Note: Descriptions are shown in the official language in which they were submitted.
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[3-LAPACHONE IS A BROAD SPECTRUM ANTI-CANCER AGENT
CROSS REFERENCE TO RELATED APPLICATIONS
This application claims the benefit under 35 U.S.C. ~ 119(e) of U.S.
Application No.
60/545,916, filed February 20, 2004.
BACKGROUND OF THE INVENTION
Cancer cells are by definition heterogeneous. For example, within a single
tissue or cell
type, multiple mutational'mechanisms' may lead to the development of cancer.
As such,
heterogeneity frequently exists between cancer cells taken from tumors of the
same tissue and
same type that have originated in different individuals. Frequently-observed
mutational
'mechanisms' associated with some cancers may differ between one tissue type
and another (e.g.,
frequently-observed mutational 'mechanisms' leading to colon cancer may differ
from
frequently-observed mechanisms leading to leukemias). It is therefore often
difficult to predict
whether a particular cancer will respond to a particular chemotherapeutic
agent. (Cancer
Medicine, 5th Edition, Bast et al. eds., B.C. Decker Inc., Hamilton, Ontario).
Surgery and radiotherapy may be curative if a cancer is found early, but
current drug
therapies for metastatic disease are mostly palliative and seldom offer a long-
term cure. Even
with the new chemotherapies entering the market, improvement in patient
survival is measured
in months rather than in years, and the need continues for new drugs effective
both in
combination with existing agents as first line therapy and as second and third
line therapies in
treatment of resistant tumors.
[3-lapachone is an agent with a reported anti-cancer activity in a limited
number of
cancers. For example, there is reported a method and composition for the
treatment of tumors,
which comprises the administration of an effective amount of ~3-lapachone, in
combination with
a taxane derivative (W000/61142). Additionally, U.S. Pat. No. 6,245,807
discloses the use of
(3-lapachone, amongst other (3-lapachone derivatives, for use in treatment of
human prostate
disease. As a single agent, (3-lapachone has also been reported to decrease
the number of
tumors, reduce tumor size, or increase the survival time, or a combination of
these, in
xenotransplant mouse models of human ovarian cancer (Li, C.J. et al., (1999)
Proc. Natl. Acad.
Sci. USA, 96(23): 13369-13374), human prostate cancer (Li, C.J. et al., (
1999) Pf~oc. Natl. Acad.
Sci. USA, 96(23): 13369-13374), human breast cancer (Li, C.J. et al., (2000)
AACR P~oc., p. 9),
and human multiple myeloma (WO 03/011224).
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SUMMARY OF THE INVENTION
The present invention provides a method of treating a cancer selected from the
group
consisting of cancers of the tongue, mouth, pharynx, and oral cavity,
esophageal cancer, stomach
cancer, gastrointestinal stromal tumor, cancer of the small intestine, anal
cancer, cancer of the
anal canal, anorectal cancer, liver cancer, intrahepatic bile duct cancer,
gallbladder cancer,
biliary cancer, cancer of other digestive organs, cancer of the larynx, bone
and joint cancer,
uterine cancer, cervical cancer, uterine corpus cancer, cancer of the vulva,
vaginal cancer,
testicular cancer, penile cancer, urinary bladder cancer, kidney cancer, renal
cancer, cancer of
the ureter and other urinary organs, ocular cancer, brain and nervous system
cancer, CNS
cancers, and thyroid cancer (collectively "a cancer of the present
invention"), comprising
administering to a subject in need thereof a therapeutically effective amount
of (3-lapachone, or a
pharmaceutically acceptable salt thereof, in combination with a
pharmaceutically acceptable
carrier, wherein said (3-lapachone, or a pharmaceutically acceptable salt
thereof, treats said
cancer selected from the group consisting of a cancer of the present
invention.
The present invention also provides a method of treating a cancer selected
from the
group consisting of a cancer of the present invention, comprising
administering to a subject in
need thereof a therapeutically effective amount of ~3-lapachone, or a
pharmaceutically acceptable
salt, prodrug, metabolite, analog or derivative thereof, in combination with a
pharmaceutically
acceptable carrier, wherein said (3-lapachone, or a pharmaceutically
acceptable salt thereof,
treats said cancer selected from the group consisting of a cancer of the
present invention.
The present invention also provides a method of treating a metastatic cancer
selected
from the group consisting of a cancer of the present invention, comprising
administering to a
subject in need thereof a therapeutically effective amount of (3-lapachone, or
a pharmaceutically
acceptable salt thereof, in combination with a pharmaceutically acceptable
carrier, wherein said
(3-lapachone, or a pharmaceutically acceptable salt thereof, treats said
metastatic cancer selected
from the group consisting of a cancer of the present invention.
The present invention also provides a method of treating a metastatic cancer
selected
from the group consisting of a cancer of the present invention, comprising
administering to a
subject in need thereof a therapeutically effective amount of ~i-lapachone, or
a pharmaceutically
acceptable salt, prodrug, metabolite, analog or derivative thereof, in
combination with a
pharmaceutically acceptable Garner, wherein said [3-lapachone, or a
pharmaceutically acceptable
salt thereof, treats said metastatic cancer selected from the group consisting
of a cancer of the
present invention.
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The present invention also provides a method for inducing cell death in a
cancer cell
comprising contacting said cancer cell with an effective amount of (3-
lapachone, or a
pharmaceutically acceptable salt thereof, wherein said cancer cell comprises a
cancer selected
from the group consisting of a cancer of the present invention, and wherein
said contacting
induces said cell death in said cancer cell.
The present invention also provides a method for inducing cell death in a
cancer cell
comprising contacting said cancer cell with an effective amount of [3-
lapachone, or a
pharmaceutically acceptable salt, prodrug, metabolite, analog or derivative
thereof, wherein said
cancer cell comprises a cancer selected from the group consisting of a cancer
of the present
invention, and wherein said contacting induces said cell death in said cancer
cell.
The present invention also provides a method of treating a cancer selected
from the
group consisting of a cancer of the present invention, comprising
administering to a subject in
need thereof a therapeutically effective amount of (3-lapachone, or a
pharmaceutically acceptable
salt, prodrug, metabolite, analog or derivative thereof, in combination with a
pharmaceutically
acceptable carrier, and activating one or more cell cycle checkpoints in a
cancer of the present
invention cells, where the (3-lapachone, or a pharmaceutically acceptable
salt, prodrug,
metabolite, analog or derivative thereof, treats the cancer selected from the
group consisting of a
cancer of the present invention.
The present invention further provides a method of treating a cancer selected
from the
group consisting of a cancer of the present invention, comprising
administering to a subject in
need thereof a therapeutically effective amount of (3-lapachone, or a
pharmaceutically acceptable
salt, prodrug, metabolite, analog or derivative thereof, in combination with a
pharmaceutically
acceptable earner, and activating one or more cell cycle checkpoint pathways
in a cancer of the
present invention cells, where the (3-lapachone, or a pharmaceutically
acceptable salt, prodrug,
metabolite, analog or derivative thereof, treats the cancer selected from the
group consisting of a
cancer of the present invention.
The present invention also provides a method of treating a cancer selected
from the
group consisting a cancer of the present invention, comprising administering
to a subject in need
thereof a therapeutically effective amount of (3-lapachone, or a
pharmaceutically acceptable salt,
prodrug, metabolite, analog or derivative thereof, in combination with a
pharmaceutically
acceptable carrier, and activating one or more cell cycle checkpoint
regulators in cells of a
cancer of the present invention, where the [3-lapachone, or a pharmaceutically
acceptable salt,
prodrug, metabolite, analog or derivative thereof, treats the cancer selected
from the group
consisting of a cancer of the present invention.
3
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WO 2005/082358 PCT/US2005/005647
The present invention further provides a method of treating a cancer selected
from the
group consisting of a cancer of the present invention, comprising
administering to a subject in
need thereof a therapeutically effective amount of (3-lapachone, or a
pharmaceutically acceptable
salt, prodrug, metabolite, analog or derivative thereof, in combination with a
pharmaceutically
acceptable carrier, and activating cell death selectively in cells of a cancer
of the present
invention, where the (3-lapachone, or a pharmaceutically acceptable salt,
prodrug, metabolite,
analog or derivative thereof, treats the cancer selected from the group
consisting of a cancer of
the present invention.
The present invention also provides a method of treating or preventing a cell
proliferative
disorder of a cancer or pre-cancer selected from the group consisting of
cancers or pre-cancers
of the tongue, mouth, pharynx, and oral cavity, esophageal cancer or pre-
cancer, stomach cancer
or pre-cancer, cancer or pre-cancer of the small intestine, anal cancer or pre-
cancer, cancer or
pre-cancer of the anal canal, anorectal cancer or pre-cancer, liver cancer or
pre-cancer,
intrahepatic bile duct cancer or pre-cancer, gallbladder cancer or pre-cancer,
biliary cancer or
pre-cancer, cancer or pre-cancer of other digestive organs, cancer or pre-
cancer of the larynx,
bone and joint cancer or pre-cancer, melanoma, skin cancer or pre-cancer,
uterine cancer or pre-
cancer, cervical cancer or pre-cancer, uterine corpus cancer or pre-cancer,
cancer or pre-cancer
of the vulva, vaginal cancer or pre-cancer, testicular cancer or pre-cancer,
penile cancer or pre-
cancer, urinary bladder cancer or pre-cancer, kidney cancer or pre-cancer,
renal cancer or pre-
cancer, cancer or pre-cancer of the ureter and other urinary organs, ocular
cancer, brain and
nervous system cancer, CNS cancers or pre-cancers, and thyroid cancer or pre-
cancer
(collectively "cancer or pre-cancer of the present invention"), comprising
administering to a
subject in need thereof a therapeutically effective amount of (3-lapachone, or
a pharmaceutically
acceptable salt, prodrug, metabolite, analog or derivative thereof, in
combination with a
pharmaceutically acceptable carrier, where the [3-lapachone, or a
pharmaceutically acceptable
salt, prodrug, metabolite, analog or derivative thereof, treats or prevents
the cell proliferative
disorder of a cancer or pre-cancer selected from the group consisting of a
cancer or pre-cancer of
the present invention.
The present invention also provides a method of treating or preventing a cell
proliferative
disorder of a cancer or pre-cancer selected from the group consisting of a
cancer or pre-cancer of
the present invention comprising administering to a subject in need thereof a
therapeutically
effective amount of [3-lapachone, or a pharmaceutically acceptable salt,
prodrug, metabolite,
analog or derivative thereof, in combination with a pharmaceutically
acceptable Garner, and
activating one or more cell cycle checkpoints in a cancer or pre-cancer of the
present invention,
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WO 2005/082358 PCT/US2005/005647
where the [3-lapachone, or a pharmaceutically acceptable salt, prodrug,
metabolite, analog or
derivative thereof, treats or prevents a cell proliferative disorder of a
cancer or pre-cancer
selected from the group consisting of a cancer or pre-cancer of the present
invention.
The present invention also provides a method of treating or preventing a cell
proliferative
disorder of a cancer or pre-cancer selected from the group consisting of a
cancer or pre-cancer of
the present invention comprising administering to a subject in need thereof a
therapeutically
effective amount of (3-lapachone, or a pharmaceutically acceptable salt,
prodrug, metabolite,
analog or derivative thereof, in combination with a pharmaceutically
acceptable carrier, and
activating one or more cell cycle checkpoint pathways in a cancer or pre-
cancer of the present
invention, where the (3-lapachone, or a pharmaceutically acceptable salt,
prodrug, metabolite,
analog or derivative thereof, treats or prevents a cell proliferative disorder
of a cancer or pre-
cancer selected from the group consisting of a cancer or pre-cancer of the
present invention.
The present invention also provides a method of treating or preventing a cell
proliferative
disorder of a cancer or pre-cancer selected from the group consisting of a
cancer or pre-cancer of
the present invention comprising administering to a subject in need thereof a
therapeutically
effective amount of [3-lapachone, or a pharmaceutically acceptable salt,
prodrug, metabolite,
analog or derivative thereof, in combination with a pharmaceutically
acceptable carrier, and
activating one or more cell cycle checkpoint regulators in a cancer or pre-
cancer of the present
invention, where the (3-lapachone, or a pharmaceutically acceptable salt,
prodrug, metabolite,
analog or derivative thereof, treats or prevents a cell proliferative disorder
of a cancer or pre-
cancer selected from the group consisting of a cancer or pre-cancer of the
present invention.
The present invention also provides a method of treating or preventing a cell
proliferative
disorder of a cancer or pre-cancer selected from the group consisting of a
cancer or pre-cancer of
the present invention comprising administering to a subject in need thereof a
therapeutically
effective amount of [3-lapachone, or a pharmaceutically acceptable salt,
prodrug, metabolite,
analog or derivative thereof, in combination with a pharmaceutically
acceptable carrier, and
activating cell death selectively in cells of a cancer or pre-cancer of the
present invention, where
the (3-lapachone, or a pharmaceutically acceptable salt, prodrug, metabolite,
analog or derivative
thereof, treats or prevents a cell proliferative disorder of a cancer or pre-
cancer selected from the
group consisting of a cancer or pre-cancer of the present invention.
BRIEF DESCRIPTION OF THE FIGURES
Figure 1 sets forth a schematic of the proposed points of action of (3-
Lapachone and
Taxol on the cell cycle.
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Figure 2 sets forth an effect of (3-Lapachone on survival of human cancer cell
lines in the
NCI60 assay in vitro.
DETAILED DESCRIPTION OF THE INVENTION
The present invention also provides a method of treating a cancer selected
from the
group consisting of a cancer of the present invention, comprising
administering to a subject in
need thereof a therapeutically effective amount of [3-lapachone, or a
pharmaceutically acceptable
salt thereof, in combination with a pharmaceutically acceptable Garner,
wherein said (3-
lapachone, or a pharmaceutically acceptable salt thereof, treats said cancer
selected from the
group consisting of a cancer of the present invention. The present invention
also provides a
method of treating a cancer selected from the group consisting of a cancer of
the present
invention, comprising administering to a subject in need thereof a
therapeutically effective
amount of (3-lapachone, or a pharmaceutically acceptable salt, prodrug,
metabolite, analog or
derivative thereof, in combination with a pharmaceutically acceptable Garner,
wherein said (3-
lapachone, or a pharmaceutically acceptable salt thereof, treats said cancer
selected from the
group consisting of a cancer of the present invention.The present invention
also provides a
method of treating or preventing a cell proliferative disorder of a cancer
selected from the group
consisting of a cancer or pre-cancer of the present invention, comprising
administering to a
subject in need thereof a therapeutically effective amount of [3-lapachone or
pharmaceutically
acceptable salts thereof, or a metabolite thereof, in combination with a
pharmaceutically
acceptable carrier. The invention also provides the use of ~3-lapachone for
the preparation of a
medicament useful for the treatment of cancer. The invention also provides the
use of [3-
lapachone for the preparation of a medicament useful for the treatment or
prevention of a cell
proliferative disorder.
While not limited by theory, the present invention includes and is based in
part on an
understanding of, and methods for, the activation of cell cycle checkpoints by
modulators of cell
cycle checkpoint activation (e.g., (i-lapachone, or a pharmaceutically
acceptable salt, prodrug,
metabolite, analog or derivative thereof). The activation of cell cycle
checkpoints in general is
referred to as Activated Checkpoint TherapyTM, or ACTTM. Briefly, many cancer
cells are
defective in their cell cycle checkpoint functions secondary to mutations in
one of their
molecular modulators, e.g., p53. It is in part, for this reason, that cancer
cells have accumulated
genetic errors during the carcinogenic process. Therapeutic agents that
activate cell cycle
checkpoint functions can selectively promote cell death in cancer cells, since
cell death appears
to be induced at least in part by the conflict between the uncontrolled-
proliferation drive in
6
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cancer cells and the checkpoint delays induced artificially. ACTTM takes
advantage of the
tendency of cell death to occur at checkpoints during the cell proliferation
cycle by activating
one or more checkpoints, thereby producing conflicting signals regarding cell
cycle progression
versus arrest. If more than one checkpoint is activated, cancer cells with
uncontrolled
proliferation signals and genetic abnormalities are blocked at multiple
checkpoints, creating
"collisions" that promote synergistic cell death.
ACTTM offers selectivity against cancer cells as compared to normal cells and
is
therefore safer than less selective therapies. First, the ACTTM method
activates but does not
disrupt cell cycle checkpoints. Second, normal cells with well-controlled
proliferation signals
can be delayed at checkpoints in a regulated fashion, resulting in no cell
death-prone collisions.
Third, normal cells with intact Gl checkpoint control are expected to arrest
in G1. Cancer cells,
on the other hand, are expected to be delayed in S-, G2-, and M-phases, since
most cancer cells
harbor Gl checkpoint defects, making cancer cells more sensitive to drugs
imposing S and M
phase checkpoints. (3-lapachone is a Gl and S phase compound, and contacting a
cell with (3-
lapachone results in activation of a Gl or S cell cycle checkpoint. Figure 1
sets forth a schematic
of the proposed points of action of [3-lapachone and Taxol on the cell cycle.
The term "modulator of cell cycle checkpoint activation," as used herein,
refers to a
compound capable of altering checkpoint activation in cells (in preferred
embodiments,
activating one or more cell cycle checkpoints), preferably by activating
checkpoint-mediated
DNA repair mechanisms, or by reinstating checkpoint activity that has been
lost due to a
malfunction or mutation in the cellular pathways that regulate cell cycle
activity. As is known in
the art, major cell cycle checkpoints occur at Gl/S phase and at the GZ/Mphase
transitions. In a
model, four major cell cycle checkpoints monitor the integrity of genetic
material. DNA
synthesis begins only past the restriction point (R point), where the cell
determines if preparation
during G1 has been satisfactory for cell cycle continuation. The second
checkpoint occurs
during replication initiation in S phase. The third and fourth checkpoints
take place in G2 phase
and M phase, respectively. Modulation of cell cycle checkpoint activation is
further discussed
in, e.g., C.J. Li et al. P~oc. Natl. Acad. Sci. USA (1999), 96(23), 13369-
13374, and Y. Li et al.
Pf~oc. Natl. Acad. Sci. USA (2003), 100(5), 2674-2678, and PCT Publication WO
00/61142
(Pardee et al.). Preferred modulators of cell cycle checkpoint activation for
use in the present
invention induce checkpoint activation (i.e., activate one or more cell cycle
checkpoints,
preferably at Gl/S phase), preferably without causing substantial DNA damage.
In addition,
certain preferred modulators of cell cycle checkpoint activation are capable
of increasing the
level or activity of E2F (more preferably E2F1) in a cell. Methods for
screening for modulators
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of cell cycle checkpoint activation, including compounds capable of elevating
E2F activity or
levels in a cell, include those that are disclosed in PCT Patent Application
No. PCT/LTS03/22631
to Li et al. In certain embodiments, preferred modulators of cell cycle
checkpoint activation are
capable of increasing the level or activity of E2F in a cell by an amount
sufficient to cause cell
death (e.g., apoptosis) if the cell is a cancerous cell. More preferred
modulators of cell cycle
checkpoint activation are capable of raising the level or activity of E2F1 in
a cell by an amount
sufficient to cause cell death (e.g., apoptosis) if the cell is a cancerous
cell. In one aspect, a
modulator of cell cycle checkpoint activation is not [3-lapachone.
Again not limited by theory, cellular response to DNA damage is regulated by
the
ATM/ATR signal transduction pathway, in which ATM and ATR are protein kinases
of the
phosphatidyl-inositol-3 kinase family (PI3K). In response to DNA damage, ATM
and ATR
phosphorylate Chk2 and Chkl respectively, which in turn activate a variety of
substrates
involved in arresting cells at the Gl/S phase of the cell cycle, as well as
inducing and activating
proteins involved in DNA repair. Chk2 has been shown to activate proteins
involved in DNA
repair including the tumor suppressor BRCA1, thereby enhancing DNA repair
capacity
following DNA damage. Chk2 has also been shown to stabilize p53 both by
directly
phosphorylating p53, and by inhibiting Mdm2, a ubiquitin ligase that targets
p53 for
degradation. Under such conditions, increased levels of p53 lead to Gl/S
arrest, DNA repair,
and apoptosis in cells with irreparable DNA damage. Again not limited by
theory, it is believed
that Chk2 is an important cell cycle regulator, which, depending on the
conditions, can induce
cell cycle arrest and DNA repair, or initiate cell death (e.g., apoptosis) if
DNA damage is too
severe. W certain embodiments, preferred modulators of cell cycle checkpoint
activation are
capable of increasing the level or activity of Chk2 in a cell by an amount
sufficient to cause cell
death (e.g., apoptosis) if the cell is a cancerous cell.
Again not limited by theory, E2F1 is one of related proteins in the E2F family
of nuclear
transcription factors, which family is critically important in regulation of
the cell cycle. E2F1 is
required for cellular proliferation by promoting passage through the G1/S
checkpoint. During
proliferation of normal cells, transcriptionally active E2F1 is liberated from
an inactive E2F1/Rb
complex following phosphorylation of Rb. E2F1 levels rise, promoting
progression through G1.
As the cell moves toward the end of S phase, E2F1 levels must decline for
progress to continue.
Sustained elevation of E2F1 at this point in the cell cycle causes activation
of the S phase
checkpoint, and subsequent cell death (e.g., by apoptosis). Thus, depending on
the phase of the
cell cycle and dynamics of E2F1 elevation, this regulatory protein may either
promote cellular
proliferation, induce cell cycle delay, DNA repair or cell death. During the
Gl phase of the cell
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cycle, phosphorylation of Rb results in dissociation of Rb-E2F1 complexes,
liberating active
E2F1, which then stimulates entry into S phase by promoting transcription of
key cell cycle
effectors. During S-phase, E2F1 must be degraded for progress to continue. In
the presence of
DNA damage, however, E2F1 levels increase rather than decrease, causing cell
cycle delay,
DNA repair, and, if damage is severe, cell death. As used herein, "E2F" is the
E2F transcription
factor family (including but not limited to E2F-1, E2F-2, E2F-3).
As used herein, "a cell cycle checkpoint pathway" refers to a biochemical
pathway that is
involved in modulation of a cell cycle checkpoint. A cell cycle checkpoint
pathway may have
stimulatory or inhibitory effects, or both, on one or more functions
comprising a cell cycle
checkpoint. A cell cycle checkpoint pathway is comprised of at least two
compositions of
matter, preferably proteins, both of which contribute to modulation of a cell
cycle checkpoint. A
cell cycle checkpoint pathway may be activated through an activation of one or
more members
of the cell cycle checkpoint pathway. Preferably, a cell cycle checkpoint
pathway is a
biochemical signaling pathway.
As used herein, "cell cycle checkpoint regulator" refers to a composition of
matter that
can function, at least in part, in modulation of a cell cycle checkpoint. A
cell cycle checkpoint
regulator may have stimulatory or inhibitory effects, or both, on one or more
functions
comprising a cell cycle checkpoint. In one aspect, a cell cycle checkpoint
regulator is a protein.
W another aspect, a cell cycle checkpoint regulator is a not a protein. In one
aspect, a cell cycle
checkpoint regulator is selected from the group consisting of ATM, ATR, Chkl,
Chk2, E2F1,
BRCAl, Rb, p53, p21, Mdm2, Cdc2, Cdc25, and 14-4-3[sigma].
I. Compositions
As used herein, the phrase "(3-lapachone" refers to 3,4-dihydro-2,2-dimethyl-
2H
naphtho[1,2-b]pyran-5,6-dione and derivatives and analogs thereof, and has the
chemical
structure:
Formula I
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Preferred derivatives and analogs are discussed below.
(3-Lapachone (3,4-dihydro-2, 2-dimethyl-2H-naphtho [1,2-b] pyran-5, 6-dione),
a simple
non-water soluble orthonapthoquinone, was first isolated in 1882 by Paterno
from the heartwood
of the lapacho tree (See Hoolcer, SC, (1936) I. Afra. Claem. Soc. 58:1181-
1190; Goncalves de
Lima, O, et al., (1962) Rev. Inst. Antibiot. Llniv. Recife. 4:3-17). The
structure of (3-Lapachone
was established by Hooker in 1896 and it was first synthesized by Fieser in
1927 (Hooker, SC,
(1936) I. Aria. Ghem. Soc. 58:1181-1190). (3-Lapachone can, for example, be
obtained by simple
sulfuric acid treatment of the naturally occurring lapachol, which is readily
isolated from
Tabebuia avellenedae growing mainly in Brazil, or is easily synthesized from
seeds of lomatia
growing in Australia (Li, CJ, et al., (1993) J. Biol. Claem. 268:22463-33464).
Methods for
formulating ~3-Lapachone or its derivatives or analogs can be accomplished as
described in U.S.
Patent No. 6,458,974 and U.S. Publication No. US-2003-0091639-Al.
As used herein, derivatives or analogs of (3-Lapachone include, for example,
3,4-dihydro-
2,2-dimethyl-3-(3-methyl-2-butenyl)-2H-naphtho[1,2-b]pyran-5,6-dione, 3,4-
dihydro-2,2-
dimethyl-2H-naphtho[1,2-b]thiopyran-5,6-dione and 3,4-dihydro-4,4-dimethyl-2H-
naphtho[1,2-
b]thiopyran-5,6-dione. Other derivatives or analogs of ~3-lapachone are
described in PCT
International Application PCT/US93/07878 (W094/04145), and U.S. Pat. No.
6,245,807. PCT
International Application PCT/LTS00/10169 (WO 00/61142), discloses (3-
lapachone, which may
have a variety of substituents at the 3- position as well as in place of the
methyl groups attached
at the 2-position. U.S. Patent Nos. 5,763,625, 5,824,700, and 5,969,163,
disclose analogs and
derivatives with a variety of substituents at the 2-, 3- and 4-positions.
Furthermore, a number of
journals report (3-lapachone analogs and derivatives with substituents at one
or more of the
following positions: 2-, 3-, 8- and/or 9-positions, (See, Sabba et al., (1984)
JMed Clzern 27:990-
994 (substituents at the 2-, 8- and 9- positions); (Portela and Stoppani,
(1996) Biochem Plaarm
51:275-283 (substituents at the 2- and 9- positions); Goncalves et al., (1998)
Molecular and
BioclZemical Parasitology 1:167-176 (substituents at the 2- and 3-
positions)). Other derivatives
or analogs of (3-lapachone have sulfur-containing hetero-rings in the "a" and
"[i" positions of
lapachone (I~urokawa S, (1970) Bulletin of Tlae Chefraical Society ofJapan
43:1454-1459;
Tapia, R.A ct al., (2000) Heterocycles 53(3):585-598; Tapia, RA et al., (1997)
Tetrahedron
Letters 38(1):153-154; Chuang, CP et al., (1996) Heterocycles 40(10):2215-
2221; Suginome H
et al., (1993) Journal of the Chemical Society, Chemical Conamutaicatioras
9:807-809; Tonholo J
et al., (1988) Jourfzal of the Brazilian. Chemical Society 9(2):163-169; and
I~rapcho AP et al.,
(1990) Journal ofMedicinal Chemistry 33(9):2651-2655).
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Further, derivatives or analogs of (3-lapachone include reduced (3-lapachone
(e.g.,
Formula la, in which R' and R" are both hydrogen) and derivatives of reduced
(3-lapachone
(see,e.g., Formula la, in which R' and R" are each independently hydrogen, Cl-
C6 alkyl, C1-C6
alkylcarbonyl, or a pharmaceutically acceptable salt).
Formula Ia
While [3-lapachone is the preferred Gl/S-phase compound for use in the
composition in
accordance with the present invention, the invention is not limited in this
respect, and (3-
lapachone derivatives or analogs, such as lapachol, and pharmaceutical
compositions and
formulations thereof are part of the present invention. Such (3-lapachone
analogs include those
recited in PCT International Application PCT/LJS93/07878 (WO 94/04145), which
discloses
compounds of the formula:
Z1
R2
Formula II
where R and Rl are each independently hydrogen, substituted and unsubstituted
aryl, substituted
and unsubstituted alkenyl, substituted and unsubstituted alkyl and substituted
or unsubstituted
alkoxy. The alkyl groups preferably have from 1 to about 15 carbon atoms, more
preferably
from 1 to about 10 carbon atoms, still more preferably from 1 to about 6
carbon atoms. The
term alkyl unless otherwise modified refers to both cyclic and noncyclic
groups, although of
course cyclic groups will comprise at least three carbon ring members.
Straight or branched
chain noncyclic alkyl groups are generally more preferred than cyclic groups.
Straight chain
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alkyl groups are generally more preferred than branched. The alkenyl groups
preferably have
from 2 to about 15 carbon atoms, more preferably from 2 to about 10 carbon
atoms, still more
preferably from 2 to 6 carbon atoms. Especially preferred alkenyl groups have
3 carbon atoms
(i.e., 1-propenyl or 2-propenyl), with the allyl moiety being particularly
preferred. Phenyl and
napthyl are generally preferred aryl groups. Alkoxy groups include those
alkoxy groups having
one or more oxygen linkage and preferably have from 1 to 15 carbon atoms, more
preferably
from 1 to about 6 carbon atoms. The substituted R and Rl groups may be
substituted at one or
more available positions by one or more suitable groups such as, for example,
alkyl groups such
as alkyl groups having from 1 to 10 carbon atoms or from 1 to 6 carbon atoms,
alkenyl groups
such as alkenyl groups having from 2 to 10 carbon atoms or 2 to 6 carbon
atoms, aryl groups
having from six to ten carbon atoms, halogen such as fluoro, chloro and bromo,
and N, O and S,
including heteroalkyl, e.g., heteroalkyl having one or more hetero atom
linkages (and thus
including alkoxy, aminoalkyl and thioalkyl) and from 1 to 10 carbon atoms or
from 1 to 6
carbon atoms.
Other (3-lapachone analogs contemplated in accordance with the present
invention
include those described in U.S. Patent No. 6,245,807, which discloses (3-
lapachone analogs and
derivatives having the structure:
R~
R
Formula III
where R and Rl are each independently selected from hydrogen, hydroxy,
sulfhydryl, halogen,
substituted alkyl, unsubstituted alkyl, substituted alkenyl, unsubstituted
alkenyl, substituted aryl,
unsubstituted aryl, substituted alkoxy, unsubstituted alkoxy, and salts
thereof, where the dotted
double bond between the ring carbons represents an optional ring double bond.
Additional ~i-lapachone analogs and derivatives are recited in PCT W
ternational
Application PCT/IJS00/10169 (W000/61142), which disclose compounds of the
structure:
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Formula IV
where Rj and R6 may be independently selected from hydroxy, C1-C6 alkyl, C1-C6
alkenyl, C1-
C6 alkoxy, C1-C6 alkoxycarbonyl, --(CHZ)ri phenyl; and R7 is hydrogen,
hydroxyl, C1-C6 alkyl,
C1-C6 alkenyl, CI-C6 alkoxy, C1-C6 alkoxycarbonyl, --(CH2)"-amino, --(CHZ)"-
aryl, --(CHZ)"-
heteroaryl, --(CHZ)n heterocycle, or --(CHZ)n phenyl, wherein n is an integer
from 0 to 10.
Other (3-lapachone analogs and derivatives are disclosed in U.S. Pat. No.
5,763,625, U.S.
Pat. No. 5,824,700, and U.S. Pat. No. 5,969,163, as well is in scientific
journal articles, such as
Sabba et al., JMed Chena 27:990-994 (1984), which discloses (3-lapachone with
substitutions at
one or more of the following positions: 2-, 8- and/or 9- positions. See also
Portela et al.,
Bioclzem Pharm 51:275-283 (1996) (substituents at the 2- and 9- positions);
Maruyama et al.,
Chem Lett 847-850 (1977); Sun et al., Tetrahedron Lett 39:8221-8224 (1998);
Goncalves et al.,
Molecular and Biochemical Parasitology 1:167-176 (1998) (substituents at the 2-
and 3-
positions); Gupta et al., Indian Journal of Chemistry 16B: 35-37 (1978); Gupta
et al., CuYr~ S'ei
46:337 (1977) (substituents at the 3- and 4- positions); DiChenna et al., JMed
Chem 44: 2486-
2489 (2001) (monoarylamino derivatives).
More preferably, analogs and derivatives contemplated by the present
application are
intended to encompass compounds having the general formula V and VI:
O
R7
Formula V Formula VI
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where the dotted double bond between the ring carbons represents an optional
ring double bond
and where R and Rl are each independently selected from hydrogen, hydroxy,
sulfhydryl,
halogen, substituted alkyl, unsubstituted alkyl, substituted alkenyl,
unsubstituted alkenyl,
substituted aryl, unsubstituted aryl, substituted alkoxy, unsubstituted
alkoxy, and salts thereof.
The alkyl groups preferably have from 1 to about 15 carbon atoms, more
preferably from 1 to
about 10 carbon atoms, still more preferably from 1 to about 6 carbon atoms.
The term alkyl
refers to both cyclic and noncyclic groups. Straight or branched chain
noncyclic alkyl groups
are generally more preferred than cyclic groups. Straight chain alkyl groups
are generally more
preferred than branched. The alkenyl groups preferably have from 2 to about 15
carbon atoms,
more preferably from 2 to about 10 carbon atoms, still more preferably from 2
to 6 carbon
atoms. Especially preferred alkenyl groups have 3 carbon atoms (i.e., 1-
propenyl or 2-
propenyl), with the allyl moiety being particularly preferred. Phenyl and
napthyl are generally
preferred aryl groups. Alkoxy groups include those alkoxy groups having one or
more oxygen
linkage and preferably have from 1 to 15 carbon atoms, more preferably from 1
to about 6
carbon atoms. The substituted R and Rl groups may be substituted at one or
more available
positions by one or more suitable groups such as, for example, alkyl groups
having from 1 to 10
carbon atoms or from 1 to 6 carbon atoms, alkenyl groups having from 2 to 10
carbon atoms or
2 to 6 carbon atoms, aryl groups having from six to ten carbon atoms, halogen
such as fluoro,
chloro and bromo, and N, O and S, including heteroalkyl, e.g., heteroalkyl
having one or more
hetero atom linkages (and thus including alkoxy, aminoalkyl and thioalkyl) and
from 1 to 10
carbon atoms or from 1 to 6 carbon atoms; and where RS and R6 may be
independently selected
from hydroxy, C1-C6 alkyl, C1-C~ alkenyl, C1-C6 alkoxy, C1-C6 alkoxycarbonyl, -
-(CHZ)"-aryl, --
(CHa)"-heteroaryl, --(CHZ)n-heterocycle, or --(CHZ)"-phenyl; and R7 is
hydrogen, hydroxyl, Cl-
C6 alkyl, C1-C6 alkenyl, C1-C6 alkoxy, Cl-C6 alkoxycarbonyl, --(CHZ)"-amino, --
(CHZ)"-aryl, --
(CHa)"-heteroaryl, --(CH2)"-heterocycle, or --(CHZ)"-phenyl, wherein n is an
integer from 0 to
10.
Preferred analogs and derivatives also contemplated by the invention include
compounds
of the following general formula VII:
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Formula VII
where Rl is (CH2)"-RZ, where n is an integer from 0-10 and R2 is hydrogen, an
alkyl, an aryl, a
heteroaromatic, a heterocyclic, an aliphatic, an alkoxy, an allyloxy, a
hydroxyl, an amine, a thiol,
an amide, or a halogen.
Analogs and derivatives also contemplated by the invention include 4-acetoxy-
(3-
lapachone, 4-acetoxy-3-bromo-(3-lapachone, 4-keto-(3-lapachone, 7-hydroxy-a-
lapachone, 7-
methoxy-(3-lapachone, 8-hydroxy-(3-lapachone, 8-methoxy-~-lapachone, 8-chloro-
(3-lapachone,
9-chloro-(3-lapachone, 8-methyl-[3-lapachone and 8,9-dimethoxy-(3-lapachone.
Preferred analogs and derivatives also contemplated by the invention include
compounds
of the following general formula VIII:
Z2
Formula VIII
where R1-R4 are each, independently, selected from the group consisting of H,
CI-C6 alkyl, C1-
C6 alkenyl, C1-C6 alkoxy, C1-C6 alkoxycarbonyl, --(CH2)"-aryl, --(CHZ)"-
heteroaryl, --(CH2)"-
heterocycle, or --(CHZ)n phenyl; or Rl and R2 combined are a single
substituent selected from
the above group, and R3 and Rq combined are a single substituent selected from
the above
groups, in which case ---- is a double bond.
Preferred analogs and derivatives also contemplated by this invention include
dunnione
and 2-ethyl-6-hydroxynaphtho[2,3-b]-furan-4,5-dione.
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Preferred analogs and derivatives also contemplated by the invention include
compounds
of the following general formula IX:
Formula IX
where Rl is selected from H, CH3, OCH3 and N02.
Additional preferred [3-lapachone analogs useful in the methods and kits of
the invention
are represented by Formula X (see also the co-owned PCT patent application
entitled "NOVEL
LAPACHONE COMPOUNDS AND METHODS OF USE THEREOF", PCT/US2003/037219,
filed November 18, 2003, and claiming priority to U.S. provisional application
no. 60/427,283,
filed November 18, 2002):
Formula X
or pharmaceutically acceptable salts thereof, or a regioisomeric mixture
thereof, wherein
Rl-R6 are each, independently, selected from the group consisting of H, OH,
substituted and
unsubstituted Cl-C~ alkyl, substituted and unsubstituted C1-C6 alkenyl,
substituted and
unsubstituted C~-C6 alkoxy, substituted and unsubstituted C1-C6
alkoxycarbonyl, substituted and
unsubstituted Cl-C~ acyl, -(CH2)ri amino, -(CHa)"-aryl, -(CH2)"-heterocycle,
and -(CH2)"-phenyl;
or one of R1 or R2 and one of R3 or R4; or one of R3 or R4 and one of RS or R6
form a fused
ring, wherein the ring has 4-8 ring members; R7-R10 are each, independently,
hydrogen,
hydroxyl, halogen, substituted or unsubstituted alkyl, substituted or
unsubstituted alkoxy, nitro,
cyano or amide; and n is an integer from 0 to 10.
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In a preferred embodiment, Rl and R2 are alkyl, R3-R6 are, independently, H,
OH,
halogen, alkyl, all~oxy, substituted or unsubstituted acyl, substituted
alkenyl or substituted alkyl
carbonyl, and R7-R10 are hydrogen. In another preferred embodiment, Rl and R2
are each
methyl and R3-R10 are each hydrogen. In another preferred embodiment, Rl-R4
are each
hydrogen, RS and R6 are each methyl and R7-R10 are each hydrogen.
Additional preferred (3-lapachone analogs useful in the methods and kits of
the invention
are represented by Formula XI (see also the co-owned PCT patent application
entitled "NOVEL
LAPACHONE COMPOUNDS AND METHODS OF USE THEREOF", PCT/US2003/037219,
filed November 18, 2003):
Formula XI
or pharmaceutically acceptable salts thereof, or a regioisomeric mixture
thereof, wherein
Rl-R4 are each, independently, selected from the group consisting of H, OH,
substituted and
unsubstituted Cl-C6 alkyl, substituted and unsubstituted C1-C6 alkenyl,
substituted and
unsubstituted C1-C6 alkoxy, substituted and unsubstituted C1-C6
alkoxycarbonyl, substituted and
unsubstituted Cl-C6 acyl, -(CHZ)"-amino, -(CH2)"-aryl, -(CHa)"-heterocycle,
and -(CH2)"-phenyl;
or one of Rl or R~ and one of R3 or R4 form a fused ring, wherein the ring has
4-8 ring
members; RS-R8 axe each, independently, hydrogen, hydroxyl, halogen,
substituted or
unsubstituted allcyl, substituted or unsubstituted alkoxy, nitro, cyano or
amide; and n is an
integer from 0 to 10. In certain embodiments of Formula XI, Rl, R2, R3, R4,
R5, R6, R7 and
R8 are not each simultaneously H.
All stereoisomers of the compounds of the instant invention are contemplated,
either in
admixture or in pure or substantially pure form. The definition of the
compounds according to
the invention embraces all possible stereoisomers (e.g., the R and S
configurations for each
asymmetric center) and their mixtures. It very particularly embraces the
racemic forms and the
isolated optical isomers having a specified activity. The racemic forms can be
resolved by
physical methods, such as, for example, fractional crystallization, separation
or crystallization of
diastereomeric derivatives or separation by chiral column chromatography. The
individual
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optical isomers can be obtained from the racemates by conventional methods,
such as, for
example, salt formation with an optically active acid followed by
crystallization. Furthermore,
all geometric isomers, such as E- and Z-configurations at a double bond, are
within the scope of
the invention unless otherwise stated. Certain compounds of this invention may
exist in
tautomeric forms. All such tautomeric forms of the compounds are considered to
be within the
scope of this invention unless otherwise stated. The present invention also
includes one or more
regioisomeric mixtures of an analog or derivative of (3-Lapachone.
As used herein, the term "salt" is a pharmaceutically acceptable salt and can
include acid
addition salts including hydrochlorides, hydrobromides, phosphates, sulphates,
hydrogen
sulphates, alkylsulphonates, arylsulphonates, acetates, benzoates, citrates,
maleates, fumarates,
succinates, lactates, and tartrates; alkali metal canons such as Na, K, Li,
alkali earth metal salts
such as Mg or Ca, or organic amine salts.
As used herein, the term "metabolite" means a product of metabolism of [3-
lapachone, or
a pharmaceutically acceptable salt, analog or derivative thereof, that
exhibits a similar activity ifz
vivo to [3-lapachone.
As used herein, the term "prodrug" means a compound of the present invention
covalently linlced to one or more pro-moieties, such as an amino acid moiety
or other water
solubilizing moiety. A compound of the present invention may be released from
the pro-moiety
via hydrolytic, oxidative, and/or enzymatic release mechanisms. In an
embodiment, a prodrug
composition of the present invention exhibits the added benefit of increased
aqueous solubility,
improved stability, and improved phannacokinetic profiles. The pro-moiety may
be selected to
obtain desired prodrug characteristics. For example, the pro-moiety, e.g., an
amino acid moiety
or other water solubilizing moiety may be selected based on solubility,
stability, bioavailability,
and/or in vivo delivery or uptake.
II. Methods of Treatment
As used herein, a "subject" can be any mammal, e.g., a human, a primate,
mouse, rat,
dog, cat, cow, horse, pig, sheep, goat, camel. In a preferred aspect, the
subject is a human.
As used herein, a "subject in need thereoF' is a subject having a cell
proliferative
disorder of the present invention, or a subject having an increased risk of
developing a cell
proliferative disorder of the present invention relative to the population at
large. In one aspect, a
subject in need thereof has a precancerous condition of the present invention.
In a preferred'
aspect, a subject in need thereof has a cancer of the present invention. In an
aspect, the subject
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may be suffering from a known (i.e., diagnosed) condition characterized by
cell
hyperproliferation (e.g., cancer).
As used herein, the term "cell proliferative disorder" refers to conditions in
which
unregulated or abnormal growth, or both, of cells can lead to the development
of an unwanted
condition or disease, which may or may not be cancerous. In one aspect, a cell
proliferative
disorder includes, for example, cancer and precancerous conditions of the
present invention. For
example, a "cell proliferative disorder of the liver" is a cell proliferative
disorder involving cells
of the liver. In one aspect, a cell proliferative disorder includes a pre-
cancer or precancerous
condition of the liver. In one aspect, a cell proliferative disorder of the
present invention
includes a non-cancerous cell proliferative disorder. In another aspect, a
cell proliferative
disorder includes a cancer of the present invention, including metastatic
lesions in other tissues
or organs distant from the primary tumor site. In one aspect, a "precancer
cell" or "precancerous
cell" is a cell manifesting a cell proliferative disorder that is a precancer
or a precancerous
condition. In another aspect, a "cancer cell" or "cancerous cell" is a cell
manifesting a cell
proliferative disorder that is a cancer. Any reproducible means of measurement
may be used to
identify cancer cells or precancerous cells. In a preferred aspect, cancer
cells or precancerous
cells are identified by histological typing or grading of a tissue sample
(e.g., a biopsy sample). In
another aspect, cancer cells or precancerous cells are identified through the
use of appropriate
molecular markers.
In a preferred aspect, the cell proliferative disorder of the present
invention is a cancer of
the present invention. In a preferred aspect, compositions of the present
invention may be used
to treat cancers of the present invention or cell proliferative disorders of
the present invention.
In one aspect, cancer includes all forms of cancer of the present invention.
Cancers to be treated
include but are not limited to sarcoma, carcinoma, and adenocarcinoma. In one
aspect, cell
proliferative disorders include hyperplasia, metaplasia, and dysplasia. In one
aspect, cell
proliferative disorders of the present invention include benign tumors.
In one aspect, a cancer of the present invention that is to be treated has
arisen in a subject
equal to or older than 30 years old, or a subject younger than 30 years old.
In one aspect, a
cancer of the present invention that is to be treated has arisen in a subject
equal to or older than
50 years old, or a subject younger than 50 years old. In one aspect, a a
cancer of the present
invention that is to be treated has arisen in a subject equal to or older than
70 years old, or a
subject younger than 70 years old. In one aspect, a cancer of the present
invention has been
typed to identify a familial or spontaneous mutation in p53, Rb, myc, or ras.
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In one aspect, a cancer of the present invention is associated with a negative
regional
lymph node biopsy. In one aspect, a cancer of the present invention is
associated with a positive
regional lymph node biopsy. W another aspect, a cancer of the present
invention has been typed
as having nodal negative status (e.g., node-negative) or nodal positive status
(e.g., node-
s positive). In another aspect, a cancer of the present invention has
metastasized to other locations
in the body. In one aspect, a cancer of the present invention is classified as
having metastasized
to a location selected from the group consisting of lymph node, stomach, bile
duct, lung, liver,
bone, and brain. In another aspect a cancer of the present invention is
classified according to a
characteristic selected from the group consisting of metastatic, limited
stage, extensive stage,
unresectable, resectable, localized, regional, local-regional, locally
advanced, distant,
multicentric, bilateral, ipsilateral, contralateral, newly diagnosed,
recurrent, and inoperable.
In one aspect, a cancer of the present invention has been staged according to
the
American Joint Committee on Cancer (AJCC) TNM classification system, where the
tumor (T)
has been assigned a stage of Tis, T1, T2, T3,T4; and where the regional lymph
nodes (I~ have
been assigned a stage of NX, N0, N1, N2, N2a, N2b, N3, N3a, N3b, or N3c; and
where distant
metastasis (M) has been assigned a stage of MX, M0, or Ml. In another aspect,
a cancer of the
present invention has been staged according to an American Joint Committee on
Cancer (AJCC)
classification as Stage 0, I, IA, IB, II, IIA, IIB, III, IIIA, IIIB, IIIC and
IV cancer. In another
aspect, a cancer of the present invention has been assigned a grade according
to an AJCC
classification as Grade GX (e.g., grade cannot be assessed), Grade 1, Grade 2,
Grade 3 or Grade
4.
In one aspect, a cancer of the present invention includes a tumor that has
been
determined to be less than or equal to about 2 centimeters in diameter. In one
aspect, a cancer of
the present invention includes a tumor that has been determined to be less
than or equal to about
3 centimeters in diameter. In another aspect, a cancer of the present
invention includes a tumor
that has been determined to be from about 3 to about 5 centimeters in
diameter. In another
aspect, a cancer of the present invention includes a tumor that has been
determined to be greater
than or equal to about 3 centimeters in diameter. In another aspect, a cancer
of the present
invention includes a tumor that has been determined to be greater than 5
centimeters in diameter.
In another aspect, a cancer of the present invention is classified by
microscopic appearance as
well differentiated, moderately differentiated, poorly differentiated, or
undifferentiated. In
another aspect, a cancer of the present invention is classified by microscopic
appearance with
respect to mitosis count (e.g., amount of cell division) or nuclear
pleiomorphism (e.g., change in
cells). In another aspect, a cancer of the present invention is classified by
microscopic
CA 02556794 2006-08-17
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appearance as being associated with areas of necrosis (e.g., areas of dying or
degenerating cells).
In one aspect, a cancer of the present invention is classified as having an
abnormal karyotype,
having an abnormal number of chromosomes, or having one or more chromosomes
that are
abnormal in appearance. In one aspect, a cancer of the present invention is
classified as being
aneuploid, triploid, tetraploid, or as having an altered ploidy. In one
aspect, a cancer of the
present invention is classified as having a chromosomal translocation, or a
deletion or
duplication of an entire chromosome, or a region of deletion, duplication or
amplification of a
portion of a chromosome.
In one aspect, a cancer of the present invention is evaluated by DNA
cytometry, flow
cytometry, or image cytometry. In one aspect, a cancer of the present
invention has been typed
as having 10%, 20%, 30%, 40%, 50%, 60%, 70%, ~0%, or 90% of cells in the
synthesis stage of
cell division (e.g., in S phase of cell division). In one aspect, a cancer of
the present invention
has been typed as having a low S-phase fraction or a high S-phase fraction.
As used herein, a "normal cell" is a cell that cannot be classified as part of
a "cell
proliferative disorder." In one aspect, a normal cell lacks unregulated or
abnormal growth, or
both, that can lead to the development of an unwanted condition or disease.
Preferably, a
normal cell possesses normally functioning cell cycle checkpoint control
mechanisms.
As used herein, "contacting a cell" refers to a condition in which a compound
or other
composition of matter is in direct contact with a cell, or is close enough to
induce a desired
biological effect in a cell.
As used herein, "monotherapy" refers to administration of a single active or
therapeutic
compound to a subject in need thereof. Preferably, monotherapy will involve
administration of
a therapeutically effective amount of an active compound. For example, [3-
lapachone
monotherapy for cancer comprises administration of a therapeutically effective
amount of (3-
lapachone, or a pharmaceutically acceptable salt, prodrug, metabolite, analog
or derivative
thereof, to a subj ect in need of treatment of cancer. Monotherapy may be
contrasted with
combination therapy, in which a combination of multiple active compounds is
administered,
preferably with each component of the combination present in a therapeutically
effective
amount. In one aspect, (3-lapachone montherapy is more effective than
combination therapy in
inducing a desired biological effect.
In one aspect, combination therapy includes [3-lapachone with taxol; [3-
lapachone with
docetaxel; (3-lapachone with vincristin; (3-lapachone with vinblastin; [3-
lapachone with
nocodazole; (3-lapachone with teniposide; [3-lapachone with etoposide; (3-
lapachone with
adriamycin; (3-lapachone with epothilone; (3-lapachone with navelbine; (3-
lapachone with
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camptothecin; (3-lapachone with daunorubicin; (3-lapachone with dactinomycin;
[3-lapachone
with mitoxantrone; (3-lapachone with amsacrine; (3-lapachone with epirubicin;
or (3-lapachone
with idarubicin. In a preferred aspect, combination therapy includes (3-
lapachone with
gemcitabine. In another aspect, combination therapy includes reduced (3-
lapachone with taxol;
reduced (3-lapachone with docetaxel; reduced (3-lapachone with vincristin;
reduced (3-lapachone
with vinblastin; reduced j3-lapachone with nocodazole; reduced (3-lapachone
with teniposide;
reduced (3-lapachone with etoposide; reduced [3-lapachone with adriamycin;
reduced (3-
lapachone with epothilone; reduced (3-lapachone with navelbine; reduced (3-
lapachone with
camptothecin; reduced [3-lapachone with daunorubicin; reduced (3-lapachone
with dactinomycin;
reduced (3-lapachone with mitoxantrone; reduced (3-lapachone with amsacrine;
reduced (3-
lapachone with epirubicin; or reduced (3-lapachone with idarubicin. In a
preferred aspect,
combination therapy includes reduced (3-lapachone with gemcitabine.
As used herein, "treating" describes the management and care of a patient for
the
purpose of combating a disease, condition, or disorder and includes the
administration of a
compound of the present invention to prevent the onset of the symptoms or
complications,
alleviating the syrnptorns or complications, or eliminating the disease,
condition or disorder.
In one aspect, treating a cancer of the present invention results in a
reduction in size of a
tumor. A reduction in size of a tumor may also be referred to as "tumor
regression." Preferably,
after treatment, tumor size is reduced by 5% or greater relative to its size
prior to treatment;
more preferably, tumor size is reduced by 10% or greater; more preferably,
reduced by 20% or
greater; more preferably, reduced by 30% or greater; more preferably, reduced
by 40% or
greater; even more preferably, reduced by 50% or greater; and most preferably,
reduced by
greater than 75% or greater. Size of a tumor may be measured by any
reproducible means of
measurement. In a preferred aspect, size of a tumor may be measured as a
diameter of the
tumor.
In another aspect, treating a cancer of the present invention results in a
reduction in
tumor volume. Preferably, after treatment, tumor volume is reduced by 5% or
greater relative to
its size prior to treatment; more preferably, tumor volume is reduced by 10%
or greater; more
preferably, reduced by 20% or greater; more preferably, reduced by 30% or
greater; more
preferably, reduced by 40% or greater; even more preferably, reduced by 50% or
greater; and
most preferably, reduced by greater than 75% or greater. Tumor volume may be
measured by
any reproducible means of measurement.
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In another aspect, treating a cancer of the present invention results in a
decrease in
number of tumors. Preferably, after treatment, tumor number is reduced by 5%
or greater
relative to number prior to treatment; more preferably, tumor number is
reduced by 10% or
greater; more preferably, reduced by 20% or greater; more preferably, reduced
by 30% or
greater; more preferably, reduced by 40% or greater; even more preferably,
reduced by 50% or
greater; and most preferably, reduced by greater than 75%. Number of tumors
may be
measured by any reproducible means of measurement. In a preferred aspect,
number of tumors
may be measured by counting tumors visible to the naked eye or at a specified
magnification. In
a preferred aspect, the specified magnification is 2x, 3x, 4x, Sx, l Ox, or
50x.
In another aspect, treating a cancer of the present invention results in a
decrease in
number of metastatic lesions in other tissues or organs distant from the
primary tumor site.
Preferably, after treatment, the number of metastatic lesions is reduced by 5%
or greater relative
to number prior to treatment; more preferably, the number of metastatic
lesions is reduced by
10% or greater; more preferably, reduced by 20% or greater; more preferably,
reduced by 30%
or greater; more preferably, reduced by 40% or greater; even more preferably,
reduced by 50%
or greater; and most preferably, reduced by greater than 75%. The number of
metastatic lesions
may be measured by any reproducible means of measurement. 111 a preferred
aspect, the number
of metastatic lesions may be measured by counting metastatic lesions visible
to the naked eye or
at a specified magnification. In a preferred aspect, the specified
magnification is 2x, 3x, 4x, Sx,
10x, or 50x.
In another aspect, treating a cancer of the present invention results in an
increase in
average survival time of a population of treated subj ects in comparison to a
population receiving
Garner alone. Preferably, the average survival time is increased by more than
30 days; more
preferably, by more than 60 days; more preferably, by more than 90 days; and
most preferably,
by more than 120 days. An increase in average survival time of a population
may be measured
by any reproducible means. In a preferred aspect, an increase in average
survival time of a
population may be measured, for example, by calculating for a population the
average length of
survival following initiation of treatment with an active compound. In an
another preferred
aspect, an increase in average survival time of a population may also be
measured, for example,
by calculating for a population the average length of survival following
completion of a first
round of treatment with an active compound.
In another aspect, treating a cancer of the present invention results in an
increase in
average survival time of a population of treated subjects in comparison to a
population of
untreated subjects. Preferably, the average survival time is increased by more
than 30 days;
23
CA 02556794 2006-08-17
WO 2005/082358 PCT/US2005/005647
more preferably, by more than 60 days; more preferably, by more than 90 days;
and most
preferably, by more than 120 days. An increase in average survival time of a
population may be
measured by any reproducible means. In a preferred aspect, an increase in
average survival time
of a population may be measured, for example, by calculating for a population
the average
length of survival following initiation of treatment with an active compound.
In an another
preferred aspect, an increase in average survival time of a population may
also be measured, for
example, by calculating for a population the average length of survival
following completion of
a first round of treatment with an active compound.
In another aspect, treating a cancer of the present invention results in
increase in average
survival time of a population of treated subjects in comparison to a
population receiving
monotherapy with a drug that is not (3-lapachone, or a pharmaceutically
acceptable salt,
metabolite, analog or derivative thereof. Preferably, the average survival
time is increased by
more than 30 days; more preferably, by more than 60 days; more preferably, by
more than 90
days; and most preferably, by more than 120 days. An increase in average
survival time of a
population may be measured by any reproducible means. In a preferred aspect,
an increase in
average survival time of a population may be measured, for example, by
calculating for a
population the average length of survival following initiation of treatment
with an active
compound. In an another preferred aspect, an increase in average survival time
of a population
may also be measured, for example, by calculating for a population the average
length of
survival following completion of a first round of treatment with an active
compound.
In another aspect, treating a cancer of the present invention results in a
decrease in the
mortality rate of a population of treated subjects in comparison to a
population receiving carrier
alone. In another aspect, treating a cancer of the present invention results
in a decrease in the
mortality rate of a population of treated subjects in comparison to an
untreated population. In a
further aspect, treating a cancer of the present invention results a decrease
in the mortality rate of
a population of treated subjects in comparison to a population receiving
monotherapy with a
drug that is not (3-lapachone, or a pharmaceutically acceptable salt,
metabolite, analog or
derivative thereof. Preferably, the mortality rate is decreased by more than
2%; more preferably,
by more than 5%; more preferably, by more than 10%; and most preferably, by
more than 25%.
In a preferred aspect, a decrease in the mortality rate of a population of
treated subjects may be
measured by any reproducible means. In another preferred aspect, a decrease in
the mortality
rate of a population may be measured, for example, by calculating for a
population the average
number of disease-related deaths per unit time following initiation of
treatment with an active
compound. In another preferred aspect, a decrease in the mortality rate of a
population may also
24
CA 02556794 2006-08-17
WO 2005/082358 PCT/US2005/005647
be measured, for example, by calculating for a population the average number
of disease-related
deaths per unit time following completion of a first round of treatment with
an active compound.
In another aspect, treating a cancer of the present invention results in a
decrease in tumor
growth rate. Preferably, after treatment, tumor growth rate is reduced by at
least 5% relative to
number prior to treatment; more preferably, tumor growth rate is reduced by at
least 10%; more
preferably, reduced by at least 20%; more preferably, reduced by at least 30%;
more preferably,
reduced by at least 40%; more preferably, reduced by at least 50%; even more
preferably,
reduced by at least 50%; and most preferably, reduced by at least 75%. Tumor
growth rate may
be measured by any reproducible means of measurement. In a preferred aspect,
tumor growth
rate is measured according to a change in tumor diameter per unit time.
In another aspect, treating a cancer of the present invention results in a
decrease in tumor
regrowth. Preferably, after treatment, tumor regrowth is less than 5%; more
preferably, tumor
regrowth is less than 10%; more preferably, less than 20%; more preferably,
less than 30%;
more preferably, less than 40%; more preferably, less than 50%; even more
preferably, less than
50%; and most preferably, less than 75%. Tumor regrowth may be measured by any
reproducible means of measurement. In a preferred aspect, tumor regrowth is
measured, for
example, by measuring an increase in the diameter of a tumor after a prior
tumor shrinkage that
followed treatment. In another preferred aspect, a decrease in tumor regrowth
is indicated by
failure of tumors to reoccur after treatment has stopped.
In another aspect, treating or preventing a cell proliferative disorder of the
present
invention results in a reduction in the rate of cellular proliferation.
Preferably, after treatment,
the rate of cellular proliferation is reduced by at least 5%; more preferably,
by at least 10%;
more preferably, by at least 20%; more preferably, by at least 30%; more
preferably, by at least
40%; more preferably, by at least 50%; even more preferably, by at least 50%;
and most
preferably, by at least 75%. The rate of cellular proliferation may be
measured by any
reproducible means of measurement. In a preferred aspect, the rate of cellular
proliferation is
measured, for example, by measuring the number of dividing cells in a tissue
sample per unit
tune.
In another aspect, treating or preventing a cell proliferative disorder of the
present
invention results in a reduction in the proportion of proliferating cells.
Preferably, after
treatment, the proportion of proliferating cells is reduced by at least 5%;
more preferably, by at
least 10%; more preferably, by at least 20%; more preferably, by at least 30%;
more preferably,
by at least 40%; more preferably, by at least 50%; even more preferably, by at
least 50%; and
most preferably, by at least 75 %. The proportion of proliferating cells may
be measured by any
CA 02556794 2006-08-17
WO 2005/082358 PCT/US2005/005647
reproducible means of measurement. In a preferred aspect, the proportion of
proliferating cells
is measured, for example, by quantifying the number of dividing cells relative
to the number of
nondividing cells in a tissue sample. In another preferred aspect, the
proportion of proliferating
cells is equivalent to the mitotic index.
In another aspect, treating or preventing a cell proliferative disorder of the
present
invention results in a decrease in size of an area or zone of cellular
proliferation. Preferably,
after treatment, size of an area or zone of cellular proliferation is reduced
by at least 5% relative
to its size prior to treatment; more preferably, reduced by at least 10%; more
preferably, reduced
by at least 20%; more preferably, reduced by at least 30%; more preferably,
reduced by at least
40%; more preferably, reduced by at least 50%; even more preferably, reduced
by at least 50%;
and most preferably, reduced by at least 75%. Size of an area or zone of
cellular proliferation
may be measured by any reproducible means of measurement. In a preferred
aspect, size of an
area or zone of cellular proliferation may be measured as a diameter or width
of an area or zone
of cellular proliferation.
hi another aspect, treating or preventing a cell proliferative disorder of the
present
invention results in a decrease in the number or proportion of cells having an
abnormal
appearance or morphology. Preferably, after treatment, the number of cells
having an abnormal
morphology is reduced by at least 5% relative to its size prior to treatment;
more preferably,
reduced by at least 10%; more preferably, reduced by at least 20%; more
preferably, reduced by
at least 30%; more preferably, reduced by at least 40%; more preferably,
reduced by at least
50%; even more preferably, reduced by at least 50%; and most preferably,
reduced by at least
75%. An abnormal cellular appearance or morphology may be measured by any
reproducible
means of measurement. In one aspect, an abnornlal cellular morphology is
measured by
microscopy, e.g., using an inverted tissue culture microscope. In one aspect,
an abnormal
cellular morphology takes the fonn of nuclear pleiomorphism.
As used herein, the term "selectively" means tending to occur at a higher
frequency in
one population than in another population. In one aspect, the compared
populations are cell
populations. In a preferred aspect, (3-lapachone, or a pharmaceutically
acceptable salt,
metabolite, analog or derivative thereof, acts selectively on a cancer or
precancer cell but not on
a normal cell. In another preferred aspect, a compound of the present
invention, or a
pharmaceutically acceptable salt, prodrug, metabolite, analog or derivative
thereof, acts
selectively to modulate ~ne molecular target (e.g., E2F-1) but does not
significantly modulate
another molecular target (e.g., Protein I~inase C). In another preferred
aspect, the invention
provides a method for selectively inhibiting the activity of an enzyme, such
as a kinase.
26
CA 02556794 2006-08-17
WO 2005/082358 PCT/US2005/005647
Yreterably, an event occurs selectively in population A relative to population
B if it occurs
greater than two times more frequently in population A as compared to
population B. More
preferably, an event occurs selectively if it occurs greater than five times
more frequently in
population A. More preferably, an event occurs selectively if it occurs
greater than ten times
more frequently in population A; more preferably, greater than fifty times;
even more
preferably, greater than 100 times; and most preferably, greater than 1000
times more frequently
in population A as compared to population B. For example, cell death would be
said to occur
selectively in cancer cells if it occurred greater than twice as frequently in
cancer cells as
compared to normal cells.
hi a preferred aspect, a compound of the present invention or a
pharmaceutically
acceptable salt, prodrug, metabolite, analog or derivative thereof, modulates
the activity of a
molecular target (e.g., E2F-1). In one aspect, modulating refers to
stimulating or inhibiting an
activity of a molecular target. Preferably, a compound of the present
invention modulates the
activity of a molecular target if it stimulates or inhibits the activity of
the molecular target by at
least 10% relative to the activity of the molecular target under the same
conditions but lacking
only the presence of said compound. More preferably, a compound of the present
invention
modulates the activity of a molecular target if it stimulates or inhibits the
activity of the
molecular target by at least 25%, at least 5~%, at least 2-fold, at least 5-
fold, at least 10-fold, at
least 20-fold, at least 50-fold, at least 100-fold relative to the activity of
the molecular target
under the same conditions but lacking only the presence of said compound. The
activity of a
molecular target may be measured by any reproducible means. The activity of a
molecular
target may be measured in vitro or in vivo. For example, the activity of a
molecular target may
be measured in vitro by an enzymatic activity assay or a DNA binding assay, or
the activity of a
molecular target may be measured in vivo by assaying for expression of a
reporter gene.
In one aspect, a compound of the present invention, or a pharmaceutically
acceptable
salt, prodrug, metabolite, analog or derivative thereof, does not
significantly modulate the
activity of a molecular target if the addition of the compound stimulates or
inhibits the activity
of the molecular target by less than 10% relative to the activity of the
molecular target under the
same conditions but lacking only the presence of said compound.
As used herein, the term "isozyme selective" means preferential inhibition or
stimulation
of a first isoform of an enzyme in comparison to a second isoform of an enzyme
(e.g.,
preferential inhibition or stimulation of a kinase isozyme alpha in comparison
to a kinase
isozyme beta). Preferably, a compound of the present invention demonstrates a
minimum of a
four fold differential, preferably a ten fold differential, more preferably a
fifty fold differential,
27
CA 02556794 2006-08-17
WO 2005/082358 PCT/US2005/005647
in the dosage required to achieve a biological effect. Preferably, a compound
of the present
invention demonstrates this differential across the range of inhibition, and
the differential is
exemplified at the ICSO, i.e., a 50% inhibition, for a molecular target of
interest.
In a preferred embodiment, administering (3-lapachone, or a pharmaceutically
acceptable
salt, prodrug, metabolite, analog or derivative thereof, to a cell or a
subject in need thereof
results in modulation (i.e., stimulation or inhibition) of an activity of a
member of the E2F
family of transcription factors (e.g., E2F-l, E2F-2, or E2F-3). As used
herein, an activity of a
member of the E2F family of transcription factors refers to any biological
function or activity
that is carried out by an E2F family member. For example, a function of E2F-1
includes binding
of E2F-1 to its cognate DNA sequences. Other functions of E2F-1 include
migrating to the cell
nucleus and activating transcription.
In one aspect, treating a cancer or a cell proliferative disorder of the
present invention
results in cell death, and preferably, cell death results in a decrease of at
least 10% in number of
cells in a population. More preferably, cell death means a decrease of at
least 20%; more
preferably, a decrease of at least 30%; more preferably, a decrease of at
least 40%; more
preferably, a decrease of at least 50%; most preferably, a decrease of at
least 75%. Number of
cells in a population may be measured by any reproducible means. In one
aspect, number of
cells in a population is measured by fluorescence activated cell sorting
(FACS). In another
aspect, number of cells in apopulation is measured by immunofluorescence
microscopy. In
another aspect, number of cells in a population is measured by light
microscopy. In another
aspect, methods of measuring cell death are as shown in Li et al., (2003)
Pr~oc Natl Acad Sci U S
A. 100(5): 2674-8. In a preferred aspect, cell death results from apoptosis.
In a preferred aspect, an effective amount of (3-lapachone, or a
pharmaceutically
acceptable salt, metabolite, analog or derivative thereof is not significantly
cytotoxic to normal
cells. A therapeutically effective amount of a compound is not significantly
cytotoxic to normal
cells if administration of the compound at a therapeutically effective amount
does not induce
apoptosis in greater than 10% of normal cells. A therapeutically effective
amount of a
compound does not significantly affect the viability of normal cells if
administration of the
compound at a therapeutically effective amount does not induce cell death in
greater than 10%
of normal cells.
In one aspect, activating refers to placing one or more compositions of matter
(e.g.,
protein or nucleic acid) in a state suitable for carrying out a desired
biological function. In one
aspect, a composition of matter capable of being activated also has an
unactivated state. In one
28
CA 02556794 2006-08-17
WO 2005/082358 PCT/US2005/005647
aspect, an activated composition of matter may have an inhibitory or
stimulatory biological
function, or both.
In one aspect, elevation refers to an increase in a desired biological
activity of a
composition of matter (e.g., a protein or a nucleic acid). In one aspect,
elevation may occur
through an increase in concentration of a composition of matter.
In one aspect, stimulation of unscheduled expression of a checkpoint molecule
by (3-
lapachone, or a pharmaceutically acceptable salt, metabolite, analog or
derivative thereof,
triggers cell death in cells with defective checkpoints, a hallmark of cancer
and pre-cancer cells.
In one aspect, contacting a cell with (3-lapachone, or a pharmaceutically
acceptable salt,
metabolite, analog or derivative thereof, stimulates unscheduled expression of
the checkpoint
molecule E2F.
In one aspect, contacting a cell with (3-lapachone, or a pharmaceutically
acceptable salt,
metabolite, analog or derivative thereof, results in activation of an E2F
checkpoint pathway.
Preferably, administering to a subject in need thereof [3-lapachone, or a
pharmaceutically
acceptable salt, metabolite, analog or derivative thereof, results in
activation of an E2F
checkpoint pathway. W a preferred aspect, E2F pathway activity is increased by
more than
10%; more than 25%; more than 50%; more than 2-fold; more than 5-fold; and
most preferably,
by more than 10-fold. In another preferred aspect, E2F activity is increased
by more than 10%;
more than 25%; more than 50%; more than 2-fold; more than 5-fold; and most
preferably, by
more than 10-fold. Methods of measuring induction of E2F activity and
elevation of E2F levels
are as shown in Li et al., (2003) Pf°oc Natl Acad Sei U S A. 100(5):
2674-8.
In one aspect, contacting a cell with (3-lapachone, or a pharmaceutically
acceptable salt,
metabolite, analog or derivative thereof, results in elevation of an E2F
transcription factor.
Preferably, administering to a subject in need thereof (3-lapachone, or a
pharmaceutically
acceptable salt, metabolite, analog or derivative thereof, results in
elevation of an E2F
transcription factor.
In one aspect, contacting a cell with (3-lapachone, or a pharmaceutically
acceptable salt,
metabolite, analog or derivative thereof, results in elevation of an E2F
transcription factor
selectively in cancer cells but not in normal cells. Preferably, administering
to a subject in need
thereof [3-lapachone, or a pharmaceutically acceptable salt, metabolite,
analog or derivative
thereof, results in elevation of an E2F transcription factor selectively in
cancer cells but not in
normal cells.
In one aspect, contacting a cell with [3-lapachone, or a pharmaceutically
acceptable salt,
metabolite, analog or derivative thereof, stimulates unscheduled activation of
an E2F
29
CA 02556794 2006-08-17
WO 2005/082358 PCT/US2005/005647
transcription factor. Preferably, administering to a subject in need thereof
~3-lapachone, or a
pharmaceutically acceptable salt, metabolite, analog or derivative thereof,
stimulates
unscheduled activation of an E2F transcription factor.
In one aspect, contacting a cell with (3-lapachone, or a pharmaceutically
acceptable salt,
metabolite, analog or derivative thereof, stimulates unscheduled activation of
an E2F
transcription factor selectively in cancer cells but not in normal cells.
Preferably, administering
to a subject in need thereof [3-lapachone, or a pharmaceutically acceptable
salt, metabolite,
analog or derivative thereof, stimulates unscheduled activation of an E2F
transcription factor
selectively in cancer cells but not in normal cells.
In normal cells with their intact regulatory mechanisms, imposed expression of
a
checkpoint molecule (e.g., as induced by contacting a cell with (3-lapachone,
or a
pharmaceutically acceptable salt, metabolite, analog or derivative thereof )
results in an
expression pattern that is not reported to be of substantial consequence. In
contrast, cancer and
pre-cancer cells have defective mechanisms, which result in unchecked or
persistent expression,
or both, of unscheduled checkpoint molecules, e.g., E2F, leading to selective
cell death in cancer
and pre-cancer cells. The present invention includes and provides for the
unchecked or
persistent expression, or both, of unscheduled checkpoint molecules by the
administration of [3-
lapachone, or a pharmaceutically acceptable salt, metabolite, analog or
derivative thereof.
In one aspect, contacting a cell with ~i-lapachone, or a pharmaceutically
acceptable salt,
metabolite, analog or derivative thereof, results in activation of one or more
cell cycle
checkpoints. Preferably, administering to a subject in need thereof (3-
lapachone, or a
pharmaceutically acceptable salt, metabolite, analog or derivative thereof,
results in activation of
one or more cell cycle checlcpoints.
In one aspect, contacting a cell with (3-lapachone, or a pharmaceutically
acceptable salt,
metabolite, analog or derivative thereof, results in activation of one or more
cell cycle
checkpoint pathways. Preferably, administering to a subject in need thereof (3-
lapachone, or a
pharmaceutically acceptable salt, metabolite, analog or derivative thereof,
results in activation of
one or more cell cycle checkpoint pathways.
In one aspect, contacting a cell with (3-lapachone, or a pharmaceutically
acceptable salt,
metabolite, analog or derivative thereof, results in activation of one or more
cell cycle
checkpoint regulators. Preferably, administering to a subject in need thereof
(3-lapachone, or a
pharmaceutically acceptable salt, metabolite, analog or derivative thereof,
results in activation of
one or more cell cycle checkpoint regulators.
CA 02556794 2006-08-17
WO 2005/082358 PCT/US2005/005647
In one aspect, contacting a cell with [3-lapachone, or a pharmaceutically
acceptable salt,
metabolite, analog or derivative thereof, induces or activates cell death
selectively in cancer
cells. Preferably, administering to a subject in need thereof (3-lapachone, or
a pharmaceutically
acceptable salt, metabolite, analog or derivative thereof, induces or
activates cell death
selectively in cancer cells. In another aspect, contacting a cell with (3-
lapachone, or a
pharmaceutically acceptable salt, metabolite, analog or derivative thereof,
induces cell death
selectively in one or more cells affected by a cell proliferative disorder of
the present invention.
Preferably, administering to a subject in need thereof (3-lapachone, or a
pharmaceutically
acceptable salt, metabolite, analog or derivative thereof, induces cell death
selectively in one or
more cells affected by a cell proliferative disorder of the present invention.
In a preferred aspect, the present invention relates to a method of treating
or preventing
cancer by administering (3-lapachone, or a pharmaceutically acceptable salt,
metabolite, analog
or derivative thereof to a subject in need thereof, where administration of
the (3-lapachone, or a
pharmaceutically acceptable salt, metabolite, analog or derivative thereof
results in one or more
of the following: accumulation of cells in Gl and/or S phase of the cell
cycle, cytotoxicity via
cell death in cancer cells but not in normal cells, antitumor activity in
animals with a therapeutic
index of at least 2, and acitvation of a cell cycle checkpoint (e.g.,
activation or elevation of a
member of the E2F family of transcription factors). As used herein,
"therapeutic index" is the
maximum tolerated dose divided by the efficacious dose.
In additional aspects, (3-lapachone, or a pharmaceutically acceptable salt,
metabolite,
analog or derivative thereof, can be administered in combination with a
chemotherapeutic agent.
Exemplary chemotheraputics with activity against cell proliferative disorders,
such as cancer,
are known to those of ordinary skill in the art, and may be found in reference
texts such as the
P7aysiciaya's Desk Refe~etzce, 59th Edition, Thomson PDR (2005). For example,
the
chemotherapeutic agent can be a taxane, an aromatase inhibitor, an
anthracycline, a microtubule
targeting drug, a topoisomerase poison drug, a targeted monoclonal or
polyconal antibody, an
inhibitor of a molecular target or enzyme (e.g., a kinase inhibitor), or a
cytidine analogue drug.
In preferred aspects, the chemotherapeutic agent can be, but is not restricted
to, tamoxifen,
raloxifene, anastrozole, exemestane, letrozole, HERCEPTIN°
(trastuzumab), GLEEVEC~
(imatanib), TAXOL~ (paclitaxel), IRESSA" (gefitinib), TARCEVATM (erlotinib),
cyclophosphamide, lovastatin, minosine, araC, 5-fluorouracil (5-FU),
methotrexate (MTX),
TAXOTERE~ (docetaxel), ZOLADEX~ (goserelin), vincristin, vinblastin,
nocodazole,
teniposide, etoposide, epothilone, navelbine, camptothecin, daunonibicin,
dactinomycin,
mitoxantrone, amsacrine, doxorubicin (adriamycin), epirubicin or idarubicin or
agents listed in
31
CA 02556794 2006-08-17
WO 2005/082358 PCT/US2005/005647
www.cancer.org/docroot/cdg/cdg_O.asp. In another aspect, the chemotherapeutic
agent can be a
cytokine such as G-CSF (granulocyte colony stimulating factor). In another
aspect, [3-
lapachone, or a pharmaceutically acceptable salt, metabolite, analog or
derivative thereof may be
administered in combination with radiation therapy. In yet another aspect, (3-
lapachone, or a
pharmaceutically acceptable salt, metabolite, analog or derivative thereof may
be administered
in combination with standard chemotherapy combinations such as, but not
restricted to, CMF
(cyclophosphamide, methotrexate and 5-fluorouracil), CAF (cyclophosphamide,
adriamycin and
5-fluorouracil), AC (adriamycin and cyclophosphamide), FEC (5-fluorouracil,
epirubicin, and
cyclophosphamide), ACT or ATC (adriamycin, cyclophosphamide, and paclitaxel),
or CMFP
(cyclophosphamide, methotrexate, 5-fluorouracil and prednisone).
In a preferred aspect, a cell proliferative disorder of the present invention,
such as cancer,
is treated by administering to a patient in need thereof a therapeutically
effective amount of (3-
lapachone, or a pharmaceutically acceptable salt, metabolite, analog or
derivative thereof, in
combination with a therapeutically effective amount of gemcitabine. GEMZAR~
(gemcitabine
HCl) is 2'-deoxy-2',2'-difluorocytidine monohydrochloride ((3-isomer), a
nucleoside analog that
exhibits antitumor activity. Gemcitabine may be used in monotherapy, or in
combination with
other agents (e.g., cisplatin, carboplatin, TAXOL~ (paclitaxel)), to treat
various cancers,
including pancreatic cancer, breast cancer, non-small cell lung cancer,
ovarian cancer, and
bladder cancer. Gemcitabine exhibits cell phase specificity, primarily killing
cells undergoing
DNA synthesis (S-phase) and also blocking the progression of cells through the
Gl/S boundary.
Without being limited by theory, it is believed that after a gemcitabine
nucleotide is incorporated
into DNA, only one additional nucleotide may be added to the growing DNA
strands. Again not
limited by theory, it is believed that DNA polymerase epsilon is unable to
remove the
gemcitabine nucleotide and repair the growing DNA strand (e.g., masked chain
termination). In
CEM T lymphoblastoid cells, gemcitabine induces internucleosomal DNA
fragmentation, one of
the characteristics of programmed cell death (e.g., apoptosis).
One skilled in the art may refer to general reference texts for detailed
descriptions of
known techniques discussed herein or equivalent techniques. These texts
include Ausubel et al.,
Curf-ent Protocols in Moleculaf° Biology, John Wiley and Sons, Inc.
(2005); Sambrook et al.,
Molecular Cloning, A Laboratory Manual (3d ed.), Cold Spring Harbor Press,
Cold Spring
Harbor, New Yorlc (2000); Coligan et al., Current Protocols in Immunology;
John Wiley &
Sons, N.Y.; Emla et al., Current Pf°otocols in Phar~rrzacology, John
Wiley & Sons, N.Y.; Fingl et
al., The Plaaf~macological Basis of Therapeutics (1975), Remington's
Plza~maceutical Sciences,
32
CA 02556794 2006-08-17
WO 2005/082358 PCT/US2005/005647
Mack Publishing Co., Easton, PA, 18th edition (1990). These texts can, of
course, also be
referred to in making or using an aspect of the invention.
A compound of the present invention, or a pharmaceutically acceptable salt,
prodrug,
metabolite, analog or derivative thereof, can be incorporated into
pharmaceutical compositions
suitable for administration. Such compositions typically comprise the compound
(i.e., including
the active compound), and a pharmaceutically acceptable excipient or Garner.
As used herein,
"pharmaceutically acceptable excipient" or "pharmaceutically acceptable
carrier" is intended to
include any and all solvents, dispersion media, coatings, antibacterial and
antifungal agents,
isotonic and absorption delaying agents, and the like, compatible with
pharmaceutical
administration. Suitable carriers are described in the most recent edition of
Remington's
Pharmaceutical Sciences, a standard reference text in the field. Preferred
examples of such
carriers or diluents include, but are not limited to, water, saline, ringer's
solutions, dextrose
solution, and 5% human serum albumin. Pharmaceutically acceptable carriers
include solid
carriers such as lactose, terra alba, sucrose, talc, gelatin, agar, pectin,
acacia, magnesium
stearate, stearic acid and the like. Exemplary liquid Garners include syrup,
peanut oil, olive oil,
water and the like. Similarly, the carrier or diluent may include time-delay
material known in the
art, such as glyceryl monostearate or glyceryl distearate, alone or with a
wax, ethylcellulose,
hydroxypropylmethylcellulose, methylmethacrylate or the like. Other fillers,
excipients,
flavorants, and other additives such as are known in the art may also be
included in a
pharmaceutical composition according to this invention. Liposomes and non-
aqueous vehicles
such as fixed oils may also be used. The use of such media and agents for
pharmaceutically
active substances is well known in the art. Except insofar as any conventional
media or agent is
incompatible with the active compound, use thereof in the compositions is
contemplated.
Supplementary active compounds can also be incorporated into the compositions.
The pharmaceutical compositions of this invention which are provided as part
of the
combination therapies may exist in the dosage form as a solid, semi-solid, or
liquid such as, e.g.,
suspensions, aerosols or the like. Preferably the compositions are
administered in unit dosage
forms suitable for single administration of precise dosage amounts. The
compositions may also
include, depending on the formulation desired, pharmaceutically-acceptable,
nontoxic carriers or
diluents, which are defined as vehicles commonly used to formulate
pharmaceutical
compositions for animal or human administration. The diluent is selected so as
not to affect the
biological activity of the combination. Examples of such diluents are
distilled water,
physiological saline, Ringer's solution, dextrose solution, and Hank's
solution. A preferred
carrier for the solubilization of (3-lapachone is hydroxypropyl beta
cyclodextrin, a water
33
CA 02556794 2006-08-17
WO 2005/082358 PCT/US2005/005647
solubilizing carrier molecule. Other water-solubilizing agents for combining
with (3-lapachone
and/or an S-phase compound, such as Poloxamer, Povidone K17, Povidone K12,
Tween 80,
ethanol, Cremophor/ethanol, polyethylene glycol 400, propylene glycol and
Trappsol, are
contemplated. Furthermore, the invention is not limited to water-solubilizing
agents, and oil-
s based solubilizing agents such as lipiodol and peanut oil, may also be used.
In addition, the pharmaceutical composition or formulation may also include
other
Garners, adjuvants, or nontoxic, nontherapeutic, nonimmunogenic stabilizers
and the like.
Effective amounts of such diluent or carrier will be those amounts which are
effective to obtain a
pharmaceutically acceptable formulation in terms of solubility of components,
or biological
activity, and the like. Liposome formulations, are also contemplated by the
present invention,
and have been described. See, e.g., U.S. Pat. No. 5,424,073.
For the purposes of the present invention, the Gl/S phase drugs or compounds,
or
derivatives or analogs thereof, and the S phase drugs or compounds, or
derivatives or analogs
thereof, described herein include their pharmacologically acceptable salts,
preferably sodium;
analogs containing halogen substitutions, preferably chlorine or fluorine;
analogs containing
ammonium or substituted armnonium salts, preferably secondary or tertiary
ammonium salts;
analogs containing alkyl, alkenyl, aryl or their alkyl, alkenyl, aryl, halo,
alkoxy, alkenyloxy
substituted derivatives, preferably methyl, methoxy, ethoxy, or phenylacetate;
and natural
analogs such as naphthyl acetate. Further, the Gl/S phase compounds or
derivatives or analogs
thereof, and the S phase compounds or derivatives or analogs thereof,
described herein may be
conjugated to a water-soluble polymer or may be derivatized with water-soluble
chelating agents
or radionuclides. Examples of water soluble polymers are, but not limited to:
polyglutamic acid
polymer, copolymers with polycaprolactone, polyglycolic acid, polyactic acid,
polyacrylic acid,
poly (2-hydroxyethyl 1-glutamine), carboxymethyl dextran, hyaluronic acid,
human serum
albumin, polyalginic acid or a combination thereof. Examples of water-soluble
chelating agents
are, but not limited to: DIPA (diethylenetriaminepentaacetic acid), EDTA,
DTTP, DOTA or
their water-soluble salts, etc. Examples of radionuclides include, but not
limited to: 111In, ~oY,
166H~' 68Ga' 99mTC, and the like.
Due to the water insolubility of [3-lapachone, pharmaceutical carriers or
solubilizing
agents may be used to provide sufficient quantities of [3-lapachone for use in
the treatment
methods of the present invention. See, e.g., U.S. Patent Publication
20030091639 to Jiang et al.,
and U.S. Patent Publication 20040001871 to Boothman et al. This publication
describes the use
of complexing agents such as cyclodextrins, including hydroxypropyl beta-
cyclodextrin
(HPBCD), to permit the solubilization of [3-lapachone at levels sufficient for
administration. See
34
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WO 2005/082358 PCT/US2005/005647
also U.S. Patent Publication 20040001871 to Bootlnnan et al. In an embodiment,
the Gl/S
phase drug, or an analog or derivative thereof, is administered with a
pharmaceutically
acceptable water solubilizing carrier molecule selected from the group
consisting of Poloxamer,
Povidone K17, Povidone I~12, Tween 80, ethanol, Cremophor/ethanol,
polyethylene glycol
(PEG) 400, propylene glycol, Trappsol, alpha-cyclodextrin or derivatives or
analogs thereof,
beta-cyclodextrin or derivatives or analogs thereof, and gamma-cyclodextrin or
derivatives or
analogs thereof.
In one aspect, a compound of the present invention, or a pharmaceutically
acceptable
salt, prodrug, metabolite, analog or derivative thereof, is administered in a
suitable dosage form
prepared by combining a therapeutically effective amount (e.g., an efficacious
level sufficient to
achieve the desired therapeutic effect through inhibition of tumor growth,
killing of tumor cells,
treatment or prevention of cell proliferative disorders, etc.) of a compound
of the present
invention, or a pharmaceutically acceptable salt, prodrug, metabolite, analog
or derivative
thereof, (as an active ingredient) with standard pharmaceutical carriers or
diluents according to
conventional procedures (i.e., by producing a pharmaceutical composition of
the invention).
These procedures may involve mixing, granulating, and compressing or
dissolving the
ingredients as appropriate to attain the desired preparation.
A pharmaceutical composition of the invention is formulated to be compatible
with its
intended route of administration. Examples of routes of administration include
parenteral, e.g.,
intravenous, intradermal, subcutaneous, oral (e.g., inhalation), transdermal
(topical), and
transmucosal administration. Although intravenous administration is preferred
as discussed
above, the invention is not intended to be limited in this respect, and the
compounds can be
administered by any means known in the art. Such modes include oral, rectal,
nasal, topical
(including buccal and sublingual) or parenteral (including subcutaneous,
intramuscular,
intravenous and intradermal) administration. For ease of administration and
comfort to the
patient, oral administration is generally preferred. However, oral
administration may require the
administration of a higher dose than intravenous administration. The skilled
artisan can
determine which form of administration is best in a particular case, balancing
dose needed
versus the number of times per month administration is necessary. Solutions or
suspensions
used for parenteral, intradermal, or subcutaneous application can include the
following
components: a sterile diluent such as water for injection, saline solution,
fixed oils, polyethylene
glycols, glycerine, propylene glycol or other synthetic solvents;
antibacterial agents such as
benzyl alcohol or methyl parabens; antioxidants such as ascorbic acid or
sodium bisulfate;
chelating agents such as ethylenediaminetetraacetic acid; buffers such as
acetates, citrates or
CA 02556794 2006-08-17
WO 2005/082358 PCT/US2005/005647
phosphates, and agents for the adjustment of tonicity such as sodium chloride
or dextrose. The
pH can be adjusted with acids or bases, such as hydrochloric acid or sodium
hydroxide. The
parenteral preparation can be enclosed in ampoules, disposable syringes or
multiple dose vials
made of glass or plastic.
Solutions or suspensions used for parenteral, intradermal, or subcutaneous
application
can include the following components: a sterile diluent such as water for
injection, saline
solution, fixed oils, polyethylene glycols, glycerine, propylene glycol or
other synthetic solvents;
antibacterial agents such as benzyl alcohol or methyl parabens; antioxidants
such as ascorbic
acid or sodium bisulfate; chelating agents such as ethylenediaminetetraacetic
acid; buffers such
as acetates, citrates or phosphates, and agents for the adjustment of tonicity
such as sodium
chloride or dextrose. The pH can be adjusted with acids or bases, such as
hydrochloric acid or
sodium hydroxide. The parenteral preparation can be enclosed in ampoules,
disposable syringes
or multiple dose vials made of glass or plastic.
A compound or pharmaceutical composition of the invention can be administered
to a
subject in many of the well-known methods currently used for chemotherapeutic
treatment. For
example, for treatment of cancers, a compound of the invention may be injected
directly into
tumors, inj ected into the blood stream or body cavities or taken orally or
applied through the
skin with patches. The dose chosen should be sufficient to constitute
effective treatment but not
so high as to cause unacceptable side effects. The state of the disease
condition (e.g., cancer,
precancer, and the like) and the health of the patient should preferably be
closely monitored
during and for a reasonable period after treatment.
The term "therapeutically effective amount," as used herein, refers to an
amount of a
pharmaceutical agent to treat, ameliorate, or prevent an identified disease or
condition, or to
exhibit a detectable therapeutic or inhibitory effect. The effect can be
detected by any assay
method known in the art. The precise effective amount for a subject will
depend upon the
subject's body weight, size, and health; the nature and extent of the
condition; and the
therapeutic or combination of therapeutics selected for administration.
Therapeutically effective
amounts for a given situation can be determined by routine experimentation
that is within the
skill and judgment of the clinician. In a preferred aspect, the disease or
condition to be treated is
cancer. In another aspect, the disease or condition to be treated is a cell
proliferative disorder.
For any compound, the therapeutically effective amount can be estimated
initially either
in cell culture assays, e.g., of neoplastic cells, or in animal models,
usually rats, mice, rabbits,
dogs, or pigs. The animal model may also be used to determine the appropriate
concentration
range and route of administration. Such information can then be used to
determine useful doses
36
CA 02556794 2006-08-17
WO 2005/082358 PCT/US2005/005647
and routes for administration in humans. Therapeutic/prophylactic efficacy and
toxicity may be
'determined by standard pharmaceutical procedures in cell cultures or
experimental animals, e.g.,
EDso (the dose therapeutically effective in 50% of the population) and LDSO
(the dose lethal to
50% of the population). The dose ratio between therapeutic and toxic effects
is the therapeutic
index, and it can be expressed as the ratio, EDSO/LDso. Pharmaceutical
compositions that exhibit
large therapeutic indices are preferred. The dosage may vary within this range
depending upon
the dosage form employed, sensitivity of the patient, and the route of
admiriistration.
Dosage and administration are adjusted to provide sufficient levels of the
active agents)
or to maintain the desired effect. Factors which may be taken into account
include the severity
of the disease state, general health of the subject, age, weight, and gender
of the subject, diet,
time and frequency of administration, drug combination(s), reaction
sensitivities, and
tolerance/response to therapy. Long-acting pharmaceutical compositions may be
administered
every 3 to 4 days, every week, or once every two weeks depending on half-life
and clearance
rate of the particular formulation.
The pharmaceutical compositions containing active compounds of the present
invention
may be manufactured in a manner that is generally known, e.~., by means of
conventional
mixing, dissolving, granulating, dragee-making, levigating, emulsifying,
encapsulating,
entrapping, or lyophilizing processes. Pharmaceutical compositions may be
formulated in a
conventional manner using one or more pharmaceutically acceptable carriers
comprising
excipients and/or auxiliaries that facilitate processing of the active
compounds into preparations
that can be used pharmaceutically. Of course, the appropriate formulation is
dependent upon the
route of administration chosen.
Pharmaceutical compositions suitable for injectable use include sterile
aqueous solutions
(where water soluble) or dispersions and sterile powders for the
extemporaneous preparation of
sterile injectable solutions or dispersion. For intravenous administration,
suitable carriers
include physiological saline, bacteriostatic water, Cremophor ELTM (BASF,
Parsippany, N.J.) or
phosphate buffered saline (PBS). In all cases, the composition must be sterile
and should be
fluid to the extent that easy syringeability exists. It must be stable under
the conditions of
manufacture and storage and must be preserved against the contaminating action
of
microorganisms such as bacteria and fungi. 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), and suitable mixtures thereof. 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. Prevention of the
37
CA 02556794 2006-08-17
WO 2005/082358 PCT/US2005/005647
action of microorganisms can be achieved by various antibacterial and
antifungal agents, for
example, parabens, chlorobutanol, phenol, ascorbic acid, thimerosal, and the
like. In many
cases, it will be preferable to include isotonic agents, for example, sugars,
polyalcohols such as
manitol, sorbitol, sodium chloride in the composition. Prolonged absorption of
the injectable
compositions can be brought about by including in the composition an agent
which delays
absorption, for example, aluminum monostearate and gelatin.
Sterile injectable solutions can be prepared by incorporating the active
compound in the
required amount in an appropriate solvent with one or a combination of
ingredients enumerated
above, as required, followed by filtered sterilization. Generally, dispersions
are prepared by
incorporating the active compound into a sterile vehicle that contains a basic
dispersion medium
and the required other ingredients from those enumerated above. In the case of
sterile powders
for the preparation of sterile injectable solutions, methods of preparation
are vacuum drying and
freeze-drying that yields a powder of the active ingredient plus any
additional desired ingredient
from a previously sterile-filtered solution thereof.
Oral compositions generally include an inert diluent or an edible
pharmaceutically
acceptable carrier. They can be enclosed in gelatin capsules or compressed
into tablets. For the
purpose of oral therapeutic administration, the active compound can be
incorporated with
excipients and used in the form of tablets, troches, or capsules. Oral
compositions can also be
prepared using a fluid carrier for use as a mouthwash, wherein the compound in
the fluid carrier
is applied orally and swished and expectorated or swallowed. Pharmaceutically
compatible
binding agents, and/or adjuvant materials can be included as part of the
composition. The
tablets, pills, capsules, troches and the like can contain any of the
following ingredients, or
compounds of a similar nature: a binder such as microcrystalline cellulose,
gum tragacanth or
gelatin; an excipient such as starch or lactose, a disintegrating agent such
as alginic acid,
Primogel, or com starch; a lubricant such as magnesium stearate or Sterotes; a
glidant such as
colloidal silicon dioxide; a sweetening agent such as sucrose or saccharin; or
a flavoring agent
such as peppermint, methyl salicylate, or orange flavoring.
For administration by inhalation, the compounds are delivered in the form of
an aerosol
spray from pressured container or dispenser, which contains a suitable
propellant, e.g., a gas
such as carbon dioxide, or a nebulizer.
Systemic administration can also be by transmucosal or transdermal means. For
transmucosal or transdermal administration, penetrants appropriate to the
barrier to be permeated
are used in the formulation. Such penetrants are generally known in the art,
and include, for
example, for transmucosal administration, detergents, bile salts, and fusidic
acid derivatives.
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WO 2005/082358 PCT/US2005/005647
Transmucosal administration can be accomplished through the use of nasal
sprays or
suppositories. For transdermal administration, the active compounds are
formulated into
ointments, salves, gels, or creams as generally known in the art.
In one aspect, the active compounds are prepared with pharmaceutically
acceptable
carriers that will protect the compound against rapid elimination from the
body, such as a
controlled release formulation, including implants and microencapsulated
delivery systems.
Biodegradable, biocompatible polymers can be used, such as ethylene vinyl
acetate,
polyanhydrides, polyglycolic acid, collagen, polyorthoesters, and polylactic
acid. Methods for
preparation of such formulations will be apparent to those skilled in the art.
The materials can
also be obtained commercially from Alza Corporation and Nova Pharmaceuticals,
Inc.
Liposomal suspensions (including liposomes targeted to infected cells with
monoclonal
antibodies to viral antigens) can also be used as pharmaceutically acceptable
carriers. These can
be prepared according to methods known to those skilled in the art, for
example, as described in
U.S. Pat. No. 4,522,811.
It is especially advantageous to formulate oral or parenteral 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 subject
to be treated; each unit
containing a predetermined quantity of active compound calculated to produce
the desired
therapeutic effect in association with the required pharmaceutical carrier.
The specification for
the dosage unit forms of the invention are dictated by and directly dependent
on the unique
characteristics of the active compound and the particular therapeutic effect
to be achieved.
In therapeutic applications, the dosages of the pharmaceutical compositions
used in
accordance with the invention vary depending on the agent, the age, weight,
and clinical
condition of the recipient patient, and the experience and judgment of the
clinician or
practitioner administering the therapy, among other factors affecting the
selected dosage.
Generally, the dose should be sufficient to result in slowing, and preferably
regressing, the
growth of the tumors and also preferably causing complete regression of the
cancer. Dosages
can range from about 0.01 mg/kg per day to about 3000 mg/kg per day. In
preferred aspects,
dosages can range from about 1 mg/kg per day to about 1000 mg/kg per day. In
an asp ect, the
dose will be in the range of about 0.1 mg/day to about 70 g/day; about 0.1
mg/day to about 25
g/day; about 0.1 mg/day to about 10 g/day; about 0.1 mg to about 3g/day; or
about 0.1 rng to
about 1 g/day, in single, divided, or continuous doses (which dose may be
adjusted for the
patient's weight in kg, body surface area in m2, and age in years).
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WO 2005/082358 PCT/US2005/005647
An effective amount of a pharmaceutical agent is that which provides an
objectively
identifiable improvement as noted by the clinician or other qualified
observer. For example,
regression of a tumor in a patient may be measured with reference to the
diameter of a tumor.
Decrease in the diameter of a tumor indicates regression. Regression is also
indicated by failure
of tumors to reoccur after treatment has stopped. As used herein, the term
"dosage effective
manner" refers to amount of an active compound to produce the desired
biological effect in a
subject or cell.
A compound of the present invention may be administered in combination with an
S
phase compound, such as an gemcitabine, in any manner found appropriate by a
clinician in
generally accepted efficacious dose ranges, such as those described in the
Physician's Desk
RefeYence, 59t~' Edition, Thomson PDR (2005)("PDR"). In general, gemcitabine
is administered
intravenously at dosages from about 10 mg/mz to about 10,000 mg/m2, preferably
from about
100 mg/mZto about 2000 mg/m2, and most preferably about 500 to about 1500
mglm2. In an
embodiment, gemcitabine is administered intravenously at a dosage from
approximately 100
mg/m2 to about 2000 mg/m2. In an embodiment, gemcitabine is administered
intravenously at a
dosage of approximately 1000 mg/m2. Dosage can be repeated, e.g., once weekly,
preferably for
about 1 to 6 weeks. It is preferred that dosages be administered over a time
period of about 30
minutes to about 6 hours, and typically over a period of about 3 hours.
The S phase drug, such as an gemcitabine, will be administered in a similar
regimen with
a G1/S phase drug, such as (3-lapachone or an analog or derivative thereof,
although the amounts
will preferably be reduced from that normally administered. It is preferred,
for example, that the
gemcitabine be administered at the same time or after the (3-lapachone has
administered to the
patient. When the gemcitabine is administered after the (3-lapachone, the
gemcitabine is
advantageously administered about 24 hours after the (3-lapachone has been
administered.
The combination therapy agents described herein may be administered singly and
sequentially, or in a cocktail or combination containing both agents or one of
the agents with
other therapeutic agents, including but not limited to, immunosuppressive
agents, potentiators
and side-effect relieving agents. As aforesaid, the therapeutic combination,
if administered
sequentially, may be more effective when the Gl/S phase drug component (e.g.,
(3-lapachone) is
administered prior to the S phase drug, e.g., gemcitabine. For example, a dose
of the G1/S phase
drug component (e.g., [3-lapachone) is administered at least one hour (more
preferably at least 2
hours, 4 hours, 8 hours, 12 hours, or 24 hours) prior to administration of a
dose of the S phase
drug, e.g., gemcitabine. In another embodiment, a dose of the G1/S phase drug
component (e.g.,
[3-lapachone) is administered at least one hour (more preferably at least 2
hours, 4 hours, ~
CA 02556794 2006-08-17
WO 2005/082358 PCT/US2005/005647
hours, 12 hours, or 24 hours) following administration of a dose of the S
phase drug, e.g.,
gemcitabine. The therapeutic agents will preferably be administered
intravenously or otherwise
systemically by injection intramuscularly, subcutaneously, intrathecally or
intraperitoneally. In
an embodiment, the S phase drug is administered simultaneously with or
following
administration of the Gl/S phase drug. In another embodiment, the S phase drug
is administered
following administration of the G1/S phase drug. In another embodiment, the S
drug is
administered within 24 hours after the G1/S phase drug is administered.
The other component of the combination therapy for combination with the S
phase drug
or compound is the G1/S phase drug, which is preferably (3-lapachone or an
analog or derivative
thereof.
(3-lapachone has been shown to have a variety of pharmacological effects. (3-
lapachone
has been shown to be a DNA repair inhibitor which sensitizes cells to DNA
damaging agents
(Boorstein, R.J., et al., (1984) Biochem. Bi~plays. Res. Cofmnun., 118:828-
834; Boothman, D.A.,
et al., (1989) J. Cancer Res., 49:605-612). (3-lapachone is generally well-
tolerated in dogs, rats,
and mice.
The present invention provides a method of treating cancer or a precancerous
condition
or preventing cancer in a subject, the method comprising administering to the
subject a
therapeutically effective amount of a pharmaceutical composition comprising (3-
lapachone, or a
derivative or analog thereof, or pharmaceutically acceptable salt thereof, or
a metabolite thereof,
and a pharmaceutically acceptable Garner such that the composition maintains a
plasma
concentration of about 0.15 ~.M to about 50 ~M and treats the cancer or
precancerous condition
or prevents the cancer. In one aspect, the plasma concentration can be about
0.1 ~M to about
100 ~.M, about 0.125 ~.M to about 75 ~M; about 0.15 p,M to about 50 ~M; about
0.175 ~,M to
about 30 ~,M; and about 0.2 ~,M to about 20 p,M. In another aspect, the
pharmaceutical
composition can maintain a suitable plasma concentration for at least a month,
at least a week, at
least 24 hours, at least 12 hrs, at least 6 hrs, at least 1 hour. In a further
aspect, a suitable plasma
concentration of the pharmaceutical composition can be maintained
indefinitely. In yet another
aspect, the subject can be exposed to the pharmaceutical composition in a AUC
(area under the
curve) range of about 0.5 ~M-hr to about 100 pM-hr, about 0.5 pM-hr to about
50 ~,M-hr, about
1 ~.M-hr to about 25 ~,M-hr, about 1 ~.M-hr to about 10 ~,M-hr; about 1.25 ~.M-
hr to about 6.75
~.M-hr, about 1.5 ~M-hr to about 6.5 ~,M-hr. The pharmaceutical composition
can be
administered at a dosage from about 2 mg/m2 to 5000 mg/m2 per day, more
preferably from
about 20 mg/m2 to 2000 mg/m2 per day, more preferably from about 20 mg/m2 to
500 mg/rn2 per
day, most preferably from about 30 to 300 mg/m~' per day. Preferably, 2 mg/ma
to 5000 rng/m2
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WO 2005/082358 PCT/US2005/005647
per day is the administered dosage for a human. In another aspect, the
pharmaceutical
composition can be administered at a dosage from about 10 to 1,000,000 ~,g per
kilogram body
weight of recipient per day; preferably about 100 to 500,000 ~,g per kilogram
body weight of
recipient per day, more preferably from about 1000 to 250,000 ~,g per kilogram
body weight of
recipient per day, most preferably from about 10,000 to 150,000 ~,g per
kilogram body weight of
recipient per day. One of ordinary skill in the art can determine the
appropriate dosage amount
in mg/mz per day or ~.g per kilogram body weight of recipient per day
depending on subject to
which the pharmaceutical composition is to be administered.
As with the use of other chemotherapeutic drugs, the individual patient will
be monitored
in a manner deemed appropriate by the treating physician. Dosages can also be
reduced if
severe neutropenia or severe peripheral neuropathy occurs, or if a grade 2 or
higher level of
mucositis is observed, using the Common Toxicity Criteria of the National
Cancer Institute.
In administering a G1/S phase compound such as (3-lapachone, the normal dose
of such
compound individually is utilized as set forth above. However, when
combination therapies are
used, it is preferable to use a lower dosage -- preferably 75% or less of the
individual amount,
more preferably 50% or less, still more preferably 40% or less. The term
"effective amount," as
used herein, refers to an amount effective to treat the disease condition in
combination with any
other active agent in a combination regimen according to the invention.
In therapeutic applications, the dosages of the agents used in accordance with
the
invention vary depending on the agent, the age, weight, and clinical condition
of the recipient
patient, and the experience and judgment of the clinician or practitioner
administering the
therapy, among other factors affecting the selected dosage. Generally, the
dose should be
sufficient to result in slowing, and preferably regressing, the growth of the
tumors and also
preferably causing complete regression of the cancer. An effective amount of a
pharmaceutical
agent is that which provides an objectively identifiable improvement as noted
by the clinician or
other qualified observer. Regression of a tumor in a patient is typically
measured with reference
to the diameter of a tumor. Decrease in the diameter of a tumor indicates
regression. Regression
is also indicated by failure of tumors to reoccur after treatment has stopped.
In preferred
embodiments, a decrease in tumor size or burden of at least 20%, more
preferably 50%, 80%,
90%, 95% or 99% is preferred.
The pharmaceutical compositions can be included in a container, pack, or
dispenser
together with instructions for administration.
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Example 1.
Exponentially growing cells are seeded at 250, 1000, or 5000 cells per well
(2.5 ml) in
six-well plates and allowed to attach for 24 hours. ~i-lapachone is dissolved
at a concentration of
20 mM in DMSO and diluted in complete media. (3-lapachone (0.5 ml) is added at
6-fold the
final concentration to a total volume of 3.0 ml/well. Control plates receive
the same volume of
DMSO alone. After a 4 hour exposure the drug is carefully removed, and drug-
free medium
may be added. Cultures are left undisturbed for 14-21 days to allow for colony
formation and
then are fixed and stained with crystal violet stain (Sigma). Colonies of
greater than 50 cells are
scored as survivors. Cells are maintained at 37 °C in S% COz in
complete humidity. See, e.g.,
Table 1.
Table 1
Tissue Origin.Cell Line LCso (wlV~
a licate Results
Pancreatic PaCa-2 1.68, 1.65, 1.52
BxPC-3 1.89, 2.09
Colon HT-29 2.19, 2.07, 1.99
HCT-116 2.29, 2.13
Prostate PC-3 2.13, 2.34
DU-145 1.10, 1.21
Ovarian SIB-OV-3 1.18, 1.29
CAOV-3 1.56, 1.44
Lun A549 2.07, 1.81, 1.79
Breast MCF-7 1.45, 1.56
Melanoma SK-MEL-28 2.40, 2.63
Example 2.
[3-lapachone is tested in the NCI in vitro screen of 60 cancer cell lines,
which allows
comparison with other anti-tumor agents under standardized conditions. The NCI
assays are
performed under standardized conditions not designed to mimic the conditions
of dosing and use
the sulforhodamine B assay as the endpoint. (3-lapachone is broadly active
against many cell
types, with LCSO (1og10 molar concentration causing 50% lethality) between -
4.5 and -5.3, and
mean of -5.07 across all cells. When compared to many FDA approved
chemotherapeutic
43
CA 02556794 2006-08-17
WO 2005/082358 PCT/US2005/005647
agents for common cancer types with publicly available data, no approved drug
exceeds the
mean of (3-lapachone across all cells and only mitoxantrone equals it. Figure
2.
4-4
