Note: Descriptions are shown in the official language in which they were submitted.
CA 02664165 2009-03-20
WO 2008/049904 PCT/EP2007/061501
-1-
USE OF MTKI 1 FOR TREATING OR PREVENTING BONE CANCER
FIELD OF THE INVENTION
The present invention is concerned with the finding that the macrocyclic
quinazoline
derivative 4,6-ethanediylidenepyrimido[4,5-b][6,1,12]benzoxadiazacyclo-
pentadecine,
17-bromo-8,9,10,11,12,13,14,19-octahydro-20-methoxy-13-methyl, described as
compound 22 in PCT publication W02004/105765, is useful in the manufacture of
a
medicament for the treatment or prevention of bone cancers and methods for
killing
bone cancer cells, including osteosarcomas, chondrosarcomas, myeloma bone
disease
and osteolytic bone metastases from other primary sites. It accordingly
provides
methods for treating, preventing, delaying or mitigating bone cancer, or for
preventing
and treating of bone loss associated with cancer metastases.
BACKGROUND OF THE INVENTION
"Bone cancer" includes primary bone cancer cells such as osteosarcoma cells,
cells
from Ewing's family of tumors, chondrosarcoma cells, malignant giant cell
tumor cells,
malignant fibrous histiocytoma cells and adamantinoma cells, as well as
secondary
bone cancer cells that have metastasized from other tissues, including breast,
lung,
prostate and kidney.
Osteosarcoma is a malignant tumor of bone, which is most prevalent in
adolescents and
young adults. Osteosarcoma accounts for approximately 5% of the tumors in
childhood
and 80% of these tumors originate around the knee. The prognosis is often poor
and
within 1 year after commencing definitive therapy, about 30% of patients
diagnosed
with osteosarcoma will develop lung metastasis. The prognosis appears to be
determined by the site of metastases and surgical resectability of the
metastatic disease,
either at diagnosis or following a variable period of chemotherapy. Patients
who have
complete surgical ablation of the primary and metastatic tumor (when confined
to the
lung) following chemotherapy may attain long-term survival, although event-
free
survival remains about 20% for patients with metastatic disease at diagnosis.
Patients
developing recurrent disease often have a poor prognosis and die within 1 year
of the
CA 02664165 2009-03-20
WO 2008/049904 PCT/EP2007/061501
-2-
development of metastatic disease.
Chemotherapy is often ineffective, resulting in a high mortality rate. Hence,
it is
important that new therapeutic approaches are evaluated for this malignant
disease.
Myeloma bone disease is a cancer of antibody-producing plasma cells in the
bone
marrow. Proliferation of the cancerous plasma cells, referred to as myeloma
cells,
causes a variety of effects, including lytic lesions (holes) in the bone,
decreased red
blood cell number, production of abnormal proteins (with attendant damage to
the
kidney, nerves, and other organs), reduced immune system function, and
elevated blood
calcium levels (hypercalcemia).
When myeloma cells are present at distinct skeletal locations, the disease is
referred to
as multiple myeloma.
Although responsible for only 1% of all cancers in the United States, with
14,600 new
cases reported in 2002, myeloma is the second most common blood cancer and may
be
increasing in prevalence, particularly among individuals under age 55
(International
Myeloma Foundation). Many different treatment options are available or in
development, but there is neither a cure nor agreement on an optimal myeloma
management regimen. Patients are treated with chemotherapy as well as symptom-
specific treatments for one or more of hypercalcemia, increased infection
risk, kidney
failure, anemia, hyperviscosity of blood, elevated stroke risk, bone
destruction and
pain, and muscle weakness. Unfortunately, dramatic reduction in the number of
myeloma cells does not necessarily translate into longer remissions or
survival times,
and therapies that were effective before a remission may not prove effective
upon
relapse of the disease.
One of the most prevalent and significant characteristics of myeloma is the
activation
of osteoclasts, multinucleated cells that absorb bone, leading to bone
thinning, lytic
bone lesions, and bone fracture. Lytic bone lesions occur in 70-80% of
multiple
myeloma patients and are frequently associated with severe bone pain and
pathologic
CA 02664165 2009-03-20
WO 2008/049904 PCT/EP2007/061501
-3-
fractures. In normal bone functioning, a balance exists between osteoclasts,
which
resorb bone, and osteoblasts, cells that produce bone. This balance is upset
in myeloma
patients, and more bone is resorbed than produced. The increased osteoclastic
bone
resorption occurs adjacent to the myeloma cells and not in areas of normal
bone
marrow, indicating that the osteoclast activation occurs by a local mechanism.
Although it is well accepted that myeloma cells activate osteoclasts, the
precise
mechanism by which this occurs is unknown. Myeloma cells, in culture, produce
or
induce production of several osteoclast-activating factors (OAFs) whose
specific roles
in vivo are yet to be determined. Recently, the chemokine macrophage
inflammatory
protein-la (MIP-la) has been implicated in osteoclast activation in vitro (S.
J. Choi et
al., Blood 96: 671-675 (2000)). Therapies addressing mechanisms involving OAFs
are
presently under development.
Currently, bone indications of multiple myeloma are treated primarily with
bisphosphonates, a class of chemicals that inhibits osteoclast activity or
osteoclast
attachment to bone surface and eventually leads to osteoclast cell death. They
may also
affect myeloma cells directly. Bisphosphonates are administered by infusion.
Third-
generation bisphosphonates are currently under development, but even improved
versions of the drugs may have potential side effects including hypocalcemia,
kidney
damage, and increased pain. -
Bisphosphonates do not completely block the bone destruction process, and
patients
eventually develop new bone lesions. An alternative therapy for bone
destruction in
multiple myeloma that can be administered orally would be highly beneficial.
Bone metastases are often associated with advanced cancer and are most common
with
breast, prostate and thyroid carcinomas and multiple myeloma (supra). Bone
metastases
are present in 65-75% of patients with advanced (metastatic) breast cancer.
Metastatic
bone lesions may be lytic or sclerotic in nature depending upon whether
increased
osteoclastic or osteoblastic activity predominates; if both processes are
equally active,
they are termed mixed lesions. Bone metastases in breast cancer patients
usually
involve osteolytic disease, where normal bone homeostasis is disrupted and
skewed
towards excessive resorption of bone (Coleman RE, Cancer Treat Rev. 27(3), 165-
76
CA 02664165 2009-03-20
WO 2008/049904 PCT/EP2007/061501
-4-
(2001)). Tumor-induced skeletal damage is mediated by osteoclasts that are
stimulated
directly or indirectly to dissolve bone by local factors (e.g. prostaglandin
E, interleukin-
1, tumor necrosis factor and procathepsin D) released by tumor cells or
associated
immune cells, or by systemic factors, such as parathyroid hormone-related
peptide. The
most frequently affected skeletal sites are the vertebrae, pelvis, ribs, femur
and skull.
Patients with bone metastases experience considerable morbidity, including
bone pain,
pathological fractures, hypercalcaemia, reduced mobility and spinal cord or
nerve root
compression. Despite the importance of these clinical problems, there are few
available treatments for bone loss associated with cancer metastasis. Thus,
there
remains a need in the art to identify new agents and methods for preventing or
treating
cancer metastasis, including osteolytic bone metastases.
SUMMARY OF THE INVENTION
The invention is directed in part to methods of treating or preventing bone
cancer, and
to methods of treating or preventing bone loss associated with cancer
metastases,
utilizing certain compounds described in WO 2004/105765, the disclosure of
which is
hereby incorporated by reference in its entirety.
In one embodiment, the present invention provides the use of the macrocyclic
quinazoline derivative 4,6-ethanediylidenepyrimido[4,5-
b][6,1,12]benzoxadiazacyclo-
pentadecine, 17-bromo-8,9,10,11,12,13,14,19-octahydro-20-methoxy-13-methyl,
described as compound 22 in PCT publication W02004/105765, in the manufacture
of
a medicament for the treatment or prevention of bone cancers and methods for
killing
bone cancer cells, including osteosarcomas, chondrosarcomas, myeloma bone
disease
and osteolytic bone metastases. It accordingly provides methods for treating,
preventing, delaying or mitigating bone cancer, or for preventing and treating
of bone
loss associated with cancer metastases.
In related embodiment, the invention provides a method of inhibiting
metastatic spread
of a cancer to skeletal system, in a mammalian subject comprising
administering to a
mammalian subject suspected of having metastatic cancer a compound of the
invention,
CA 02664165 2009-03-20
WO 2008/049904 PCT/EP2007/061501
-5-
in an amount effective to inhibit metastatic spread of the cancer to the
skeletal system;
and a method for treating bone cancer comprising administering to a mammalian
subject diagnosed with a cancer a composition comprising a compound of the
invention, in an amount effect to reduce growth or neoplastic spread of the
bone cancer.
It will be appreciated that any reduction in the rate of cancer growth or
spread (which
can prolong life and quality of life) is indicative of successful treatment.
In preferred
embodiments, cancer growth is halted completely. In still more preferred
embodiments,
cancers shrink or are eradicated entirely. Preferred subjects for treatment
are human
subjects, but other animals, especially murine, rat, bovine, porcine, primate,
and other
model systems for cancer treatment, are contemplated. Metastatic cancers as
used
herein are contemplated to include a variety of cancers can metastasize to the
bone, but
the most common metastasizing cancers are breast, lung, renal, multiple
myeloma,
thyroid and prostate. By way of example, other cancers that have the potential
to
metastasize to bone include but are not limited to adenocareinoma, blood cell
malignancies, including leukemia and lymphoma; head and neck cancers;
gastrointestinal cancers, including stomach cancer, colon cancer, colorectal
cancer,
pancreatic cancer, liver cancer; malignancies of the female genital tract,
including
ovarian carcinoma, uterine endometrial cancers and cervical cancer; bladder
cancer ;
brain cancer, including neuroblastoma; sarcoma, osteosarcoma; and skin cancer,
including malignant melanoma and squamous cell cancer. The present invention
especially contemplates prevention and treatment of tumor-induced osteolytic
lesions in
bone
In one variation of the foregoing methods of treatment, the compounds are
administered along with a second cancer therapeutic agent. The second agent
can be
any chemotherapeutic agent, radioactive agent, radiation, nucleic acid
encoding a
cancer therapeutic agent, antibody, protein, and/or other anti-lymphangiogenic
agent or
an anti-angiogenic agent. The second agent may be administered before, after,
or
concurrently with the compounds of the invention.
In one variation, the subject to be treated has been diagnosed with an
operable tumor,
and the administering step is performed before, during, or after the tumor is
resected
CA 02664165 2009-03-20
WO 2008/049904 PCT/EP2007/061501
-6-
from the subject. Compound treatment in conjunction with tumor resection is
intended
to reduce or eliminate regrowth of tumors from cancer cells that fail to be
resected.
Stated more generically, the invention provides a method of treating or
preventing bone
cancer, and to methods of treating or preventing bone loss associated with
cancer
metastases comprising the step of administering to a mammal (including, but
not
limited to humans, rats, canines, bovines, porcines, and primates) in need
thereof a
compound of the invention.
BRIEF DESCRIPTION OF THE DRAWINGS
Figure 1: Effect of treatrnent on spontaneous lifting behaviour of the left
hind paw.
The data is presented as the percentage of the time the paw was raised over an
observation period of 4 minutes. The vehicle group was dosed by oral gavage,
daily
with a 20% HPCD solution at pH 4Ø Compound 1 was dosed once daily at its
maximum tolerated dose (MTD) of 200mg per kg, Iressa was also dosed at its
maximum tolerated dose, by oral gavage, of 50mg per kg daily for 14 days.
Figure 2: Representative reconstructions from Ct s of the ipsilateral left
hindlimbs
showing osteolytic bone destruction in the tumor inoculated animals.
Figure 3: Dose dependent inhibition of breast tumor growth in a bone cancer
metastasis
model. MDA B231
DETAILED DESCRIPTION OF THE INVENTION
WO-2004/105765 describes the preparation, formulation and pharmaceutical
properties
of macrocyclic quinazoline derivatives of formula (I) as multi targeted kinase
inhibitors
(MTKIs).
CA 02664165 2009-03-20
WO 2008/049904 PCT/EP2007/061501
-7-
X2 3 Ri
/'
y 2'
J\ 51
z 6' RZ
X 5
6/ N3 R3
R4-~ I '/J
7 2 (I)
8 N1
It has now been found that one compound in the aforementioned class, i.e 4,6-
ethanediylidenepyrimido[4,5-b][6,1,12]benzoxadiazacyclo-pentadecine, 17-bromo-
8,9,10,11,12,13,14,19-octahydro-20-methoxy-13-methyl, described as compound 22
in
the aforementioned PCT publication, herein also referred to as MTKI 1 or
Compound
1, has clinical activity in bone cancer models and accordingly provide the use
of these
compounds for the preparation of a pharmaceutical composition for treating
bone
cancer, including primary bone cancers and bone metastases as defined
hereinbefore.
The present invention also concerns a method of treating tumor-induced
osteolytic
lesions in bone of a mammal, comprising the step of administering a
therapeutically
effective amount of a compound according to the invention to said mammal.
Accordingly, in one aspect the present invention provides the use of 4,6-
ethanediylidenepyrimido[4,5-b][6,1,12]benzoxadiazacyclo-pentadecine, 17-bromo-
8,9,10,11,12,13,14,19-octahydro-20-methoxy-13-methyl or a pharmaceutically
acceptable acid or base addition salt thereof, in the manufacture of a
medicament for
the treatment or prevention of bone cancer, including primary bone cancers and
bone
metastases as defined hereinbefore.
A further aspect of the present invention is directed to a method for the
treatment of
prevention of bone cancer in a mammalian subject, comprising administering a
therapeutically effective amount of 4,6-ethanediylidenepyrimido[4,5-
b] [6,1,12]benzoxadiazacyclo-pentadecine, 17-bromo-8,9,10,11,12,13,14,19-
octahydro-
CA 02664165 2009-03-20
WO 2008/049904 PCT/EP2007/061501
-8-
20-methoxy-13-methyl or a pharmaceutically acceptable acid or base addition
salt
thereof, to a mammalian subject in need of such treatment.
The pharmaceutically acceptable acid or base addition salts as mentioned
hereinabove
are meant to comprise the therapeutically active non-toxic acid and non-toxic
base
addition salt forms which MTKI 1 is able to form. The basic properties can be
converted
in their pharmaceutically acceptable acid addition salts by treating said base
form with
an appropriate acid. Appropriate acids comprise, for example, inorganic acids
such as
hydrohalic acids, e.g. hydrochloric or hydrobromic acid; sulfuric; nitric;
phosphoric and
the like acids; or organic acids such as, for example, acetic, propanoic,
hydroxyacetic,
lactic, pyruvic, oxalic, malonic, succinic (i.e. butanedioic acid), maleic,
fumaric, malic,
tartaric, citric, methanesulfonic, ethanesulfonic, benzenesulfonic, p-
toluenesulfonic,
cyclamic, salicylic, p-aminosalicylic, pamoic and the like acids.
The acidic properties may be converted in their pharmaceutically acceptable
base
addition salts by treating said acid form with a suitable organic or inorganic
base.
Appropriate base salt forms comprise, for example, the ammonium salts, the
alkali and
earth alkaline metal salts, e.g. the lithium, sodium, potassium, magnesium,
calcium
salts and the like, salts with organic bases, e.g., the benzathine, N-methyl-D-
glucamine,
hydrabamine salts, and salts with amino acids such as, for example, arginine,
lysine and
the like.
The terms acid or base addition salt also comprise the hydrates and the
solvent addition
forms which MTKI 1 is able to form. Examples of such forms are e.g., hydrates,
alcoholates and the like.
In particular, the present invention is. concerned with a use of the
dihydrobromide salt
of 4,6-ethanediylidenepyrimido[4,5-b][6,1,12]benzoxadiazacyclo-pentadecine, 17-
bromo-8,9,10,11,12,13,14,19-octahydro-20-methoxy-13-methyl, i.e., 17-bromo-
8,9,10,11,12,13,14,19-octahydro-20-methoxy-13-methyl-4,6-
ethanediylidenepyrimido
[4,5-b][6,1,12]benzoxadiazacyclopentadecine dihydrobromide, in any of the
aforementioned uses for MTKI 1.
CA 02664165 2009-03-20
WO 2008/049904 PCT/EP2007/061501
-9-
In a further embodiment, the present invention provides the use of the
aforementioned
MTKI 1 for the preparation of a pharmaceutical composition for the prevention
and/or
treatment of bone cancers.
The present invention also concerns a method of preventing and/or treating
bone cancer
in a mammal, comprising the step of administering a therapeutically effective
amount
of the aforementioned MTKI 1 to said mammal.
In a further embodiment, the present invention provides the use of MTKI 1 for
the
preparation of a pharmaceutical composition for the prevention and/or
treatment of
bone loss.
The present invention also concerns a method for preventing and/or treating of
bone
loss associated with cancer metastases in a mammal, comprising the step of
administering a therapeutically effective amount of MTKI 1 to said mammal.
Accordingly, in a further aspect, the most preferred compounds for use in
accordance
with the present invention are those selected from the group consisting of
compounds
having the following structure:
HN Br HNaBr
and
O N O N
O N O N .2 HBr
The compounds according to the invention can be prepared and formulated into
pharmaceutical compositions by methods known in the art and in particular
according
to the methods described in the published patent specification WO-2004/105765
mentioned herein and incorporated by reference.
CA 02664165 2009-03-20
WO 2008/049904 PCT/EP2007/061501
-10-
A suitable preparation of the preferred compound used in this invention, taken
from
WO-2004/105765, follows:
Example 1
a) Preparation of 1-pentanol, 5-[[(4-bromo-2-nitrophenyl)methyl]amino]-
(intermediate
1)
A solution of 4-bromo-2-nitro- benzaldehyde,(0.013 mol), 5-amino-l-pentanol
(0.013 mol) and titanium, tetrakis (2-propanolato) (0.014 mol) in EtOH (15 ml)
was
stirred at RT for 1 hour, then the reaction mixture was heated to 50 C and
stirred for
30 min. The mixture was cooled to RT and NaBH4 (0.013 mol) was added
portionwise.
The reaction mixture was stirred overnight and then poured out into ice water
(50 ml).
The resulting mixture was stirred for 20 min., the formed precipitate was
filtered off
(giving Filtrate (I)), washed with H20 and stirred in DCM (to dissolve the
product and
to remove it from the Ti-salt). The mixture was filtered and then the filtrate
was dried
(MgSO4) and filtered, finally the solvent was evaporated. Filtrate (I) was
evaporated
until EtOH was removed and the aqueous concentrate was extracted 2 times with
DCM. The organic layer was separated, dried (MgSO4), filtered off and the
solvent was
evaporated, yielding 3.8 g (93 %) of intermediate 1.
Example 2
a) Preparation of 1-pentanol, 5-[[(4-bromo-2-nitrophenyl)methyl]methylamino]-
(intermediate 2)
A solution of intermediate 50 (0.0047 mol), formaldehyde (0.025 mol) and
titanium, tetrakis (2-propanolato) (0.0051 mol) in EtOH (150 ml) was heated to
50 C
and stirred for 1 hour, then NaBH4 (0.026 mol) was added portionwise at RT.
The
reaction mixture was stirred overnight and then quenched with water (100 ml).
The
resulting mixture was stirred for 1 hour; the formed precipitate was filtered
off and
washed. The organic filtrate was concentrated, then the aqueous concentrate
was
extracted with DCM and dried. The solvent was evaporated and the residue was
filtered
over silica gel (eluent: DCM/CH3OH from 98/2 to 95/5). The product fractions
were
collected and the solvent was evaporated, yielding 0.5 g of intermediate 2.
CA 02664165 2009-03-20
WO 2008/049904 PCT/EP2007/061501
-11-
b) Preparation of 1-pentanol, 5-[[(4-bromo-2-nitrophenyl)methyl]methylamino]-,
acetate (ester) (intermediate 3)
A solution of intermediate 2 (0.0015 mol) and pyridine (0.015 mol) in acetic
anhydride (8 ml) was stirred overnight at RT, then the solvent was evaporated
and co-
evaporated with toluene, yielding intermediate 3.
c) Preparation of 1-pentanol, 5-[[(2-amino-4-bromophenyl)methyl]methylamino]-,
acetate (ester) (intermediate 4)
A mixture of intermediate 3 (0.0015 mol) in THF (50 ml) was hydrogenated
with Pt/C 5% (0.5 g) as a catalyst in the presence of thiophene solution (0.5
ml) [H179-
034]. After uptake of H2 (3 equiv.), the catalyst was filtered off and the
filtrate was
evaporated, yielding 0.5 g of intermediate 4.
d) Preparation of 6-quinazolinol, 4-[[2-[[[5-
(acetyloxy)pentyl]methylamino]methyl]-5-
bromophenyl] amino] -7-methoxy-, acetate (ester) (intermediate 5)
A mixture of intermediate 4(0.0015 mol) and 4-chloro-7-methoxy-6-
quinazolinol acetate (ester) (0.0015 mol) in 2-propanol (30 ml) was heated to
80 C and
the reaction mixture was stirred for 1 day. The solvent was evaporated under
reduced
pressure and the residue was used as such in the next reaction step, yielding
0.83 g of
intermediate 5.
e) Preparation of 6-quinazolinol, 4-[[5-bromo-2-[[(5-
hydroxypentyl)methylamino]methyl]phenyl] amino]-7-methoxy- (intermediate 6)
A solution of intermediate 5 (0.0015 mol) in methanol (25 ml) was stirred at
RT
and a solution of K2CO3 (0.003 mol) in H20 (2.5 ml) was added, then the
reaction
mixture was heated to 60 C and stirred for 18 hours. The solvent was
evaporated and
H20 (20 ml) was added, then the mixture was neutralized with acetic acid and
the
formed precipitate was filtered off. The filtrate was concentrated under
reduced
pressure and the concentrate was extracted with DCM, filtered, then dried
(MgSO4) and
the mixture was concentrated under reduced pressure, yielding 0.5 g (70 %) of
intermediate 6.
CA 02664165 2009-03-20
WO 2008/049904 PCT/EP2007/061501
-12-
Example 3 a)Preparation of 4,6-ethanediylidenepyrimido[4,5-b]
[6,1,12]benzoxadiazacyclo-
pentadecine, 17-bromo-8,9,10,11,12,13,14,19-octahydro-20-methoxy-13-methyl-
(compound MTKII)
A solution of intermediate 6 (0.0011 mol) in THF (50 ml) was stirred at RT and
tributylphosphine (0.0016 mol) was added, then 1,1'-(azodicarbonyl)bis-
piperidine
(0.0016 mol) was added and the reaction mixture was stirred for 2 hours. The
solvent
was evaporated until 1/3 of the initial volume. The resulting precipitate was
filtered off
and washed. The filtrate was evaporated and the residue was purified by RP
high-
performance liquid chromatography. The product fractions were collected and
the
organic solvent was evaporated. The aqueous concentrate was extracted 2 times
with
DCM and the organic layer was dried (MgSO4), then filtered off. The solvent
was
evaporated and the residue was dried (vac.) at 50 C, yielding 0.004 g (0.8 %)
of
compound MTKIl .
To prepare the aforementioned pharmaceutical compositions, a therapeutically
effective
amount of the particular compound, optionally in addition salt form, as the
active
ingredient is combined in intimate admixture with a pharmaceutically
acceptable
carrier, which may take a wide variety of forms depending on the form of
preparation
desired for administration. These pharmaceutical compositions are desirably in
unitary
dosage form suitable, preferably, for systemic administration such as oral,
percutaneous, or parenteral administration; or topical administration such as
via
inhalation, a nose spray, eye drops or via a cream, gel, shampoo or the like.
For
example, in preparing the compositions in oral dosage form, any of the usual
pharmaceutical media may be employed, such as, for example, water, glycols,
oils,
alcohols and the like in the case of oral liquid preparations such as
suspensions
(including nanosuspensions), syrups, elixirs and solutions; or solid carriers
such as
starches, sugars, kaolin, lubricants, binders, disintegrating agents and the
like in the
case of powders, pills, capsules and tablets. Because of their ease in
administration,
tablets and capsules represent the most advantageous oral dosage unit form, in
which
case solid pharmaceutical carriers are obviously employed. For parenteral
compositions, the carrier will usually comprise sterile water, at least in
large part,
though other ingredients, for example, to aid solubility, may be included.
Injectable
CA 02664165 2009-03-20
WO 2008/049904 PCT/EP2007/061501
-13-
solutions, for example, may be prepared in which the carrier comprises saline
solution,
glucose solution or a mixture of saline and glucose solution. Injectable
solutions
containing compounds of formula (I) may be formulated in an oil for prolonged
action.
Appropriate oils for this purpose are, for example, peanut oil, sesame oil,
cottonseed
oil, corn oil, soy bean oil, synthetic glycerol esters of long chain fatty
acids and
mixtures of these and other oils. Injectable suspensions may also be prepared
in which
case appropriate liquid carriers, suspending agents and the like may be
employed. In
the compositions suitable for percutaneous administration, the carrier
optionally
comprises a penetration enhancing agent and/or a suitable wettable agent,
optionally
combined with suitable additives of any nature in minor proportions, which
additives
do not cause any significant deleterious effects on the skin. Said additives
may
facilitate the administration to the skin and/or may be helpful for preparing
the desired
compositions. These compositions may be administered in various ways, e.g., as
a
transdermal patch, as a spot-on or as an ointment. As appropriate compositions
for
topical application there may be cited all compositions usually employed for
topically
administering drugs e.g. creams, gels, dressings, shampoos, tinctures, pastes,
ointments,
salves, powders and the like. Application of said compositions may be by
aerosol, e.g.
with a propellent such as nitrogen, carbon dioxide, a freon, or without a
propellent such
as a pump spray, drops, lotions, or a semisolid such as a thickened
composition which
can be applied by a swab. In particular, semisolid compositions such as
salves, creams,
gels, ointments and the like will conveniently be used.
It is especially advantageous to formulate the aforementioned pharmaceutical
compositions in dosage unit form for ease of administration and uniformity of
dosage.
Dosage unit form as used in the specification and claims herein refers to
physically
discrete units suitable as unitary dosages, each unit containing a
predetermined quantity
of active ingredient calculated to produce the desired therapeutic effect in
association
with the required pharmaceutical carrier. Examples of such dosage unit forms
are
tablets (including scored or coated tablets), capsules, pills, powder packets,
wafers,
injectable solutions or suspensions, teaspoonfuls, tablespoonfuls and the
like, and
segregated multiples thereof.
CA 02664165 2009-03-20
WO 2008/049904 PCT/EP2007/061501
-14-
Preferably, a therapeutically effective amount of the pharmaceutical
composition
comprising a compound according to the invention, is administered orally or
parenterally. Said therapeutically effective amount is the amount that
effectively
prevents metastasis and/or growth or reduces the size of a variety of
neoplastic disorders
or cell proliferative disorders (supra) in patients. On the basis of the
current data, it
appears that a pharmaceutical composition comprising a compound of the present
invention, and in particular
4,6-ethanediylidenepyrimido[4,5-b][6,1,12]benzoxadiazacyclo-pentadecine, 1 7-
bromo-
8,9,10,11,12,13 ,14,19-octahydro-20-methoxy-13 -methyl (MTIKII, or Compound 1)
as
the active ingredient can be administered orally in an amount of from 10 mg to
several
(1 to 5) grams daily, either as a single dose or subdivided into more than one
dose,
including, e.g., two, three or even four times daily. A preferred amount
ranges from 500
to 4,000 mg daily. A particularly, preferred dosage for such a compound is in
the range
of 750 mg to 3,000 mg daily. It will be appreciated that the amount of a
compound
according to the present invention, also referred to here as the active
ingredient, which is
required to achieve a therapeutic effect will, of course, vary with, the route
of
administration, the age and condition of the recipient, and the particular
disorder or
disease being treated. The optimum dosage amounts and regimen can be readily
determined by those skilled in the art using conventional methods and in view
of the
information set out herein. This treatment can be given either continuously or
intermittently, including, e.g., but not limited to, cycles of 3-4 weeks with
treatment
given for 1-21 days per cycle or other schedules shown to be efficacious and
safe.
One illustrative formulation is as follows:
Example 4: Formulation:
The product MTKI1 can be prepared as a 10-mg/mL oral solution, pH 2. It
contains an excipient, Captisol (chemical name: sulfobutyl ether-(3-
cyclodextrin,
SBE-(3-CD), citric acid, Tween 20, HCI, and NaOH in purified water. The
formulation can be stored refrigerated (2-8 C; 36-46 F) and allowed to warm
to room
temperature for maximally 1 hour prior to dose preparation.
The product MTKI1 can also be prepared as 50-mg, 100-mg and 300-mg oral
immediate release capsules, containing the active chemical entity MTKI1,
lactose
CA 02664165 2009-03-20
WO 2008/049904 PCT/EP2007/061501
-15-
monohydrate (200 mesh), sodium lauryl sulphate and magnesium stearate in hard
gelatin capsules, sizes 3, 4 and 00, respectively. The capsules may also
contain any or
all of the following ingredients: gelatin, red iron oxide and titanium oxide.
The above MTKI 1 may be used in combination with one or more other cancer
treatments. Such combinations could encompass any established antitumor
therapy, such
as, but not limited to, chemotherapies, irradiation, and target based
therapies such as
antibodies and small molecules (such as for example bisphosphonates, taxanes,
anthracyclines, capecitabine, Herceptin, docetaxel, satraplatin, cetuximab,
avastin,
aromatase inhibitors and methothrexate). These therapies may be combined in
systemic
therapy, or local instillation/administration (e.g. intrathecally), depending
on optimum
efficacy/safety requirements.
The MTKI 1 and the further anti-cancer agent may be administered
simultaneously
(e.g. in separate or unitary compositions) or sequentially in either order. In
the latter
case, the two compounds will be administered within a period and in an amount
and
manner that is sufficient to ensure that an advantageous or synergistic effect
is
achieved. It will be appreciated that the preferred method and order of
administration
and the respective dosage amounts and regimens for each component of the
combination will depend on the particular MTKI and further anti-cancer agents
being
administered, their route of administration, the particular tumor being
treated and the
particular host being treated. The optimum method and order of administration
and the
dosage amounts and regimen can be readily determined by those skilled in the
art using
conventional methods and in view of the information set out herein.
EXPERIMENTAL DATA
The unique physico-chemical properties of MTKI 1, also referred to herein as
Compound 1, has resulted in an extremely favourable tissue distribution
profile
including the ability to cross the intact blood brain barrier whilst still
retaining good
cellular activity and oral bioavailability. Here, we further demonstrate that
this
preferential tissue distribution to the bone marrow compartment results in
significant
anti tumoral activity using experimental models of bone metastases.
CA 02664165 2009-03-20
WO 2008/049904 PCT/EP2007/061501
-16-
NCTC2472 fibrosarcoma (ATCC Rockvile, MD USA) or MDA-MB231 breast cancer
cells (Dr. Yoneda, Univ. of Michigan, USA) (bone homing variant) were injected
into
the tibia of nude mice, the hole sealed and tumor growth observed at
predefined times
(Verrneirsch, H et al Pharmacol Biochem Behav. 2004 Oct; 79(2):243-51).
Spontaneous `paw lifting' was used as a pain response indicator whilst Ct and
histology was use to demonstrate osteolytic anti tumor activity and tumor
growth.
Methods
Animal model
Male C3H/HeNCr1 mice for the NCTC2472 mouse fibrosarcoma cells (20-25 g,
Charles River, Sulzfeld, Germany) or female NMRI Nude mice for the MDA MB 231
human breast cancer cells (Janvier, France) were used. Induction of bone
cancer was
carried out as previously described (Schwei et al., 1999 1). Induction of
general
anaesthesia was performed under 4%o isoflurane in a mixture of 30% 02 and 70%
air
(1000 ml/ min). Anaesthesia was then maintained at 2.5 % isoflurane for the
duration
of the surgical procedure. The left hind paw was shaved and disinfected with
povidone-
iodine followed by 70% ethanol. A superficial 1 cm incision was made over the
knee
overlaying the patella. The patella ligament was cut, exposing the condyles of
the distal
femur. A 23-gauge needle was inserted at the level of the intercondylar notch
and the
intramedullary canal of the femur to create a cavity for injection of the
cells. Tumour
cells (2.5x 106 cells/20 l) were then injected into the bone cavity using a
0.3 ml
syringe. To prevent leakage of cells outside the bone, the injection site was
sealed with
dental acrylic (Paladur, Heraeus Kulzer, GmbH, Wehrheim, Germany) and the
wound
closed with skin stitches. For the sham-operated group, an identical procedure
was
followed except that medium without cells was injected.
Drug treatment:
Treatment was initiated on day 1 following tumor cell induction. Mice were
treated
once daily (Q1D) with either vehicle (20% Hydroxypropyl-(3-cyclodextrine, pH
4.0) or
CA 02664165 2009-03-20
WO 2008/049904 PCT/EP2007/061501
-17-
vehicle formulated to give a dose of 200 or 50 mg/kg of Compound 1
respectively by
gavage (p.o.) administered in a volume of 10 ml/kg body weight. Mice were
treated up
to 18 days after bone tumor induction.
Pain Assesment:
Pain behaviours (see below) were evaluated in the group of sham and bone
tumour
mice and were behaviourally tested during a 2-week period prior to and 7, 9,
12 and 14
days after tumour inoculation. At the end of the experiment the femur of the
left hind
limb was sampled and used for CT scanning as described in Vermeirsh et al.,
(2004) 2.
Spontaneous lifting behavior: Animals were habituated to the laboratory room
at least
30 minutes in a transparent acrylic cylinder of 20 cm diameter and thereafter
observed
during 4 minutes for spontaneous lifting behaviour of the left hind paw.
Evaluation of bone destruction:
Bone analysis was carried out on ipsilateral left hind limbs prior to and 7,
12, 15 and 18
days following cell injection. Limbs were fixed in 10% phosphate-buffered
formalin
and transfered to a plastic cuvette filled with 70% ethanol for scanning using
the
SkyScan microtomograph (Skyscan 1067 , Skyscan, Aartselaar, Belgium). For
medium resolution measurement, the X-ray beam was collimated to a diameter of
18
mm, line spacing and point resolution were set at 0.254 and 0.127 mm,
respectively.
After standardized reconstruction, the datasets for each bone were re-sampled
using
computer software (Ant, 3D-creator vs. 2.2e, Skyscan, Aartselaar, Belgium) so
that the
medial axis of the bone was centrally oriented for each bone. Scans were
processed and
a two- and three-dimensional morphometric analysis was performed on a 5 mm
femur
bone segment at proximal end of the patellar trochlea using free software
(CTanalyzer
vs. 1.02, Skyscan, Aartselaar, Belgium).
1. Schwei MJ, Honore P, Rogers SD, Salak-Johnson JL, Finke MP,
Ramnaraine ML, et al. Neurochemical and cellular reorganization of the
spinal cord in a murine model of bone cancer pain. J Neurosci
1999;19:10886 -97.
CA 02664165 2009-03-20
WO 2008/049904 PCT/EP2007/061501
-18-
2. Vermeirsch, H., Nuydens, R., Salmon, P.L., and Meert, T.F. Pharmacol.
Biochem. Behav. 2004. 79: 243-251.
Results
Pain Assessment
Animals were habituated to the laboratory room at least 30 minutes in a
transparent
acrylic cylinder of 20 cm diameter and thereafter observed during 4 minutes
for
spontaneous lifting behaviour of the left hind paw. The data is presented as
the
percentage of the time the paw was raised over this period of time. The
vehicle group
was dosed by oral gavage, daily with a 20% HPCD solution at pH 4Ø Compound 1
was dosed once daily at its maximum tolerated dose (MTD) of 200mg per kg,
Iressa
was also dosed at its maximum tolerated dose, by oral gavage, of 50mg per kg
daily for
14 days. The vehicle treated group of animals displayed detectable paw lifting
behaviours starting seven days post tumor cell inoculation. The percentage of
the time
the animals paws were raised during the observation period increased at both
day 9, 12
and 14 post inoculation at which time the paw was raised 80% of the time.
Dosing
animals with Compound 1 at its MTD was found to reduce the time the animals
did not
use their left paws quite dramatically so that at day 14, spontaneous paw
lifting
behaviour was only detected to occur -8% of the time. The reference compound
used
in this study, Iressa when dosed at its MTD also led to a statistically
significant
reduction in spontaneous paw lifting behaviour, however the effect was far
less
extensive with spontaneous paw lifting being observed more than 35% of the
time.
Evaluation of bone destruction
Representative reconstructions from Ct s of the ipsilateral left hindlimbs
showing
osteolytic bone destruction in the tumor inoculated animals (Figure 2).
Considerable
bone loss was observed in the vehicle and Iressa treated groups whilst
significantly less
bone destruction can be seen in the Compound 1 treated animals. The sham
operated
animals showed no signs of osteolytic activity.
CA 02664165 2009-03-20
WO 2008/049904 PCT/EP2007/061501
-19-
Animal model
In the bone cancer metastasis model MDA B231 bone homing clone cells were
inoculated into the tibia as described. After 42 days, animals were sacrificed
and the
amputated paws placed in fixative. The legs were de-calcified and sections cut
to
determine levels of bone destruction. The vehicle treated animals were
observed to
have large tumor mass (encircled area in Figure 3) that has expanded out of
the initial
site of inoculation (black arrow in Figure 3) and in the process resulted in
significant
destruction of the bone (See Figure 2). The amount of tumor growth and the
amount of
bone destruction was seen to be dose dependently reduced with the highest dose
of
Compound 1 tested in this study, l 00mg per kg, po, qd, showing no signs of
tumor cell
not any signs bone destruction. The latter is also apparent from the
histological
sections in Figure 3. In vehicle treated animals a large tumor mass has
extended into
the femur (grey arrow), where in animals treated with l 00mg per kg, po, qd,
of MTKI
1, detectable tumor is hardly present (grey arrow) and close to the site of
inoculation
(black arrow).
While the foregoing specification teaches the principles of the present
invention, with
examples provided for the purpose of illustration, it will be understood that
the practice
of the invention encompasses all of the usual variations, adaptations and/or
modifications as come within the scope of the following claims and their
equivalents.
