Note : Les descriptions sont présentées dans la langue officielle dans laquelle elles ont été soumises.
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TREATMENT OF CANCER WITH 2 DEOXYGALACTOSE
CROSS-REFERENCE TO RELATED APPLICATIONS
The present application claims the benefit of priority under 35 U.S.C.
119(e) to U.S.
Provisional Patent Application No. 60/775,630, entitled Treatment of Cancer
with 2-
Deoxygalactose, filed February 21, 2006, the disclosure of which is
incorporated herein by
reference in its entirety.
BACKGROUND OF THE 1NVENTION
[0001] The term'"cancer" generally refers to one of a group ofmore than 100
diseases caused by
the uncontrolled growth and spread of abnormal cells that can take the form of
solid tumors,
lymphomas, and non-solid cancers such as leukemia. Normal cells reproduce
until maturation is
attained and then only:as necessary for replacement. Benign tumors, or benign
hyperplasia,
involve an overgrowth of cells without spread to other organs. Cancer cells,
conversely, grow
and divide endlessly, crowding out neafby cells and eventually spreading to
other parts of the
body. Cancer cells that develop at one site can grow rapidly into a malignant
tumor, invading and
destroying nearby tissues. Malignant cancer tumor cells eventually
metastasize, or spread to
other parts of the body via the bloodstream or lymphatic system, where the
cells begin
multiplying and developing into new tumors. This sort of tumor progression
makes cancer
dangerously fatal. Although there have been great improvements in diagnosis,
general patient
care, surgical techniques, and local and systemic adjuvant therapies, most
deaths from cancer are
-still-due-to metastases-and resistance to conventional therapies.
[0002] The vast majority of drug-mediated conventional cancer therapies rely
on the use of drugs
that act as selective poisons for dividing cells. These drugs are effective,
because cancer cells
generally divide moxe frequently than normal cells. However, for a variety of
reasons, such drugs
almost inevitably do not kill all cells in a tumor. One reason is that not all
cancer cells divide
more frequently than all normal cells. Another is that specific proteins can
confer drug resistance
to a cancer cell, creating multiple drug resistance ('"MDR") phenotype.
Another is the very nature
of the tumor, particularly its vascular architecture. As a tumor grows, it
requires a blood supply
and growth of new vasculature. The new vasculature that supports the tumor
growth is often
disordered, leaving significant portions of the turn.or under-vascularized and-
even the
vascularized portions subject to intermittent vascular blockage. The
vasculature delivers not only
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oxygen but also most anti-cancer drugs to cells. Thus, the hypoxic regions of
tumors are
typically under-supplied with anti-cancer drugs.Oxygen is critical for
supplying energy to a cell
in the form of ATP produced by mitochondrial action, and cell growth and
division are energy
intensive processes. In addition, oxygen is required for the cytotoxic action
of some anti-cancer
drugs and radiation therapies. A cell's only other source of ATP in the
amounts needed to support
the cell is from anaerobic glycolysis. Given the demand for ATP caused by cell
division and the
hypoxic nature of tumors, it is therefore not surprising that many cancers
exhibit, relative to
normal cells, increased glucose transport and glycolysis. This attribute of
cancer cells was
described, for example, in Dickens, 1943, Cancer Research 3:73, which reported
"the typical
intact cancer cell exhibits an unusual ability to utilize glucose by the
process of anaerobic
glycolysis through lactate".
[0003] Today, this unusual ability of cancer cells to utilize glucose is
exploited to image tumors
in the diagnostic technique of PET scanning, which utilizes a radioactively
labeled glucose
analog 18F-2-deoxy-D-glucose ("FDG") that preferentially accumulates in cancer
cells relative
to most normal cells. See Som et al., 1980, "A fluorinated glucose analog,
2fluoro-2-deoxy-D-
glucose (F-18): nontoxic tracer for rapid tumor detection", J. Nucl. Med. 21:
670-675. Scientists
have questioned whether the increased glycoslysis in cancer cells relative to
normal cells would
also allow the use of metabolic poisons of anaerobic glycolysis to target
cancer cells
preferentially. 2-Deoxy-D-glucose ("2-DG"; see Bergmann, 1922, Deutsch. Chem.
Ges. 56:158-
60; Cramer, 1952, Franklin Inst. 253:277-80; Japan patent publication No. 54-
041384) is a
metabolic poison (see McDonald, 1952, Cancer Research 351-353) that inhibits
glycolysis in
cancer cells (see Woodward, 1954, Cancer Res. 14:599605).
[0004] 2-DG is believed to inhibit glycolysis by accumulating in the cell,
into which it is
transported by one or more glucose transporters. Once in the cell, hexokinase
converts 2-DG to
2-DG-6-phosphate ("2DG6P"), which cannot be converted to fructose-6-phosphate,
a substrate
required for glycolysis, and which cannot leave the cell unless the phosphate
group is removed
by glucose-6-phosphatase (abundant in liver cells) or non-specific,
intracellular phosphatases.
When intracellular concentrations of 2DG6P reach a certain threshold amount,
glycolysis is
effectively shut down, and the cell dies from a lack of energy in the form of
ATP. 2-DG may
have other cytotoxic effects as well. It can act as a trap for phosphate and
so reduce intracellular
ATP, first in its conversion to 2DG6P and then its conversion to uridyl
diphosphate-2-DG
("UDP2DG"). The formation of the latter compound UDP2DG acts as a trap for
uridyl, thus
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inhibiting nucleic acid synthesis. In addition, 2DG6P may inhibit the pentose
cycle that generates
ribose and deoxyribose and so could inhibit DNA synthesis even beyond the
inhibition caused by
the phosphate, ATP, and uridyl trapping attendant upon 2DG uptake by a cell.
There is some
evidence that 2-DG can be converted to 2-DG-lphosphate ("2DG1P.' ; see Colwell
et ai., 1996,
Int. J. Biochem. Celt. Biol. 28(1): 115-121) and then be incorporated into
glycogen. In addition,
there is evidence that 2-DG is incorporated into glycolipids and glycoproteins
(via UDP2DG)
and so interfere with their normal processing and function (see Steiner et
at., '1974, Biochem.
Biophys. Res. Comrn.61(2): 795). '
[0005] 2-DG has been shown to retard tumor growth in animal models (see
Sokoloff, 1955,
A.M.A. Arch. Of Path. 729-732, and Ball, 1957, Cancer Res. 17:235-39) and was
administered-to humans as early as the 1950s (see Landau, 1958, J. Natl. Canc.
Inst. 21:485-
494). Cell-based, animal, and human studies of 2-DG conducted since the 1950s
have, with some
exceptions, continued to indicate that 2-DG has an impact on cancer cells,
alone or in
combination with radiation or other chemotherapy. Thus, Laszlo et al., Feb.
1960, J. Natl.
Canc. Inst. 24(2):267-281, reported cancer studies in mice in which "some
evidence of
interference with the normal course of the diseases, either by impairment of
local tumor growth
or by prolongation of host survival, or both" was seen. See also Haberkorn,
Nov. 1992, J. Nucl.
Med. 33(11):1981-87, and Malaisse, Mar. 1998, Cancer Lett. 125:45-49.
Likewise, Purohit,
Mar. 1982, Int J. Radiat. Oncol. Biol. Phys. 8:495-99, reported 2-DG inhibited
cancer cell
growth, with more pronounced inhibition under hypoxic conditions and in
combination with X-
irradiation. See also Dwarakanath, Mar. 1999, Int. J. Radiat. Oncol. Biol.
Phys. 43(5):1125-33;
Dwarkanath, Jul. 2001, Int. J. Radiat. Oncol. Biot. Phys. 50(4):1051-61; and
Yeung, 11
Dec. 2001, PCT WO 02/58741, which reports thatr8F-2-DG could be administered
to treat
cancer, .at doses significantly higher than those used for diagnostic imaging.
Combination studies
of 2-DG with other cytotoxiins and anti-cancer drugs include those described
in Lampidis, 2 Mar.
2001, PCT WO 01/82926, which states that 2DG, oxamate, and various analogs are
selectively
toxic toward anaerobic cells and can be used to increase the efficacy of
standard cancer
chemotherapeutic and.radiation regimens. The reference Pitha, 21 Mar. 200.2,
U.S patent
publication No. 20020035071, 'discloses a set of compounds that purportedly
mimic the effect of
2-DG without the toxicities attributed to that compound (including hunger,
sweating, ataxia, and
convulsions). [0006] 2-Deoxy-D-galactose ("2-DGaI") can be considered an
analog of 2-DG, in
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that it has a closely related structure, differing only in the position of the
hydroxyl group at C-
4.2DGa1 can in fact be converted in the body to 2-DG by a process that
involves conversion of 2-
DGaI first to 2-DGal-l-phosphate, then to uridyl diphosphate 2-
DGaI("UDP2DGal"), then to
UDP2DG, then to 2-DG-1-phosphate, then to 2-DG-6-phosphate, and finally to 2-
DG. This
bioconversion closely follows the pathways for the natural sugars galactose
and glucose.
Galactose is metabolized in the glycolytic pathway by conversion first to
galactose-lphosphate,
then to uridine diphosphate galactose ("UDPGaI"), then to UDP-glucose
("UDPGIu"; the
precursor to glycogen), then to glucose-l-phosphate, and then to glucose-
6phosphate, the initial
substrate in the glycolytic pathway. However, galactose and glucose have very
different roles in
the body and are metabolized quite differently in some respects. First,
galactose is metabolized
first by galactokinase to form galacatose-l-phosphate, whereas glucose is
metabolized first by
hexokinase to glucose-6-phosphate. Second UDPGaI is a direct substrate used in
the
glycosylation of proteins; UDPGIu is not. Third, the liver is the major organ
of galactose
metabolism, and galactose is cleared rapidly from the blood so that, if
galactose is ingested, the
concentration entering the liver via the portal vein is much higher than that
leaving via the
hepatic vein. Fourth, the accumulation of galactose-l-phosphate results in the
clinical symptoms
of the disease condition known as galactosemia, which is fatal unless
galactose is avoided and
can have severe long term complications even so.
[0007] In other respects, 2-DG and 2-DGaI are similar. Both compounds are
actively transported
across membranes in insulin responsive tissues (see Landau et al., Jun. 1958,
Am. J. Physiol.
193(3): 461-465). Neither compound is actively transported by the intestine,
although passive
diffusion clearly enables absorption of large amounts of the compounds (see
Wilson and Landau,
1960, Am. J. Physiol. 198: 99-102). Both 2-DG and 2-DGaI have been shown to
inhibit
glycolysis in tumor cell lines and types (see Laszlo et al., Sep. 1958,1 Natl.
Cancer Inst.
21(3): 475-483). In addition, both 2-DG and 2-DGaI were reported to cause
"inhibition of tumor
growth ... and a modest prolongation of survival time . . . " in animal
studies, although the results
reported indicated that 2-DGaI was less efficacious in the model systems
employed (see Laszlo
et al., Feb. 1960, J Natl. Canc. Inst. 24(2):267-28 1, supra). Interestingly
as well, the 2-
fluoro-derivative of 2-DGaI has been reported to have properties that might
make it a useful
cancer imaging andlor therapeutic agent (see Ishiwata et al., 1989, Nuel. Med.
Biol. 16(3):
247-254; Grun et al., 1990, AdU. Enzyme Regul. 30: 231-242; Grun et al., 31
May 1990,
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Eur. J. Biochem.. 190(1): 11-19; Fukuda et al., 1986, Eur. J. Nucl. Med. I
I(111: 444-
448; and Paul et al., 1989, Int. J. Rad. Appl. Instrum. B. 16(.S): 449-453).
Also, 2-DGaI
has been shown to trap phosphorous and'uridylate and to block glycolysis, all
leading to a
depletion of ATP, in cancer cells (see Smith and Keppler, 1977, Eur. J.
Biochem. 73: 8392,
and Keppler et al., 1985, Adv. En.z. Reg. 23: 61-79).
[00081 However, after'more than five decades of study, the benefits of 2-DG or
analogs thereof
such as 2-DGaI in cancer therapy are still uncertain, and neither 2-DG, 2-
DGaI, nor any analog
compound has been approved for the treatment of cancer in the United States or
Europe. Given
the many reported studies that suggest 2-DG and its analogs may have a role to
play in cancer
therapy, perhaps in combination with other treatment regimens, there remains a
need for methods
of treating cancer with 2-DG or its analogs. The present invention meets that
and other needs.
SUMMARY OF TIiF INVENTION
[0009] In a first aspect, the present invention provides a method of treating
cancer, which
method comprises administering to a mammal a therapeutically effective dose of
2-DGal or a 2-
DGaI analog. In one embodiment, the therapeutically effective dose is a dose
in the range of
about 1 mg/kg (patient weight) to about 5 g/kg of 2-DGaI or a 2-DGaI analog.
In another
embodiment, the therapeutically effective dose is a dose in the range of about
10 mg/kg to about
I g/kg. In another embodiment, the therapeutically effective dose is about 50
mg/kg to about 500
mg/kg. In one embodiment, 2-DGal or a 2-DGal analog is administered in a
single oral dose of
from about 5 to about 25 grams. In one embodiment, the 2-DGaI or 2-DGaI analog
is
administered orally once (qday), twice (bid), three times (tid), or four times
(qid) a day or once
every other day (qod) or once a week (qweek), and treatment is continued fo'r
a period ranging
.from three days to two weeks or longer. In one embodiment, the treatment is
continued for one to
three months. In another embodiment, the treatment is continued for a year.
[0010] In a second aspect, the present invention provides a pharmaceutically
acceptable
formulation of 2-DGal and 2-DGal analog useful in the methods of the present
invention. In one
embodiment, the formulation is crystalline in nature, and the 2-DGaI or 2-DGal
analog is
packaged in a sachet that is decanted into and dissolved in a liquid for 'oral
administration to the
patient. In other formulations, the 2-DGaI or 2-DGaI analog is not crystalline
but can be
amorphous in nature. In another embodiment, the 2-DGal or 2-DGaI analog is
formulated as a
tablet or pill containing 2-DGal or a 2-DGal analog in an amount in the range
of about 250 mg to
about 2 g.
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[0011] In a third aspeat, the present invention provides a method of treating
or preventing the
reoccurrence of liver or brain cancer, which method comprises administering to
a mammal a
therapeutically effective dose of 2-DGaI or a 2-DGaI analog.
[0012] In a fourth aspect, the present invention provides a method of treating
or preventing
cancer, which method comprises administering to a human or other mammal a
therapeutically
effective dose of 2-DGa1 or a 2-DGaI analog in combination with another anti-
cancer agent
and/or radiation therapy. In one embodiment, the cancer is a liver cancer, a
brain cancer, or a
multi-drug resistant cancer or a cancer that is otherwise refractory to
treatment. Iri one
embodiment, the cancer is a brain cancer, and the 2-DGaI is administered
concurrently with
radiation therapy. In another embodiment, the 2-DGaI or 2-DGaI analog is co-
administered with
one or more of the following agents: 2-deoxy-D-glucose, 3-bromopyruvate, and a
cytotoxic
agent.
[0013] These and other aspects and embodiments of the invention are described
in more detail in
the detailed description, examples, and claims that follow.
DETAILED DESCRIPTION OF THE INVENTION
[0014] The present invention provides methods of treating cancer by
administering a
therapeutically effective dose of 2-DGaI or a 2-DGal analog, alone or in
combination with other
anti-cancer therapies, including surgical resection, radiation therapy, and
drug therapy.'To aid in
the appreciation of the invention, this description is divided into the
following topics: (i)
therapeutically effective administration of 2-DGaI or a 2-DGaI analog; (ii) co-
administration
with other anti-cancer agents; (iii) treating particular cancers; and (iv)
formulation and packaging
of a 2-DGaI and 2-DGal analogs.
Therapeutically effective administration of 2-DGaI and 2-DGaI analogs
[0015] The present invention provides methods and compositions for the
treatment of cancer
with 2-DGaI and 2-DGal analogs. In accordance with the methods of the present
invention, a 2-
DGaI analog is any D-galactose analog other than 2-DGal that does not have a
hydroxyl group at
the 2 position of the galactose ring and that is capable of being
phosphorylated in vivo in
mammalian cells when present in the cell in monomer form. Lgalactose and its L-
analogs are not
2-DGaI analogs for purposes of the present invention. A galactose analog
includes D-lyxo-
hexos-2-ulose, D-arabino-hexos-2-ulose, and 5-thio-galactose. An analog of
galactose or 2-DGaI
can have a fluorine in place of a hydrogen at any position on the galactose
ring; thus, 2-fluoro-2-
deoxv-D-galactose (2-FDGal) and 2-difluoro2-deoxy-D-galactose are 2-DGal
analogs. An
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analog of galactose or 2-DGaI can have an amino group in place of a hydroxyl
group at any
position on the glucose ring other than the 6 position; thus, 2-amino-2-deoxy-
D-galactose (2-
galactosamine) is a 2-DGaI analog. Other illustrative 2-DGaI analogs include
di, tri, and other
oligosaccharides that contain 2-DGaI and/or a 2-DGaI analog.
[00 16] In accordance with the methods of the present invention, 2-DGaI or a 2-
Dgal analog must
be administered in a dose in the range of about 1 mg to about 1 g of 2-DGaI or
a 2-DGaI analog
per kg of body weight of the patient to be treated, and more than one dose
must be administered.
While the patient will most often be a human patient, those of skill in the
art will appreciate that
the methods and compositions of the invention can be used to treat cancer in
any mammal. In
one embodiment, the 2-DGaI or 2-DGaI analog is administered in a dose in the
range of about 10
mg to about 750 mg of 2-DGaI or 2-DGal analog per kg of body weight of the
patient to be
treated. In another embodiment, the 2-DGaI or 2-DGaI analog is administered in
a dose in the
range of about 100 mg to about 500 mg of 2-DGaI or 2-DGaI analog per kg of
body weight of
the patient to be treated. In certain other embodiments, the 2-DGaI or 2-DGaI
analog is
administered in a dose of about 50 to 250 mg of 2-DGaI or 2-DGaI analog per kg
of body weight
of the patient to be treated. In one embodiment, from about 10 to about 25 g
of 2-DGaI or a 2-
DGaI analog are administered orally per day (in one or more doses) for the
treatment of cancer.
[0017] The therapeutically effective dose of 2-DGaI or a 2-DGaI analog is
administered daily, or
once every other day, or once a week to the patient, and multiple
administrations of the drug are
employed. Depending on the dose selected by the practitioner and the
convenience of the patient,
the entire dose may be administered once daily, or the dose may be
administered in multiple
smaller doses through the course of a day. For example, the dose may be
divided into two
smaller doses and administered twice daily, or divided into three smaller
doses and administered
thrice daily. Alternatively, the dose may be combined and given every other
day, or even less
frequently, but in any event, the dose is repeatedly administered over a
period of time.
[0018] For optimum treatment benefit, the administration of the
therapeutically effective dose is
continued for multiple days, typically for at least five consecutive days, and
often for at least a
week and often for several weeks or more. Thus, a patient may be administered
2-DGaI or a 2-
DGaI analog in accordance with the present methods for a week, a month, two
months, three
months, six months, or a year or longer. For preventive applications,
treatment may continue
indefinitely throughout the life of the patient. As is well understood in the
art for other cancer
therapeutic drugs, treatment with 2-DGaI or a 2-DGaI analog may be suspended
temporarily if
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toxicity is observed or for the convenience of the patient without departing
from the scope of the
invention. In re-treatment regimens, the dose is adjusted to reflect patient
tolerance of the prior
treatment.
[0019] In one preferred embodiment of the invention, the 2-DGal or a 2-DGaI
analog is
administered orally, and multiple doses are administered over an extended
period of time. Using
this therapeutically effective dosing and administration regimen,
practitioners of skill in the art
can significantly improve treatment outcomes with all currently used cancer
therapies, including
surgical resection, radiation therapy, and drug therapies. In one important
aspect, the present
invention provides new methods for treating cancer using existing anti-cancer
drugs as well as
other compounds, as discussed in the following section.
Co-administration with other anti-cancer agents
[00201 In accordance with the methods of the invention, 2-DGa1 or a 2-DGaI
analog can be co-
administered in combination with other anti-cancer and antineoplastic agents.
[0021] In some embodiments, 2-DGaI or a 2-DGaI analog is administered prior to
the initiation
of any other cancer therapy, and treatment is continued throughout the course
of the other
therapy. In other embodiments, 2-DGa1 or a 2-DGaI analog is administered after
the initiation or
completion of the other cancer therapy. In other embodiments, 2-DGaI or a
2DGaI analog is first
administered at the initiation of the other cancer therapy.
[0022] In one important embodiment, 2-DGaI or a 2-DGaI analog is administered
with another
anti-cancer agent that is more effective when ATP levels in the cancer cell
are low. 2DGaI and 2-
DGaI analogs act in part by reducing the ATP available to the cancer cell.
Thus, in one aspect of
the invention, 2-DGaI or a 2-DGaI analog is administered once in an amount
effective for
reducing ATP levels in the tumor and administered again only after ATP levels
begin to rise
again; thereafter, 2-DGaI or a 2-DGaI analog is administered to maintain ATP
at a low level in
the tumor. As but one example, the DNA damage induced by radiation therapy and
by certain
drug therapies requires ATP to repair. Consequently, administration of 2-DGaI
or a 2-DGaI
analog in accordance with the methods of the present invention can improve
patient outcomes
when conducted concurrently with such therapies. In one embodiment, the
present invention
provides a method of treating brain cancer, which method comprises
administering a
therapeutically effective dose of 2-DGaI or a 2-DGaI analog as described
herein in combination
with a radiation therapy regimen that has been used for the treatment of brain
cancer.
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[0023] In addition to the c'ombination of 2-DGal or a 2-DGaI analog with
agents that are more
effective when ATP levels are reduced, the present invention provides a
variety of synergistic
combinations of 2-DGal or a 2-DGaI analog and other anticancer drugs. Those of
skill in the art
can readily determine in accordance~ with the invention what anti-cancer drugs
can be used
"synergistically" with 2-DGaI or a 2-DGaI analog. For example, the reference
Vendetti,
"Relevance of Transplantable Animal-Tumor Systems to the Selection of New
Agents for
Clinical Trial," Pharmacological Basis of Cancer Chemotherapy, Williams and
Wilkins,
Baltimore, 1975, and Simpson Herren et al., 1985, "Evaluation of In Vivo Tumor
Models for
Predicting Clinical Activity for Anticancer Drugs," Proc. Am. Assoc. Cancer
Res. 26: 330, each
of which is incorporated herein by reference, describe methods to aid in the
determination of
whether two drugs act synergistically. While synergy is not required for
therapeutic benefit in
accordance with the methods of the invention, synergy is preferred, and two
drugs can be said to
possess therapeutic synergy if a combination dose regimen of the two drugs
produces a
significantly better tumor cell kill than either of the single agents at
optimal or maximum
tolerated doses. The "degree of synergy" can be defined as net logs of tumor
cell kill by the
optimum combination regimen minus net logs of'tumor cell kill by the optimal
dose of the most
active single agent. Differences in cell kill of greater than ten-fold (one
log) are considered
conclusively indicative of therapeutic synergy. [0024] Even where 2-DGaI or a
2-DGaI analog
acts synergistically or merely additively with another anti-cancer agent, 2-
DGaI or a 2-DGaI
analog will, at least in, some embodiments, be administered prior to the
initiation of therapy with
the other drug or drugs and will, in any event, typically continue throughout
the course of
treatment with the other drug or drugs. In a related aspect, the present
invention also provides
new methods of using known anti-cancer therapies, particularly drug-based anti-
cancer therapies,
in which 2-Dgal or a 2-DGaI analog is administered concurrently with a therapy
in which the
active agent is delivered at a lower dose, and optionally-for longer periods,
than is currently
practiced. Such "low dose" therapies can involve, for example, administering
an anti-cancer
drug, including but not limited to paclitaxel, doxorubicin, cisplatin, or
carboplatin, at a lower
than approved dose and for a longer period of time together with 2-DGal or a 2-
DGaI analog
administered in accordance with the methods of the present invention. These
methods can be
used to improve patient outcomes over currently practiced therapies by more
effectively killing
cancer cells or stopping cancer cell growth as well as diminishing unwanted
side effects of the
other therapy.
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[0025] When employed in combination with one of these agents, the dosages of
the additional
agent are either the standard dosages employed for those agents or are
adjusted downward from
levels employed when that agent is used alone. Thus, the administration of a 2-
DGa1 or a 2-
DGaI analog in accordance with the methods of the invention can allow the
physician to treat
cancer with existing drugs, but at a lower concentration or dose than is
currently used, thus
ameliorating the toxic side effects of such drugs. The determination of the
exact dosages for a
given patient varies, dependent upon a number of factors including the drug
combination
employed, the particular disease being treated, and the condition and prior
history of the patient,
but is within the skill of the ordinarily skilled artisan in view of the
teachings herein.
[0026] Specific dose regimens for known and approved anti-neoplastic agents
are given, for
example, in the, product descriptions found in the current edition of the
Physician's Desk
Reference, Medical Economics Company, Inc., Oradell, N.J. Illustrative dosage
regimens for
certain anti-cancer drugs are also provided below. Those of skill in the art
will recognize that
many of the known anti-cancer drugs discussed herein are routinely used in
combination with
other drugs. In accordance with the methods of the present invention, 2-DGaI
or a 2-DGaI analog
can be co-administered in such multiple drug treatment regimens. [0027] Cancer
drugs can be
classified generally as alkylators, anthracyclines, antibiotics, aromatase
inhibitors,
biphosphonates, cyclo-oxygenase inhibitors, estrogen receptor modulators,
folate antagonists,
inorganic arsenates, microtubule inhibitors, metabolic inhibitors, modifiers,
nitrosoureas,
nucleoside analogs, osteoclast inhibitors, platinum containing compounds,
retinoids,
topoisomerase 1 inhibitors, topoisomerase 2 inhibitors, and tyrosine kinase
inhibitors. -In
accordance with the methods of the present invention, 2-DGaI or a 2-DGaI
analog can be co-
administered with any anti-cancer drug from any of these classes or can be
administered prior to
or after treatment with any such drug or combination of such drugs. In one
embodiment,
however, the anti-cancer drug co-administered with 2DGa1 or a 2-DGaI analog is
not a
topoisomerase inhibitor.
[0028] Alkylators useful in the practice of the present invention include,but
are not limited to
busulfan (Myleran, Busulfex), chlorambucil (Leukeran), cyclophosphamide
(Cytoxan, Neosar),
melphalan, L-PAM (Alkeran), dacarbazine (DTIC-Dome), and temozolamide
(Temodar). In
accordance with the methods of the present invention 2-DGaI or a 2-DGaI analog
is co-
administered with an alkylator to treat cancer. In one embodiment, the cancer
is chronic
rnveln¾enous leukemia, multiple myeloma, or anaplastic astrocytoma. As one
example, the
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compound 2-bis[(2-Chloroethyl) amino]tetrahydro-2H-1,3,2-oxazaphosphorine, 2-
oxide, also
commonly known as cyclophosphamide, is an alkylator used in the treatment of
Stages III and
IV malignant lymphomas, multiple myeloma, leukemia, mycosis fungoides,
neuroblastoma,
ovarian adenocarcinoma, retinoblastoma, and carcinoma of the breast.
Cyclophosphamideis
administered for induction therapy in doses of 1500-1800 mg/rn2 that are
administered
intravenously in divided doses over a period of three to five days; for
maintenance therapy, 350-
550 mg/m2 are administered every 7-10 days, or 110-185 mg/m2 are administered
intravenously
twice weekly. In accordance with_the methods of the invention, 2-DGaI or a 2-
DGaI analog is
co-administered with cyclosphosphaniide administered at such doses.
[0029] Anthracyclines useful in the practice of the present invention include
but are not limited
to doxorubicin (Adriamycin, Doxil, Rubex), mitoxantrone (Novantrone),
idarubicin (Idamycin),
vdlrubicin (Valstar), and epirubicin (Ellence). In accordance with the methods
of the present
invention 2-DGaI or a 2-DGal analog is co-administered with an anthracycline
to treat cancer. In
one embodiment, the cancer is acute nonlymphocytic leukemia, Kaposi's sarcoma,
prostate
cancer, bladder cancer, metastatic carcinoma of the ovary, and breast cancer.
As one example the
compound (8S, l OS)-10-[(3-Amino-2,3,6-trideoxy-.alpha.-L-
lyxohexopyranosyl)oxy]-8-
glycoloyl-7,8,9,10-tetrahydro-6,8,11-trihydroxy-l-methoxy-
5,12naphthacenedione, more
commonly known as doxorubicin, is a cytotoxic anthracycline antibiotic
isolated from cultures of
Streptomyces peucetius var. caesius. Doxorubicin has been used successfully to
produce
regression in disseminated neoplastic conditions such as acute lymphoblastic
leukemia, acute
rnyeloblastic leukemia, Wilm's tumor, neuroblastoma, soft tissue and bone
sarcomas, breast
carcinoma, ovarian carcinoma, transitional cell bladder carcinoma, thyroid
carcinoma,
lymphomas of both Hodgkin and non-Hodgkin types, bronchogenic carcinoma, and
gastric
carcinoma. Doxorubicin is typically administered in a dose in the range of 30-
75 mg/m2 as a
single intravenous injection administered at 21-day intervals; weekly
intravenous injection at
doses of 20 mg/m2; or 30 mg/m2 doses on each of three successive days repeated
every four
weeks. In accordance with the methods of the invention, 2-DGaI or a 2-DGa1
analog is co-.
administered starting prior to and continuing after the administration of
doxorubicin at such
doses.
[0030] Antibiotics useful in the practice of the present invention include but
are not limited to
dactinomycin, actinomycin D (Cosmegen), bleomycin (Blenoxane), and
daunorubicin,
daiinnmycin (Cerubidine, DanuoXorne). In accordance with the methods of the
present -invention
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2-DGaI or a 2-DGal analog is co-administered with an antibiotic to treat
cancer. In one
embodiment, the cancer is acute lymphocytic leukemia, other leukemias, and
Kaposi's sarcoma.
[0031] Aromatase inhibitors useful in the practice of the present invention
include but are not
limited to anastrozole (Arimidex) and letroazole (Femara). In accordance with
the methods of the
present invention 2-DGaI or a 2-DGaI analog is co-administered with an
aromatase inhibitor to
treat cancer. In one embodiment, the cancer is breast cancer.
[00321 Biphosphonate inhibitors useful in the practice of the present
invention include but are
not limited to zoledronate (Zometa). In accordance with the methods of the
present invention 2-
DGal. or a 2-DGal analog is co-administered with a biphosphonate inhibitor to
treat cancer. In
one embodiment, the cancer is multiple myeloma, bone metastases from solid
tumors, or prostate
cancer.
[0033] Cyclo-oxygenase inhibitors useful in the practice of the present
invention include but are
not limited to celecoxib (Celebrex). In accordance with the methods of the
present invention 2-
DGaI or a 2-DGaI analog is co-administered with a cyclo-oxygenase inhibitor to
treat cancer. In
one embodiment, the cancer is colon cancer or a pre-cancerous condition known
as familial
adenomatous polyposis.
[0034] Estrogen receptor modulators useful in the practice of the present
invention include but
are not limited to tamoxifen (Nolvadex) and fulvestrant (Faslodex). In
accordance with the
methods of the present invention 2-DGal or a 2-DGa.I analog is co-administered
with an estrogen
receptor modulator to treat cancer. In one embodiment, the cancer is breast
cancer or the
treatment is administered to prevent the occurrence or reoccurrence of breast
cancer. [0035]
Folate antagonists useful in the practice of the present invention include but
are inot limited to
methotrexate and tremetrexate. In accordance with the methods of the present
invention 2-DGaI
or a 2-DGaI analog is co-administered with a folate antagonist to treat
cancer. In one
embodiment, the cancer is osteosarcoma. Antifolate drugs have been used in
cancer
chemotherapy for over thirty years. As one example, the compound N-[4-
[[(2,4diamino-6-
pteridinyl)methyl metliylamino]benzoyl]-L-glutamic acid, conunonly known as
methotrexate, is
an antifolate drug that has been used in the treatment of gestational
choriocarcinoma and in the
treatment of patients with chorioadenoma destruens and hydatiform mole. It is
also useful in the
treatment of advanced stages of malignant lymphoxna and in the treatm.ent of
advanced cases of
mycosis fungoides. 5-Methyl-6-[[(3,4,5trimethoxyphenyl)-amino]methyl]-2,4-
auinazolinediamine is another antifolate drug and is commonly known as
trimetrexate.
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Methotrexate is administered as follows. For choriocarcinoma, intramuscular
injections of doses
of 15 to 30 mg daily for a five-day course, such courses repeated as needed
with rest period of
one or more weeks interposed between courses of therapy. For leukemias, twice
weekly
intramuscular injections in doses of 30 mg/m2. For mycosis fungoides, weekly
intramuscular
injections of doses of 50 mg or, altematively, of 25 mg twice weekly. In
accordance with the
methods of the invention, 2DGaI or a 2-DGaI analog is co-administered with
methotrexate
administered at such doses.
[0036] Inorganic arsenates useful in the practice of the present invention
include but are not
limited to arsenic trioxide (Trisenox). In accordance with the methods of the
present invention 2-
DGaI or a 2-DGaI analog is co-administered with an inorganic arsenate to treat
cancer. In one
embodiment, the cancer is refractory APL.
[0037] Metabolic inhibitors useful in the practice of the present invention
include any agent that
interferes with glycoly.sis, such as, for example, 2-deoxy-D-glucose; 2-fluoro-
2-deoxy-Dglucose;
2-deoxyfructose; lonidamine; and 3-bromopyruvate. In accordance with the
methods of the
present invention 2-DGaI or a 2-DGal analog is co-administered with a
metabolic inhibitor, alone
or in combination with another cytotoxic or other anti-cancer agent, to treat
cancer. The present
invention also provides novel compounds of the invention and pharmaceutical
compositions
comprising such compounds useful in'these methods. [0038] Thus, in one
embodiment, the
present invention provides disaccharides, trisaccharides, and longer
oligosaccharides comprising
at least a 2-DGal or a 2-DGaI analog and another pentose or hexose sugar,
including but not
.limited to a pentose or hexose..sugar that is itself an inhibitor of
glycolysis or of DNA synthesis
or some other aspect of nucleotide metabolism. In the trisaccharides and
longer oligosaccharides,
the 2-DGaI or 2-DGal analog content can be varied, such as, for example 33% or
66%, as may
be convenient for optimal delivery. The linkages of the monosaccharides to one
another in such
disaccharides, trisaccharides, and longer oligosaccharides can include at
least one 1, 4 linkage,
such as in 2DGa1-beta(1,4)-2-DG, that is cleavable by lactase. Other novel
compounds of the
invention are conjugates of 2-DGal or a 2-DGal analog and 3-bromopyruvate
joined by an ester
linkage. Other novel compounds of the invention that are useful metabolic
inhibitors alone or in
combination with 2-DGal or a 2-DGal analog include a.disaccharide,
trisaccharide, or other
oligosaccharide that is 2-DG bound to 2-DG in a 1,4 alpha linkage (2-deoxy-
maltose), and that
would be cleavable by.maltase, and that is 2-DG bound to 2-deoxyfructose to
form the 2-deoxy
analop- of sucrose.
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[0039] Microtubule inhibitors.(as used herein, a "microtubule inhibitor" is
any agent that
interferes with the assembly or disassembly of microtubules) useful in the
practice of the present
invention include but are not limited to vincristine (Oncovin), vinblastine
(Velban), paclitaxel
(Taxol, Paxene), vinorelbine (Navelbine), docetaxel (Taxotere), epothilone B
or D or a derivative
of either, and discodermolide or its derivatives. In accordance with the
methods of the present
invention 2-DGaI or a 2-DGaI analog is co-administered with a microtubule
inhibitor to treat
cancer. In one embodiment, the cancer is ovarian cancer, breast cancer, non-
small cell lung
cancer, Kaposi's sarcoina, and metastatic cancer of breast or ovary origin. As
one example, the
compound 22-oxo-vincaleukoblastine, also commonly known as vincristine, is an
alkaloid
obtained from the common periwinkle plant (Vinca rosea, Linn.) and is useful
in the treatment of
acute leukemia. It has also been shown to be useful in combination with other
oncolytic agents in
the treatment of Hodgkin's disease, lymphosarcoma, reticulum-cell sarcoma,
rhabdomyosarcoma, neuroblastoma, and Wilm's tumor. Vincristine is administered
in weekly
intravenous doses of 2: mg/m2 for children and 1.4 mg/m2 for adults. In
accordance with the
methods of the invention, 2-DGaI or a 2-DGaI analog is co-administered with
vincristine
administered at such doses.
[0040] Modifiers usefiil in the practice of the present invention include but
are not limited to
Leucovorin (Welleovorin), which is used with other drugs such as 5-
fluorouracil to treat
colorectal cancer. In accordance with the methods of the present invention 2-
DGaI or a 2-DGaI
analog is co-administered with a modifier and another anti-cancer agent to
treat cancer. In one
embodiment, the cancer is colon cancer. In one embodiment, themodifier is a
compound that
increases the ability of a cell to uptake glucose, including but not limited
to the compound N-
hydroxyurea. N-hydroxyurea has been reported to enhance the ability of a cell
to uptake 2-
deoxyglucose (see the reference Smith et al., 1999, Cancer Letters 141: 85,
incorporated herein
by reference), and adrriinistration of N-hydroxyurea at levels reported to
increase 2-DG uptake or
to treat leukemia together with administration of 2-DGaI or a 2-DGaI analog as
described herein
is one embodiment of the therapeutic methods provided by the invention. In
another such
embodiment, 2-DGaI or a 2-DGaI analog is co-administered with nitric oxide or
a nitric oxide
precursor, such as an organic nitrite or a spercnineNONOate, to treat cancer,
as the latter
compounds stimulate the uptake of glucose and so stimulate the uptake of 2-
DGaI or a 2-DGaI
analog.
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[0041] Nitrosoureas useful in the practice of the present invention include
but are not limited to
procarbazine (Matulane), lomustine, CCNU (CeeBU), carmustine (BCNU, BiCNU,
Gliadel
Wafer), and estramustine (Emcyt). In accordance with the methods of the
present invention 2-
DGaI or a 2-DGaI analog is co-administered with a nitrosourea to treat cancer.
In one
embodiment, the cancer is prostate cancer or glioblastoma, including recurrent
glioblastoma
multiforme.
[0042] Nucleoside analogs useful in the practice of the present invention
include but are not
limited to mercaptopurine, 6-MP (Purinethol), fluorouracil, 5-FU (Adrucil),
thioguanine, 6TG
(Thioguanine), hydroxyurea (Hydrea), cytarabine (Cytosar-U, DepoCyt),
floxuridine (FUDR),
fludarabine (Fludara), pentostatin (Nipent), cladribine (Leustatin, 2-CdA),
gemcitabine
(Gemzar), and capecitabine (Xeloda). In accordance with the methods of the
present invention 2-
DGaI or a 2-DGaI analog is co-administered with a nucleoside analog to treat
cancer. In one
embodiment, the cancer is B-cell lymphocytic leukemia (CLL), hairy cell
leukemia,
adenocarcinoma of the pancreas, metastatic breast cancer, non-small cell lung
cancer, and
metastatic colorectal carcinoma. As one example, the compound 5-
fluoro2,4(1H,3H)-
pyrimidinedione, also commonly known as 5-fluorouracil, is an antimetabolite
nucleoside analog
effective in the palliative management of carcinoma of the colon, rectum,
breast, stomach, and
pancreas in patients who are considered incurable by surgical or other means.
5-Fluorouracil is
administered in initial therapy in doses of 12 mg/mz given intravenously once
daily for 4
successive days with the daily dose not exceeding 800 mg. If no toxicity is
observed at any time
during the course of the therapy, 6 mg/kg are given intravenously on the 6th,
8th, 10th, and 12th
days. No therapy is given on the 5th, 7th, 9th, or I lth days. In poor risk
patients or those who are
not in an adequate nutritional state, a daily dose of 6 mg/kg is administered
for three days, with
the daily dose not exceeding 400 mg. If no toxicity is observed at any time
during the treatment,
3 mg/kg may be given on the 5th, 7th, and 9th days. No therapy is given on the
4th, 6th, or 8th
days. A sequence of injections on either schedule constitutes a course of
therapy. In accordance
with the methods of the invention, 2-DGaI or a 2-DGaI analog is co-
administered with 5-FU
administered at such doses or with the prodrug form Xeloda with
correspondingly adjusted
doses. As another example, the compound 2-aznino-l,7-dihydro-6H-purine-6-
thione, also
commonly known as 6-thioguanine, is a nucleoside analog effective in the
therapy of acute non-
pymphocytic Ieukemias. 6-Thioguanine is orally administered in doses of about
2 mg/kg of body
weiaht ner day. The total daily dose may be given at one time. If after four
weeks of dosage at
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this level there is no improvement, the dosage may be cautiously increased to
3 mg/kg/day. In
accordance with the methods of the invention, 2-DGal -or a 2-DGaI*analog is co-
administered
with 6-TG administered at such doses.
[0043] Osteoclast inhibitors useful in the practice of the present invention.
include but are not
limited to pamidronate (Aredia). In accordance with the methods of the present
invention 2-DGaI
or a 2-DGaI analog is co-administered with an osteoclast inhibitor to treat
cancer. In one
embodiment, the cancer is osteolytic bone metastases ofbreast cancer, and one
or more
additional anti-cancer agents are also co-administered with 2-DGa1 or a 2-DGaI
analog.
[0044] Platinum compounds useful in the practice of the present invention
include but are not
limited to cisplatin (Platinol) and carboplatin (Paraplatin). In accordance
with the methods of the
present invention 2-DGaI or a 2-DGa1 analog is co-administered with a platinum
compound to
treat cancer. In one embodiment, the cancer is metastatic testicular cancer,
metastatic ovarian
cancer, ovarian carcinoma, and transitional cell bladder cancer. As one
example, the compound
cis-Diamminedichloroplatinum (II), commonly known as cisplatin, is useful in
the palliative
treatment of metastatic testicular and ovarian tumors, and for the treatment
of transitional cell
bladder cancer which is not amenable to surgery or radiotherapy. Cisplatin,
when used for
advanced bladder cancer, is administered in intravenous injections of doses of
50-70 mg/m2 once
every three to four weeks. In accordance with the methods of the present
invention, 2-DGaI or a
2-DGaI analog is co-administered with cisplatin administered at these doses.
One or more
additional anti-cancer agents can be co-administered with the platinum
compound and 2-DGaI or
a 2-DGa1 analog. As one exa.mple, Platinol, Blenoxane, and Velbam may be co-
administered
with 2-DGaI or a 2-DGaI analog. As another example, Platinol and Adiriamycin
may be co-
administered with 2-DGal or a 2-DGaI analog.
[0045] Retinoids useful in the practice of the present invention include but
are not limited to
tretinoin, ATRA (Vesanoid), alitretinoin (Panretin), and bexarotene
(Targretin). In accordance
with the methods of the present invention 2-DGa1 or a 2-DGaI analog is
coadministered with a
retinoid to treat cancer. In one embodiment, the cancer is acute promyelocytic
leukemia (APL),
Kaposi's sarcoma, or T-cell lymphoma.
[0046] Topoisomerase I inhibitors useful in the practice of the present
invention include but are
not limited to topotecan (Hycamtin) and irinotecan (Camptostar). In accordance
with the
methods of the present invention 2-DGal or a 2-DGaI analog is co-administered
with a
tonc,isomerase 1 inhibitor to treat cancer. In one embodiment, the cancer is
metastatic carcinoma
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of the ovary, colon, or rectum, or small cell lung cancer. As noted above,
however, in one
embodiment of the present invention, administration of 2-DGaI or a 2-DGaI
analog either
precedes or follows, or both, administration of a topoisomerase 1 inhibitor
but is not conducted
concurrently therewith.
[0047] Topoisomerase 2 inhibitors useful in the practice of the present
invention include but are
not limited to etoposid:e, VP-16 (Vepesid), teniposide, VM-26 (Vumon), and
etoposide
phosphate (Etopophos). In accordance with the methods of the present invention
2DGaI or a 2-
DGaI analog is co-administered with a topoisomerase 2 inhibitor to treat
cancer. In one
embodiment, the cancer is refractory testicular tumors, refractory acute
lymphoblastic leukemia
(ALL), or small cell lung cancer. As noted above, however, in one embodiment
of the present
invention, administration of 2-DGaI or a 2-DGaI analog either precedes or
follows, or both,
administration of a topoisomerase 2 inhibitor but is not conducted
concurreintly therewith.
[0048] Tyrosine kinase inhibitors useful in the practice of the present
invention include but are
not limited to imatinib' (Gleevec). In accordance with the methods of the
present invention 2-
DGaI or a 2-DGaI analog is co-administered with a tyrosine kinase inhibitor to
treat cancer. In
one embodiment, the cancer is CML or metastatic or unresectable malignant
gastrointestinal
stromal tumors.
[0049] Thus, the present invention provides methods of treating cancer in
which 2-DGaI or a 2-
DGaI analog or an acetylated, benzylated, or other modified and prodrug
versions thereof and
one or more additional anti-cancer agents are adniinistered to a patient.
Specific embodiments of
such other anti-cancer agents include without limitation 5-metliyl-6-[[(3,4,5-
trirnethoxyphenyl)amino]-methyl]-2,4-quinazolinediarnine or a
pharniaceutically acceptable salt
thereof, (8S,1OS)-10-(3-amino-2,3,6-trideoxy-alpha-L-lyxo-hexopyranosyl)oxy]-
8glycoloyl-
7,8,9,10-tetrahydro-6,8,11-trihydroxy-l-methoxy-5,12-naphthacenedione or a
phacmaceutically
acceptable salt thereof; 5-fluoro-2,4(1H,3H)-pyrimidinedione or a
pharmaceutically acceptable
salt thereof; 2-amino-1,7-dihydro-6H-purine-6-thione or a pharmaceutically
acceptable salt
thereof; 22-oxo-vincaleukoblastine or a pharmaceutically acceptable salt
thereof; 2-bis[(2-
chloroethyl)amino]tetrahydro-2H-1,3,2-oxazaphosphorine, 2-oxide, or a
pharmaceutically
acceptable salt thereof; N-[4-[[(2,4-diamino-6-pteridinyl)methyl]-
methylamino]benzoyl]-L-
glutamic acid, or a pharmaceutically acceptable salt thereof; or cis-
diamminedichloroplati.num
(II). The methods of tlie present invention are generally applicable to all
cancers but have
narticularly significant therapeutic benefit in the treatment of solid
tu.inors, which are
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characterized by extensive regions of hypoxic tissue. Particular cancers that
can be treated with
the methods of the present invention are discussed in the following section.
Treating ,particular cancers
[0050] The methods and compositions of the invention may be used to treat any
cancer, whether
malignant or benign, as well as any precancerous condition, including but not
limited to
hyperplasias such as benign prostatic hyperplasia. In one important
embodiment, the invention
provides methods of treating particular types of malignant cancer, including
but not limited to
non-small cell lung cancer, head and neck cancers, prostate cancer, colon
cancer, and breast
cancer in humans and other mammals. These methods comprise administering an
antineoplastically effective amount of 2-DGal or a 2-DGaI analog or a
pharmaceutically
acceptable salt thereof either alone or in combination with an
antineoplastically effective amount
of one or more additional anti-cancer compounds. For purposes of illustration
and not limitation,
the methods of the invention for treating particular types of cancer are
described below.
[0051] In addition to the cancers listed above, the methods and compositions
of the present
invention can also be used to treat common cancers such as bladder cancer,
colorectal cancer,
endometrial cancer, leukemia, lung cancer, lymphoma, melanoma, and ovarian
cancer, as well as
less common cancers, including but not limited to acute lymphocytic leukemia,
adult acute
myeloid leukemia, adult non-Hodgkin's lymphoma, brain tumors, cervical
cancers, childhood
cancers, childhood sarcoma, chronic lymphocytic leukemia, chronic myeloid
leukemia,
esophageal cancer, hairy cell leukemia, kidney cancer, liver cancer, multiple
myeloma,
neuroblastoma, oral cancer, pancreatic cancer, primary central nervous system
lymphoma, skin
cancer, and small-cell lung cancer. Childhood cancers amenable to treatment by
the methods and
with the compositions of the present invention include but are not limited to
brain stem glioma,
cerebellar astrocytoma, cerebral astrocytoma, ependymoma, Ewing's sarcoma and
family of
tumors, germ cell tumor - extracranial, Hodgkin's disease, ALL, AML, liver
cancer,
medulloblastoma, neuroblastoma, non-Hodgkin's lymphoma, osteosarcoma,
malignant fibrous
histiocytoma of bor-e, retinoblastoma, rhabdomyosarconia, soft tissue sarcoma,
supratentorial
primitive neuroectodermal and pineal tumors, unusual childhood cancers, visual
pathway and
hypothalamic gliorna, aind Wilms's tumor and other childhood kidney tumors.
[0052] The methods and compositions of the present invention can also be used
to treat cancers
that have originated in or metastasized to the bone, brain, breast, digestive
and gastrointestinal
svstems. endocrine system, eye, genitourinary tract, germ cells, gynecological
system, head and
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neck, hematologic system, blood, lung, respiratory system, thorax,
musculoskeletal system, and
skin.
[0053] In one preferred embodiment of the invention, 2-DGaI or a 2-DGaI analog
is
administered to treat non-small-cell lung cancer. Current treatment regiments
for non-small-cell
lung cancer include without limitation administration of Gemcitabine (Eli
Lilly,
difluorodeoxycytidine), vinorelbine, paclitaxel, docetaxel, cisplatin,
carboplatin, or Irinotecan
(camptothecin-11) as single agents; and administration of etoposide and
cisplatin, Vindesine
(deacetyl vinblastine carboxamide) and cisplatin, paclitaxel and carboplatin,
Gemcitabine and
carboplatin, docetaxel and cisplatin, vinorelbine and cisplatin, or Irinotecan
and cisplatin in
combination therapies. See Bunn, 15 Sep. 2002, J. Clin. One. 20(l8s): 23-33,
incorporated herein
by reference. In accordance with the methods of the present invention, 2-DGaI
or a 2-DGal
analog can be co-administered in such therapeutic regimens to improve patient
outcomes.
[0054] In another preferred embodiment of the invention, 2-DGaI or a 2-DGaI
analog is
administered to treat prostate cancer. In one embodiment, 2-DGaI or a 2-DGa1
analog is
administered with prednisone to treat prostate cancer. The present invention
also provides
pharmaceutical formulations comprising prednisone admixed with 2-DGal or a 2-
DGaI analog in
amounts effective for the treatment of prostate cancer and suitable for oral
administration. In
another embodiment, 2-DGaI or a 2-DGaI analog is administered with prednisone
and
mitoxanthrone for the treatment of prostate cancer. In another embodiment,
2DGaI or a 2-DGal
analog is administered with TaxotereTm (Aventis, docetaxel) for the treatment
of prostate cancer.
[0055] In another preferred embodiment of the invention, 2-DGal or a 2-DGaI
analog is
administered to treat liver cancer, including but not limited to any hepatic
tumor, hepatocellular
carcinoma, or hepatoma, alone or in combination with other anti-cancer agents.
While not to be
bound by theory, this aspect of the invention may be particularly effective
due to the increased
uptake, relative to other cells, of 2-DGaI and certain 2-DGaI analogs,
including but not limited to
2-F-DGaI and 2-difluoro-DGaI, by liver cells (see Keppler et al., 1985, Adv.
Enz. Reg. 23: 61-
79, supra), and to their retention, once inside the liver cancer cell, due to
(i) the presence of
galactokinase that converts them to their 1-phosphate derivatives (see Bauer
et al., Jun. 1980,
Cancer Res. 40: 2026-2032); and (ii) the decreased expression of glucose-6-
phosphatase
("G6Pase") in liver cancer cells (see Ashmore et al., Sep. 1958, Cancer Res.
.18(8): 974-979, and
Landau et al., Nov. - Dec. 1962, Cancer 15(6): 1188-1196). In one embodiment,
the hepatoma to
be treated is relatively undifferentiated, and/or the level of G6Pase in the
tumor is measured prior
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to administration of the 2-DGal or 2-DGaI analog, with the treatment decision
based on the
amount of G6Pase measured. Tumors that have no or very low levels of G6Pase
are deemed to
be especially sensitive to treatment with 2-DGaI or a 2-DGaI analog in
accordance with the
methods of the invention.
[0056] In another preferred embodiment of the invention, 2-DGaI or a 2-DGaI
analog is
administered to treat brain cancer, alone or in combination with other anti-
cancer agents and/or
radiation therapy. While not to be bound by theory, this aspect of the
invention may be
particularly effective due to the increased uptake, relative to other cells,
of 2-DGaI and certain 2-
DGaI analogs, including but not limited to 2-F-DGaI and 2-difluoro-DGaI, by
brain cells.
[0057] In other preferred embodiments of the invention, 2-DGaI or a 2-DGaI
analog is
administered to treat a cancer selected from the group consisting of cancers
arising from
leukocytes, fibroblasts; kidney cells, skeletal muscle cells, intestinal
mucosa cells, testicular
cells, and ovarian cells; alone or in combination with other anti-cancer
agents. While not to be
bound by theory, this aspect of the invention may be particularly effective
due to the increased
uptake, relative to other cells, of 2-DGal and certain 2-DGal analogs,
including but not limited to
2-F-DGaI and 2-difluoro-DGaI, by such cells.
[0058] In another aspect, the present invention provides a method for
determining whether
administration of 2-DGaI or a 2-DGaI derivative is effective against a
particular tumor or to
follow the progress of such therapy, which_method comprises subjecting the
patient being treated
to PET scanning using, for example, either 18F-2-DG or18F-2-DGaI as the
imaging agent.
[0059] Because the methods of the present invention are applicable to a wide
variety of cancers
in a wide variety of tissues and organ systems, those of skill in the art will
appreciate that the
active compound can be formulated in a wide variety of formulations, with the
particular
formulation selected by the practitioner of skill in the art to best treat the
patient. Formulations
provided by the present invention are discussed in the following section.
Formulation aridpackagin of 2-DGaI or a 2-DGa1 analog
[0060] The present invention provides a pharmaceutically acceptable
formulation of 2DGal or a
2-DGaI useful in the methods of the present invention. In one embodiment, the
fonnulation is
crystalline in nature, arid the 2-DGaI or 2-DGaI analog is packaged in a
sachet that is decanted
into a potable liquid for oral administration to the patient. In this
embodiment, the liquid can be a
syrup or, more conveniently, a commonly consumed liquid, such as water, fruit
juice, or cola. In
some embodiments, the liquid will be glucose-free. In another embodiment, the
2-DGaI or 2-
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DGaI analog is formulated as a tablet or pill containing 2-DGaI or a 2-DGat
analog in an amount
in the range of about 100 mg to about 10 g. In some embodiments, each tablet
or pill contains
about 1 g to about 5 g of 2-DGaI or a 2-DGaI analog.
[0061] A decided practical advantage of the compounds of the present invention
is that the
compounds can be administered in any convenient manner such as by the oral,
intravenous,
intramuscular, topical, or subcutaneous routes.
[0062] Thus, 2-DGai or a 2-DGaI analog can be orally administered, for
example, with an inert
diluent or with an assimilable edible carrier, including but not liniited to
any type of food, or it
can be enclosed in hard or soft shell gelatin capsules, or compressed into
tablets, or incorporated
directly with the food of the diet. For oral therapeutic administration, 2-
DGaI or a 2-DGaI analog
can be incorporated with excipients and used in the form of ingestible
tablets, buccal tablets,
troches, capsules, elixirs, suspensions, syrups, wafers, and the like. Such
compositions and
preparations contain enough of the active agent to deliver the therapeutically
active doses
described above.
[0063] The tablets, troches, pills, capsules, and the like may also contain
the following: a binder
such as gum tragacanth, acacia, corn starch, or gelatin; excipients such as
dicalcium phosphate; a
disintegrating agent such as corn starch, potato starch, alginic acid, and the
like; a lubricant such
as magnesium stearate; a sweetening agent such as saccharin; and/or a
flavoring agent such as
peppermint, oil of wintergreen, or cherry flavoring. When the dosage unit form
is a capsule, it
can contain, in addition to materials of the above types, a liquid carrier.
Various other materials
can be present as coatings or to otherwise modify the physical form of the
dosage unit. For
instance, tablets, pills, or capsules can be coated with shellac. A'syrup or
elixir can contain the
active compound, a sweetening agent, methyl and propylparabens as
preservatives, and a
flavoring such as cherry or orange flavor. Of course, any material used in
preparing any dosage
unit form should be pharmaceutically pure and substantially non-toxic in the
arnounts employed.
In addition, the active compound can be incorporated into sustained-release
preparations and
' formulations. The 2-DGal or 2-DGaI analog can also be administered
parenterally or
intraperitoneally. A solution of the active compound can be prepared in water
suitably mixed
with a surfactant such as hydroxypropylcellulose. Dispersioins can also be
prepared in glycerol,
liquid polyethylene glycols and mixtures thereof, and in oils. Under ordinary
conditions of
storage and use, these preparations contain a preservative to prevent the
growth of
microorganisms. [0064] The pharmaceutical forms suitable for injectable use
include sterile
21
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WO 2007/098183 PCT/US2007/004465
aqueous solutions or dispersions and sterile powders for the extemporaneous
preparation of
sterile injectable solutions or dispersions. In all cases, the form must be
sterile and, in final form,
must be fluid to the extent that easy syringability exists. It must be stale
under the conditions of
manufacture and storage and must be preserved against the contaminating action
of
microorganisms such as bacteria and fungi.
[0065] The pharmaceutical forms suitable for topical use include oil and water
emulsions and
liposomal formulations, as well as lotions, creams, and ointments commonly
used for topical
administration of drugs. The topical formulation optionally includes one or
more additional anti-
cancer agents to be_co-administered with the 2-DGaI or 2-DGaI analog. [0066]
The carrier can
be a solvent or dispersion medium containing, for example, water, ethanol,
polyol, for example,
glycerol, propylene glycol, and liquid polyethylene glycol, and the like,
suitable mixtures
thereof, and vegetable oils. The proper fluidity can be maintained, for
example, by the use of a
coating such as lecithin, by the maintenance of the required particle size in
the case of dispersion
and by the use of surfactants. The prevention of the action of microorganisms
can be brought
about by various anti-bacterial and anti-fungal agents, for example, parabens,
chlorobutanol,
phenol, sorbic acid, thimerosal, and the like. In many cases, it will be
preferable to include
isotonic agents, for example sodium chloride. Prolonged absorption of the
injectable
compositions can be brought about-by the use in the compositions of agents
delaying absorption,
for example, aluminum monostearate and gelatin.
[0067] Sterile injectable solutions are prepared by incorporating the active
compound in the
required amount in the appropriate solvent with various of the other
ingredients enumerated
above, as required, followed by filtered sterilization. Generally, dispersions
are prepared by
incorporating the various sterilized active ingredient into a sterile vehicle
which contains the
basic dispersion medium and the required other ingredients from those
enumerated above. In the
case of sterile powders.foir the preparation of sterile injectable solutions,
the preferred methods of
preparation are vacuum drying and the freeze drying technique, which yield a
powder of the
active ingredient plus any additional desired ingredient from previously
sterile filtered solution
thereof.
[0068] As used herein, a "pharmaceutically acceptable carrier" includes any
and all solvents,
dispersion media, coatings, antibacterial and antifungal agents, isotonic
agents, absorption
delaying agents, and the like. The use of such media and agents for
pharmaceutical active '
substances is well known in the art. Except insofar as any conventional media
or agent is
22
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incompatible with the active ingredient, its use in the therapeutic
compositions of the invention is
contemplated. Supplementary active ingredients can be incorporated into the
compositions of the
invention.
[0069] The present invention also provides slow release forms of 2-DGaI or a 2-
DGaI analog. In
one embodiment, the slow release form is a pharmaceutical formulation in which
the 2-DGaI or a
2-DGaI analog is embedded in or coated by a material from which the 2DGaI or a
2-DGaI
analog is released over an extended period of time. The present invention also
provides slow
release forms of 2-DGaI or a 2-DGal analog in which an acid labile
polyethylene glycol (PEG)
moiety is attached to the 2-DGaI or a 2-DGaI analog, preferably at the
hydroxyl groups at the 4
and 6 positions. Such a slow release form can be readily synthesized by first
treating PEG with
Des Martin periodinane and reacting the resulting aldehyde 2-DGaI or a 2-DGal
analog. The
resulting compound is a novel compound of the invention, having the structure
shown below for
2-DG, with the exception that 2-DGaI or a 2-DGaI analog of choice replaces 2-
DG in the
structure shown (the structure shown is the 4epimer of the 2-DGaI compound of
the invention).
0 oH
A~ O
OH
[0070] It is essentially advantageous to formulate parental and other
compositions in dosage unit
form for ease of administration and uniformity of dosage. Dosage unit form as
used herein refers
to physically discrete units suited as unitary dosages for the mammalian
subjects to be treated;
each unit containing a predetermined quantity of active material calculated to
produce the
desired therapeutic effect in association with the required pharmaceutical
carrier. The
specification for the novel dosage unit forms of the invention are dictated by
and directly
dependent on the patient and cancer to be treated and can vary from patient to
patient and cancer
to cancer, but generally, a dosage unit form contains from about 100 mg to
about 10 g of 2-DGaI
or a 2-DGal analog. Typical unit forms can contain about 0.5 to about 5 g of 2-
DGaI or a 2-DGaI
analog.
[0071] The present invention having been described in detail in the preceding
sections, the
following examples are provided to illustrate certain aspects of, but not to
limit, the invention.
23
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EXAMPLE I
Growth Inhibitory Activity of 2-Deoxy-D-Qalactose Against Human Heuatocellular
Carcinoma
Cell Lines
[0072] In this example, 2-DGal (or a 2-DGaI analog) can be demoristrated to
inhibit cell lines
obtained from hepatocellular carcinomas. The cells are cultured at 37 degrees
C in a humidified
atmosphere containing 5% C02. For passaging, cells grown in 75 cm2 flasks
(6070%
confluency) are washed with PBS and dissolved from the flasks with trypsin
(Gibco BRL) before
being plated. Ten thousand cells are plated into each well of a 96-well
microtiter plate in 100 ul
of medium. The cells are then incubated for 24 h at 37 degrees C in a
humidified atmosphere
containing 5% C02 prior to exposure to the 2-DG analog or other anti-cancer
agent.
[0073] Cultures are treated with a range of concentrations of 2-DGaI or a 2-
DGaI analog
solubilized in cell culture medium, diluted IOOx to the desired concentration,
continuously for 72
h. Docetaxel (TaxotereT', Aventis Pharmaceuticals, Inc.) solubilized in 100%
DMSO and diluted
I 000x to the desired concentration, is used as a positive control. The
sulforhodamine B (SRB)
assay, a dye-based method for determining cell number by virtue of SRB binding
to basic amino
acids of cellular macromolecules, is employed to assess the growth inhibitory
activity of 2-DGal
or a 2-DGal analog.
[0074] Treatment with 2-DGaI or a 2-DGaI analog inhibits the growth of
hepatocellular 30
carcinoma cell lines. These results demonstrate that liver tumor cells -are
sensitive to 2-DGal or
a 2-DGaI analog.
EXAMPLE 2
Evaluation of 2-DGaI or a 2-DGaI Analogas a Sinngle Agent and in Combination
with Cisplatin
[0075] The efficacy of 2-DGaI or a 2-DGaI analog in combination with cisplatin
can be
compared with cisplatin alone in a tumor growth delay study in a mouse
xenograft model. Small
pieces of tumor, such as a liver tumor serially passaged in nude mice, are
implanted
subcutaneously in nude mice, and the tumors allowed to grow to 25 mrn2. Mice
are then
randomized to receive no treatment, treatment with 2-DGaI or a 2-DGaI analog
alone, treatment
with cisplatin alone, or treatment with both 2-DGaI or a 2-DGaI analog and
cisplatin. Cisplatin is
dosed i.p. at 1 mg/kg on days I to 5.2-DGaI or a 2-DGaI analog is dosed orally
at 750 mg/kg, =
twice daily, for the duration of the experiment. Each mouse is sacrificed when
its tumor volume
reaches 1000 mg.
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[0076] The results show that treatment with 2-DGaI or a 2-DGa1 analog in
combination with
cisplatin delays tunior growth significantly more than treatment with
cisplatin alone.
EXAMPLE 3
Oral Formulations of 2-DGaI or a 2-DGaI Analo~
[0077] This example illustrates the preparation of representative
pharmaceutical formulations for
oral administration.
A. 2-DGal or a 2-DGa1 analog is dispensed into -hard-shell gelatin capsules
containing
between 100 mg and 1 g of 2-DGaI or a 2-DGal analog; optionally, about 0.5%
(weight/weight)
magnesium stearate can be added. In addition, a mixture of 2-DGaI or a 2-DGaI
analog and
lactose can be used in the capsule.
B. 2-DGaI or a 2-DGaI analog (20.0% - 89.9% wt./wt., depending on whether
lactose is
present, and how much); magnesium stearate (0.9%); starch (8.6%); optionally
lactose (0 -
69.6%) and PVP (polyvinylpyrrolidine; 0.9%) are, with the exception of the
magnesium stearate,
combined and granulated using water as a granulating liquid. The formulation
is then dried,
mixed with the magnesium stearate and formed into tablets with a
tableting.machine.
C. 2-DGaI or a 2-DGal analog is dissolved in a mixture of propylene glycol,
polyethylene
glycol 400, and polysorbate 80; water is added; and the resulting mixture is
dispensed into
bottles.
D. A mixture of 2-DGal or a 2-DGal analog (20% to 60% wt./wt.), peanut oil
(38% - 78%),
and 2.0% (wt_/wt.) Span 60 is prepared, melted, mixed, and filled into soft
elastic capsules.
EXAMPLE 4
2-DGal or a 2-DGal Analog Formulation for Insufflation
[0078] This example illustrates the preparation of a representative
pharmaceutical formulation
for insufflation. Micronized 2-DGaI or a 2-DGal analog with or without
micronized lactose is
milled and packaged in an insufflator equipped with a dosing pump.
[0079] While the present invention has been described with reference to the
specific
embodiments thereof, it should be understood by those skilled in the art that
various changes
may be made and equivalents may be substituted without departing from the true
spirit and scope
of the invention. In addition, many modifications may be made to adapt a
particular situation,
material, composition of matter, process, process step or steps, to the
objective, spirit and scope
of the present invention. All such modifications are intended to be within the
scope of the claims
annPnriPd hereto. All patents and publications cited above are hereby
incorporated by reference.
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EXAMPLE 5
LIVER TOXICITY OF 2-FLUORO-2-DEOXYGALACTOSE
[0080] The objective of this study was to evaluate the liver toxicity of 2-
Fluoro-2-
deoxygalactose when administered via intraperitoneal or intravenous injection
to CD 1 mice. The
study was initiated on January 11, 2007, and consisted of three groups with
three (3) male mice
per group. On Day 1, each mouse GROUP was administered either about 0.2 ml of
10% 2 Fluoro
2 Deoxygalactose, intraperitoneally, about 0.2 ml of 10% 2 Fluoro 2
Deoxyglactose,
intravenously, or about 0.2 ml.of sterile 0.9% sodium chloride for injection,
USP, intravenously.
Criteria for evaluation included clinical observations and limited serum
chemistry parameters
(alanine aminotransferase [ALT], aspartate aminotransferase [AST), and total
bilirubin). On Day
2, all animals were euthanized, blood was collected, and livers were collected
and stored for
potential histopathological evaluation.
[0081) Clinical observations were performed at least once daily. There were no
unscheduled
deaths during this study and there were no adverse test article related
clinical findings.
B. The issue with the study is that one of the control male animals had very
high levels of AST
and ALT c/w illness. Based on the historical controls see Table I, group 1 and
animal 2 had an
ALT of 421 and an AST of 398. These are the highest of any animals and not
consistent with the
other two animals in the group receiving sodium chloride. It is believed that
this one control
animal, a classic CD-1 mouse, was sick and not typical for the strain.
C. The intravenous and intraperitoneal data is very consistent with a
treatment effect. The
intravenous group, Table I, group 3 has one animal with elevated enzymes at 24
hours and two
that has returned to normal. Given the more limited exposure and clearance of
the compound,
one would expect this pattern when compared with the intrapertioneal
administration where one
will have more prolonged exposure, close to IV, but typically a lower Cmax
(but above the
threshold) but with prolonged exposure. When this group is examined, the data
is consistent with
glactosamine with a 2-3X rise in ALT and AST over normal animals (based on
using the two
expected historical controls from Group 1). The data and effects in the
intravenous and
intraperitoneal group are consistent with expected data and the magnitude of
damage at single
administration makes sense.
D. The gross pathology finding of a single animal in the intraperitonneal
administered group is
also c/w a biological effect. It is believed that changes in the control group
would not be
exnected and changes are- very unlikely-in the intravenous group. In addition,
it is believed that
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there should be a less number of observations in the intravenous group as
compared with the
intrapertioneal group. It is believed that the fact that there was any damage
observed in the
intraperitonneal group is a sign consistent with a biological effect.
[0082) Study in which 2 fluoro 2 deoxygalactose was administered the CD 1 mice
intraperitoneally demonstrated increases in concentration of transanxinases in
accord with the
expectation that the compound would damage liver tissue with continued dosing.
If there was
administration of galactose before providing a deoxy glalactose it is believed
to be possible to
protect normal liver parenchyma while damaging liver tumor (either within the
liver itself or for
tissue outsisde of the liver). While studies are not yet completed in terms of
specific sites of
administration, the procedure outlined derives from the demonstration that
galactose markedly
inhibits the uptake of 2-Fluoro-2-deoxygalactose by normal liver tissue. An
example of the
situation would in an individual who has primary liver cancer with or without
metastatic disease.
This individual would be administered unlabelled galactose to the point where
unlabelled
galactose reaches the concentration in the hepatic vein that it exceeds the
amount that can be
absorbed in normal liver. Until that point, the concentration of galactose
would be negligible.
Thus, we believe to have demonstrated that giving many grams of galactose can
be taken up by
liver before the liver begins to utilize galactose significantly. Therefore,
the subject, for example,
receives a treatment of 2 2-Fluoro-2-deoxygalactose at a dose which would
normally be taken up
by liver,but it would not occur if the subject had previously ingested, in the
GI tract, high
concentrations of galactose. The normal liver would then be protected from 2-
Fluoro-2-
deoxygalactose. On the other hand since the blood supply to a tumor is
arterial the tumor would
be exposed to the 2-Fluoro-2-deoxygalactose infused cir presented to the tumr
tissue_ Again, it is
believed that the liver would be protected because the galactose given in the
GI tract would
compete with the 2 deoxy 2 fluoro galactose that would be attempted to be
metabolized by the
liver.
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TablO I
Individual Serum Chemistry Values
Lead Optimization Toxicity Study oE 2-Fluoro-2-deoxygalactose Administered
Intraperitoneally
Arnd Intravenously In CD-1 Mice
-------------------------------------------------------------------------------
-------------
Gxoup Animal
Sex Number Day Alt Ast Tbili
Number (U/L) (U/L) (mg/dL)
im 1 2 26 37 0.2
^----- ---^ ----- ------ -------
2 2 421 398 0.3
-^-----
3 2 32 68 0.2
Nominal Dose: Group 1- 0.9% Sodium Chloride for injection, USP, intravenously
Group 2 - 10% 2-Fluoro-2-deoxygalactose, intraperitoneally
Group 3 - 10% 2-Fluoro-2-deoxygalactose, intravenously
KEY FOR SERUM CHEMISTRY VALUES
Alanine aminotransferase (Alt)
Aspartate aminotransferase (Ast)
Total ba.lirubin (Tbili.)
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Table x
IndividuaL Serum Chemistry Values
Lead Optimization Toxicity Study of 2-Fluoro-2-deoxygalactose Administered
intraperitoneally
And intravenously In CD-1 Mice
--------------------------=-----------------------------------^-------------^--
--------^---------
Group Animal
Sex Number Day Alt Ast Tbili
Number (U/L) (U/L) (mg/dL)
2m 4 2 109 234 0.2
------ ---- ----- ----- -------
2 73 114 0.2
------ ---- ----- --^-- -------
6 2 72 118 0.2
Nominal Dose: Group 1 - 0.9% Sodium Chloride for Injection, USP, intravenously
Group 2 - 10% 2-Fluoro-2-deoxygalactose, intraperitoneally
Group 3 - 10% 2-Fluoro-2-deoxygalactose, intravenously
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Table I
Individual Serum Chemistry Values
Lead Optimization Toicicity Study of 2-Fluoro-2-deoxygalactose Administered
Intraperitoneally
And intravenously In.CD-1 Mice
----------------------^-------------------------------------------------------
..----=---------------
Group Animal
Sex . Number Day Alt Ast Tbili
Number (U/L) (U/L)= (mg/dL)
3m 7 2 45 112 0.2
------ ---- ----- ----- -------
8 2 124 116 0.2
------ ---- ----- ----- -------
9 2 30 52 0.1
Nominal Dose: Group 1 - 0.9% Sodium Chloride for Injection, USP, intravenously
Group 2 - 10% 2-Fluoro-2-deoxygalactose, intraperitoneally
Group 3 10% 2-Fluoro-2-deoxygalactose, intravenously
