Note: Descriptions are shown in the official language in which they were submitted.
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1
Compositions Comprising a Taxoid and a Benzimidazole Carbamate for the
Treatment of Cancer
Technical Field
The present invention relates generally to methods and compositions for the
treatment of
tumours.
Background of the invention
Paciltaxel and docetaxel are members of the taxoid family of anti-mitotic
drugs widely used
as chemotherapeutic agents. Taxoids promote microtubule polymerization and
stabilization, Their
anti-mitotic properties are derived from their ability to bind tubulin and
disrupt microtubule dynamics
thereby inducing mitotic arrest and cell death.
Paclitaxel has activity against a broad band of tumour types, including
breast, ovarian, lung,
head and neck cancers, Paclitaxel also has activity in other malignancies that
are refractory to
conventional chemotherapy, including previously-treated lymphoma and small
cell lung cancers
and oesophageal, gastric, endometrial, bladder and germ cell tumours (Mekhail
and Markman,
2002; Yamazaki et al., 1998). It is one of the most unique, and successful,
chemotherapeutic
agents currently used in the clinic for cancer treatment However major
problems associated with
paclitaxel therapy exist One of these is toxicity. Common toxicities of
paclitaxel include total
alopecia, hypersensitivity reactions, bone marrow suppression (principally
neutropenia), arthralgia,
myaiglas, and peripheral neuropathy (Markman, 2003). The development of
tolerance or drug
resistance in tumour cells to paclitaxel is also a significant factor
hindering the ongoing efficacy of
paclitaxel treatment. To ovemome resistance, typically the dosages of
paclitaxel administered are
increased thus leading to the development of side effects.
Accordingly, there is a clear need for alternative improved strategies for
taxoid-based cancer
treatments.
Another group of agents that target microiyubule,s are the benzimidazole
carbamates that
have an opposing mode of action to the taxoids in that they inhibit
microtubule polymerization
rather than polymerize tubulin (Lacey, 1990; Lacey and Gill, 1994).
Benzimidazole carbamates
include albendazole, a broad spectrum anthelmintic used clinically for the
treatment of a number of
parasitic infections (Horton, 2000), The present inventors have previously
found that albendazole
has an anti-proliferative effect on a range of cancer cell lines in vitro and
on cancers in animal
models and clinical studies (WO 02/076454).
As disclosed herein, the present inventors have now surprisingly found that
cancer cells
highly resistant to paclitaxel and partially resistant to vincristine and
colchicine are in fact
hypersensitive to the anti-proliferative effects of albendazole. Further,
albendazole potentiates the
CA 02849147 2014-04-16
2
effect of paclitaxel in human cancer cells, both in paclitaxel-sensitive and
paclitaxel-resistant cell
lines, such that used in combination these drugs have an additive or
synergistic effect in inhibiting
cancer cell proliferation.
Summary of the Invention
s According to a first aspect of the present invention there is provided a
method for the treatment of a
'tumour in a subject, the method comprising administering to the subject an
effective amount of it
least one taxoid and an effective amount of at least one benzimidazole
carbamate compound of
formula l; =
R2
Ri ___________________________
to
wherein RI Is selected from 1-1, substituted or unsubstituted, straight or
branch chain alkyl,
alkenyl, alkanylalkyl, cycloalkyl, cycloalkylalkyl, Cycloalkenylalkyl, aryl,
arylalkyl, -SR7, -SORB, -
S02R9, -SCN, Er(CH2)nBRio, -0(0)-Rii or ¨OR, COORI3, -NO2, NR136COORi3b,
isothiocyartato, or
¨CM where R7 to RI3b are each independently selected from H, substituted or
unsubstituted,
is straight or branch chain alkyl, alkenyl, alkenylalkyl, cycloalkyl,
cycloalkylalkyl, cycloalkenylalkyl,
aryl, arylalkyl, 8 and B' are independently selected from 0, S, S(0) or 502
and n is 1 to 4;
R2 is selected from H, or substituted or unsubstituted alkyl;
R3 is selected from H, substituted or unsubstituted, straight or branch chain
alkyl, alkenyl,
alkenylalkyl, cycloalkyl, cycloalkylalkyl, cycloalkenyialkyl, aryl, arylalkyl,
6- or 6- membered
20 heterocyclic ring the heteroatom(s) of which are selected from one or
more of 0, S and/or N, -5R14,
-0Ris, -SORie, -SO2R17, -SCN, NR23000Rzi,
where R18 to NI are each
independently selected from H, substituted or unsubstituted, straight or
branch chain alkyl, alkenyl,
alkenylalkyl, cycloalkyl, cycloalkylalkyl, cycloalkenylalkyl, aryl or
arylalkyl;
or a metabolite, derivative or analogue thereof; and
25 wherein the administration of the taxoid and benzimidazolecarbamate
has an additive and/or
synergistic anti-tumour effect
The Ri substitution may occur in the 5 or 6 position,
The benzirnidazole carbamate compound may be a compound of Formula II;
NHCOOR2i
401 IN
11
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112005/001839
3
wherein Ri is selected from H, substituted or unsubstituted, straight or
branch chain alkyl,
alkenyl, alkenylalkyl, cycloalkyl, cycloalkylalkyl, cycloalkenyialkyl, aryl,
arylalkyl, -SR7, -SORB, -
SO2R9, -SCN, Bs(CH2)nBRio, -0(0)-Rii or ¨0R12, COOR13, -NO2, Nii13aCOOR13b,
isothiocyanato,
or ¨ON where R7 to R13b are each independently selected. from H, substituted
or unsubstituted,
straight or branch chain alkyl, alkenyl, alkenylalkyl, cycloalkyl,
cycloalkyralkyl, cycloalkenylalkyl,
aryl, arylalkyl, B and B' are independently selected from 0, 5, S(0) or S02
and n is 1 to 4;
R21 is H, substituted or unsubstituted, straight or branch chain alkyl,
alkenyl, alkenylalkyl,
cycloalkyl, cycloalkylalkyl, cycloakenylalkyl, aryl or arylalkyl.
The benzimidazole carbamate compound may be a compound of Formula III:
=1110
RIn
wherein Ri is selected from H, substituted or unsubstituted, straight or
branch chain alkyl,
alkenyl, alkenylalkyl, cycloalkyl, cycloalkylalkyl, cycloalkenylalkyl, aryl,
arylalkyl, SR7, -SORB, -
SO2R9, -SCN, BICH2)nBRio, -0(0)-Rti or --OR12, C00R13, -NO2, NRi3eCOORiab,
isothkxyanato,
or ¨ON where R7 to R13b are each independently selected from H, substituted or
unsubstituted,
straight or branch chain alkyl, alkenyl, alkenylalkyl, cycloalkyl,
cycloalkylalkyl, cycloalkenylalkyl,
aryl, arylalkyl, B and B' are independently selected from 0, S, S(0) or SO2
and n is 1 to 4.
The benzimidazole carbamate compound may be selected from the group consisting
of
albendazole, albendazole suiphoxide, mebendazole, flubendazole,
triclabendazole,
oxfenbendazole, luxabendazole, cambendazole, oxibendazoie, parbendazoie,
thiabendazole,
cyclobendazole, dribendazole, etibendazole and fenbendazole.
In one embodiment the benzimidazole carbamate compound is albendazole, or a
metabolite,
derivative or analogue thereof.
The taxoid *may be paclitaxel, docataxel, or a metabolite, derivative or
analogue thereof.
The tumour may be a liver, ovarian, colorectal, lung, small cell lung, breast,
prostate,
pancreatic, renal, gastric, endometrial, oesophageal, head or neck tumour,
peritoneal
carcinomatosis, leukaemia, lymphoma, sarcoma or secondary metastases thereof.
The tumour may be insensitive to treatment with one or more antimitotic drugs,
The one or
more antimitotic drugs may be selected from a taxoid, a Vince alkaloid and a
colchicinold. In one
embodiment the tumour is a taxoid-insensitive tumour, According to this
embodiment, the amount
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r3A461.1.0%.
4
of taxoid administered may be an amount otherwise ineffective to treat the
tumour if
administered alone.
The taxoid and the benzimidazole carbamate compound may be administered
simultaneously or sequentially. Accordingly, the taxoid and the .
benzimidazole carbamate
s compound may be
present in a single pharmaceutical composition or in separate eompositions.
The taxoid and the benzimldazole carbamate compound may be administered
systemically.
According to a second aspect of the present invention there is provided a
method for the
treatment of a tumour in a subject, the method comprising administering to the
subject an effective
amount of paclitaxel and an effective amount of albendazole wherein the
administration of the
paclitaxel and albendazole has an additive and/or synergistic anti-tumour
effect.
The paclitaxel and albendazole may be administered simultaneously or
sequentially.
Accordingly, the paclitaxel and albendazole may be present in a single
pharmaceutical composition
or in separate compositions. The taxoid and the benzimidazole carbamate
compound may be
administered systemically.
1S According to a
third aspect of the present invention there is provided a method for the
treatment of a taxoid-insensitive tumour in a subject, the method comprising
systemically
administering to the subject paclitaxel and an effective amount of albendazole
wherein the
administration of the paclitaxel and albendazole has an additive and/or
synergistic anti-tumour
effect.
20 The amount of
paclitaxel administered may be an amount otherwise ineffective to treat the
tumour if administered alone..
Aboording to a fourth aspect of the present invention there is .provided a
pharmaceutia.al
composition for the treatment and/or prevention of cancer comprising a
synergistic combination of
at least one taxoid and at least one benzimidazole carbamate compound of
formula I:
F(12
N 4
I I
wherein Ri is selected from H, substituted or unsubstituted, straight or
branch .chain alkyl,
alkenyi, alkenylalkyl, cycloalkyl, cycloalkylalkyl, cycloalkenylalkyl, aryl,
arylalkyl, -SRa, -SON, -
SOzRe, -SCN, BI(CH2)nBR1a, -C(0)-Ra or ¨0Re, C00% -NO2, NilisaCOORI3a,
isothiocyanato, or
30 --CN where Rr to
Ri3i) are each independently selected from H, substituted or unsubstituted,
straight or branch chain alkyl, alkinyl, alkenylalkyl, cycloalkyl,
cycloalkylalkyl, .cycloalkenylalkyl,
aryl, arylalkyl, B and B' are independently selected from O,S, S(0) or S02 and
n is Ito 4;
R2 Is selected from H, or substituted or unsubstituted alkyl;
R3 is selected from H, substituted or unsubstituted, straight or branch chain
alkyl, alkenyi,
35 alkenylalkyl,
cycloalkyl, cycloalkyialkyi, cycloalkenylalkyl, aryl, arylalkyl, 5- or 6-
membered
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heterocyclic ring the heteroatom(s) of which are selected from one or more of
0, S and/or N, -SRI4
-0R18, -S0R18, -S02R17, -SCN, -O(0)-R18, -0R19, NR2000OR2i, where R15 to R21
are each
independently selected from H, substituted or unsubstituted, straight or
branch chain alkyl, alkenyl,
alkenylalkyl, cycloalkyl, cycloalkylalkyl, cycloalkenylalkyl, aryl or
arylalkyl; -
5 or a metabolite, derivative or
analogue thereof.
The Ri substitution may.occur in the 5 or 6 position.
The benzimidazole carbamate compound may be a compound of Formula II:
H
el N NHCOOR2i
1
N
R1
ii
wherein Ri is selected from H, substituted or unsubstituted, straight or
branch chain alkyl,
alkenyl, alkenylalkyl, cycloalkyl, cycloalkylalkyl, cycloalkenylalkyl, aryl,
arylalkyl, -SR7, -SOR8, -
S02R9, -SON, B'(O1-12)nBRio, -C(0)-Rii or ¨0R12, COOR13, -NO2, NRi3aCOOR13b,
isothiocyanato,
or ¨ON where R7 to R13b are each independently selected from H, substituted or
unsubstituted,
straight or branch chain alkyl, alkenyl, alkenylalkyl, cycloalkyl,
cycloalkylalkyl, cycloalkenylalkyl,
aryl, arylalkyl, B and B' are independently selected from 0, S, S(0) or SO2
and n is 1 to 4;
R21 is H, substituted or unsubstituted, straight or branch chain alkyl,
alkenyl, alkenylalkyl,
cycloalkyl, cycloalkylalkyl, cycloakenylalkyl, aryl or arylalkyl.
The benzimidazole carbamate compound may be a compound of Formula Ill:
zo
1 I
40i H
I
N N
Ri
iti
wherein Ri Is selected from H, substituted or unsubstituted, straight or
branch chain alkyl,
alkenyl, alkenylalkyl, cycloalkyl, cycloalkylalkyl, cycloalkenylalkyl, aryl,
arylalkyl, -SR7, -SORB, -
S02R9, -SON, IT(CH2)nBRi0, -O(0)-Rii or ¨0R12, 000R13, -NO2, NR13aCOORI3b,
isothiocyanato,
or ¨ON where RT to R13b are each independently selected from H, substituted or
unsubstituted,
straight or branch chain alkyl, alkenyl, alkenylalkyl, cycloalkyl,
cycloalkylalkyl, cycloalkenylalkyl,
aryl, arylalkyl, B and B' are independently selected from 0, S, S(0) or SO2
and n is 1 to 4.
CA 02849147 2014-04-16
Nevuivms LI ral4SUM, AAP.",
6
The benzimidazole carbamate compound may be selected from the group consisting
of
albendazole, albendazole .sulphoxide, mebendazole, flubendazole,
biclabendazole,
oxfenbendazole, luxabendazole, carnbendazole, oxibendazole, parbendazole,
thiabenciazole,
dyclobenclazole, dribendazoie, etibendazole and fenbendazole.
In pne embodiment the benzimidazole carbarnate compound is albendazole, or a
metabolite,
derivative DT analogue thereof.
The taxoid may be pacittaxel, docataxel, or a metabolite, derivative or
analogue thereof.
The composition may, further comprise one or more pharmaceutically acceptable
carriers,
adjuvants or diluents.
According to a fifth aspect. of the present Invention there is provided a
pharmaceutical
composition comprising a synergistic combination of paclitaxel and
albenclazole.
The composition may include one or more pharmaceutically acceptable carriers,
adjuvants or
diluents.
According to a sixth aspect of the present invention there is provided a
composition for the
is treatment of a tumour in a subject, the composition comprising a
synergistic combination of at least
one taxced and at least one benzimidazole carbamate compound of formula I.
The tumour may be a liver, ovarian, colorectal, lung, small cell lung, breast,
prostate,
pancreatic, renal, gastric, endometrial, oesophageal, head or neck tumour,
peritoneal
carcinomatosis, leukaemia, lymphoma, sarcoma or secondary metastases thereof.
20 The tumour may be insensitive to treatment with one or more
antimitotic drugs. The one or
more antimitotic drugs may be selected from a taxoid, a Vinc,a alkaloid and a
colchicinoid. In one
embodiment the tumour is a taxoid-insensitive tumour.
According to a seventh aspect of the present Invention there is provided a
composition for
the treatment of a taxoid-insensitive tumour in a subject, the composition
comprising paciltaxel and
2.5 al bendazole .
According to an eighth aspect of the present invention there Is provided a
method for the
treatment of a tumour in a subject, the method comprising administering to the
subject an effective
amount of a composition according to the third, fourth or fifth aspect
According to .a ninth aspect of the present invention there is provided a use
of at least one
30 taxoid and at least one benzimidazole carbamate compound of formula
I for the manufacture of a
medicament for the treatment of a tumour in a subject wherein used said
medicament has an
additive and/or synergistic anti tumour effect on the patient.
According to a tenth aspect of the present invention there is provided a
method for the
treatment of a tumour in a subject, wherein the tumour is insensitive to one
or more ant-mitotic
35 drugs, the method comprising administering to the subject an
effective amount of at least one
benzimidazole carbamate compound of formula I:
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RI2
R1 ______________________________
wherein Ri is selected from H, substituted or unsubstituted, straight or
branch chain alkyl,
alkenyl, alkenylalkyl, cycloalkyl, cycloalkylalkyl, cycloalkenylalkyl, aryl,
arylalkyl, -SR7, -SORB, -
SO2R9, -SCN, BICH2),BRio, -0(0)-Rii or ¨0R12, 000R13, -NO2, NR13a000R13b,
isothiocyanato, or
¨ON where R7 to R13b are each independently selected from H, substituted or
unsubstituted,
straight or branch chain alkyl, alkenyl, alkenylalkyl, cycloalkyl,
cycloalkylalkyl, cycloalkenylalkyl,
aryl, arylalkyl, B and B' are independently selected from 0, S, S(0) or SO2
and n is 1 to 4;
R2 is selected from H, or substituted or unsubstituted alkyl;
R3 is selected from H, substituted or unsubstituted, straight or branch chain
alkyl, alkenyl,
alkenylalkyl, cycloalkyl, cycloalkylalkyl, cycloalkenylalkyl, aryl, arylalkyl,
5- or 6- membered
heterocyclic ring the heteroatom(s) of which are selected from one or more of
0, S and/or N, -SR=14,
-0R15, -
SO2R17, -SON, -0(0)-Ria, -0R19, NR20000R21, where R15 to R21 are each
independently selected from H, substituted or unsubstituted, straight or
branch chain alkyl, alkenyl,
alkenylalkyl, cycloalkyl, cycloalkylalkyl, cycloalkenylalkyl, aryl or
arylalkyl;
or a metabolite, derivative or analogue thereof.
The tumour may be insensitive to one or more of a taxoid, a Vinca alkaloid
arid a
colchicinoid. The taxoid may be paclitaxel. The Vinca alkaloid may be
vincristine. The colchicinoid
may be colchicine. In one embodiment the tumour is insensitive to at least
paclitaxel,
According to an eleventh aspect of the present invention there is provided the
use of at least
one benzimidazoie carbamate compound of formula I for the manufacture of a
medicament for the
treatment of a tumour insensitive to at least one anti-mitotic drug.
In the aspects and embodiments of the invention described above the subject is
typically
human.
Also contemplated within the above aspects and embodiments are isomers,
including
stereoisomers and geometric Isomers of the compounds of Formula I, II and III,
as well as
tautomeric forms thereof.
Definitions
In the context of this specification, the term "comprising' means "including
principally, but not
necessarily solely'. Furthermore, variations of the word 'comprising', such as
"comprise" and
"comprises", have correspondingly varied meanings.
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As used herein the terms "treating" and "treatment" refer to any and all uses
which remedy a
condition or symptoms, prevent the establishment of a condition or disease, or
otherwise prevent,
hinder, retard, or reverse the progression of a condition or disease or other
undesirable symptoms
In any way whatsoever.
As used herein the term "effective amount" includes within its meaning a non-
toxic but
sufficient amount of an agent or compound to provide the desired effect. The
exact amount
required will vary from subject to subject depending on factors such as the
species being treated,
the age and general condition of the subject, the severity of the condition
being treated, the
particular agent being administered and the mode of administration and so
forth, Thus, it is not
to possible to specify an exact "effective amount". However, for any given
case, an appropriate
"effective amount" may be determined by one of ordinary skill in the art using
only routine
experimentation.
As used herein the term "insensitive" refers to a tumour or portion thereof
which is refractory,
to some degree, to treatment with a particular therapeutic agent. The term
"'insensitive" therefore
is is used to describe tumours otherwise referred to as resistant, for
example paclitaxel-resistant.
However this term is not limited to tumours which show complete or even
significant levels of
resistance to the therapeutic agent in question, but rather includes within
its scope tumours that
retain sensitivity to the agent but which display a diminished responsiveness
to the agent when
compared to sensitive tumours.
20 The term "alkyl" as used herein, includes within its meaning
monovalent, saturated, straight
and branched chain hydrocarbon radicals.
The term "alkenyr as used herein, includes within its meaning, monovalent,
straight and
branched chain hydrocarbon radicals having at least one double bond.
The term 'aryl" as used herein, includes within its meaning monovalent,
single, polynuclear,
25 conjugated and fused aromatic hydrocarbon radicals.
Brief Description of the Drawings
The present invention will now be described, by way of example only, with
reference to the
following drawings:
Figure 1: Cytotoxic activity of albendazole and paclitaxel in inhibiting
proliferation of OVCAR-
30 3 cells in vitro. (A) Dose-response inhibition of cell proliferation by
albendazole alone. (B) Dose-
related inhibition of proliferation by paclitaxei and potentiation of this
effect when cells were co-
incubated with 0.25 pM albendazole. Cell proliferation was measured using a
sulforhodamine B
assay, Results are presented as the % of control (vehicle treated cells).
Figure 2: Cytotoxic activity of albendazole and pacfitaxel in inhibiting
proliferation of SKOV-3
35 cells in vitro. (A) Dose-response inhibition of cell proliferation by
albendazole alone. (B) Dose-
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9
related inhibition of proliferation by paclitaxel and potentiation of this
effect when cells were co-
incubated with 0.25 pM albendazole. Cell proliferation was measured using a
sulforhodamine B
assay. Results are presented as the % of control (vehicle treated cells).
Figure 3: Cytotoxic activity of albendazole and paclitaxel in inhibiting
proliferation of 1A9
cells in vitro. (A) Dose-response inhibition of cell proliferation by
albendazole alone. (B) Dose-
related inhibition of proliferation by paclitaxel alone, (C) Combined effect
of co- incubation of 1A9
cells with varying doses of paclitaxel and 0.1 pM albendazole. Cell
proliferation was measured
using a sulforhodamine B assay. Results are presented as the % of control
(vehicle treated cells).
Figure 4: Cytotoxic activity of albendazole and paclitaxel in inhibiting
proliferation of
to 1A9PTX22 cells in vitro. (A) Dose-response inhibition of cell
proliferation by albendazole alone. (B)
Dose-related inhibition of proliferation by paclitaxel alone. (C) Combined
effect of co- incubation of
1A9 cells with varying doses of paclitaxel and 0.1 pM albendazole. Cell
proliferation was measured
using a sulforhodamine B assay. Results are presented as the % of control
(vehicle treated cells).
Figure 5: Cytotoxic activity of vincristine (A) and colchicine (B) in
inhibiting proliferation of
1A9 and 1A9PTX22 cells in vitro. Concentrations of vincristine (A) shown are
0.1 nM, 0.5 nM, 1
nM, 5 nM, 10 nM, 50 nM and 100 nM, Concentrations of colchicine (B) shown are
0.1 nM, 0.5 nM,
1 nM, 5 nM, 10 nM, 50 nM, 100 nM, 500 nM and 1000 nM. Cell proliferation was
measured using a
sulforhodamine B assay - the percentage of cells alive (cell growth) was
calculated by defining the
optical density of untreated cells (control) as 100%. Values represent the
mean SD of 8 replicate
experiments, each repeated at least twice.
Best Mode of Performing the Invention
Dose-related toxicity and resistance are significant factors limiting the
adoption and efficacy
of treatment of patients with cancer using antmitotic drugs such as
paclitaxel. The strategy of
increasing the dose of paclitaxel to overcome resistance merely exacerbates
the problems of
paclitaxel toxicity and the occurrence of side effects, while also potentially
promoting the
development of increased drug resistance. There is a clear need for strategies
to maximise the
benefits of paclitaxel, and other taxoid, treatments of tumours to increase
drug efficacy without the
need to increase dosages.
As disclosed herein, the present inventors have now found that albendazole
potentiates the
effect of paclitaxel in inhibiting proliferation of human cancer cells. The
effect is observed both in
paclitaxel-sensitive and paclitaxel-resistant human cancer cells. The
inventors have also
demonstrated that the paclitaxel-resistant tumour cells are hypersensitive to
albendazole, a
member of a different class of anti-tubulin agents. This is a particularly
surprising finding as one
skilled in the art would expect that a cell resistant to one type of anti-
tubulin agent would be equally
resistant to another. The inventors have demonstrated that this is not
necessarily the case,
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Accordingly, in the clinical setting the addition of albendazole to paclitaxel
may lead to a
reduction of the paclitaxel dose required to produce an antitumour effect.
Furthermore, in paclitaxel
resistant tumours, the use of albendazole, either in the presence or absence
of a paclitaxel
treatment regimen may lead to tumour responsiveness and thus a beneficial
therapeutic effect
5 previously unattainable. As exemplified herein, typically in paclitaxel-
resistant cell lines an
approximately 50-fold increase in paclitaxel concentration is required to
achieve a similar response
to that observed in paclitaxel-sensitive cells. However the inventors
demonstrate here that in the
presence of albendazole this paclitaxel concentration can be reduced by about
25 to 50-fold to
achieve a similar response to that observed in paclitaxel-sensitive cells.
That is, the addition of
10 albendazole to an otherwise ineffective dose of paclitaxel can produce a
beneficial response.
Accordingly, one aspect of the present invention provides a method for the
treatment of a
tumour in a subject, the method comprising administering to the subject an
effective amount of at
least one taxoid and an effective amount of at least one benzimidazole
carbamate compound of
formula I:
R2
R3
'=///
________________________________ 1
Ri ______________________________
wherein Ri is selected from H, substituted or unsubstituted, straight or
branch chain alkyl,
alkenyl, alkenylalkyl, cycloalkyl, cycloalkylalkyl, cycloalkenylalkyl, aryl,
arylalkyl, -SR7, -SORB, -
S02R9, -SCN, Bi(CH2).BRi0, -C(0)-Rji or ¨0R12, COOR13, -NO2, NR13.000Ri3b,
isothiocyanato, or
--CN where R7 to R13b are each independently selected from H, substituted or
unsubstituted,
straight or branch chain alkyl, alkenyl, alkenylalkyl, cycloalkyl,
cycloalkylalkyl, cycloalkenylalkyl,
aryl, arylalkyl, B and B' are independently selected from 0, S, S(0) or SO2
and n is 1 to 4;
R2 is selected from H, or substituted or unsubstituted alkyl;
R3 is selected from H, substituted or unsubstituted, straight or branch chain
alkyl, alkenyl,
alkertylalkyl, cycloalkyl, cycloalkylalkyl, cycloalkenylalkyl, aryl,
arylalkyl, 5- or 6- membered
heterocyclic ring the heteroatom(s) of which are selected from one or more of
0, S andior N, -SR14,
-0R15, -SOR16, -SON, -S02R17, -C(0)-R19, -0R19, NR20000R2i, where R15 to R21
are each
independently selected from H, substituted or unsubstituted, straight or
branch chain alkyl, alkenyl,
alkenylalkyl, cycloalkyl, cycloalkylalkyl, cycloalkenylalkyl, aryl or
arylalkyl; or an analogue,
metabolite or derivative thereof.
Typically the benzimidazole carbamate compound is a compound of formula II
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11
H
R1 opo
N N HCOO R21
IN
ir
wherein R1 is selected from H, substituted or unsubstituted, straight or
branch chain alkyl,
alkenyl, alkenylalkyl, cycloalkyl, cycloalkylalkyl, cycloalkenylalkyl, aryl,
arylalkyl, -SR7, -SORa, -
SO2R9, -SCN, B'(CH2)nBRio, -C(0)-R11 or --0R12, COOR13, -NO2, NRisaCOORub,
isothiocyanato, ,
or ¨CN where R7 to Rub are each independently selected from H, substituted or
unsubstituted,
straight or branch chain alkyl, alkenyl, alkenylalkyl, cycloalkyl,
cycloalkylalkyl, cycloalkenylalkyl,
aryl, arylalkyl, B and B' are independently selected from 0, S, S(0) or S02
and n is 1 to 4;
R21 is H, substituted or unsubstituted, straight or branch chain alkyl,
alkenyt, alkenylalkyl,
cycloalkyl, cycloalkylalkyl, cycloakenylalkyl, aryl or arylalkyl,
or of formula Ill
1 11
Hs./t N
N
110 i
N
RI
HI
wherein RI is selected from H, substituted or unsubstituted, straight or
branch chain alkyl,
alkenyl, alkenylalkyl, cycloalkyl, cycloalkylalkyl, cycloalkenylalkyl, aryl,
arylalkyl, -SR7, -SOR8, -
S02%, -SCN, 131(CH2)nBRio, -C(0)-Ril or ¨0R12, COORta, -NO2, NRuaCOORub,
isothlocyanato,
or ¨ON where R7 to R13b are each independently selected from H, substituted or
unsubstituted,
straight or branch chain alkyl, alkenyl, alkenylalkyl, cycloalkyl,
cycloaikylalkyl, cycloalkenylalkyl,
aryl, arylalkyl, B and B' are independently selected from 0, S, S(0) or S02
and n is 1 to 4.
Those skilled in the art will readily appreciate that isomers, including'
stereoisomers and
geometric isomers, of the above described benzimidazole carbarnate compounds
may exists and
the use of such isomers are included within the scope of the present
invention. Further, the use of
tautomeric forms of the above compounds is also contemplated. For example the
substituted
benzimidazole group may exist in a number of tautomeric forms, including where
the Ri substituent
is in any one of the 4 to 7 positions.
As disclosed herein, the inventors have also found that cancer cells highly
resistant to
paclitaxel and partially resistant to vincristine and colchicine are
hypersensitive to the anti-
proliferative effects of albendazole. This opens previously unrealised avenues
for the treatment of
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12
tumours resistant or insensitive to anti-mitotic drugs by using benzimidazole
carbamates such as
albendazole. At present resistance is combated by increasing the dosage of the
agent to which
resistance has developed, thereby increasing the risk of side effects such as
toxicity and leading to
the development of greater resistance and failure of the therapy. The clinical
application of
benzimidazole carbamates in the treatment of tumours resistant to drugs such
as paclitaxel
overcomes these inherent deficiencies in the prior art approach.
Accordingly, an aspect of the present invention provides a method for the
treatment of a
tumour in a subject, wherein the tumour is insensitive to one or more anti-
mitotic drugs, the method
comprising administering to the subject an effective amount of at least one
benzimidazole
carbamate compound of formula 1, 11 or 111 as defined above. For example, the
tumour may show
complete or partial resistance to one or more of the following: taxanes, Vinca
alkaloids and
colchicinoids, or derivatives or analogues thereof.
Albendazole or a metabolite, derivative or analogue thereof (such as
albendazole sulphoxide
or albendazole sulfone) is one benzimidazole carbamate particularly useful in
the methods and
compositions of the present invention. However it will be readily appreciated
by those skilled in the
art that other benzimidazole carbamates may also be employed. For example
other suitable
benzimidazole carbamates include, but are not limited to, mebendazole,
flubendazoie,
triclabendazole, mdenbendazole, luxabendazole, cambendazole, oxibendazole,
parbendazole,
thiabendazole, cyclobendazole, dribendazole, ethendazole and fenbendazole.
Typically in methods and compositions of the invention employing the use of
taxoids, the
taxoid is paclitaxel or a metabolite, derivative or analogue thereof, or
doclitaxel or a metabolite,
derivative or analogue thereof. However it will be readily appreciated by
those skilled in the art that
other taxoids may also be employed. For example a large number of derivatives
of paclitaxel and
docataxel are currently in the experimental phase or in clinical trial. It
will be understood by those
skilled in the art that such derivatives are within the scope of the methods
and compositions of the
invention.
The present invention further provides pharmaceutical compositions comprising
at least one
taxoid and at least one benzimidazole carbamate compound of formula I.
Typically these
compositions are used according to the methods of the invention.
11 will be readily appreciated by those skilled in the art that according to
the methods of the
present invention each component of the combination may be administered at the
same time, or
sequentially in any order, or at different times, so as to provide the desired
therapeutic effect.
When administered separately, it may be preferred for the components to be
administered by the
same route of administration, although it is not necessary for this to be so.
Alternatively, the
components may be formulated together in a single dosage unit as a combination
product.
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13
The methods of the present invention may further comprise the administration
of one or more
cortic,osteroids and/or antihistamines (such as diphenydramine), for example
as a pre-treatment, to
counteract the risk of the patient having an adverse reaction to the taxoid.
Similarly, the methods of
the invention may comprise the administration of one or more potentiators of
the effect of the taxoid
s and/or benzimidazole carbamate compound on the tumour to be treated. Such
administration may
be concomitant with the administration of either or both of the taxoid and the
benzimidazole
carbamate compound, For example, a suitable potentiator of benzimidazole
carbamates is an
isoquinoline such as praziquantel.
Tumours
io Those skilled in the art will readily appreciate that the methods and
compositions of the
present invention find application in the treatment of any tumour type
amenable to treatment with
taxoids and benzimidazole carbamates independently. For example the tumours
which may be
treated using methods and compositions of the present invention include liver,
ovarian, colorectal,
lung, small cell lung, breast, prostate, pancreatic, renal, gastric,
endometrial, oesophageal, head or
is neck tumours, peritoneal caninomatosis, leukaemia, lymphomas, sarcomas
or secondary
metastases thereof,
Further those skilled in the art will appreciate that the present invention
finds particular
application in the treatment of taxoid resistant tumours, such as paclitaxel
resistant tumours, as well
as tumours resistant to treatment with Vince alkaloids or colchicinoids.
20 Compositions and routes of administration
According to the methods of present invention compounds and compositions may
be
administered by any suitable route, either systemically, regionally or
locally. The particular route of
administration to be used in any given circumstance will depend on a number of
factors, including
the nature of the tumour to be treated, the severity and extent of the tumour,
the required dosage of
25 the particular compounds to be delivered and the potential side-effects
of the compounds.
For example, in circumstances where it is required that appropriate
concentrations of the
desired compounds are delivered directly to the site In the body to be
treated, administration may
be regional rather than systemic. Regional administration provides the
capability of delivering very
high local concentrations of the desired compounds to the required site and
thus is suitable for
30 achieving the desired therapeutic or preventative effect whilst avoiding
exposure of other organs of
the body to the compounds and thereby potentially reducing side effects.
By way of example, administration according to embodiments of the invention
may be
achieved by any standard routes, including intracavitary, intravesical,
intramuscular, intraarterial,
intravenous, subcutaneous, topical or oral, Intracavitary administration may
be intraperitoneal or
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14
intrapieural, In particular embodiments, administration may be via intravenous
infusion or
intraperitoneal administration.
In general, suitable compositions may be prepared according to methods which
are known to
those of ordinary skill in the art and may include pharmaceutically acceptable
diluents, adjuvants
s and/or excipients. The diluents, adjuvants and excipients must be
'acceptable" in terms of being
compatible with the other ingredients of the composition, and not deleterious
to the recipient
thereof.
Examples of pharmaceutically acceptable diluents are demineralised or
distilled water; saline
solution; vegetable based oils such as peanut oil, safflower oil, olive oil,
cottonseed oil, maize oil,
sesame oils such as peanut oil, safflower oil, olive oil, cottonseed oil,
maize oil, sesame oil, arachis
oil or coconut oil; silicone oils, including polysiloxanes, such as methyl
polysiloxane, phenyl
polysiloxane and methylphenyl polysolpoxane; volatile silicones; mineral oils
such as liquid paraffin,
soft paraffin or squalane; cellulose derivatives such as methyl cellulose,
ethyl cellulose,
carboxymethylcellulose, sodium carboxymethylcellulose or
hydroxypropylmethylcellulose; lower
is alkanois, for example ethanol or iso-propanol; lower aralkanols; lower
polyalkylene glycols or lower
alkylene glycols, for example polyethylene glycol, polypropylene glycol,
ethylene glycol, propylene
glycol, 1,3-butylene glycol or glycerin; fatty acid esters such as isopropyl
palmftate, isopropyl
myristate or ethyl oleate; polyvinylpyrridone; agar; carrageenan; gum
tragacanth or gum acacia,
and petroleum jelly. Typically, the carrier or carriers will form torn 1% to
99.9% by weight of the
compositions,
For administration as an injectable solution or suspension, non-toxic
parenterally acceptable
diluents or carriers can include, Ringers solution, medium chain triglyceride
(MCT), isotonic saline,
phosphate buffered saline, ethanol and 1,2 propylene glycol. For example, a
commonly used
carrier or vehicle for paclitaxel is Cremapho7EL. Paclitaxel is typically
prepared in 50% Cremaphor
EL and 50% ethanol.
Some examples of suitable carriers, diluents, excipients and adjuvants for
oral use include
peanut oil, liquid paraffin, sodium
carboxymethylcellulose' , methylcellulose,
hydroxypropylmethylcellulose, sodium alginate, gum acacia, gum tragacanth,
dextrose, sucrose,
sorbitol, mannitol, gelatine and lecithin. In addition these oral formulations
may contain suitable
flavouring and colourings agents. When used in capsule form the capsules may
be coated with
compounds such as glyc;eryl monostearate or glyceryl distearate which delay
disintegration.
Adjuvants typically include *emollients, emulsifiers, thickening agents,
preservatives,
bactericides and buffering agents.
Solid forms for oral administration may contain binders acceptable in human
and veterinary
pharmaceutical practice, sweeteners, disintegrating agents, diluents,
flavourings, coating agents,
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preservatives, lubricants and/or time delay agents. Suitable binders include
gum acacia, gelatine,
corn starch, gum tragacanth, sodium
alginate, carboxymethylcellulose,
hydroxypropylmethylcellulose or polyethylene glycol. - Suitable sweeteners
include sucrose,
lactose, glucose, aspartame or saccharine. Suitable disintegrating agents
include corn starch,
5 methylcellulose, polyvinylpyrrolidone, guar gum, xanthan gum, bentonile,
alginic acid or agar.
Suitable diluents include lactose, sorbitol, mannitol, dextrose, kaolin,
cellulose, calcium carbonate,
calcium silicate or dicalcium phosphate. Suitable flavouring agents include
peppermint oil, oil of
wintergreen, cherry, orange or raspberry flavouring. Suitable coating agents
include polymers or
copolymers of acrylic acid and/or methacrylic acid and/or their esters, waxes,
fatty alcohols, zein,
to shellac or gluten. Suitable preservatives include sodium benzoate,
vitamin E, alpha-tocopherol,
ascorbic acid, methyl paraben, propyl paraben or sodium bisulphite. Suitable
lubricants include
magnesium stearate, stearic acid, sodium oleate, sodium chloride or talc.
Suitable time delay
agents include glyceryi monostearate or glyceryl distearate.
Liquid forms for Oral administration may contain, in addition to the above
agents, a liquid
15 carrier. Suitable liquid carriers include water, oils such as olive oil,
peanut oil, sesame oil,
sunflower oil, safflower oil, arachis oil, coconut oil, liquid paraffin,
ethylene glycol, propylene glycol,
polyethylene glycol, ethanol, propanol, isopropanol, glycerol, fatty alcohols,
trigtycerides or
mixtures thereof,
Suspensions for oral administration may further comprise dispersing agents
and/or
suspending agents. Suitable suspending agents. include sodium
carboxymethylcellulose,.
methyloellulose, hydroxypropylmethylcellulose, poly-vinyl-pyrrolidone, sodium
alginate or acetyl
alcohol. Suitable dispersing agants include lecithin, polyoxyethylene esters
of fatty acids such as
stearic acid, polyoxyethylene sorbitol mono- or di-oleate, -stearate or -
laurate, polyoxyethylene
sorbitan mono- or di-oleate, -stearate or -laurate and the like.
Emulsions for oral administration may further comprise one or more emulsifying
agents.
Suitable emulsifying agents include dispersing agents as exemplified above or
natural gums such
as guar gum, gum acacia or gum tragacanth.
Methods for preparing parenterally administrable compositions are apparent to
those skilled
in the art, and are described in more detail in, for example, Remington's
Pharmaceutical Science,
15th ed., Mack Publishing Company, Easton, Pa.
The composition may incorporate any suitable surfactant such as an anionic,
cationic or
non-ionic surfactant such as sorbitan esters or polyoxyethylene derivatives
thereof. Suspending
agents such as natural gums, cellulose derivatives or inorganic materials such
as silicaceous
silicas, and other ingredients such as lanolin, may also be included.
=
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The compositions may also be administered in the form of liposomes. Liposomes
are
generally derived from phospholipids or other lipid substances, and are formed
by mono- or multi-
lamellar hydrated liquid crystals that are dispersed in an aqueous medium. Any
non-toxic,
physiologically acceptable and metabolisable lipid capable of forming
liposomes can be used. The
compositions in liposome form may contain stabilisers, preservatives,
excipients and the like. The
preferred lipids are the phospholipids and the phosphatidyl cholines
(lecithins), both natural and
synthetic. Methods to form liposomes are known in the art, and in relation to
this specific reference
is made to... Prescott, Ed., Methods in Cell Biology, Volume XIV, Academic
Press, New York, N.Y.
(1976), p. 33 ef seq.
to The effective dose level of the administered compound for any particular
subject will depend
upon a variety of factors including; the type of tumour being treated and the
stage of the tumour;
the activity of the compound employed; the composition employed; the age, body
weight, general
health, sex and diet of the patient the time of administration; the route of
administration; the rate of
sequestration of compounds; the duration of the treatment; drugs used in
combination or
is coincidental with the treatment, together with other related factors
well known in medicine.
One skilled in the art would be able, by routine experimentation, to determine
an effective,
non-toxic dosage which would be required to treat applicable conditions. These
will most often be
determined on a case-by-case basis.
For compositions of the present inventiOn, the taxoid may be present in the
composition in a
20 concentration of at least about 1pM, The concentration of the taxoid in
the composition may be
from about 1pM to about 50 nM, from about 0.01 nM to about 10 nM, from about
0.05 nM to about
5 nM, or from about 0.1 nM to about 1 nM.
The benzimidazole carbamate compound may be present in the composition in a
concentration of at least about 0.005 M. The concentration of the
benzimidazole carbamate in
25 the composition may be from about 0.005 M to about 10 jIM, from about
0.01 1.iM to about 1 p,M,
from about 0.1 1.1,M to about 0.5 pcM, or from about 0.1 M to about 0.25 pcM,
Generally, an effective dosage of a composition for administration to a
patient is expected to
be in the range of about 0.01mg to about 150mg per. kg body weight per 24
hours; typically, about
0.1mg to about 150mg per kg body weight per 24 hours; about 01 mg to about
100mg per kg body
30 weight per 24 hours; about 0,5mg to about 100mg per kg body weight per
24 hours; or about
1,0mg to about 100mg per kg body weight per 24 hours. More typically, an
effective dose range is
expected to be in the range of about 5mg to about 50mg per kg body weight per
24 hours,
Alternatively, an effective dosage may be up to about 5000mg/m2. Generally, an
effective
dosage is expected to be in the range of about 10 to about 5000mg/m2,
typically about 10 to about
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17
2500mgim2, about 25 to about 2000mg1m2, about 50 to about 1500mglm2, about 50
to about
1000mg/m2, or about 75 to about 600mg/m2.
Further, it will be apparent to one of ordinary skill in the art that the
optimal quantity and
spacing of indMdual dosages will be determined by the nature and extent of the
condition being
treated, the form, route and site of administration, and the nature of the
particular individual being
treated. Also, such optimum conditions can be determined by conventional
techniques.
It will also be apparent to one of ordinary skill in the art that the optimal
course of treatment,
such as, the number of doses of the composition given per day for a defined
number of days, can
be ascertained by those skilled in the art using conventional course of
treatment determination
to tests.
The present invention will now be further described in greater detail by
reference to the
following specific examples, which should not be construed as in any way
limiting the scope of the
invention.
Examples
Example 1. Paclitaxel and albendazole administration to human ovarian
carcinoma cells
Human ovarian carcinoma cell lines OVCAR-3, SKOV-3, 1A9 and 1A9PTX22 were
used,
OVCAR-3 and SKOV-3 cells were obtained from the American Type Culture
Collection (ATCC) and
maintained on RPMI medium and McCoy5A medium respectively according to ATCC
instructions.
1A9 is a clpne of the human ovarian carcinoma cell line, A2780 (Sackett et
al., 1997). 1A9PTX22 is
a paclitaxel resistant subcAone of 1A9 cells, isolated as an individual clone
in a single step selection
by exposing 1A9 cells to 5 ng/m1 paclitaxel in the presence of 5 pg/m1
verapamil, a Pgp antagonist
(Giannakakou et al., 1997), Cells were maintained in 15 nglml paclitaxel and 5
pg/ml verapamil
continuously.
Sulforhodamine B (SRB) colorimetric assay was used to study the effect of
albendazole,
paclitaxel or their combination on tumour cell cytotoxicity in vitro.
Cells were harvested from exponential phase cultures by trypsinization,
counted and plated
in 96-well plates. Optimal seeding densities for each cell line were
determined to ensure
exponential growth during a 5 -day assay. Seeding densities were 5000, 1000,
400 and 3500 Palls
per well for OVCAR-3, SKOV-3, 1A9 and 1A9PTX22 cells, respectively. Cells
plated in 96-well
tissue culture plates were treated with 100 pl cell culture medium containing
various concentrations
of albendazole, paclitaxel or a combination of the two, Both albendazole and
paclitaxel were
originally made up in absolute ethanol and subsequently diluted with cell
culture medium to give the
desired drug concentrations with a final ethanol concentration of 1%.
Treatment media were
replaced on alternate days. At the end of the treatment period (5 days), wells
were assayed for
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cellular protein content, The SRB assay was performed according to the method
described by
Skehan et af. (1990) and Papazisis et a/. (1997), with minor modifications.
The culture medium
was aspirated prior to fixation of the cells by the addition of 100 p110% cold
trichloroacetic acid.
After an hour incubation at 4 C, cells were washed five times with water. The
cells were then
stained with 200 p1 0.4% SRB dissolved in 1% acetic acid for at least 15 min
and subsequently
washed four times with 1% acetic acid to remove unbound stain. The plates were
left to dry at
room temperature and bound protein stain was solubilized with 100 p110 rnM un-
buffered Ttis base
[tris(hydroxymethyl) aminomethane)] before reading the optical density (OD) at
570 nm.
For each of the four cell lines used in this study, sensitivity of each line
was initially tested by
to incubating cells with various concentrations of albendazole or
paclitaxel for 5 days. The SRB assay
was performed at the end of the treatment period to determine cell response to
drug treatment.
Data from OVCAR-3 cells are presented in Figure 1, where response to various
doses of
albendazole is depicted in Figure 1A and response to various doses of
paclitaxel alone or in
combination with albendazole (0.25 pM) is presented in Figure 1B. As can be
seen from Figure
18, the addition of 0.25 pM albendazole to the paclitaxel containing
incubation medium led to a
complete halt of cell proliferation.
SKOV-3 cells (Figure 2A and 28) responded to albendazole, paclitaxel or the
combination in
a similar fashion. A dose-response inhibition of cell proliferation by
paclitaxel was profoundly
potentiated upon co-incubation with albendazole. Treatment of these cells with
0.01 nM paclitaxel
zo alone led to 14.5 % inhibition of cell proliferation and with 0.25 pM
albendazole alone to a 48.4 %
reduction. Upon co- incubation with paclitaxel (0,01nM) and albendazole (0.25
piVI) there was
69.4% reduction in cell proliferation (p<0.001 compared to paclitaxel alone).
This inhibitory effect was more intense in 1A9 cells (Figure 3), where
treatment with a low
albendazole concentration of 0.1 M alone (Figure 3A) or treatment with 0.1nM
paclitaxel alone
(Figure 3B) led to 11.8% and 21.8% inhibition respectively. Co-incubation of
the cells with the two
drugs at these concentrations led to 87.7% inhibition of proliferation
(p<0.001) (Figure 3C).
Treatment of 1A9P1X22, a paclitaxel resistant sub-line of 1A9, with various
concentrations of
albendazole revealed the extra sensitivity of these cells to albendazole on
one hand (Figure 4A)
and confirmed the resistant nature of the cells to paclitaxel on the other
(Figure 48). From these
results it can be seen that, compared to the parent line (1A9), the paclitaxel
resistant sub-line
1A9PTX22 is even more sensitive to albendazole. The addition of a low
concentration (0.1 p,M) of
albendazole to medium containing various concentrations of paclitaxel had a
dramatic effect on cell
proliferation (Figure 4C). Whereas cell proliferation was not affected by the
addition of 1.0 nM or
5.0 nM paclitaxel alone, at the same paclitaxel concentrations, co-incubation
with albendazole (0.1
M) led to 79.1% inhibition of cell proliferation at 1.0 nM paclitaxel and
99.1% inhibition of cell
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19
proliferation at 5.0 nM paclitaxel. Treatment with albendazole alone at 0.1
p.M had a 53% inhibitory
effect on proliferation of these cells.
Example 2- Cross resistance in paclitaxel resistant cell line
Taxoids, such as paclitaxel, are one class of antimitotic drugs that target
tubulin. A further
s well-characterized class of tubulin-binding drugs are the Vince
alkaloids, exemplified by vincristine,
vinbiastine and vinoreibine. The Vinca alkaloids interfere with a cells
ability to properly form the
mitotic spindle by preventing the normal polymerization of microtubules. They
have importance in
the treatment of leukemia, lymphomas, small cell lung cancer, and other
malignancies. A third
class of anti-tubulin drugs, exemplified by colchicine, is comprised of a
structurally diverse
collection of small molecules that are related by the fact that all bind to a
common site on tubulln
known as the colchicine site and prevent the normal polymerization of
microtubules.
As described above in Example 1, the present inventors have shown that the
paclitaxel
resistant cell line 1A9PTX22 displays increased sensitivity to the
antiproliferative effects of
albendazole compared to the paclitaxel sensitive parent line 1A9. To
investigate the properties of
Is 1A9PTX22 cells further the inventors investigated the level of
sensitivity of these cells to
representatives of two other classes of antimitotic drugs, namely vincristine
and colchicine.
1A9 and 1A9PTX22 cells plated in 96-well tissue culture plates were treated
with 100 1.11 cell
culture medium containing various concentrations of vincristine and colchicine
for 72 hours and
SRB assays conducted as described above. As for paclitaxel (see Example 1),
both vincristine and
colchicine were originally made up in absolute ethanol and subsequently
diluted with cell culture
medium to give the desired drug concentrations with a final ethanol
concentration of 1%. The
results are illustrated in Figures 5A (vincristine) and 5B (colchicine), The
degree of sensitivity of
1A9PTX22 cells to both vincristine and oplchicine is significantly reduced
when compared to that of
the parent cell line 1A9. The partial resistance of 1A9PTX22 is apparent for
vincristine at
concentrations of 5 nM and above and for colchicine at 100 nM and above
(p<0.001).
Example 3- Compositions for treatment
In accordance with the best mode of performing the invention provided herein,
specific
preferred compositions are outlined below. The following are to be construed
as merely illustrative
examples of compositions and not as a limitation of the scope of the present
invention in any way.
Example 3(A). Composltion for parenteral administration
A composition for parenteral injection could be prepared to contain 0.05 .mg
to 5 g of
albendazole and 0,05 mg to 5 g of paclitaxel in 10
mls to 2 litres of 0.1-10%
carboxymethylcellulose,
Similarly, a composition for intravenous infusion may comprise 250 ml of
sterile Ringer's
solution, and 0.05 mg to 5 g of albendazole and 0.05 mg to 5 g of paciltaxel.
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Example 3(B)- Composition for oral administration
A composition ,of a suitable agent in the form of a capsule may be prepared by
filling a
standard two-piece hard gelatin capsule with 500 mg of albendazole, in
powdered form, 500 mg of
paclitaxel, 100 mg of lactose, 35 mg of talc and 10 mg of magnesium stearate.
5 References
Giannakakou, P., Sackett, D. L., Kang, Y. K., Zhan, Z., Buters, J. T, Fojo,
T., and Poruchynsky, M.
S. Paclitaxel-resistant human ovarian cancer cells have mutant beta-tubulins
that exhibit
impaired paclitaxel-driven polymerization. Journal of Biological Chemistry,
272: 17118-
17125, 1997.
la Horton, J. Albendazole: a review of anthelmintic efficacy and safety in
humans. Parasitology, 121,
2000.
Lacey, E. Mode of action of benzimidazoles, Parasitol, Today, 6: 112-115,
1990.
Lacey, E. and Gill, J. H. Biochemistry of benzimidazole resistance. Acta
Twice, 56: 245-262,
1994.
15 Markman, M. Management of toxicities associated with the administration
of taxanes, Expert
Opinion on Drug Safety, 2: 141-146, 2003.
Mekhail, T. M. and Markman, M. Paclitaxel in cancer therapy. Expert Opinion on
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3:755-766, 2002.
Papazisis, K T., Geromichalos, G. D., Dimitriadis, K. A., and Kortsaris, A. H.
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20 sulforhodamine B calorimetric assay. Journal of Immunological Methods,
208: 151-158,
=1997.
Sackett, D. L., Giannakakou, P,, Poruchynsky, M., and Fojo, A. Tubulin from
paclitaxel-resistant
cells as a probe for novel antimicrotubule agents. Cancer Chemotherapy &
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40:228-232, 1997.
Skehan, P., Storeng, R., Scudiero, D., Monks, A., McMahon, J., Vistica, D.,
Warren, J. T., Bokesch,
H., Kenney, S., and Boyd, M. R. New colorimetric cytotoxicity assay for
anticancer-drug
screening. Journal of the National Cancer Institute, 82: 1107-1112, 1990.
Yamazaki, S., Sekine, I., and Saijo, N. [Paclitaxel (taxol): a review of its
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