dosing, twice weekly dosing, or once weekly dosing.
[0292] In another embodiment of this aspect of the invention, the method of
increasing
overall survival in breast cancer patients comprises a composition of Formula
I in a dosing
regimen administered twice weekly, once weekly, or alternate weekly.
[0293] In another embodiment of this aspect of the invention, the method of
increasing
overall survival in breast cancer patients, the dosing regimen of a
composition of Formula I is
administered twice weekly or once weekly.
[0294] In another embodiment of this aspect of the invention, the dosing
regimen of a
composition of Formula I is administered only twice weekly.
[0295] Another embodiment of this aspect of the invention relates to the
method of
increasing overall survival in breast cancer patients which comprises
administering a
composition of Formula I in a dosing regimen sufficient to block TIE2 kinase
in the tumor
microenvironment in combination with one or more agents taken from an anti-
tubulin agent, a
DNA alkylating agent, a DNA synthesis-inhibiting agent, a DNA intercalating
agent, an anti-
estrogen agent, an anti-HER2 agent, a kinase inhibitor, an anti-angiogenic
agent, or an
immunomodulating agent.
[0296] In another embodiment of this aspect of the invention, the method of
increasing
overall survival in breast cancer patients comprises administering a
composition of Formula I in
combination with paclitaxel.
[0297] In another embodiment of this aspect of the invention, the method of
increasing
overall survival in breast cancer patients comprises administering a
composition of Formula I in
combination with paclitaxel protein-bound particles for injectable suspension.
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[0298] In another embodiment of this aspect of the invention, the method of
increasing
overall survival in breast cancer patients comprises administering a
composition of Formula I in
combination with docetaxel.
[0299] In another embodiment of this aspect of the invention, the method of
increasing
overall survival in breast cancer patients comprises the administration of a
composition of
Formula I in combination with eribulin.
[0300] In another embodiment of this aspect of the invention, the method of
increasing
overall survival in breast cancer patients comprises the administration of a
composition of
Formula I in combination with ixabepilone.
[0301] In another embodiment of this aspect of the invention, the method of
increasing
overall survival in breast cancer patients comprises the administration of a
composition of
Formula I in combination with vinorelbine.
[0302] In another embodiment of this aspect of the invention, the method of
increasing
overall survival in breast cancer patients comprises the administration of a
composition of
Formula I in combination with capecitabine.
[0303] In another embodiment of this aspect of the invention, the method of
increasing
overall survival in breast cancer patients comprises the administration of a
composition of
Formula I in combination with gemcitabine.
[0304] In another embodiment of this aspect of the invention, the method of
increasing
overall survival in breast cancer patients comprises the administration of a
composition of
Formula I in combination with 5-fluorouracil.
[0305] In another embodiment of this aspect of the invention, the method of
increasing
overall survival in breast cancer patients comprises the administration of a
composition of
Formula I in combination with methotrexate.
[0306] In another embodiment of this aspect of the invention, the method of
increasing
overall survival in breast cancer patients comprises the administration of a
composition of
Formula I in combination with cyclophosphamide.
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[0307] In another embodiment of this aspect of the invention, the method of
increasing
overall survival in breast cancer patients comprises the administration of a
composition of
Formula I in combination with cisplatin.
[0308] In another embodiment of this aspect of the invention, the method of
increasing
overall survival in breast cancer patients comprises the administration of a
composition of
Formula I in combination with carboplatin.
[0309] In another embodiment of this aspect of the invention, the method of
increasing
overall survival in breast cancer patients comprises administering of a
composition of Formula I
in combination with doxorubicin.
[0310] In another embodiment of this aspect of the invention, the method of
increasing
overall survival in breast cancer patients comprises administering of a
composition of Formula I
in combination with pegylated liposomal doxorubicin.
[0311] In another embodiment of this aspect of the invention, the method of
increasing
overall survival in breast cancer patients comprises administering of a
composition of Formula I
in combination with epirubicin.
[0312] In yet another embodiment of this aspect of the invention, the
method of increasing
overall survival in breast cancer patients comprises the administration of a
composition of
Formula I in combination with tamoxifen.
[0313] In another embodiment of this aspect of the invention, the method of
increasing
overall survival in breast cancer patients comprises the administration of a
composition of
Formula I in combination with fulvestrant.
[0314] In another embodiment of this aspect of the invention, the method of
increasing
overall survival in breast cancer patients comprises the administration of a
composition of
Formula I in combination with anastrozole.
[0315] In another embodiment of this aspect of the invention, the method of
increasing
overall survival in breast cancer patients comprises the administration of a
composition of
Formula I in combination with letrozole.
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[0316] In another embodiment of this aspect of the invention, the method of
increasing
overall survival in breast cancer patients comprises the administration of a
composition of
Formula I in combination with exemestane.
[0317] In another embodiment of this aspect of the invention, the method of
increasing
overall survival in breast cancer patients comprises the administration of a
composition of
Formula I in combination with trastuzumab.
[0318] In another embodiment of this aspect of the invention, the method of
increasing
overall survival in breast cancer patients comprises the administration of a
composition of
Formula I in combination with ado-trastuzumab emtansine.
[0319] In another embodiment of this aspect of the invention, the method of
increasing
overall survival in breast cancer patients comprises administering a
composition of Formula I in
combination with pertuzumab.
[0320] In another embodiment of this aspect of the invention, the method of
increasing
overall survival in breast cancer patients comprises the administration of a
composition of
Formula I at doses sufficient to block TIE2 kinase in tumor microenvironment
TIE2-expressing
macrophages administered in combination with lapatinib.
[0321] In another embodiment of this aspect of the invention, the method of
increasing
overall survival in breast cancer patients comprises the administration of a
composition of
Formula I at doses sufficient to block TIE2 kinase in tumor microenvironment
TIE2-expressing
macrophages administered in combination with everolimus.
[0322] In another embodiment of this aspect of the invention, the method of
increasing
overall survival in breast cancer patients comprises the administration of a
composition of
Formula I at doses sufficient to block TIE2 kinase in tumor microenvironment
TIE2-expressing
macrophages administered in combination with temsirolimus.
[0323] In another embodiment of this aspect of the invention, the method of
increasing
overall survival in breast cancer patients comprises the administration of a
composition of
Formula I at doses sufficient to block TIE2 kinase in tumor microenvironment
TIE2-expressing
macrophages administered in combination with the CDK4/6 inhibitor LY2835219.
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[0324] In another embodiment of this aspect of the invention, the method of
increasing
overall survival in breast cancer patients comprises the administration of a
composition of
Formula I at doses sufficient to block TIE2 kinase in tumor microenvironment
TIE2-expressing
macrophages administered in combination with the CDK4/6 inhibitor LEE011.
[0325] In another embodiment of this aspect of the invention, the method of
increasing
overall survival in breast cancer patients comprises the administration of a
composition of
Formula I at doses sufficient to block TIE2 kinase in tumor microenvironment
TIE2-expressing
macrophages administered in combination with the CDK4/6 inhibitor PD 0332991.
[0326] In another embodiment of this aspect of the invention, the method of
increasing
overall survival in breast cancer patients comprises the administration of a
composition of
Formula I in combination with bevacizumab.
[0327] In another embodiment of this aspect of the invention, the method of
increasing
overall survival in breast cancer patients comprises administering a
composition of Formula I in
combination with an anti-CTLA-4 agent.
[0328] In another embodiment of this aspect of the invention, the method of
increasing
overall survival in breast cancer patients comprises administering a
composition of Formula I in
combination with ipilimumab.
[0329] In another embodiment of this aspect of the invention, the method of
increasing
overall survival in breast cancer patients comprises administering a
composition of Formula I in
combination with an anti-PD-1 agent.
[0330] In another embodiment of this aspect of the invention, the method of
increasing
overall survival in breast cancer patients comprises administering a
composition of Formula I in
combination with lambrolizumab.
[0331] In another embodiment of this aspect of the invention, the method of
increasing
overall survival in breast cancer patients comprises administering a
composition of Formula I in
combination with an anti- anti-PD L-1 agent.
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In another embodiment of this aspect of the invention, the method of
increasing overall survival
in breast cancer patients comprises administering a composition of Formula I
in combination
with MPDL3280A.
Methods for increasing overall survival in breast cancer patients:
[0332] In another aspect of the invention, a method of increasing overall
survival in breast
cancer patients comprises the administration of a composition of Formula I at
doses sufficient to
block TIE2 kinase in the tumor microenvironment, wherein a dosing regimen of
the composition
of Formula I is intermittent non-daily dosing administration, including
alternate daily dosing,
every third daily dosing, twice weekly dosing, or once weekly dosing.
[0333] In one embodiment of this aspect of the invention, a method of
increasing overall
survival in breast cancer patients comprises the administration of a
composition of Formula I at
doses sufficient to block TIE2 kinase in the tumor microenvironment, wherein a
dosing regimen
of the composition of Formula I is twice weekly, once weekly, or alternate
weekly,
administration.
[0334] In another embodiment of this aspect of the invention, a method of
increasing overall
survival in breast cancer patients comprises the administration of a
composition of Formula I at
doses sufficient to block TIE2 kinase in the tumor microenvironment, wherein a
dosing regimen
of the composition of Formula I is twice weekly or once weekly administration.
[0335] In another embodiment of this aspect of the invention, a method of
increasing overall
survival in breast cancer patients comprises the administration of a
composition of Formula I at
doses sufficient to block TIE2 kinase in the tumor microenvironment, with a
dosing regimen of
the composition of Formula I is twice weekly administration.
[0336] In another embodiment of this aspect of the invention, a method of
increasing overall
survival in breast cancer patients comprises the administration of a
composition of Formula I at
doses sufficient to block TIE2 kinase in the tumor microenvironment
administered in
combination with one or more agents taken from an anti-tubulin agent, a DNA
alkylating agent,
a DNA synthesis-inhibiting agent, a DNA intercalating agent, an anti-estrogen
agent, an anti-
HER2 agent, a kinase inhibitor, an anti-angiogenic agent, or an
immunomodulating agent.
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[0337] In another embodiment of this aspect of the invention, the method of
increasing
overall survival in breast cancer patients administering a composition of
Formula I in
combination with paclitaxel.
[0338] In another embodiment of this aspect of the invention, the method of
increasing
overall survival in breast cancer patients comprises administering a
composition of Formula I in
combination with paclitaxel protein-bound particles for injectable suspension.
[0339] In another embodiment of this aspect of the invention, the method of
increasing
overall survival in breast cancer patients comprises administering a
composition of Formula I in
combination with docetaxel.
[0340] In another embodiment of this aspect of the invention, the method of
increasing
overall survival in breast cancer patients comprises the administration of a
composition of
Formula I in combination with eribulin.
[0341] In another embodiment of this aspect of the invention, the method of
increasing
overall survival in breast cancer patients comprises the administration of a
composition of
Formula I in combination with ixabepilone.
[0342] In another embodiment of this aspect of the invention, the method of
increasing
overall survival in breast cancer patients comprises the administration of a
composition of
Formula I in combination with vinorelbine.
[0343] In another embodiment of this aspect of the invention, the method of
increasing
overall survival in breast cancer patients comprises the administration of a
composition of
Formula I in combination with capecitabine.
[0344] In another embodiment of this aspect of the invention, the method of
increasing
overall survival in breast cancer patients comprises the administration of a
composition of
Formula I in combination with gemcitabine.
[0345] In another embodiment of this aspect of the invention, the method of
increasing
overall survival in breast cancer patients comprises the administration of a
composition of
Formula I in combination with 5-fluorouracil.
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[0346] In another embodiment of this aspect of the invention, the method of
increasing
overall survival in breast cancer patients comprises the administration of a
composition of
Formula I in combination with methotrexate.
[0347] In another embodiment of this aspect of the invention, the method of
increasing
overall survival in breast cancer patients comprises the administration of a
composition of
Formula I in combination with cyclophosphamide.
[0348] In another embodiment of this aspect of the invention, the method of
increasing
overall survival in breast cancer patients comprises the administration of a
composition of
Formula I in combination with cisplatin.
[0349] In another embodiment of this aspect of the invention, the method of
increasing
overall survival in breast cancer patients comprises the administration of a
composition of
Formula I in combination with carboplatin.
[0350] In another embodiment of this aspect of the invention, the method of
increasing
overall survival in breast cancer patients comprises administering of a
composition of Formula I
in combination with doxorubicin.
[0351] In another embodiment of this aspect of the invention, the method of
increasing
overall survival in breast cancer patients comprises administering of a
composition of Formula I
in combination with pegylated liposomal doxorubicin.
[0352] In another embodiment of this aspect of the invention, the method of
increasing
overall survival in breast cancer patients comprises administering of a
composition of Formula I
in combination with epirubicin.
[0353] In yet another embodiment of this aspect of the invention, the
method of increasing
overall survival in breast cancer patients comprises the administration of a
composition of
Formula I in combination with tamoxifen.
[0354] In another embodiment of this aspect of the invention, the method of
increasing
overall survival in breast cancer patients comprises the administration of a
composition of
Formula I in combination with fulvestrant.
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[0355] In another embodiment of this aspect of the invention, the method of
increasing
overall survival in breast cancer patients comprises the administration of a
composition of
Formula I in combination with anastrozole.
[0356] In another embodiment of this aspect of the invention, the method of
increasing
overall survival in breast cancer patients comprises the administration of a
composition of
Formula I in combination with letrozole.
[0357] In another embodiment of this aspect of the invention, the method of
increasing
overall survival in breast cancer patients comprises the administration of a
composition of
Formula I in combination with exemestane.
[0358] In another embodiment of this aspect of the invention, the method of
increasing
overall survival in breast cancer patients comprises the administration of a
composition of
Formula I in combination with trastuzumab.
[0359] In another embodiment of this aspect of the invention, the method of
increasing
overall survival in breast cancer patients comprises the administration of a
composition of
Formula I in combination with ado-trastuzumab emtansine.
[0360] In another embodiment of this aspect of the invention, the method of
increasing
overall survival in breast cancer patients comprises administering a
composition of Formula I in
combination with pertuzumab.
[0361] In another embodiment of this aspect of the invention, the method of
increasing
overall survival in breast cancer patients comprises the administration of a
composition of
Formula I at doses sufficient to block TIE2 kinase in tumor microenvironment
TIE2-expressing
macrophages administered in combination with lapatinib.
[0362] In another embodiment of this aspect of the invention, the method of
increasing
overall survival in breast cancer patients comprises the administration of a
composition of
Formula I at doses sufficient to block TIE2 kinase in tumor microenvironment
TIE2-expressing
macrophages administered in combination with everolimus.
[0363] In another embodiment of this aspect of the invention, the method of
increasing
overall survival in breast cancer patients comprises the administration of a
composition of
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Formula I at doses sufficient to block TIE2 kinase in tumor microenvironment
TIE2-expressing
macrophages administered in combination with temsirolimus.
[0364] In another embodiment of this aspect of the invention, the method of
increasing
overall survival in breast cancer patients comprises the administration of a
composition of
Formula I at doses sufficient to block TIE2 kinase in tumor microenvironment
TIE2-expressing
macrophages administered in combination with the CDK4/6 inhibitor LY2835219.
[0365] In another embodiment of this aspect of the invention, the method of
increasing
overall survival in breast cancer patients comprises the administration of a
composition of
Formula I at doses sufficient to block TIE2 kinase in tumor microenvironment
TIE2-expressing
macrophages administered in combination with the CDK4/6 inhibitor LEE011.
[0366] In another embodiment of this aspect of the invention, the method of
increasing
overall survival in breast cancer patients comprises the administration of a
composition of
Formula I at doses sufficient to block TIE2 kinase in tumor microenvironment
TIE2-expressing
macrophages administered in combination with the CDK4/6 inhibitor PD 0332991.
[0367] In another embodiment of this aspect of the invention, the method of
increasing
overall survival in breast cancer patients comprises the administration of a
composition of
Formula I in combination with bevacizumab.
[0368] In another embodiment of this aspect of the invention, the method of
increasing
overall survival in breast cancer patients comprises administering a
composition of Formula I in
combination with an anti-CTLA-4 agent.
[0369] In another embodiment of this aspect of the invention, the method of
increasing
overall survival in breast cancer patients comprises administering a
composition of Formula I in
combination with ipilimumab.
[0370] In another embodiment of this aspect of the invention, the method of
increasing
overall survival in breast cancer patients comprises administering a
composition of Formula I in
combination with an anti-PD-1 agent.
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[0371] In another embodiment of this aspect of the invention, the method of
increasing
overall survival in breast cancer patients comprises administering a
composition of Formula I in
combination with lambrolizumab.
[0372] In another embodiment of this aspect of the invention, the method of
increasing
overall survival in breast cancer patients comprises administering a
composition of Formula I in
combination with an anti- anti-PD L-1 agent.
[0373] In another embodiment of this aspect of the invention, the method of
increasing
overall survival in breast cancer patients comprises administering a
composition of Formula I in
combination with MPDL3280A.
Methods for treating breast cancer patients in a neoadjuvant setting prior to
surgical resection
of tumor:
[0374] Another aspect of the invention relates to a method of treating
breast cancer patients
in a neoadjuvant setting prior to surgical resection of tumor, comprising
administering to a
patient in need thereof and effective amount of a composition of Formula I, a
dosing regimen of
the composition of Formula I is sufficient to block TIE2 kinase in the tumor
microenvironment.
[0375] In another embodiment of this aspect of the invention, the method of
treating breast
cancer patients in a neoadjuvant setting prior to surgical resection of tumor
comprises
administering to a patient in need thereof an effective amount of a
composition of Formula I in a
dosing regimen sufficient to block TIE2 kinase in the tumor microenvironment.
[0376] In another embodiment of this aspect of the invention, a method of
treating breast
cancer patients in a neoadjuvant setting prior to surgical resection comprises
the administration
of a composition of Formula I at doses sufficient to block TIE2 kinase in the
tumor
microenvironment, with a dosing regimen of the composition of Formula I being
administered
daily.
[0377] In another embodiment of this aspect of the invention, the method of
treating breast
cancer patients in a neoadjuvant setting prior to surgical resection comprises
administering to a
patient in need thereof an effective amount of a composition of Formula I in a
dosing regimen
sufficient to block TIE2 kinase in the tumor microenvironment, with a dosing
regimen of the
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composition of Formula I administered in an intermittent non-daily manner,
including alternate
daily dosing, every third daily dosing, twice weekly dosing, or once weekly
dosing.
[0378] In another embodiment of this aspect of the invention, the method of
treating breast
cancer patients in a neoadjuvant setting prior to surgical resection comprises
administering to a
patient in need thereof an effective amount of a composition of Formula I in a
dosing regimen
administered twice weekly, once weekly, or alternate weekly.
[0379] In another embodiment of this aspect of the invention, the method of
treating breast
cancer patients in a neoadjuvant setting prior to surgical resection comprises
the administration
of a composition of Formula I in a dosing regimen administered twice weekly or
once weekly.
[0380] In another embodiment of this aspect of the invention, the dosing
regimen of a
composition of Formula I is administered twice weekly.
[0381] In another embodiment of this aspect of the invention, the method of
treating breast
cancer patients in a neoadjuvant setting prior to surgical resection comprises
the administration
of a composition of Formula I in dosing regimen sufficient to block TIE2
kinase in tumor
microenvironment TIE2-expressing macrophages in combination with one or more
agents taken
from an anti-tubulin agent, a DNA alkylating agent, a DNA synthesis-inhibiting
agent, a DNA
intercalating agent, an anti-estrogen agent, an anti-HER2 agent, a kinase
inhibitor, an anti-
angiogenic agent, or an immunomodulating agent.
[0382] In another embodiment of this aspect of the invention, the method of
treating breast
cancer patients in a neoadjuvant setting prior to surgical resection of tumor
comprises
administering a composition of Formula I in combination with paclitaxel.
[0383] In another embodiment of this aspect of the invention, the method of
treating breast
cancer patients in a neoadjuvant setting prior to surgical resection of tumor
comprises
administering a composition of Formula I in combination with paclitaxel
protein-bound particles
for injectable suspension.
[0384] In another embodiment of this aspect of the invention, the method of
treating breast
cancer patients in a neoadjuvant setting prior to surgical resection of tumor
comprises
administering a composition of Formula I in combination with docetaxel.
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[0385] In another embodiment of this aspect of the invention, the method of
treating breast
cancer patients in a neoadjuvant setting prior to surgical resection of tumor
comprises the
administration of a composition of Formula I in combination with eribulin.
[0386] In another embodiment of this aspect of the invention, the method of
treating breast
cancer patients in a neoadjuvant setting prior to surgical resection of tumor
comprises the
administration of a composition of Formula I in combination with ixabepilone.
[0387] In another embodiment of this aspect of the invention, the method of
treating breast
cancer patients in a neoadjuvant setting prior to surgical resection of tumor
comprises the
administration of a composition of Formula I in combination with vinorelbine.
[0388] In another embodiment of this aspect of the invention, the method of
treating breast
cancer patients in a neoadjuvant setting prior to surgical resection of tumor
comprises the
administration of a composition of Formula I in combination with capecitabine.
[0389] In another embodiment of this aspect of the invention, the method of
treating breast
cancer patients in a neoadjuvant setting prior to surgical resection of tumor
comprises the
administration of a composition of Formula I in combination with gemcitabine.
[0390] In another embodiment of this aspect of the invention, the method of
treating breast
cancer patients in a neoadjuvant setting prior to surgical resection of tumor
comprises the
administration of a composition of Formula I in combination with 5-
fluorouracil.
[0391] In another embodiment of this aspect of the invention, the method of
treating breast
cancer patients in a neoadjuvant setting prior to surgical resection of tumor
comprises the
administration of a composition of Formula I in combination with methotrexate.
[0392] In another embodiment of this aspect of the invention, the method of
treating breast
cancer patients in a neoadjuvant setting prior to surgical resection of tumor
comprises the
administration of a composition of Formula I in combination with
cyclophosphamide.
[0393] In another embodiment of this aspect of the invention, the method of
treating breast
cancer patients in a neoadjuvant setting prior to surgical resection of tumor
comprises the
administration of a composition of Formula I in combination with cisplatin.
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[0394] In another embodiment of this aspect of the invention, the method of
treating breast
cancer patients in a neoadjuvant setting prior to surgical resection of tumor
comprises the
administration of a composition of Formula I in combination with carboplatin.
[0395] In another embodiment of this aspect of the invention, the method of
treating breast
cancer patients in a neoadjuvant setting prior to surgical resection of tumor
comprises
administering of a composition of Formula I in combination with doxorubicin.
[0396] In another embodiment of this aspect of the invention, the method of
treating breast
cancer patients in a neoadjuvant setting prior to surgical resection of tumor
comprises
administering of a composition of Formula I in combination with pegylated
liposomal
doxorubicin.
[0397] In another embodiment of this aspect of the invention, the method of
treating breast
cancer patients in a neoadjuvant setting prior to surgical resection of tumor
comprises
administering of a composition of Formula I in combination with epirubicin.
[0398] In yet another embodiment of this aspect of the invention, the
method of treating
breast cancer patients in a neoadjuvant setting prior to surgical resection of
tumor comprises the
administration of a composition of Formula I in combination with tamoxifen.
[0399] In another embodiment of this aspect of the invention, the method of
treating breast
cancer patients in a neoadjuvant setting prior to surgical resection of tumor
comprises the
administration of a composition of Formula I in combination with fulvestrant.
[0400] In another embodiment of this aspect of the invention, the method of
treating breast
cancer patients in a neoadjuvant setting prior to surgical resection of tumor
comprises the
administration of a composition of Formula I in combination with anastrozole.
[0401] In another embodiment of this aspect of the invention, the method of
treating breast
cancer patients in a neoadjuvant setting prior to surgical resection of tumor
comprises the
administration of a composition of Formula I in combination with letrozole.
[0402] In another embodiment of this aspect of the invention, the method of
treating breast
cancer patients in a neoadjuvant setting prior to surgical resection of tumor
comprises the
administration of a composition of Formula I in combination with exemestane.
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[0403] In another embodiment of this aspect of the invention, the method of
treating breast
cancer patients in a neoadjuvant setting prior to surgical resection of tumor
comprises the
administration of a composition of Formula I in combination with trastuzumab.
[0404] In another embodiment of this aspect of the invention, the method of
treating breast
cancer patients in a neoadjuvant setting prior to surgical resection of tumor
comprises the
administration of a composition of Formula I in combination with ado-
trastuzumab emtansine.
[0405] In another embodiment of this aspect of the invention, the method of
treating breast
cancer patients in a neoadjuvant setting prior to surgical resection of tumor
comprises
administering a composition of Formula I in combination with pertuzumab.
[0406] In another embodiment of this aspect of the invention, the method of
treating breast
cancer patients in a neoadjuvant setting prior to surgical resection of tumor
comprises the
administration of a composition of Formula I at doses sufficient to block TIE2
kinase in tumor
microenvironment TIE2-expressing macrophages administered in combination with
lapatinib.
[0407] In another embodiment of this aspect of the invention, the method of
treating breast
cancer patients in a neoadjuvant setting prior to surgical resection of tumor
comprises the
administration of a composition of Formula I at doses sufficient to block TIE2
kinase in tumor
microenvironment TIE2-expressing macrophages administered in combination with
everolimus.
[0408] In another embodiment of this aspect of the invention, the method of
treating breast
cancer patients in a neoadjuvant setting prior to surgical resection of tumor
comprises the
administration of a composition of Formula I at doses sufficient to block TIE2
kinase in tumor
microenvironment TIE2-expressing macrophages administered in combination with
temsirolimus.
[0409] In another embodiment of this aspect of the invention, the method of
treating breast
cancer patients in a neoadjuvant setting prior to surgical resection of tumor
comprises the
administration of a composition of Formula I at doses sufficient to block TIE2
kinase in tumor
microenvironment TIE2-expressing macrophages administered in combination with
the CDK4/6
inhibitor LY2835219.
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[0410] In another embodiment of this aspect of the invention, the method of
treating breast
cancer patients in a neoadjuvant setting prior to surgical resection of tumor
comprises the
administration of a composition of Formula I at doses sufficient to block TIE2
kinase in tumor
microenvironment TIE2-expressing macrophages administered in combination with
the CDK4/6
inhibitor LEE011.
[0411] In another embodiment of this aspect of the invention, the method of
treating breast
cancer patients in a neoadjuvant setting prior to surgical resection of tumor
comprises the
administration of a composition of Formula I at doses sufficient to block TIE2
kinase in tumor
microenvironment TIE2-expressing macrophages administered in combination with
the CDK4/6
inhibitor PD 0332991.
[0412] In another embodiment of this aspect of the invention, the method of
treating breast
cancer patients in a neoadjuvant setting prior to surgical resection of tumor
comprises the
administration of a composition of Formula I in combination with bevacizumab.
[0413] In another embodiment of this aspect of the invention, the method of
treating breast
cancer patients in a neoadjuvant setting prior to surgical resection of tumor
comprises
administering a composition of Formula I in combination with an anti-CTLA-4
agent.
[0414] In another embodiment of this aspect of the invention, the method of
treating breast
cancer patients in a neoadjuvant setting prior to surgical resection of tumor
comprises
administering a composition of Formula I in combination with ipilimumab.
[0415] In another embodiment of this aspect of the invention, the method of
treating breast
cancer patients in a neoadjuvant setting prior to surgical resection of tumor
comprises
administering a composition of Formula I in combination with an anti-PD-1
agent.
[0416] In another embodiment of this aspect of the invention, the method of
treating breast
cancer patients in a neoadjuvant setting prior to surgical resection of tumor
comprises
administering a composition of Formula I in combination with lambrolizumab.
[0417] In another embodiment of this aspect of the invention, the method of
treating breast
cancer patients in a neoadjuvant setting prior to surgical resection of tumor
comprises
administering a composition of Formula I in combination with an anti- anti-PD
L-1 agent.
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[0418] In another embodiment of this aspect of the invention, the method of
treating breast
cancer patients in a neoadjuvant setting prior to surgical resection of tumor
comprises
administering a composition of Formula I in combination with MPDL3280A.
Methods for treating breast cancer patients in a neoadjuvant setting prior to
surgical
resection:
[0419] In another aspect of the invention, a method of treating breast
cancer patients in a
neoadjuvant setting prior to surgical resection comprises the administration
of a composition of
Formula I at doses sufficient to block TIE2 kinase in the tumor
microenvironment, with a dosing
regimen of the composition of Formula I administered as intermittent non-daily
dosing,
including alternate daily dosing, every third daily dosing, twice weekly
dosing, or once weekly
dosing.
[0420] In one embodiment of this aspect of the invention, a method of
treating breast cancer
patients in a neoadjuvant setting prior to surgical resection comprises the
administration of a
composition of Formula I at doses sufficient to block TIE2 kinase in the tumor
microenvironment, with a dosing regimen of the composition of Formula I
administered twice
weekly, once weekly, or alternate weekly.
[0421] In another embodiment of this aspect of the invention, a method of
treating breast
cancer patients in a neoadjuvant setting prior to surgical resection comprises
the administration
of a composition of Formula I at doses sufficient to block TIE2 kinase in the
tumor
microenvironment, with a dosing regimen of the composition of Formula I
administered twice
weekly or once weekly.
[0422] In another embodiment of this aspect of the invention, a method of
treating breast
cancer patients in a neoadjuvant setting prior to surgical resection comprises
the administration
of a composition of Formula I at doses sufficient to block TIE2 kinase in the
tumor
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microenvironment, with a dosing regimen of the composition of Formula I
administered twice
weekly.
[0423] In another embodiment of this aspect of the invention, a method of
treating breast
cancer patients in a neoadjuvant setting prior to surgical resection comprises
the administration
of a composition of Formula I at doses sufficient to block TIE2 kinase in the
tumor
microenvironment administered in combination with one or more
[0424] agents taken from an anti-tubulin agent, a DNA alkylating agent, a
DNA synthesis-
inhibiting agent, a DNA intercalating agent, an anti-estrogen agent, an anti-
HER2 agent, a kinase
inhibitor, an anti-angiogenic agent, or an immunomodulating agent.
[0425] In another embodiment of this aspect of the invention, the method of
treating breast
cancer patients in a neoadjuvant setting prior to surgical resection of tumor
comprises
administering a composition of Formula I in combination with paclitaxel.
[0426] In another embodiment of this aspect of the invention, the method of
treating breast
cancer patients in a neoadjuvant setting prior to surgical resection of tumor
comprises
administering a composition of Formula I in combination with paclitaxel
protein-bound particles
for injectable suspension.
[0427] In another embodiment of this aspect of the invention, the method of
treating breast
cancer patients in a neoadjuvant setting prior to surgical resection of tumor
comprises
administering a composition of Formula I in combination with docetaxel.
[0428] In another embodiment of this aspect of the invention, the method of
treating breast
cancer patients in a neoadjuvant setting prior to surgical resection of tumor
comprises the
administration of a composition of Formula I in combination with eribulin.
[0429] In another embodiment of this aspect of the invention, the method of
treating breast
cancer patients in a neoadjuvant setting prior to surgical resection of tumor
comprises the
administration of a composition of Formula I in combination with ixabepilone.
[0430] In another embodiment of this aspect of the invention, the method of
treating breast
cancer patients in a neoadjuvant setting prior to surgical resection of tumor
comprises the
administration of a composition of Formula I in combination with vinorelbine.
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[0431] In another embodiment of this aspect of the invention, the method of
treating breast
cancer patients in a neoadjuvant setting prior to surgical resection of tumor
comprises the
administration of a composition of Formula I in combination with capecitabine.
[0432] In another embodiment of this aspect of the invention, the method of
treating breast
cancer patients in a neoadjuvant setting prior to surgical resection of tumor
comprises the
administration of a composition of Formula I in combination with gemcitabine.
[0433] In another embodiment of this aspect of the invention, the method of
treating breast
cancer patients in a neoadjuvant setting prior to surgical resection of tumor
comprises the
administration of a composition of Formula I in combination with 5-
fluorouracil.
[0434] In another embodiment of this aspect of the invention, the method of
treating breast
cancer patients in a neoadjuvant setting prior to surgical resection of tumor
comprises the
administration of a composition of Formula I in combination with 5-
methotrexate.
[0435] In another embodiment of this aspect of the invention, the method of
treating breast
cancer patients in a neoadjuvant setting prior to surgical resection of tumor
comprises the
administration of a composition of Formula I in combination with
cyclophosphamide.
[0436] In another embodiment of this aspect of the invention, the method of
treating breast
cancer patients in a neoadjuvant setting prior to surgical resection of tumor
comprises the
administration of a composition of Formula I in combination with cisplatin.
[0437] In another embodiment of this aspect of the invention, the method of
treating breast
cancer patients in a neoadjuvant setting prior to surgical resection of tumor
comprises the
administration of a composition of Formula I in combination with carboplatin.
[0438] In another embodiment of this aspect of the invention, the method of
treating breast
cancer patients in a neoadjuvant setting prior to surgical resection of tumor
comprises
administering of a composition of Formula I in combination with doxorubicin.
[0439] In another embodiment of this aspect of the invention, the method of
treating breast
cancer patients in a neoadjuvant setting prior to surgical resection of tumor
comprises
administering of a composition of Formula I in combination with pegylated
liposomal
doxorubicin.
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[0440] In another embodiment of this aspect of the invention, the method of
treating breast
cancer patients in a neoadjuvant setting prior to surgical resection of tumor
comprises
administering of a composition of Formula I in combination with epirubicin.
[0441] In yet another embodiment of this aspect of the invention, the
method of treating
breast cancer patients in a neoadjuvant setting prior to surgical resection of
tumor comprises the
administration of a composition of Formula I in combination with tamoxifen.
[0442] In another embodiment of this aspect of the invention, the method of
treating breast
cancer patients in a neoadjuvant setting prior to surgical resection of tumor
comprises the
administration of a composition of Formula I in combination with fulvestrant.
[0443] In another embodiment of this aspect of the invention, the method of
treating breast
cancer patients in a neoadjuvant setting prior to surgical resection of tumor
comprises the
administration of a composition of Formula I in combination with anastrozole.
[0444] In another embodiment of this aspect of the invention, the method of
treating breast
cancer patients in a neoadjuvant setting prior to surgical resection of tumor
comprises the
administration of a composition of Formula I in combination with letrozole.
[0445] In another embodiment of this aspect of the invention, the method of
treating breast
cancer patients in a neoadjuvant setting prior to surgical resection of tumor
comprises the
administration of a composition of Formula I in combination with exemestane.
[0446] In another embodiment of this aspect of the invention, the method of
treating breast
cancer patients in a neoadjuvant setting prior to surgical resection of tumor
comprises the
administration of a composition of Formula I in combination with trastuzumab.
[0447] In another embodiment of this aspect of the invention, the method of
treating breast
cancer patients in a neoadjuvant setting prior to surgical resection of tumor
comprises the
administration of a composition of Formula I in combination with ado-
trastuzumab emtansine.
[0448] In another embodiment of this aspect of the invention, the method of
treating breast
cancer patients in a neoadjuvant setting prior to surgical resection of tumor
comprises
administering a composition of Formula I in combination with pertuzumab.
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[0449] In another embodiment of this aspect of the invention, the method of
treating breast
cancer patients in a neoadjuvant setting prior to surgical resection of tumor
comprises the
administration of a composition of Formula I at doses sufficient to block TIE2
kinase in tumor
microenvironment TIE2-expressing macrophages administered in combination with
lapatinib.
[0450] In another embodiment of this aspect of the invention, the method of
treating breast
cancer patients in a neoadjuvant setting prior to surgical resection of tumor
comprises the
administration of a composition of Formula I at doses sufficient to block TIE2
kinase in tumor
microenvironment TIE2-expressing macrophages administered in combination with
everolimus.
[0451] In another embodiment of this aspect of the invention, the method of
treating breast
cancer patients in a neoadjuvant setting prior to surgical resection of tumor
comprises the
administration of a composition of Formula I at doses sufficient to block TIE2
kinase in tumor
microenvironment TIE2-expressing macrophages administered in combination with
temsirolimus.
[0452] In another embodiment of this aspect of the invention, the method of
treating breast
cancer patients in a neoadjuvant setting prior to surgical resection of tumor
comprises the
administration of a composition of Formula I at doses sufficient to block TIE2
kinase in tumor
microenvironment TIE2-expressing macrophages administered in combination with
the CDK4/6
inhibitor LY2835219.
[0453] In another embodiment of this aspect of the invention, the method of
treating breast
cancer patients in a neoadjuvant setting prior to surgical resection of tumor
comprises the
administration of a composition of Formula I at doses sufficient to block TIE2
kinase in tumor
microenvironment TIE2-expressing macrophages administered in combination with
the CDK4/6
inhibitor LEE011.
[0454] In another embodiment of this aspect of the invention, the method of
treating breast
cancer patients in a neoadjuvant setting prior to surgical resection of tumor
comprises the
administration of a composition of Formula I at doses sufficient to block TIE2
kinase in tumor
microenvironment TIE2-expressing macrophages administered in combination with
the CDK4/6
inhibitor PD 0332991.
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[0455] In another embodiment of this aspect of the invention, the method of
treating breast
cancer patients in a neoadjuvant setting prior to surgical resection of tumor
comprises the
administration of a composition of Formula I in combination with bevacizumab.
[0456] In another embodiment of this aspect of the invention, the method of
treating breast
cancer patients in a neoadjuvant setting prior to surgical resection of tumor
comprises
administering a composition of Formula I in combination with an anti-CTLA-4
agent.
[0457] In another embodiment of this aspect of the invention, the method of
treating breast
cancer patients in a neoadjuvant setting prior to surgical resection of tumor
comprises
administering a composition of Formula I in combination with ipilimumab.
[0458] In another embodiment of this aspect of the invention, the method of
treating breast
cancer patients in a neoadjuvant setting prior to surgical resection of tumor
comprises
administering a composition of Formula I in combination with an anti-PD-1
agent.
[0459] In another embodiment of this aspect of the invention, the method of
treating breast
cancer patients in a neoadjuvant setting prior to surgical resection of tumor
comprises
administering a composition of Formula I in combination with lambrolizumab.
[0460] In another embodiment of this aspect of the invention, the method of
treating breast
cancer patients in a neoadjuvant setting prior to surgical resection of tumor
comprises
administering a composition of Formula I in combination with an anti- anti-PD
L-1 agent.
[0461] In another embodiment of this aspect of the invention, the method of
treating breast
cancer patients in a neoadjuvant setting prior to surgical resection of tumor
comprises
administering a composition of Formula I in combination with MPDL3280A.
[0462] Another aspect of the invention relates to a method of treating
ovarian cancer as TIE2
pathway signaling has been shown to contribute to ovarian cancer progression
(Karlan et al, J.
Clinical Oncology (2012) 30: 362-370). The method comprises administering to a
patient in
need thereof an effective amount of a composition of Formula I in a dosing
regimen sufficient to
block TIE2 kinase in the tumor microenvironment.In one embodiment of this
aspect of the
invention, a composition of Formula I is administered as a single agent.
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[0463] In another embodiment of this aspect of the invention, a composition
of Formula I is
administered in combination with paclitaxel + carboplatin, doxetaxel +
carboplatin, paclitaxel +
cisplatin, or other taxane + platinum drug regimens.
[0464] Another aspect of the invention relates to a method of treating
hepatocellular
carcinoma as TIE2 pathway signaling has been shown to contribute to
hepatocellular cancer
progression and as a diagnostic marker (Matsubara et al, Hepatology (2013) 57:
1416-1425;
Mitsuhashi et al, Hepatology (2003) 37: 1105-1113; Tanaka et al, J. Clin
Invest (1999) 103: 341-
345). The method comprises administering to a patient in need thereof an
effective amount of a
composition of Formula I in a dosing regimen sufficient to block TIE2 kinase
in the tumor
mi cro environment .
[0465] In one embodiment of this aspect of the invention, a composition of
Formula I is
administered as a single agent.
[0466] In another embodiment of this aspect of the invention, a composition
of Formula I is
administered in combination with a kinase inhibitor including sorafenib,
crizotinib, cabozantinib,
sunitinib, pazopanib, sorafenib, regorafenib, or axitinib.
[0467] Another aspect of the invention relates to a method of treating
glioma as TIE2
pathway signaling has been shown to contribute to glioma cancer progression
(Liu et al,
Oncotarget (2010) 1: 700-709; Brunckhorst et al, Cancer Research (2010) 70:
7283-7293). The
method comprises administering to a patient in need thereof an effective
amount of a
composition of Formula I in a dosing regimen sufficient to block TIE2 kinase
in the tumor
mi cro environment .
[0468] In one embodiment of this aspect of the invention, a composition of
Formula I is
administered as a single agent.
[0469] In another embodiment of this aspect of the invention, a composition
of Formula I is
administered in combination with radiotherapy.
[0470] In another embodiment of this aspect of the invention, a composition
of Formula I is
administered in combination with temozolomide therapy.
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[0471] In another embodiment of this aspect of the invention, a composition
of Formula I is
administered in combination with radiotherapy and temozolomide therapy.
[0472] In another embodiment of this aspect of the invention, a composition
of Formula I is
administered in combination with bevacizumab therapy.
[0473] In another embodiment of this aspect of the invention, a composition
of Formula I is
administered in combination with radiotherapy and bevacizumab therapy.
[0474] In another embodiment of this aspect of the invention, a composition
of Formula I is
administered in combination with temozolomide therapy and bevacizumab therapy.
[0475] Another aspect of the invention relates to a method of treating
melanoma as TIE2
pathway signaling has been shown to contribute to melanoma progression
(Helfrich et al, Clin
Cancer Res (2009) 15: 1384-1392; Peinado et al, Nature Medicine (2012) 18: 883-
891). The
method comprises administering to a patient in need thereof an effective
amount of a
composition of Formula I in a dosing regimen sufficient to block TIE2 kinase
in the tumor
mi cro environment .
[0476] In one embodiment of this aspect of the invention, a composition of
Formula I is
administered as a single agent.
[0477] In another embodiment of this aspect of the invention, a composition
of Formula I is
administered in combination with vemurafenib.
[0478] In another embodiment of this aspect of the invention, a composition
of Formula I is
administered in combination with dabrafenib.
[0479] In another embodiment of this aspect of the invention, a composition
of Formula I is
administered in combination with dabrafenib and trametinib.
[0480] In another embodiment of this aspect of the invention, a composition
of Formula I is
administered in combination with temozolomide.
[0481] In another embodiment of this aspect of the invention, a composition
of Formula I is
administered in combination with dacarbazine.
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[0482] In another embodiment of this aspect of the invention, a composition
of Formula I is
administered in combination with ipilimumab.
[0483] In another embodiment of this aspect of the invention, a composition
of Formula I is
administered in combination with labrolizumab or nivolumab.
[0484] In another embodiment of this aspect of the invention, a composition
of Formula I is
administered in combination with MPDL3280A.
[0485] In another embodiment of this aspect of the invention, a composition
of Formula I is
administered in combination with imatinib.
[0486] Another aspect of the invention relates to a method of treating
colorectal cancer as
TIE2 pathway signaling has been shown to contribute to colorectal cancer
progression (Ahmad
et al, Cancer (2001) 92: 1138-1143; Hashizume et al, Cancer Research (2010)
70: 2213-2223).
The method comprises administering to a patient in need thereof an effective
amount of a
composition of Formula I in a dosing regimen sufficient to block TIE2 kinase
in the tumor
mi cro environment .
[0487] In one embodiment of this aspect of the invention, a composition of
Formula I is
administered as a single agent.
[0488] In another embodiment of this aspect of the invention, a composition
of Formula I is
administered in combination with mFOLFOX6 therapy (oxaplatin + leucovorin + 5-
fluorouracil).
[0489] In another embodiment of this aspect of the invention, a composition
of Formula I is
administered in combination with mFOLFOX6 therapy and bevacizumab.
[0490] In another embodiment of this aspect of the invention, a composition
of Formula I is
administered in combination with mFOLFOX6 therapy and panitumumab.
[0491] In another embodiment of this aspect of the invention, a composition
of Formula I is
administered in combination with mFOLFOX6 therapy and cetuximab.
[0492] In another embodiment of this aspect of the invention, a composition
of Formula I is
administered in combination with capecitabine.
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[0493] In another embodiment of this aspect of the invention, a composition
of Formula I is
administered in combination with capecitabine and bevacizumab.
[0494] In another embodiment of this aspect of the invention, a composition
of Formula I is
administered in combination with FOLFIRI therapy (irinotecan + leucovorin + 5-
fluorouracil).
[0495] In another embodiment of this aspect of the invention, a composition
of Formula I is
administered in combination with FOLFIRI therapy and bevacizumab.
[0496] In another embodiment of this aspect of the invention, a composition
of Formula I is
administered in combination with FOLFIRI therapy and aflibercept.
[0497] In another embodiment of this aspect of the invention, a composition
of Formula I is
administered in combination with FOLFIRI therapy and cetuximab.
[0498] In another embodiment of this aspect of the invention, a composition
of Formula I is
administered in combination with FOLFIRI therapy and panitumumab.
[0499] In another embodiment of this aspect of the invention, a composition
of Formula I is
administered in combination with panitumumab.
[0500] In another embodiment of this aspect of the invention, a composition
of Formula I is
administered in combination with panitumumab and irinotecan.
[0501] In another embodiment of this aspect of the invention, a composition
of Formula I is
administered in combination with cetuximab.
[0502] In another embodiment of this aspect of the invention, a composition
of Formula I is
administered in combination with cetuximab and irinotecan.
[0503] Another aspect of the invention relates to a method of treating
acute myeloid
leukemia as TIE2 pathway signaling has been shown to contribute to acute
myeloid leukemia
progression (Muller et al, Leukemia Research (2002) 26: 163-168; Hou et al,
Leukemia Research
(2008) 32: 904-912). The method comprises administering to a patient in need
thereof an
effective amount of a composition of Formula I in a dosing regimen sufficient
to block TIE2
kinase in the tumor microenvironment.
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[0504] In one embodiment of this aspect of the invention, a composition of
Formula I is
administered as a single agent.
[0505] In another embodiment of this aspect of the invention, a composition
of Formula I is
administered in combination with daunorubicin and cytarabine.
[0506] In another embodiment of this aspect of the invention, a composition
of Formula I is
administered in combination with idarubicin and cytarabine.
[0507] In another embodiment of this aspect of the invention, a composition
of Formula I is
administered in combination with mitoxantrone and cytarabine.
[0508] In another embodiment of this aspect of the invention, a composition
of Formula I is
administered in combination with cytarabine.
[0509] In another embodiment of this aspect of the invention, a composition
of Formula I is
administered in combination with 5-azacytabine.
[0510] In another embodiment of this aspect of the invention, a composition
of Formula I is
administered in combination with decitabine.
[0511] In another embodiment of this aspect of the invention, a composition
of Formula I is
administered in combination with quizartinib.
[0512] Another aspect of the invention relates to a method of treating
cancer. The method
comprises administering an effective amount of a composition of Formula I to a
patient in need
thereof. In one embodiment the patient overexpresses Tunica interna
endothelial cell kinase 2
(TIE2) and the cancer is selected from breast cancer, colorectal cancer,
hepatocellular carcinoma,
head and neck cancer, bladder cancer, ovarian cancer, gliomas, angiosarcomas,
melanomas, or
acute myeloid leukemia.
[0513] In certain embodiments of the invention of the treatment regimen,
the composition of
Formula I is administered at a frequency of daily.
[0514] In other embodiments of the invention of the treatment regimen, the
composition of
Formula I is administered at a frequency of non-daily intermittent.
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[0515] In other embodiments of the invention of the treatment regimen, the
composition of
Formula I is administered at a frequency of three times weekly.
[0516] In other embodiments of the invention of the treatment regimen, the
composition of
Formula I is administered at a frequency of two times weekly.
[0517] In other embodiments of the invention of the treatment regimen, the
composition of
Formula I is administered at a frequency of one time weekly.
[0518] In other embodiments of the invention of the treatment regimen, the
composition of
Formula I is administered at a frequency of one time every two weeks.
[0519] In other embodiments the cancer is metastatic, triple negative
breast cancer (estrogen
receptor negative, progesterone receptor negative, HER2 negative).
[0520] In other embodiments the cancer is estrogen positive (ER) and HER2
receptor kinase
negative (HER2-) breast cancer.
[0521] In other embodiments the cancer is inflammatory breast cancer.
[0522] In another embodiment, the method comprises the treatment of
preventing or
reducing one or more of primary tumor growth, tumor invasiveness, cancer
intravasation, cancer
dissemination, metastasis, and tumor immunotolerance. In certain embodiments
the method
increases patient survival rates.
Formulations, Administration, Dosing, and Treatment Regimens
[0523] The present invention includes the described salts of Formulae I
and/or II (and/or
additional agents) in various formulations. Any composition (and/or additional
agents) described
herein can take the form of solutions, suspensions, emulsion, drops, tablets,
pills, pellets,
capsules, capsules containing liquids, powders, sustained-release
formulations, suppositories,
emulsions, aerosols, sprays, suspensions, or any other form suitable for use.
In one embodiment,
the composition is in the form of a capsule (see, e.g., U.S. Patent No.
5,698,155). Other
examples of suitable pharmaceutical excipients are described in Remington 's
Pharmaceutical
Sciences 1447-1676 (Alfonso R. Gennaro eds., 19th ed. 1995), incorporated
herein by reference.
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[0524] Where necessary, the salts herein described can also include a
solubilizing agent.
Also, the agents can be delivered with a suitable vehicle or delivery device
as known in the art.
Combination therapies outlined herein can be co-delivered in a single delivery
vehicle or
delivery device. Compositions for administration can optionally include a
local anesthetic such
as, for example, lignocaine to lessen pain at the site of the injection.
[0525] In one embodiment, the salts of Formulae I and/or II (and/or
additional agents)
described herein is formulated in accordance with routine procedures as a
composition adapted
for a mode of administration.
[0526] In certain embodiments, routes of administration include, for
example: intradermal,
intramuscular, intraperitoneal, intravenous, subcutaneous, intranasal,
epidural, oral, sublingual,
intranasal, intracerebral, intravaginal, transdermal, rectally, by inhalation,
or topically,
particularly to the ears, nose, eyes, or skin. In some embodiments, the
administering is effected
orally or by parenteral injection. The mode of administration can be left to
the discretion of the
practitioner, and depend in-part upon the site of the medical condition. In
most instances,
administration results in the release of any agent described herein into the
bloodstream.
[0527] In specific embodiments, it may be desirable to administer locally
to the area in need
of treatment or blocking.
[0528] In one embodiment, the salts (and/or additional agents) described
herein is formulated
in accordance with routine procedures as a composition adapted for oral
administration to
humans. Compositions for oral delivery can be in the form of tablets,
lozenges, aqueous or oily
suspensions, granules, powders, emulsions, capsules, syrups, or elixirs, for
example. Orally
administered compositions can comprise one or more agents, for example,
sweetening agents
such as fructose, aspartame or saccharin; flavoring agents such as peppermint,
oil of wintergreen,
or cherry; coloring agents; and preserving agents, to provide a
pharmaceutically palatable
preparation. Moreover, where in tablet or pill form, the compositions can be
coated to delay
disintegration and absorption in the gastrointestinal tract, thereby providing
a sustained action
over an extended period of time. Selectively permeable membranes surrounding
an osmotically
active driving the salt of Formula I or II (and/or additional agents)
described herein are also
suitable for orally administered compositions. In these latter platforms,
fluid from the
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environment surrounding the capsule is imbibed by the driving composition,
which swells to
displace the agent or agent composition through an aperture. These delivery
platforms can
provide an essentially zero order delivery profile as opposed to the spiked
profiles of immediate
release formulations. A time-delay material such as glycerol monostearate or
glycerol stearate
can also be useful. Oral compositions can include standard excipients such as
mannitol, lactose,
starch, magnesium stearate, sodium saccharin, cellulose, and magnesium
carbonate. In one
embodiment, the excipients are of pharmaceutical grade. Suspensions, in
addition to the active
compositions, may contain suspending agents such as, for example, ethoxylated
isostearyl
alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline
cellulose, aluminum
metahydroxide, bentonite, agar-agar, tragacanth, etc., and mixtures thereof.
[0529] Dosage forms suitable for parenteral administration (e.g.
intravenous, intramuscular,
intraperitoneal, subcutaneous and intra-articular injection and infusion)
include, for example,
solutions, suspensions, dispersions, emulsions, and the like. They may also be
manufactured in
the form of sterile solid compositions (e.g. lyophilized composition), which
can be dissolved or
suspended in sterile injectable medium immediately before use. They may
contain, for example,
suspending or dispersing agents known in the art.
[0530] The dosage of the salt of Formulae I and/or II (and/or additional
agents) described
herein as well as the dosing schedule can depend on various parameters,
including, but not
limited to, the disease being treated, the subject's general health, and the
administering
physician's discretion. Any agent described herein, can be administered prior
to (e.g., 5 minutes,
15 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 12
hours, 24 hours, 48
hours, 72 hours, 96 hours, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6
weeks, 8 weeks, or 12
weeks before), concurrently with, or subsequent to (e.g., 5 minutes, 15
minutes, 30 minutes, 45
minutes, 1 hour, 2 hours, 4 hours, 6 hours, 12 hours, 24 hours, 48 hours, 72
hours, 96 hours, 1
week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 8 weeks, or 12 weeks after)
the
administration of an additional therapeutic agent, to a subject in need
thereof. In various
embodiments any agent described herein is administered 1 minute apart, 10
minutes apart, 30
minutes apart, less than 1 hour apart, 1 hour apart, 1 hour to 2 hours apart,
2 hours to 3 hours
apart, 3 hours to 4 hours apart, 4 hours to 5 hours apart, 5 hours to 6 hours
apart, 6 hours to 7
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hours apart, 7 hours to 8 hours apart, 8 hours to 9 hours apart, 9 hours to 10
hours apart, 10 hours
to 11 hours apart, or 11 hours to 12 hours apart.
[0531] The dosage of the salt of Formula I or II (and/or additional agents)
described herein
can depend on several factors including the severity of the condition, whether
the condition is to
be treated or prevented, and the age, weight, and health of the subject to be
treated. Additionally,
pharmacogenomic (the effect of genotype on the pharmacokinetic,
pharmacodynamic or efficacy
profile of a therapeutic) information about a particular subject may affect
dosage used.
Furthermore, the exact individual dosages can be adjusted somewhat depending
on a variety of
factors, including the specific combination of the agents being administered,
the time of
administration, the route of administration, the nature of the formulation,
the rate of excretion,
the particular disease being treated, the severity of the disorder, and the
anatomical location of
the disorder. Some variations in the dosage can be expected.
[0532] Generally, when orally administered to a mammal, the dosage of any
composition of
Formula I (and/or additional agents) described herein may be 0.001 mg/kg/day
to 100
mg/kg/day, 0.01 mg/kg/day to 50 mg/kg/day, or 0.1 mg/kg/day to 10 mg/kg/day.
When orally
administered to a human, the dosage of any agent described herein is normally
0.001 mg to 1500
mg per day, 1 mg to 600 mg per day, or 5 mg to 30 mg per day. In some
embodiments, the
dosage of the salt (or agent) ranges from 57 mg to 1200 mg per day. In other
embodiments, the
dosage of the agents or salt ranges from 100 mg to 200 mg per day.
[0533] For administration of the salts of Formulae I or II (and/or
additional agents) described
herein by parenteral injection, the dosage is normally 0.1 mg to 250 mg per
day, 1 mg to 20 mg
per day, or 3 mg to 5 mg per day. Injections may be given up to four times
daily. Generally,
when orally or parenterally administered, the dosage of any agent described
herein is normally
0.1 mg to 1500 mg per day, or 0.5 mg to 10 mg per day, or 0.5 mg to 5 mg per
day. A dosage of
up to 3000 mg per day can be administered.
[0534] Administration of the salts (and/or additional agents) described
herein can,
independently, be one to four times daily. Specifically, administration of the
salt can be once a
day at a dosing regimen of the salt is from about 50 mg to 1500 mg. Suitable
daily dosage for
the prophylactic effects sought is 57-1200 mg/day. If administered twice
daily, a suitable dosage
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is 100 mg to 200mg of the salt. Administration of the salt may also be
intermittently non-daily.
In particular, administration of the salt may be done one to four times per
month or one to six
times per year or once every two, three, four or five years. In certain
embodiments
administration of the salt is done weekly or bi-weekly. When administered
weekly or bi-weekly,
a suitable salt dosing regimen ranges from 50-200 mg/ per administration. In
certain weekly or
bi-weekly administrations, dosage is 200-400 mg/ per administration. Yet other
mode of weekly
or bi-weekly administration include 400-500 mg/ per administration, 500-600
mg/ per
administration, 600-700 mg/ per administration, 700-800 mg/ per
administration, 800-900 mg/
per administration, 900-1000 mg/ per administration, 1000-1100 mg/ per
administration, or
1100-1200 mg/ per administration. Administration can be for the duration of
one day or one
month, two months, three months, six months, one year, two years, three years,
and may even be
for the life of the subject. Chronic, long-term administration will be
indicated in many cases.
The dosage may be administered as a single dose or divided into multiple
doses. In general, the
desired dosage should be administered at set intervals for a prolonged period,
usually at least
over several weeks or months, although longer periods of administration of
several months or
years or more may be needed.
Examples
Example 1. Biochemical inhibition of unphosphorylated TIE2 (uTIE2) by 1-(3-
tert-buty1-
1-(quinolin-6-y1)-1H-pyrazol-5-y1)-3-(2-fluoro-4-(2-(methylcarbamoyl)pyridin-4-
yloxy)phenyl)urea
Biochemical assay for uTIE2 (Seq. ID No. 1)
[0535] Activity of uTIE2 kinase was determined by following the production
of ADP from
the kinase reaction through coupling with the pyruvate kinase/lactate
dehydrogenase system
(e.g., Schindler et al. Science (2000) 289: 1938-1942). In this assay, the
oxidation of NADH
(thus the decrease at A340nm) was continuously monitored
spectrophotometrically. The reaction
mixture (100 L) contained TIE2 (SignalChem) (5.6 nM), BSA (0.004% (w/v)),
polyEY (1.5
mg/ml), MgC12 (15 mM), DTT (0.5 mM), pyruvate kinase (4 units), lactate
dehydrogenase (7
units), phosphoenol pyruvate (1 mM), and NADH (0.28 mM) and ATP (1.5 mM) in 90
mM Tris
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buffer containing 0.2% octyl-glucoside and 1% DMSO, pH 7.5. The inhibition
reaction was
started by mixing serial diluted test composition with the above reaction
mixture. The
absorption at 340 nm was monitored continuously for 6 hours at 30 C on a
plate reader
(BioTek). The reaction rate was calculated using the 5 to 6 h time frame.
Percent inhibition was
obtained by comparison of reaction rate with that of a control (i.e. with no
test composition). IC50
values were calculated from a series of percent inhibition values determined
at a range of
inhibitor concentrations using software routines as implemented in the
GraphPad Prism software
package. The composition 1 -(3-tert-buty1-1 -(quinolin-6-y1)- 1H-pyrazol-5-y1)-
3 -(2 -fluoro-4-(2 -
(methylcarbamoyOpyridin-4-yloxy)phenyOurea para-toluene sulfonic acid salt
(compound 1 as
described in figures) exhibited an IC50 value of 3.5 nM.
uTIE2 protein sequence used for screening (Seq. ID No. 1)
QLKRANVQRRMAQAFQNVREEPAVQFNSGTLALNRKVKNNPDPTIYPVLDWNDIKFQ
DVIGEGNFGQVLKARIKKDGLRMDAAIKRMKEYASKDDHRDFAGELEVLCKLGHHPNI
INLLGACEHRGYLYLAIEYAPHGNLLDFLRKSRVLETDPAFAIANSTASTLSSQQLLHFA
ADVARGMDYLSQKQFIHRDLAARNILVGENYVAKIADFGLSRGQEVYVKKTMGRLPV
RWMAIESLNYSVYTTNSDVWSYGVLLWEIVSLGGTPYCGMTCAELYEKLPQGYRLEKP
LNCDDEVYDLMRQCWREKPYERPSFAQILVSLNRMLEERKTYVNTTLYEKFTYAGIDC
SAEEAA
Example 2. Biochemical inhibition of phosphorylated TIE2 (pTIE2) by 1-(3-tert-
butyl-1-
(quinolin-6-y1)-1H-pyrazol-5-y1)-3-(2-fluoro-4-(2-(methylcarbamoyl)pyridin-4-
yloxy)phenyl)urea
Biochemical assay for pTIE2 (Seq. ID No. 2)
[0536] Activity of pTIE2 kinase was determined by following the production
of ADP from
the kinase reaction through coupling with the pyruvate kinase/lactate
dehydrogenase system
(e.g., Schindler et al. Science (2000) 289: 1938-1942). In this assay, the
oxidation of NADH
(thus the decrease at A340nm) was continuously monitored
spectrophotometrically. The reaction
mixture (100 L) contained TIE2 (Life Technologies) (6 nM), BSA (0.004%
(w/v)), polyEY (1.5
mg/ml), MgC12 (15 mM), DTT (0.5 mM), pyruvate kinase (4 units), lactate
dehydrogenase (7
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units), phosphoenol pyruvate (1 mM), and NADH (0.28 mM) and ATP (1.5 mM) in 90
mM Tris
buffer containing 0.2% octyl-glucoside and 1% DMSO, pH 7.5. The inhibition
reaction was
started by mixing serial diluted test composition with the above reaction
mixture. The
absorption at 340 nm was monitored continuously for 6 hours at 30 C on a
plate reader
(BioTek). The reaction rate was calculated using the 2 to 3 h time frame.
Percent inhibition was
obtained by comparison of reaction rate with that of a control (i.e. with no
test composition). IC50
values were calculated from a series of percent inhibition values determined
at a range of
inhibitor concentrations using software routines as implemented in the
GraphPad Prism software
package. When tested the compositions 1 -(3 -tert-butyl- 1 -(quino lin-6 -y1)-
1 H-pyrazo 1-5 -y1)-3 -(2-
fluoro-4-(2-(methylcarbamoyl)pyridin-4-yloxy)phenyl)urea para-toluene sulfonic
acid salt and 1-
(3 -tert-butyl- 1 -(quino lin- 6-y1)- 1 H-pyrazol-5 -y1)-3 -(2 -fluoro- 4-(2 -
(methylc arb amoyOpyri din-4-
yloxy)phenyOurea bis-hydrochloric acid salt exhibited >50% inhibition of pTIE2
kinase at <0.1
iuM concentration. 1 -(3 -tert-butyl- 1 -(quinolin-6-y1)- 1 H-pyrazo 1-5
-y1)-3 -(2- fluoro- 4-(2 -
(methylcarbamoyOpyridin-4-yloxy)phenyOurea para-toluene sulfonic acid salt
exhibited an IC50
value of 4.2 nM. 1 -(3 -tert-butyl- 1 -(quinolin-6-y1)- 1 H-pyrazo 1-5 -
y1)-3 -(2 -fluor -4- (2-
(methylcarbamoyOpyridin-4-yloxy)phenyOurea bis-hydrochloric acid salt
exhibited an IC50
value of 2.2 nM.
pTIE2 protein sequence used for screening (Seq. ID No. 2)
PVLDWNDIKFQDVIGEGNFGQVLKARIKKDGLRMDAAIKRMKEYASKDDHRDFAGELE
VLCKLGHHPNIINLLGACEHRGYLYLAIEYAPHGNLLDFLRKSRVLETDPAFAIANSTAS
TLS S Q QLLHFAADVARGMDYL S QKQFIHRDLAARNILVGENYVAKIADF GL S RGQ EVY
VKKTMGRLPVRWMAIESLNYSVYTTNSDVWSYGVLLWEIVSLGGTPYCGMTCAELYE
KLPQGYRLEKPLNCDDEVYDLMRQCWREKPYERP SFAQILVSLNRMLEERKT
Example 3. Cellular inhibition of TIE2 in CHO cells by 1-(3-tert-buty1-1-
(quinolin-6-y1)-
1H-pyrazol-5-y1)-3-(2-flu oro-4-(2-(methylc arb amoyl)pyridin-4-yloxy)p he
nyl)u re a
CHO-K1 Cell Culture
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[0537] CHO-K1 cells (catalog #CCL-61) were obtained from the American Type
Culture
Collection (ATCC, Manassas, VA). Briefly, cells were grown in F 12K medium
supplemented
with 10% characterized fetal bovine serum (Invitrogen, Carlsbad, CA), 100
units/mL penicillin
G, 100 lag/m1 streptomycin, and 0.29 mg/mL L-glutamine (Invitrogen, Carlsbad,
CA) at 37
degrees Celsius, 5% CO2, and 95% humidity. Cells were allowed to expand until
reaching 70-
95% confluence at which point they were subcultured or harvested for assay
use.
TIE2-transfected CHO K1 Phospho-TIE2 Western Blot Assay
[0538] CHO K1 cells (1 x 105 cells/well) were added to a 24-well tissue-
culture treated plate
in 1 mL of RPMI1640 medium supplemented with 10% characterized fetal bovine
serum and 1X
non-essential amino acids (Invitrogen, Carlsbad, CA). Cells were then
incubated overnight at 37
degrees Celsius, 5% CO2, and 95% humidity. Medium was aspirated, and 0.5 mL of
medium
was added to each well. Transfection-grade plasmid DNA (TIE2 gene Gateway
cloned into
pcDNA3.2Tm/V5-DEST expression vector, Invitrogen, Carlsbad, CA) was diluted to
5 lag/mL in
room temperature Opti-MEMO I Medium without serum (Invitrogen, Carlsbad, CA).
Two iaL of
Lipofectamine LTX Reagent (Invitrogen, Carlsbad, CA) was added per 0.5 jag of
plasmid DNA.
The tube was mixed gently and incubated for 25 minutes at room temperature to
allow for DNA-
Lipofectamine LTX complex formation. 100 iaL of the DNA-Lipofectamine LTX
complex was
added directly to each well containing cells and mixed gently. Twenty-four
hours post-
transfection, medium containing DNA-Lipofectamine complexes was aspirated,
cells were
washed with PBS, and RPMI1640 medium supplemented with 10% characterized fetal
bovine
serum (Invitrogen, Carlsbad, CA), and 1X non-essential amino acids
(Invitrogen, Carlsbad, CA)
was added. Test composition (1 -(3-tert-buty1-1-(quinolin-6-y1)-1H-pyrazol-5-
y1)-3-(2-fluoro -4-
(2-(methylcarbamoyl)pyridin-4-yloxy)phenyOurea para-toluene sulfonic acid
salt) or DMSO was
added to the wells (0.5% final DMSO concentration). The plates were then
incubated for 4 hours
at 37 degrees Celsius, 5% CO2, and 95% humidity. Following the incubation, the
media was
aspirated and the cells were washed with PBS. The cells were lysed using MPER
lysis buffer
(Pierce, Rockford, IL) containing Halt Phosphatase and Protease Inhibitors
(Pierce, Rockford,
IL) and Phosphatase inhibitor cocktail 2 (Sigma, St. Louis, MO) at 4 C for 10
minutes with
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shaking. Cleared lysates were separated by SDS-PAGE on a 4-12% Novex NuPage
Bis-Tris gel
(Invitrogen, Carlsbad, CA) and then transferred to PVDF (Invitrogen, Carlsbad,
CA). After
transfer, the PVDF membrane was blocked with BSA (Santa Cruz Biotechnology,
Santa Cruz,
CA) and then probed with an antibody for phospho-TIE2 (Cell Signaling
Technology, Beverly,
MA). A secondary anti-rabbit antibody conjugated to horseradish peroxidase
(Cell Signaling
Technology, Beverly, MA) was used to detect phospho-TIE2. ECL Plus (GE
Healthcare,
Piscataway, NJ), a substrate for horseradish peroxidase that generates a
fluorescent product, was
added. Fluorescence was detected using a Storm 840 phosphorimager (GE
Healthcare,
Piscataway, NJ) in fluorescence mode. The 160 kDa phospho-TIE2 band was
quantified using
ImageQuant software (GE Healthcare, Piscataway, NJ). Data was analyzed using
Prism software
(GraphPad Software, San Diego, CA) to calculate IC50 values. The composition 1-
(3-tert-butyl-
1-(quino lin-6-y1)-1H-pyrazol-5-y1)-3-(2-fluoro -4-(2-(methylc arb amoyl)pyri
din-4-
yloxy)phenyOurea para-toluene sulfonic acid salt exhibited an IC50 value of
2.0 nM.
Example 4. Cellular inhibition of TIE2 in CHO cells after inhibitor wash-out
by 1-(3-tert-
buty1-1-(quinolin-6-y1)-1H-pyrazol-5-y1)-3-(2-fluoro-4-(2-
(methylcarbamoyl)pyridin-4-
yloxy)phenyl)urea
CHO-K1 Cell Culture
[0539] CHO-K1 cells (catalog #CCL-61) were obtained from the American Type
Culture
Collection (ATCC, Manassas, VA). Briefly, cells were grown in F 12K medium
supplemented
with 10% characterized fetal bovine serum (Invitrogen, Carlsbad, CA), 100
units/mL penicillin
G, 100 jug/m1 streptomycin, and 0.29 mg/mL L-glutamine (Invitrogen, Carlsbad,
CA) at 37
degrees Celsius, 5% CO2, and 95% humidity. Cells were allowed to expand until
reaching 70-
95% confluence at which point they were subcultured or harvested for assay
use.
TIE2-transfected CHO K1 Phospho-TIE2 Western Blot Composition Washout Assay
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[0540] CHO K1 cells (1 x 105 cells/well) were added to a 24-well tissue-
culture treated plate
in 1 mL of RPMI1640 medium supplemented with 10% characterized fetal bovine
serum and 1X
non-essential amino acids (Invitrogen, Carlsbad, CA). Cells were then
incubated overnight at 37
degrees Celsius, 5% CO2, and 95% humidity. Medium was aspirated, and 0.5 mL of
medium
was added to each well. Transfection-grade plasmid DNA (TIE2 gene Gateway
cloned into
pcDNA3.2Tm/V5-DEST expression vector, Invitrogen, Carlsbad, CA) was diluted to
5 g/mL in
room temperature Opti-MEMO I Medium without serum (Invitrogen, Carlsbad, CA).
Two L of
Lipofectamine LTX Reagent (Invitrogen, Carlsbad, CA) was added per 0.5 ng of
plasmid DNA.
The tube was mixed gently and incubated for 25 minutes at room temperature to
allow for DNA-
Lipofectamine LTX complex formation. 100 1._, of the DNA-Lipofectamine LTX
complex was
added directly to each well containing cells and mixed gently. Approximately
18-24 hours post-
transfection, medium containing DNA-Lipofectamine complexes was aspirated,
cells were
washed with PBS, and RPMI1640 medium supplemented with 10% characterized fetal
bovine
serum (Invitrogen, Carlsbad, CA), and 1X non-essential amino acids
(Invitrogen, Carlsbad, CA)
was added. Test composition (1 -(3-tert-buty1-1-(quinolin-6-y1)-1H-pyrazol-5-
y1)-3-(2-fluoro -4-
(2-(methylcarbamoyl)pyridin-4-yloxy)phenyOurea para-toluene sulfonic acid
salt) or DMSO was
added to the wells (0.5% final DMSO concentration). The plates were then
incubated for 2 hours
at 37 degrees Celsius, 5% CO2, and 95% humidity. Following the incubation, the
media was
aspirated and the cells were washed three times with 1 mL media to wash out
free composition.
Next, 1 mL fresh media was added and cells were incubated for specific times
points prior to
lysis (i.e., 0, 1, 2, 4, 6, and 24 hours). The cells were lysed using MPER
lysis buffer (Pierce,
Rockford, IL) containing Halt Phosphatase and Protease Inhibitors (Pierce,
Rockford, IL) and
Phosphatase inhibitor cocktail 2 (Sigma, St. Louis, MO) at 4 C for 10 minutes
with shaking.
Cleared lysates were separated by SDS-PAGE on a 4-12% Novex NuPage Bis-Tris
gel
(Invitrogen, Carlsbad, CA) and then transferred to PVDF (Invitrogen, Carlsbad,
CA). After
transfer, the PVDF membrane was blocked with BSA (Santa Cruz Biotechnology,
Santa Cruz,
CA) and then probed with an antibody for phospho-TIE2 (Cell Signaling
Technology, Beverly,
MA). A secondary anti-rabbit antibody conjugated to horseradish peroxidase
(Cell Signaling
Technology, Beverly, MA) was used to detect phospho-TIE2 ECL Plus (GE
Healthcare,
Piscataway, NJ), a substrate for horseradish peroxidase that generates a
fluorescent product, was
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added. Fluorescence was detected using a Storm 840 phosphorimager (GE
Healthcare,
Piscataway, NJ) in fluorescence mode. PVDF membranes were stripped and then re-
probed with
total TIE2 antibody (Santa Cruz Biotechnology, Inc., Dallas, TX) as above. The
160 kDa
phospho-TIE2 and total TIE2 bands were quantified using ImageQuant software
(GE Healthcare,
Piscataway, NJ). Phospho-TIE2 levels were normalized to total TIE2 levels, and
data was plotted
using Prism software (GraphPad Software, San Diego, CA). When incubated with
TIE2-
transfected CHO K1 cells for 2 hours at 0.1 IVI ¨ 1 IVI prior to being
washed out, Composition
1-(3-tert-buty1-1-(quino lin-6-y1)-1H-pyrazol-5-y1)-3-(2-fluoro-4-(2-(methylc
arb amoyOpyri din-4-
yloxy)phenyOurea para-toluene sulfonic acid salt disclosed herein exhibited
>50% inhibition of
phospho-TIE2 levels for >24 hours.
Example 5. Cellular inhibition of TIE2 in HUVEC cells by 1-(3-tert-buty1-1-
(quinolin-6-
y1)-1H-pyrazol-5-y1)-3-(2-fluo ro-4-(2-(methylc arb a moyl)pyridin-4-yloxy)p
he nyl)u re a
HUVEC Cell Culture
[0541] HUVEC (Human umbilical vein endothelial cells; Catalog #CRL-1730)
cells were
obtained from the American Type Culture Collection (ATCC, Manassas, VA).
Briefly, cells
were grown in EGM-2 (Lonza, Walkersville, MD) at 37 degrees Celsius, 5%CO2,
and 95%
humidity. Cells were allowed to expand until reaching 90-95% saturation at
which point they
were subcultured or harvested for assay use.
HUVEC Phospho-TIE2 Western Blot Assay
[0542] HUVEC cells (2.5 x 105 cells/well) were added to a 24-well tissue-
culture treated
plate in 1 mL of EGM-2 culture medium (Lonza, Walkersville, MD). Cells were
then incubated
overnight at 37 degrees Celsius, 5% CO2, and 95% humidity. Media was then
aspirated and 1
mL EBM-2 basal medium (Lonza, Walkersville, MD) supplemented with 2% FBS
(Invitrogen,
Carlsbad, CA) was added. Test composition or DMSO was added to the wells (0.5%
final
DMSO concentration). The plates were then incubated for 4 hours at 37 degrees
Celsius, 5%
CO2, and 95% humidity. During the incubation, histidine-tagged angiopoietin 1
(ANG1) growth
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factor (R&D Systems, Minneapolis, MN) was added to a anti-polyhistidine
antibody (R&D
Systems, Minneapolis, MN) for 30 minutes at room temperature to generate
multimers of ANG1.
Following the four hour incubation of composition, cells were stimulated with
800 ng/mL of the
ANG l/anti-polyhistidine antibody complex mixture for 15 minutes. The media
was aspirated and
the cells were washed with PBS. The cells were lysed using MPER lysis buffer
(Pierce,
Rockford, IL) containing Halt Phosphatase and Protease Inhibitors (Pierce,
Rockford, IL) and
Phosphatase inhibitor cocktail 2 (Sigma, St. Louis, MO) at 4 C for 10 minutes
with shaking.
Cleared lysates were separated by SDS-PAGE on a 4-12% Novex NuPage Bis-Tris
gel
(Invitrogen, Carlsbad, CA) and then transferred to PVDF (Invitrogen, Carlsbad,
CA). After
transfer, the PVDF membrane was blocked with BSA (Santa Cruz Biotechnology,
Santa Cruz,
CA) and then probed with an antibody for phospho-TIE2 (Cell Signaling
Technology, Beverly,
MA). A secondary anti-rabbit antibody conjugated to horseradish peroxidase
(Cell Signaling
Technology, Beverly, MA) was used to detect phospho-TIE2. ECL Plus (GE
Healthcare,
Piscataway, NJ), a substrate for horseradish peroxidase that generates a
fluorescent product, was
added. Fluorescence was detected using a Storm 840 phosphorimager (GE
Healthcare,
Piscataway, NJ) in fluorescence mode. The 160 kDa phospho-TIE2 band was
quantified using
ImageQuant software (GE Healthcare, Piscataway, NJ). Data was analyzed using
Prism software
(GraphPad Software, San Diego, CA) to calculate IC50 values. The composition 1-
(3-tert-butyl-
1-(quino lin-6-y1)-1H-pyrazol-5-y1)-3-(2-fluoro -4-(2-(methylc arb amoyl)pyri
din-4-
yloxy)phenyOurea para-toluene sulfonic acid salt disclosed herein exhibited an
IC50 value of
0.018 nM.
Example 6. Inhibition of angiopoietin 1 (ANGI) or angiopoietin 2 (ANG2)
stimulated
capillary tube formation by 1-(3-tert-buty1-1-(quinolin-6-y1)-1H-pyrazol-5-y1)-
3-(2-fluoro-4-
(2-(methylca rb am oyl)pyridin-4-yloxy)p he nyl)u re a
HMVEC Cell Culture
[0543] HMVEC (Human microvascular endothelial cells; Catalog #PCS-110-010)
cells were
obtained from the American Type Culture Collection (ATCC, Manassas, VA).
Briefly, cells
were grown in EGM-2 MV (Lonza, Walkersville, MD) at 37 degrees Celsius, 5%
CO2, and 95%
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humidity. Cells were allowed to expand until reaching 90-95% saturation at
which point they
were subcultured or harvested for assay use.
HMVEC Capillary Tube Formation Assay
[0544] HMVEC cells (1.5 x 104 cells/well) mixed with test composition or
DMSO control
and the appropriate growth factor (ANG1 or ANG2) or control were added to a 96-
well tissue-
culture treated plate coated with growth-factor reduced Matrigel in 0.1 mL of
EBM-2 basal
medium (Lonza, Walkersville, MD). Cells were then incubated for 18 hours at 37
degrees
Celsius, 5% CO2, and 95% humidity. Media was then gently aspirated and wells
were gently
washed with 0.1 mL EBM-2 basal medium. Media was again aspirated and 1 iuM
Calcein AM
solution (Invitrogen, Carlsbad, CA) in basal medium was added to each well to
fluorescently
label live cells. Cells were then incubated for 30 minutes at 37 degrees
Celsius, 5% CO2, and
95% humidity. Media was aspirated and wells were gently washed with phosphate-
buffered
saline twice. Images of each well were acquired with a fluorescent microscope
and processed
using ImagePro Analyzer (Media Cybernetics, Inc., Rockville, MD) using an
automated macro
that measures total capillary tube length. Data was analyzed using Prism
software (GraphPad
Software, San Diego, CA) to calculate IC50 values. The composition 1-(3-tert-
buty1-1-(quinolin-
6-y1)-1H-pyrazol-5-y1)-3-(2-fluoro-4-(2-(methylcarbamoyl)pyridin-4-
yloxy)phenyOurea para-
toluene sulfonic acid salt disclosed herein exhibited an IC50 value of 6.9 nM
for inhibition of
ANG1-stimulated HMVEC capillary tube formation. The composition of Formula I
disclosed
herein exhibited an IC50 value of 34 nM for inhibition of ANG2-stimulated
HMVEC capillary
tube formation.
Example 7. Inhibition of in vivo primary tumor growth and invasiveness in the
murine
PyMT breast cancer model by 1-(3-tert-buty1-1-(quinolin-6-y1)-1H-pyrazol-5-y1)-
3-(2-
fluoro-4-(2-(methylcarbamoyl)pyridin-4-yloxy)phenyl)urea as a single agent and
in
combination with paclitaxel
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PyMT syngeneic breast cancer model primary tumor growth
[0545] The
PyMT syngeneic breast cancer implant mouse model was used to evaluate in vivo
activity of compound 1. Briefly, 1 x 106 cells (dissociated from PyMT tumor
fragments) in 0.1
mL total volume were implanted into the fourth mammary fat pad on the left
side of female mice
(FVB/NJ, JAXWEST:RB05 mice from Jackson Labs). A total of 10 mice were
implanted in
each group. Molecular Imaging, Inc.'s Animal Care and Use Committee approved
all the
experimental protocols and conducted experiments in compliance with all the
laws, regulations
and guidelines of the National Institutes of Health (NIH). Treatment was
initiated by oral
administration (gavage) of Compound 1 twice daily or vehicle (0.4%
hydroxypropylmethylcellulose in water) and/or intravenous administration (IV)
of paclitaxel
every five days or vehicle (10% ethanol, 10% Cremophor EL and 80% saline)
according to
individual body weight on the day of treatment at 0.2 mL per 20 g when tumor
size reached
approximately 850 mg.
Animals were dosed for 21 days. Body weights and tumor
measurements were recorded three times weekly. Tumor burden (mg) was estimated
from caliper
measurements by the formula for the volume of a prolate ellipsoid assuming
unit density as:
Tumor burden (mg) = (L x W2)/2, where L and W are the respective orthogonal
tumor length and
width measurements (mm),In the PyMT model, both Compound 1 and paclitaxel
groups
evidenced tumor growth inhibition. Compound 1 in combination with paclitaxel
demonstrated
additive activity (Figure 1). These data evidence in vivo activity by Compound
1 and show
correlation to enzymatic and cell data.
Example 8. Inhibition of in vivo primary tumor macrophage accumulation in the
murine
PyMT breast cancer model by 1-(3-tert-buty1-1-(quinolin-6-y1)-1H-pyrazol-5-y1)-
3-(2-
fluoro-4-(2-(methylcarbamoyl)pyridin-4-yloxy)phenyl)urea as a single agent and
in
combination with paclitaxel
PyMT syngeneic breast cancer model primary tumor macrophage accumulation
[0546] The
PyMT syngeneic breast cancer implant mouse model was used to evaluate in vivo
activity of compound 1. Briefly, 1 x 106 cells (dissociated from PyMT tumor
fragments) in 0.1
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mL total volume were implanted into the fourth mammary fat pad on the left
side of female mice
(FVB/NJ, JAXWEST:RB05 mice from Jackson Labs). A total of 10 mice were
implanted in
each group. Molecular Imaging, Inc.'s Animal Care and Use Committee approved
all the
experimental protocols and conducted experiments in compliance with all the
laws, regulations
and guidelines of the National Institutes of Health (NIH). Treatment was
initiated by oral
administration (gavage) of Compound 1 twice daily or vehicle (0.4%
hydroxypropylmethylcellulose in water) and/or intravenous administration (IV)
of paclitaxel
every five days or vehicle (10% ethanol, 10% Cremophor EL and 80% saline)
according to
individual body weight on the day of treatment at 0.2 mL per 20 g when tumor
size reached
approximately 850 mg.
Animals were dosed for 21 days. Body weights and tumor
measurements were recorded three times weekly.
[0547] At
the end of study, tumors were excised and placed in formalin. Formalin-fixed
tumor samples were then placed in paraffin blocks. Formalin fixed-paraffin
embedded tissue
slides were deparaffinized with xylene and hydrated to distilled water through
a graduated series
of alcohol rinses. Antigenic retrieval was performed using Dako's PT Link
Module with a
Tris/EDTA buffer target retrieval solution at 95 C for 20 minutes. Once the
slides cooled down,
they were loaded onto a Dako AutostainerPlusLink for immunohistochemical
staining at room
temperature with F4/80 and CD31 antibodies, to stain for macrophages and
endothelial cells,
respectively. Endogenous peroxidase and alkaline phosphatase activity in the
tissues was
quenched with a Dual Endogenous Enzyme Block solution (Dako, S2003) for 5
minutes. Non-
specific protein binding was blocked with serum free Protein Block (Dako,
X0909) for 5
minutes. The rat anti mouse CD31 primary antibody was then incubated on the
experimental
tissue sections for 30 minutes at an immunogenic concentration of 1:100. The
primary antibody
was then conjugated with a rabbit anti rat immunoglobulin secondary antibody
(Dako, E0468).
The secondary antibody was then amplified with a goat anti rabbit peroxidase
labeled polymer
(Dako, K4003) for 30 minutes. Enzymatic staining was developed with substrate-
chromogen
DAB+ (Dako, K3468) for 5 minutes. Excess rat IgG components were further
blocked with
Rodent Block Rat (Biocare Medical, RBR962H) for 5 minutes. The rat anti mouse
F4/80 primary
antibody was incubated on the experimental tissue sections for 30 minutes. The
F4/80 was then
conjugated with a rabbit anti rat immunoglobulin secondary antibody (Dako,
E0468). The
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secondary antibody was then amplified with a goat anti rabbit alkaline
phosphatase labeled
polymer, (Biocare RALP525) for 30 minutes. Enzymatic staining was developed
with substrate
chromogen WARP Red (Biocare WR806). The counter staining was performed with
automation
hematoxylin for 10 minutes. The tissue slides were then air dried and cleared
to xylene for glass
cover slipping. Slides were scored for F4/80 staining using a scale of 0, no
visible staining; 1,
weak staining; 2, moderate staining; 3 strong staining.In the PyMT model,
Compound 1
evidenced a decrease in macrophage accumulation at the primary tumor, whereas
paclitaxel did
not decrease macrophage accumulation. Compound 1 in combination with
paclitaxel
demonstrated similar activity to Compound 1 single-agent treatment (Figure 2).
These data
evidence in vivo activity by Compound 1 and show correlation to enzymatic and
cell data.
Example 9. Inhibition of in vivo primary tumor TIE2 cell accumulation in the
murine
PyMT breast cancer model by 1-(3-tert-buty1-1-(quinolin-6-y1)-1H-pyrazol-5-y1)-
3-(2-
fluoro-4-(2-(methylcarbamoyl)pyridin-4-yloxy)phenyl)urea as a single agent and
in
combination with paclitaxel
PyMT syngeneic breast cancer model primary tumor macrophage accumulation
[0548] The
PyMT syngeneic breast cancer implant mouse model was used to evaluate in vivo
activity of compound 1. Briefly, 1 x 106 cells (dissociated from PyMT tumor
fragments) in 0.1
mL total volume were implanted into the fourth mammary fat pad on the left
side of female mice
(FVB/NJ, JAXWEST:RB05 mice from Jackson Labs). A total of 10 mice were
implanted in
each group. Molecular Imaging, Inc.'s Animal Care and Use Committee approved
all the
experimental protocols and conducted experiments in compliance with all the
laws, regulations
and guidelines of the National Institutes of Health (NIH). Treatment was
initiated by oral
administration (gavage) of Compound 1 twice daily or vehicle (0.4%
hydroxypropylmethylcellulose in water) and/or intravenous administration (IV)
of paclitaxel
every five days or vehicle (10% ethanol, 10% Cremophor EL and 80% saline)
according to
individual body weight on the day of treatment at 0.2 mL per 20 g when tumor
size reached
approximately 850 mg.
Animals were dosed for 21 days. Body weights and tumor
measurements were recorded three times weekly.
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[0549] At the end of study, tumors were excised and placed in formalin.
Formalin-fixed
tumor samples were then placed in paraffin blocks. The experimental formalin
fixed-paraffin
embedded tissue slides were deparaffinized with xylene and hydrated to
distilled water through a
graduated series of alcohol rinses. Antigenic retrieval was performed using
Dako's PT Link
Module with a Citrate pH 6 buffer target retrieval solution at 95 C for 20
minutes. Once the
slides cooled down, they were loaded onto a Dako AutostainerPlusLink for
immunohistochemical staining at room temperature with both TIE2 and CD31
antibodies.
Endogenous peroxidase and alkaline phosphatase activity in the tissues was
quenched with a
Dual Endogenous Enzyme Block solution (Dako, S2003) for 5 minutes. Non-
specific protein
binding was blocked with serum free Protein Block (Dako, X0909) for 5 minutes.
The rat anti
mouse CD31 primary antibody was then incubated on the experimental tissue
sections for 30
minutes at an immunogenic concentration of 1:100. The primary antibody was
then conjugated
with a rabbit anti rat immunoglobulin secondary antibody (Dako, E0468). The
secondary
antibody was then amplified with a goat anti rabbit peroxidase labeled polymer
(Dako, K4003)
for 30 minutes. Enzymatic staining was developed with substrate-chromogen DAB+
(Dako,
K3468) for 5 minutes. Excess protein components were further blocked with
Protein Block
(Dako, X0909) for 5 minutes. The rabbit anti TIE2 primary antibody was
incubated on the
experimental tissue sections for 30 minutes. The TIE2 antibody was then
conjugated with an
alkaline phosphatase labeled goat anti rabbit polymer for 30 minutes.
Enzymatic staining was
developed with substrate chromogen WARP Red (Biocare WR806). The counter
staining was
performed with automation hematoxylin for 10 minutes. The tissue slides were
then air dried and
cleared to xylene for glass cover slipping. Slides were scored for TIE2
staining using a scale of
0, no visible staining; 1, weak staining; 2, moderate staining; 3 strong
staining.In the PyMT
model, Compound 1 evidenced a decrease in TIE2-expressing cell accumulation at
the primary
tumor, whereas paclitaxel did not decrease TIE2-expressing cell accumulation.
Compound 1 in
combination with paclitaxel demonstrated enhanced activity compared to
Compound 1 single-
agent treatment (Figure 3). These data evidence in vivo activity by Compound 1
and show
correlation to enzymatic and cell data.
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Example 10. Inhibition of in vivo lung metastases in the murine PyMT breast
cancer model
by 1-(3-tert-buty1-1-(quinolin-6-y1)-1H-pyrazol-5-y1)-3-(2-fluoro-4-(2-
(methylcarbamoyl)pyridin-4-yloxy)phenyl)urea as a single agent and in
combination with
paclitaxel
PyMT syngeneic breast cancer model lung metastasis evaluation
[0550] The
PyMT syngeneic breast cancer implant mouse model was used to evaluate in vivo
activity of compound 1. Briefly, 1 x 106 cells (dissociated from PyMT tumor
fragments) in 0.1
mL total volume were implanted into the fourth mammary fat pad on the left
side of female mice
(FVB/NJ, JAXWEST:RB05 mice from Jackson Labs). A total of 10 mice were
implanted in
each group. Molecular Imaging, Inc.'s Animal Care and Use Committee approved
all the
experimental protocols and conducted experiments in compliance with all the
laws, regulations
and guidelines of the National Institutes of Health (NIH). Treatment was
initiated by oral
administration (gavage) of Compound 1 twice daily or vehicle (0.4%
hydroxypropylmethylcellulose in water) and/or intravenous administration (IV)
of paclitaxel
every five days or vehicle (10% ethanol, 10% Cremophor EL and 80% saline)
according to
individual body weight on the day of treatment at 0.2 mL per 20 g when tumor
size reached
approximately 850 mg.
Animals were dosed for 21 days. Body weights and tumor
measurements were recorded three times weekly.
[0551] At
the end of study, lung tissues were excised and placed in formalin. Formalin-
fixed
lung samples were then placed in paraffin blocks. Each lung block had three
slides with two
levels per slide cut and stained with Hematoxylin and Eosin. Metastatic lung
nodules were
counted via microscopy.In the PyMT model, both Compound 1 and paclitaxel
evidenced a
similar decrease in lung metastasis. Compound 1 in combination with paclitaxel
demonstrated
additive activity compared to single-agent treatments (Figure 4). These data
evidence in vivo
activity by Compound 1 and show correlation to enzymatic and cell data.
Example 11. Inhibition of in vivo lung metastases in the murine PyMT breast
cancer model
by 1-(3-tert-buty1-1-(quinolin-6-y1)-1H-pyrazol-5-y1)-3-(2-fluoro-4-(2-
CA 02929715 2016-05-04
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(methylcarbamoyl)pyridin-4-yloxy)phenyl)urea dosed intermittently (non-daily)
in
combination with paclitaxel
PyMT syngeneic breast cancer model lung metastasis evaluation
[0552] The PyMT syngeneic breast cancer implant mouse model was used to
evaluate in vivo
activity of compound 1. Briefly, 1 x 106 cells (dissociated from PyMT tumor
fragments and
stored frozen in cell-freezing medium) in 0.1 mL total volume were implanted
into the fourth
mammary fat pad on the left side of female mice (FVB/NJ, JAXWEST:RB05 mice
from Jackson
Labs). A total of three mice were implanted in each group. Molecular Imaging,
Inc.'s Animal
Care and Use Committee approved all the experimental protocols and conducted
experiments in
compliance with all the laws, regulations and guidelines of the National
Institutes of Health
(NIH). Treatment was initiated by oral administration (gavage) of Compound 1
twice weekly or
vehicle (0.4% hydroxypropylmethylcellulose in water) and/or intravenous
administration (IV) of
paclitaxel every five days or vehicle (10% ethanol, 10% Cremophor EL and 80%
saline)
according to individual body weight on the day of treatment at 0.2 mL per 20 g
when tumor size
reached approximately 600 mg. Animals were dosed for 12 days. Body weights and
tumor
measurements were recorded three times weekly.
[0553] At the end of study, lung tissues were excised and placed in
formalin. Formalin-fixed
lung samples were then placed in paraffin blocks. Each lung block had three
slides with two
levels per slide cut and stained with Hematoxylin and Eosin. Metastatic lung
nodules were
counted via microscopy.In the PyMT model, paclitaxel evidenced a decrease in
lung metastasis.
Compound 1 in combination with paclitaxel demonstrated additive activity
compared to single-
agent treatment (Figure 5). These data evidence in vivo activity by Compound 1
and show
correlation to enzymatic and cell data.
Example 12. Inhibition of in vivo lung metastases in the murine PyMT breast
cancer model
by 1-(3-tert-buty1-1-(quinolin-6-y1)-1H-pyrazol-5-y1)-3-(2-fluoro-4-(2-
(methylcarbamoyl)pyridin-4-yloxy)phenyl)urea dosed intermittently (non-daily)
in
combination with eribulin
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PyMT syngeneic breast cancer model lung metastasis evaluation
[0554] The PyMT syngeneic breast cancer implant mouse model was used to
evaluate in vivo
activity of compound 1. Briefly, 1 x 106 cells (dissociated from PyMT tumor
fragments and
stored frozen in cell-freezing medium) in 0.1 mL total volume were implanted
into the fourth
mammary fat pad on the left side of female mice (FVB/NJ, JAXWEST:RB05 mice
from Jackson
Labs). A total of three mice were implanted in each group. Molecular Imaging,
Inc.'s Animal
Care and Use Committee approved all the experimental protocols and conducted
experiments in
compliance with all the laws, regulations and guidelines of the National
Institutes of Health
(NIH). Treatment was initiated by oral administration (gavage) of Compound 1
twice weekly or
vehicle (0.4% hydroxypropylmethylcellulose in water) and/or intravenous
administration (IV) of
eribulin three times weekly or vehicle (80% saline) according to individual
body weight on the
day of treatment at 0.2 mL per 20 g when tumor size reached approximately 600
mg. Animals
were dosed for 12 days. Body weights and tumor measurements were recorded
three times
weekly.
[0555] At the end of study, lung tissues were excised and placed in
formalin. Formalin-fixed
lung samples were then placed in paraffin blocks. Each lung block had three
slides with two
levels per slide cut and stained with Hematoxylin and Eosin. Metastatic lung
nodules were
counted via microscopy.In the PyMT model, eribulin evidenced a decrease (or
increase at low
dose) in lung metastasis. Compound 1 in combination with eribulin demonstrated
additive
activity compared to single-agent treatment (Figure 6). These data evidence in
vivo activity by
Compound 1 and show correlation to enzymatic and cell data.
Example 13. Increase in overall survival in the murine PyMT breast cancer
model by 1-(3-
tert-buty1-1-(quinolin-6-y1)-1H-pyrazol-5-y1)-3-(2-fluoro-4-(2-
(methylcarbamoyl)pyridin-4-
yloxy)phenyl)urea dosed intermittently (non-daily) in combination with
eribulin
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PyMT syngeneic breast cancer model survival evaluation
[0556] The PyMT syngeneic breast cancer implant mouse model was used to
evaluate in vivo
activity of compound 1. Briefly, 1 x 106 cells (dissociated from PyMT tumor
fragments and
stored frozen in cell-freezing medium) in 0.1 mL total volume were implanted
into the fourth
mammary fat pad on the left side of female mice (FVB/NJ, JAXWEST:RB05 mice
from Jackson
Labs). A total of ten mice were implanted in each group. Molecular Imaging,
Inc.'s Animal
Care and Use Committee approved all the experimental protocols and conducted
experiments in
compliance with all the laws, regulations and guidelines of the National
Institutes of Health
(NIH). Treatment was initiated by oral administration (gavage) of Compound 1
once or twice
weekly or vehicle (0.4% hydroxypropylmethylcellulose in water) and/or
intravenous
administration (IV) of eribulin three times weekly or vehicle (80% saline)
according to
individual body weight on the day of treatment at 0.2 mL per 20 g when tumor
size reached
approximately 850 mg. Tumors were then resected three days after treatment
began. Animals
were then dosed for the duration of the survival experiment. Body weights and
tumor
measurements were recorded three times weekly. In the PyMT model, eribulin at
0.1 mg/kg
evidenced no increase in survival. Compound 1 in combination with eribulin
demonstrated
significant increases in survival (Figure 7). These data evidence in vivo
activity by Compound 1
and show correlation to enzymatic and cell data.
[0557] The present invention is not to be limited in scope by the specific
embodiments
disclosed in the examples which are intended as illustrations of a few aspects
of the invention
and any embodiments that are functionally equivalent are within the scope of
this invention.
Indeed, various modifications of the invention in addition to those shown and
described herein
will become apparent to those skilled in the art and are intended to fall
within the scope of the
appended claims.
EQUIVALENTS
[0558] Those skilled in the art will recognize, or be able to ascertain,
using no more than
routine experimentation, numerous equivalents to the specific embodiments
described
specifically herein. Such equivalents are intended to be encompassed in the
scope of the
following claims.
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