U.S. patent number RE45,471 [Application Number 14/195,592] was granted by the patent office on 2015-04-14 for pharmaceutical composition or combination drug.
This patent grant is currently assigned to Nanocarrier Co., Ltd.. The grantee listed for this patent is Nanocarrier Co., Ltd.. Invention is credited to Iulian Bobe, Mitsunori Harada, Takashi Shimizu.
United States Patent |
RE45,471 |
Harada , et al. |
April 14, 2015 |
**Please see images for:
( Certificate of Correction ) ** |
Pharmaceutical composition or combination drug
Abstract
A pharmaceutical composition or combination drug, which
contains, as active ingredients, (a) a coordination compound
composed of a block copolymer represented by the following formula
I or formula II and cisplatin, and (b) gemcitabine hydrochloride.
##STR00001## In the formulae I and II, R.sub.1, A, R.sub.2,
R.sub.3, m and n are as defined in the description.
Inventors: |
Harada; Mitsunori (Kashiwa,
JP), Bobe; Iulian (Kashiwa, JP), Shimizu;
Takashi (Kashiwa, JP) |
Applicant: |
Name |
City |
State |
Country |
Type |
Nanocarrier Co., Ltd. |
Kashiwa-shi, Chiba |
N/A |
JP |
|
|
Assignee: |
Nanocarrier Co., Ltd.
(Kashiwa-shi, JP)
|
Family
ID: |
40912598 |
Appl.
No.: |
14/195,592 |
Filed: |
March 3, 2014 |
PCT
Filed: |
January 13, 2009 |
PCT No.: |
PCT/JP2009/050608 |
371(c)(1),(2),(4) Date: |
July 27, 2010 |
PCT
Pub. No.: |
WO2009/096245 |
PCT
Pub. Date: |
August 06, 2009 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
Issue Date |
|
Reissue of: |
12864863 |
Jan 13, 2009 |
8436044 |
May 7, 2013 |
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Foreign Application Priority Data
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Jan 28, 2008 [JP] |
|
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2008-016688 |
|
Current U.S.
Class: |
514/492; 424/617;
514/49; 424/78.17; 424/649; 423/22 |
Current CPC
Class: |
A61K
45/06 (20130101); C08L 77/04 (20130101); A61P
35/00 (20180101); C08G 69/40 (20130101); A61K
31/7068 (20130101); A61K 47/595 (20170801); A61K
47/60 (20170801); A61K 31/282 (20130101); A61K
47/50 (20170801); A61P 43/00 (20180101); A61K
33/243 (20190101); A61K 31/282 (20130101); A61K
2300/00 (20130101); A61K 31/7068 (20130101); A61K
2300/00 (20130101); A61K 33/24 (20130101); A61K
2300/00 (20130101); C08K 5/3445 (20130101) |
Current International
Class: |
A61K
31/282 (20060101); A61K 31/7068 (20060101); A61K
33/24 (20060101); A61K 45/06 (20060101); C08G
69/40 (20060101); C08K 5/3445 (20060101) |
References Cited
[Referenced By]
U.S. Patent Documents
Foreign Patent Documents
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1 695 991 |
|
Aug 2006 |
|
EP |
|
05-000955 |
|
Jan 1993 |
|
JP |
|
08-310970 |
|
Nov 1996 |
|
JP |
|
2008-501652 |
|
Jan 2008 |
|
JP |
|
WO 02/26241 |
|
Apr 2002 |
|
WO |
|
WO 2007/035783 |
|
Mar 2007 |
|
WO |
|
Other References
Reagan-Shaw et al. (FASEB Journal 2007;22:659-661). cited by
examiner .
Examination Report for corresponding European Application No. 09
705 599.0, dated Feb. 6, 2012, 4pp. cited by applicant .
Jassem, Jacek, et al.; "A phase II study of gemcitabine plus
cisplatin in patients with advanced non-small cell lung cancer:
clinical outcomes and quality of life"; Lung Cancer 35; 2002; pp.
73-79. cited by applicant .
Xiong, Jian Ping, et al.; "Phase II trial of low-dose gemcitabine
in prolonged infusion and cisplatin for advanced non-small cell
lunch cancer"; Lung Cancer; 2008; pp. 208-214. cited by applicant
.
International Search Report, dated Mar. 10, 2009, corresponding to
PCT/JP2009/050608. cited by applicant .
Bergman, et al., "Synergistic Interaction between Cisplatin and
Gemcitabine in Vitro," Clinical Cancer Research, vol. 2, No. 3,
Mar. 1996, pp. 521-530. cited by applicant .
Extended European Search Report for European Application No.
09705599.0, dated Aug. 29, 2011, 8pp. cited by applicant .
Lee, Nam-Su et al.; "Combination of Gemcitabine and Cisplatin as
First-Line Therapy in Advance Non-Small-Cell Lung Cancer" Cancer
Research and Treatment; 2004; pp. 173-177. cited by applicant .
Nishiyama, Nubuhiro et al.; "Preparation and characterization of
size-controlled polymeric micelle containing
cis-dichlorodiammineplatinum (II) in the core"; Journal of
Controlled Release; 2001; pp. 83-94. cited by applicant .
Uchino, H. et al.; "Cisplatin-incorporating polymeric micelles
(NC-6004) can reduce nephrotoxicity and neurotoxicity of cisplatin
in rats"; British Journal of Cancer; 2005; pp. 678-687. cited by
applicant .
Yokoyama, Masayuki et al.; "Introduction of cisplatin into
polymeric micelle"; Journal of Controlled Release; 1996; pp.
351-356. cited by applicant .
European Office action dated Oct. 22, 2012 for corresponding
European Patent Application No. 09 705 599.0, 4pp. cited by
applicant .
Heinemann, V.; "Increased survival using platinum analog combined
with gemcitabine as compared to single-agent gemcitabine in
advanced pancreatic cancer: pooled analysis of two randomized
trials, the GERCOR/GISCAD intergroup study and a German multicenter
study"; Annals of Oncology 18; 2007; pp. 1652-1659. cited by
applicant .
Kato, Toshiki et al.; "Carbohydrate antigen 19-9-positive prostatic
ductal adenocarcinoma effectively treated with cisplatin and
gemcitabine"; Interventional Journal of Urology; 2007; 14; pp.
1103-1106. cited by applicant.
|
Primary Examiner: Arnold; Ernst V
Attorney, Agent or Firm: Christie, Parker & Hale,
LLP
Claims
The invention claimed is:
1. A pharmaceutical composition or a combination drug .Iadd.for
treatment of cancer in humans.Iaddend., comprising as active
ingredients thereof, (a) a coordination compound composed of a
block copolymer represented by the following formula I or formula
II and cisplatin coordinated in the block copolymer: ##STR00004##
wherein, R.sub.1 independently represents a hydrogen atom or an
alkyl group that may be substituted by a functional group or
substituent, A independently represents NH, CO,
R.sub.5(CH.sub.2).sub.pR.sub.6 or a direct bond, R.sub.5 represents
O, OCO, OCONH, NHCO, NHCOO, NHCONH, CONH or COO, R.sub.6 represents
NH or CO, p represents an integer of 1 to 6, R.sub.2 independently
represents a hydrogen atom, alkaline metal, alkyl group or aralkyl
group, R.sub.3 independently represents a hydrogen atom, hydroxyl
group or hydrophobic residue, m independently represents an integer
of 40 to 450, and n independently represents an integer of 20 to
80, and (b) gemcitabine hydrochloride which is separate from any
polymeric carrier wherein the cisplatin .[.coordination compound.].
is at an amount .[.of.]. .Iadd.equivalent to .Iaddend.at least 5
mg/kg .Iadd.in a mouse, .Iaddend.and the gemcitabine hydrochloride
is at an amount .[.of.]. .Iadd.equivalent to .Iaddend.at least 75
mg/kg .Iadd.in a mouse.Iaddend..
2. The pharmaceutical composition or combination drug according to
claim 1, wherein the block copolymer is represented by the formula
I and R.sub.2 represents a hydrogen atom or alkaline metal.
3. .[.The.]. .Iadd.A .Iaddend.pharmaceutical composition or
combination drug .[.according to claim 1, which is for treatment of
a cancer selected from the group consisting of lung cancer,
prostate cancer, pancreatic cancer, colorectal cancer and breast
cancer.]. .Iadd.for treatment of lung-cancer, prostate cancer,
pancreatic cancer, colorectal cancer and/or breast cancer in
humans, comprising as active ingredients thereof, (a) a
coordination compound composed of a block copolymer represented by
the following formula I or formula II and cisplatin coordinated in
the block copolymer: ##STR00005## wherein, R.sub.1 independently
represents a hydrogen atom or an alkyl group that may be
substituted by a functional group or substituent, A independently
represents NH, CO, R.sub.5(CH.sub.2).sub.pR.sub.6 or a direct bond,
R.sub.5 represents O, OCO, OCONH, NHCO, NHCOO, NHCONH, CONH or COO,
R.sub.6 represents NH or CO, p represents an integer of 1 to 6,
R.sub.2 independently represents a hydrogen atom, alkaline metal,
alkyl group or aralkyl group, R.sub.3 independently represents a
hydrogen atom, hydroxyl group or hydrophobic residue, m
independently represents an integer of 40 to 450, and n
independently represents an integer of 20 to 80, and (b)
gemcitabine hydrochloride which is separate from any polymeric
carrier, wherein the cisplatin is at an amount equivalent to at
least 5 mg/kg in a mouse, and the gemcitabine hydrochloride is at
an amount equivalent to at least 75 mg/kg in a mouse.Iaddend..
4. A kit for the treatment of cancer comprising the pharmaceutical
or combination drug according to any of claim 1, comprising: (i) at
least one coordination compound composed of a block copolymer
represented by the formula I or formula II and cisplatin, (ii)
gemcitabine hydrochloride, and (iii) regimen instructions
indicating simultaneous or continuous administration according to
the type of cancer so that the pharmaceutical or combined agent is
sequentially administered to a patient at a predetermined
interval.
5. A kit for the treatment of cancer comprising the pharmaceutical
or combination drug according to claim 2, comprising: (i) at least
one coordination compound composed of a block copolymer represented
by the formula I or formula II and cisplatin, (ii) gemcitabine
hydrochloride, and (iii) regimen instructions indicating
simultaneous or continuous administration according to the type of
cancer so that the pharmaceutical or combined agent is sequentially
administered to a patient at a predetermined interval.
6. A kit for the treatment of cancer comprising the pharmaceutical
or combination drug according to claim .[.3.]. .Iadd.1.Iaddend.,
comprising: (i) at least one coordination compound composed of a
block copolymer represented by the formula I or formula II and
cisplatin, (ii) gemcitabine hydrochloride, and (iii) regimen
instructions indicating simultaneous or continuous administration
according to the type of cancer so that the pharmaceutical or
combined agent is sequentially administered to a patient at a
predetermined interval.
.Iadd.7. A pharmaceutical composition or a combination drug for
treatment of cancer in humans, comprising as active ingredients
thereof, (a) a coordination compound composed of a block copolymer
represented by the following formula I or formula II and cisplatin
coordinated in the block copolymer: ##STR00006## an alkyl group
that may be substituted by a functional group or substituent, A
independently represents NH, CO, R.sub.5(CH.sub.2).sub.pR.sub.6 or
a direct bond, R.sub.5 represents O, OCO, OCONH, NHCO, NHCOO,
NHCONH, CONH or COO, R.sub.6 represents NH or CO, p represents an
integer of 1 to 6, R.sub.2 independently represents a hydrogen
atom, alkaline metal, alkyl group or aralkyl group, R.sub.3
independently represents a hydrogen atom, hydroxyl group or
hydrophobic residue, m independently represents an integer of 40 to
450, and n independently represents an integer of 20 to 80, and (b)
gemcitabine hydrochloride which is separate from any polymeric
carrier, wherein the cisplatin is at an amount of about 10
mg/m.sup.2 to about 400 mg/m.sup.2 and the gemcitabine
hydrochloride is at an amount of about 100 mg/m.sup.2 to about 1300
mg/m.sup.2..Iaddend.
.Iadd.8. A pharmaceutical composition or a combination drug for
treatment of cancer in humans, comprising as active ingredients
thereof, (a) a coordination compound composed of a block copolymer
represented by the following formula I or formula II and cisplatin
coordinated in the block copolymer: ##STR00007## an alkyl group
that may be substituted by a functional group or substituent, A
independently represents NH, CO, R.sub.5(CH.sub.2).sub.pR.sub.6 or
a direct bond, R.sub.5 represents O, OCO, OCONH, NHCO, NHCOO,
NHCONH, CONH or COO, R.sub.6 represents NH or CO, p represents an
integer of 1 to 6, R.sub.2 independently represents a hydrogen
atom, alkaline metal, alkyl group or aralkyl group, R.sub.3
independently represents a hydrogen atom, hydroxyl group or
hydrophobic residue, m independently represents an integer of 40 to
450, and n independently represents an integer of 20 to 80, and (b)
gemcitabine hydrochloride which is separate from any polymeric
carrier, wherein the cisplatin is at an amount of at least about 15
mg/m.sup.2 and the gemcitabine hydrochloride is at an amount of at
least about 225 mg/m.sup.2..Iaddend.
Description
CROSS-REFERENCE TO RELATED APPLICATION
This application is a National Phase Patent Application and claims
the priority of International Application Number PCT/JP2009/050608,
filed on Jan. 13, 2009, which claims priority of Japanese Patent
Application Number 2008-016688, filed on Jan. 28, 2008.
TECHNICAL FIELD
The present invention relates to a pharmaceutical composition or
combination drug for the treatment of cancer comprising or
combining a coordination compound composed of a block copolymer and
cisplatin, and gemcitabine hydrochloride.
BACKGROUND ART
Cancer is currently the leading cause of death. However, despite
extensive research having been conducted on a wide range of
treatment methods, such as surgical intervention or radiotherapy,
cancer has yet to be overcome, and considerable amounts of funding
and time are being devoted to cancer research even now.
In particular, chemotherapy is one of the major forms of treatment,
and numerous anticancer drugs have been researched and developed.
For example, various anticancer drugs are known, including
alkylating agents, platinum preparations, metabolic antagonists and
plant alkaloids.
Gemcitabine hydrochloride (Gemzar.RTM.) is a fluorinated
pyrmidine-based anticancer drug that is classified as a metabolic
antagonist, and as a result of being metabolized in cells, is
converted to active nucleotides in the form a diphosphate and
triphosphate that are known to demonstrate cytocidal action by
directly or indirectly inhibiting DNA synthesis. Gemcitabine
hydrochloride has been approved in Japan for use in pancreatic
cancer, non-small-cell lung cancer and biliary tract cancer, and
has been approved overseas for use in breast cancer, urinary
bladder cancer, ovarian cancer and cervical cancer.
On the other hand, extensive research has recently been conducted
on drug delivery systems (DDS), which constitute a technology for
delivering a drug to be administered to a specific site where the
drug is to act in the body while controlling the duration of
administration and dosage of the drug for the purpose of enhancing
efficacy and reducing adverse side effects. Specific examples of
DDS means that are used include methods using liposomes, emulsions
or nanoparticles as drug carriers, methods in which drugs are
enclosed in polymeric carriers such as high molecular weight
synthetic polymer micelles, and methods in which drugs are
covalently bonded to synthetic polymers or natural
polysaccharides.
Among these methods, WO 02/262414A1 discloses a coordination
compound in which cisplatin is coordination-bonded to a carboxy
anion of a block copolymer composed of poly(ethylene glycol) and
poly(glutamic acid). This coordination compound forms polymer
micelles in an aqueous medium, has been reported to be able to
reduce nephrotoxicity attributable to cisplatin in animal studies
(Br. J. Cancer, 19, 93(6), 678-87 (2005)), and is currently at the
stage of clinical studies.
However, despite research and development of various anticancer
drugs, cancer has yet to be overcome, and since there are
limitations on dosage due to potent toxicity on normal cells in the
case of treatment using a single anticancer drug, and from the
viewpoint of response rate and adverse side effects, current
treatment cannot be said to be adequate with the exception of some
cancers. Thus, numerous attempts have been made using concomitant
therapy combining various anticancer drugs. For example, A. M.
Bergman, et al., Clin. Cancer Res., 2, 521-530 (1996) reports
concomitant effects of gemcitabine hydrochloride and cisplatin.
DISCLOSURE OF THE INVENTION
The present invention was achieved for the purpose of demonstrating
higher response rates while diminishing adverse side effects, which
present problems in current chemotherapy.
The present invention includes the following aspects: [1] a
pharmaceutical composition or combination drug, which contains, as
active ingredients, (a) a coordination compound composed of a block
copolymer represented by the following formula I or formula II and
cisplatin, and (b) gemcitabine hydrochloride.
##STR00002##
In the formulae I and II, R.sub.1 independently represents a
hydrogen atom or an alkyl group that may be substituted by a
functional group or substituent, A independently represents NH, CO,
R.sub.5(CH.sub.2).sub.pR.sub.6 or a direct bond, R.sub.5 represents
O, OCO, OCONH, NHCO, NHCOO, NHCONH, CONH or COO, R.sub.6 represents
NH or CO, p represents an integer of 1 to 6, R.sub.2 independently
represents a hydrogen atom, alkaline metal, alkyl group or aralkyl
group, R.sub.3 independently represents a hydrogen atom, hydroxyl
group or hydrophobic residue, m independently represents an integer
of 40 to 450, and n independently represents an integer of 20 to
80; [2] the pharmaceutical composition or combination drug
according to [1], wherein the block copolymer is represented by the
formula I and R.sub.2 represents a hydrogen atom or alkaline metal;
[3] the pharmaceutical composition or combination drug according to
[1], which is for treatment of a cancer selected from the group
consisting of lung cancer, prostate cancer, pancreatic cancer,
colorectal cancer and breast cancer; and, [4] a kit for the
treatment of cancer comprising the pharmaceutical or combined agent
according to [1] to [3]above, which includes,: (i) at least one
coordination compound composed of a block copolymer represented by
the formula I or formula II and cisplatin, (ii) gemcitabine
hydrochloride, and (iii) regimen instructions indicating
simultaneous or continuous administration according to the type of
cancer (so that the pharmaceutical or combined agent is
sequentially administered to a patient at a predetermined
interval).
It was surprisingly found that, by using a pharmaceutical
composition or combined agent comprising a coordination component
composed of a block copolymer represented by the formula I or
formula II and cisplatin, and gemcitabine hydrochloride, higher
degrees of synergistic effects and safety are able to be achieved
than pharmaceutical compositions or combination drug comprising
cisplatin and gemcitabine hydrochloride.
BRIEF DESCRIPTION OF THE DRAWINGS
FIG. 1 is a graph indicating in vitro growth inhibitory effects of
combinations of CDDP and gemcitabine hydrochloride on human
prostate cancer PC-3 cells (mean.+-.SD of 3 wells). In the graph,
.largecircle. indicates cell growth due to changes in the
concentration of CDDP alone, while black diamonds, black triangles,
black squares and black circles represent cell growth in the case
of changing the concentration of gemcitabine hydrochloride with the
concentrations of CDDP fixed at 0.2 .mu.g/mL, 0.6 .mu.g/mL, 1.9
.mu.g/mL and 5.6 .mu.g/mL, respectively. .DELTA. indicates cell
growth due to changes in the concentration of gemcitabine
hydrochloride alone. The vertical axis represents the cell growth
rate, while the horizontal axis represents gemcitabine
hydrochloride concentration. CDDP concentration is indicated in the
case of CDDP alone (.largecircle.).
FIG. 2 is a graph indicating the GI.sub.50 values of the
combination of CDDP and gemcitabine hydrochloride against human
prostate cancer PC-3 cells. The vertical axis represents
gemcitabine hydrochloride concentration, while the horizontal axis
represents CDDP concentration.
FIG. 3 is a graph indicating in vitro growth inhibitory effects of
combinations of CDDP and gemcitabine hydrochloride on
CDDP-resistant human lung cancer MOR/CPR cells (mean.+-.SD of 3
wells). In the graph, .largecircle. indicates cell growth due to
changes in the concentration of CDDP alone, while black diamonds,
black triangles, black squares and black circles represent cell
growth in the case of changing the concentration of gemcitabine
hydrochloride with the concentrations of CDDP fixed at 0.2
.mu.g/mL, 0.6 .mu.g/mL, 1.9 .mu.g/mL and 5.6 .mu.g/mL,
respectively. .DELTA. indicates cell growth due to changes in the
concentration of gemcitabine hydrochloride alone. The vertical axis
represents the cell growth rate, while the horizontal axis
represents gemcitabine hydrochloride concentration. CDDP
concentration is indicated in the case of CDDP alone
(.largecircle.).
FIG. 4 is a graph indicating the GI.sub.50 values of the
combination of CDDP and gemcitabine hydrochloride against
CDDP-resistant human lung cancer MOR/CPR cells. The vertical axis
represents Gemcitabine hydrochloride, while the horizontal axis
represents CDDP concentration.
FIG. 5 is a graph indicating in vitro growth inhibitory effects of
combinations of a CDDP coordination compound and gemcitabine
hydrochloride on CDDP-resistant human lung cancer MOR/CPR cells
(mean.+-.SD of 3 wells). In the graph, .largecircle. indicates cell
growth due to changes in the concentration of the coordination
compound alone, while black diamonds, black triangles, black
squares and black circles represent cell growth in the case of
changing the concentration of gemcitabine hydrochloride with the
concentrations of the CDDP coordination compound fixed at 3.1
.mu.g/mL, 9.3 .mu.g/mL, 27.8 .mu.g/mL and 83.3 .mu.g/mL,
respectively. .DELTA. indicates cell growth due to changes in the
concentration of gemcitabine hydrochloride alone. The vertical axis
represents the cell growth rate, while the horizontal axis
represents gemcitabine hydrochloride concentration. CDDP
concentration is indicated in the case of CDDP alone
((.largecircle.).
FIG. 6 is a graph indicating the GI.sub.50 values of the
combination of a CDDP coordination compound or CDDP and gemcitabine
hydrochloride against CDDP-resistant human lung cancer MOR/CPR
cells. The vertical axis represents gemcitabine hydrochloride
concentration, the horizontal axis represents the CDDP coordination
compound or CDDP concentration, black circles indicate the CDDP
coordination compound and black squares indicate CDDP.
FIG. 7 is a graph indicating tumor reduction effects of concomitant
use of a cisplatin coordination compound and gemcitabine
hydrochloride in nude mice xenografted with human prostate cancer
PC-3 (mean.+-.SE). * indicates a control (untreated), black
squares, black diamonds and black circles indicate concomitant
administration of a CDDP coordination compound at 5 mg/kg and
gemcitabine hydrochloride at 33 mg/kg, 50 mg/kg and 75 mg/kg,
respectively, .diamond-solid. indicates administration of a CDDP
coordination compound alone at 5 mg/kg, .largecircle. indicates
administration of CDDP alone at 3.3 mg/kg, black triangles indicate
concomitant administration of CDDP at 3.3 mg/kg and gemcitabine
hydrochloride at 50 mg/kg, and .DELTA. and .quadrature. indicate
administration of gemcitabine hydrochloride alone at 50 mg/kg and
75 mg/kg, respectively. The vertical scale represents the relative
tumor volume to a value of 100% for the tumor volume at the start
of administration, while the horizontal scale represents the day
after the start of administration.
FIG. 8 is a graph indicating changes in body weight during
concomitant use of a cisplatin coordination compound and
gemcitabine hydrochloride in nude mice xenografted with human
prostate cancer PC-3 (mean.+-.SE). Each symbol is used similarly in
FIG. 7. The vertical scale represents the relative body weight to a
value of 100% for the body weight at the start of administration,
while the horizontal scale represents the day after the start of
administration.
FIG. 9 is a graph indicating tumor reduction effects of concomitant
use of a cisplatin coordination compound and gemcitabine
hydrochloride in nude mice xenografted with cisplatin-resistant
lung cancer MOR/CPR (mean.+-.SE). .largecircle. indicates a control
(untreated), black diamonds indicate administration of a CDDP
coordination compound alone at 5 mg/kg, black squares indicate
concomitant administration of gemcitabine hydrochloride at 75 mg/kg
and a CDDP coordination compound at 5 mg/kg, black triangles
indicate administration of CDDP alone at 3.3 mg/kg, black circles
indicate concomitant administration of CDDP at 3.3 mg/kg and
gemcitabine hydrochloride at 75 mg/kg (note that the fourth
administration was not carried out), and .quadrature. indicates
administration of gemcitabine hydrochloride alone at 75 mg/kg. The
vertical scale represents the relative tumor volume to a value of
100% for the tumor volume at the start of administration, while the
horizontal scale represents the day after the start of
administration.
FIG. 10 is a graph indicating changes in body weight during
concomitant use of a cisplatin coordination compound and
gemcitabine hydrochloride in nude mice xenografted with
cisplatin-resistant lung cancer MOR/CPR (mean.+-.SE). Each symbol
is used similarly in FIG. 9. The vertical scale represents the
relative body weight to a value of 100% for the body weight at the
start of administration, while the horizontal scale represents the
day after the start of administration.
FIG. 11 is a graph indicating in vitro growth inhibitory effects of
combinations of a CDDP coordination compound and gemcitabine
hydrochloride on human pancreatic cancer BxPC3 cells (mean.+-.SD of
3 wells). In the graph, .largecircle. indicates cell growth due to
changes in the concentration of the CCDP coordination compound
alone, while black circles, black triangles, black squares and
black stars represent cell growth in the case of changing the
concentration of gemcitabine hydrochloride with the concentrations
of the CDDP coordination compound (in terms of CDDP) fixed at
0.11.mu.g/mL, 0.34 .mu.g/mL, 1.0 .mu.g/mL and 3.1 .mu.g/mL,
respectively. .DELTA. indicates cell growth due to changes in the
concentration of gemcitabine hydrochloride alone. The vertical axis
represents the cell growth rate, while the horizontal axis
represents gemcitabine hydrochloride concentration. Concentration
converted to CDDP concentration is indicated in the case of the
CDDP coordination compound alone (.largecircle.).
FIG. 12 is a graph indicating the GI.sub.50 values of the
combination of a CDDP coordination compound and gemcitabine
hydrochloride against pancreatic cancer BxPC-3 cells. The vertical
axis represents hydrochloride concentration, while the horizontal
axis represents CDDP coordination compound concentration (in terms
of CDDP).
FIG. 13 is a graph indicating in vitro growth inhibitory effects of
combinations of a CDDP coordination compound and gemcitabine
hydrochloride on human breast cancer MDA-MB-231 cells (mean.+-.SD
of 3 wells). In the graph, .largecircle. indicates cell growth due
to changes in the concentration of the CCDP coordination compound
alone, while black circles, black triangles, black squares and
black stars represent cell growth in the case of changing the
concentration of gemcitabine hydrochloride with the concentrations
of the CDDP coordination compound (in terms of CDDP) fixed at 1.0
.mu.g/mL, 3.1 .mu.g/mL, 9.3 .mu.g/mL and 28 .mu.g/mL, respectively.
.DELTA. indicates cell growth due to changes in the concentration
of gemcitabine hydrochloride alone. The vertical axis represents
cell growth rate, while the horizontal axis represents gemcitabine
hydrochloride concentration. Concentration converted to CDDP
concentration is indicated in the case of the CDDP coordination
compound alone (.largecircle.).
FIG. 14 is a graph indicating the GI.sub.50 values of the
combination of a CDDP coordination compound and gemcitabine
hydrochloride against human breast cancer MDA-MB-231 cells. The
vertical axis represents the gemcitabine hydrochloride
concentration, while the horizontal axis represents CDDP
coordination compound concentration (in terms of CDDP).
FIG. 15 is a graph indicating in vitro growth inhibitory effects of
combinations of a CDDP coordination compound and gemcitabine
hydrochloride on human colorectal cancer LS174T cells (mean.+-.SD
of 3 wells). In the graph, .largecircle. indicate cell growth due
to changes in the concentration of the CCDP coordination compound
alone, while black circles, black triangles, black squares and
black stars represent cell growth in the case of changing the
concentration of gemcitabine hydrochloride with the concentrations
of the CDDP coordination compound (in terms of CDDP) fixed at 0.11
.mu.g/mL, 0.34 .mu.g/mL, 1.0 .mu.g/mL and 3.1 .mu.g/mL,
respectively. .DELTA. indicate cell growth due to changes in the
concentration of gemcitabine hydrochloride alone. The vertical axis
represents the cell growth rate, while the horizontal axis
represents gemcitabine hydrochloride concentration. Concentration
converted to CDDP concentration is indicated in the case of the
CDDP coordination compound alone (.largecircle.).
FIG. 16 is a graph indicating the GI.sub.50 values of the
combination of a CDDP coordination compound and gemcitabine
hydrochloride against human colorectal cancer LS174T cells. The
vertical axis represents gemcitabine hydrochloride concentration,
while the horizontal axis represents CDDP coordination compound
concentration (in terms of CDDP).
BEST MODE FOR CARRYING OUT THE INVENTION
In the present invention, the block copolymer represented by
formula I or formula II is preferably that represented by formula
I. An alkyl group represented by R.sub.1 in formula I or formula II
refers to a C.sub.1-6 alkyl group, and examples of functional
groups or substituents include an optionally protected hydroxyl
group, carboxyl group, aldehyde group, amino group, mercapto group
and maleimido group. Although there are no particular limitations
on A since it varies according to the method used to synthesize the
block copolymer, it represents R.sub.5(CH.sub.2).sub.pR.sub.6 in
the case of formula I, wherein R.sub.5 preferably represents O,
R.sub.6 preferably represents NH and p preferably represents an
integer of 1 to 6, or represents CO or a direct bond in the case of
formula II. R.sub.2 preferably represents a hydrogen atom or
alkaline metal in either case. R.sub.3 represents a hydrogen atom,
C.sub.8-16 alkylcarbonyl, phenylacetyl, diphenylacetyl or
pyrenesulfonyl in the case of formula I, represents a hydroxyl
group or a hydrophobic residue selected from the group consisting
of a C.sub.8-16 alkyl, benzyl, benzhydryl, adamantyl and
cholesteryl in the case of formula II, particularly preferably
represents a hydrogen atom in the case of formula I, and
particularly preferably represents a hydroxyl group in the case of
formula II. m independently represents an integer of 40 to 450,
preferably an integer of 60 to 410 and particularly preferably an
integer of 110 to 340. n independently represents an integer of 20
to 80 and particularly preferably an integer of 30 to 50.
Although there are no particular limitations on the synthesis
method of the aforementioned block copolymer provided it yields the
desired block copolymer, the block copolymer can be obtained by,
for example, using Me0-PEG-CH.sub.2CH.sub.2CH.sub.2-NH.sub.2 as an
initiator and adding N-carboxy-.gamma.-benzyl-L-glutamic anhydride
(BLG-NCA) to a desired degree of polymerization (degree of
polymerization is indicated by the number of amino acid units,
namely the value of n in formula I and formula II) in a dehydrated
organic solvent, allowing to react therein, and removing the benzyl
group by alkaline hydrolysis.
The combination drug in the present invention refers to the
combination of (a) a component composed of a coordination compound
composed of a block copolymer represented by formula I or formula
II and cisplatin, and (b) a component composed of gemcitabine
hydrochloride, wherein the component (a) and the component (b) are
administered simultaneously or at different times (or
consecutively).
The present invention includes a method for treating cancer
comprising administration of the aforementioned component (a) and
the aforementioned component (b) to a patient either simultaneously
or at different times (or consecutively). Furthermore, in this
case, the order in which the component (a) and the component (b)
are administered is suitably selected according to the type of
cancer. Moreover, the present invention also includes a use of the
component (a) and the component (b) for producing the
pharmaceutical composition or pharmaceutical combined agent for the
treatment of cancer, a kit for the treatment of cancer comprising
the component (a) and the component (b), and a use of the component
(a) and the component (b) for producing the kit.
The pharmaceutical composition of the present invention is one
containing the aforementioned component (a) and the aforementioned
component (b), and may be a pharmaceutical composition in which the
component (a) and the component (b) are used as is as active
ingredients thereof, or may be a pharmaceutical composition in
which is used a preparation containing the component (a) as an
active ingredient and a preparation containing the component (b) as
an active ingredient. In addition, the pharmaceutical composition
of the present invention may also be one in which either one of the
component (a) or the component (b) is used as is while the other is
used after having been formulated into a preparation in advance.
Examples of preparations of the pharmaceutical composition of the
present invention include liquid preparations and freeze-dried
preparations, with freeze-dried preparations being particularly
preferable.
In addition, in the combination drug of the present invention, a
combination drug in which each component is formulated separately
in advance, or in other words, a preparation containing the
aforementioned component (a) as an active ingredient thereof and a
preparation containing the aforementioned component (b) as an
active ingredient thereof, are normally administered simultaneously
or at different times (or consecutively).
Commonly used diluents, excipient, isotonic agents, pH adjusters
and the like can be used to formulate the pharmaceutical
composition or pharmaceutical combined agent of the present
invention.
The administration route of the pharmaceutical composition or
pharmaceutical combination drug of the present invention is
preferably intravenous injection.
The dosage of the pharmaceutical composition or combination drug of
the present invention is suitably selected according to the
administration method, age and gender of the patient, patient
status and other conditions. Although not limited thereto, the
amount of the cisplatin coordination compound of component (a)
contained as cisplatin in a preparation used for a single
administration in the case of a pharmaceutical composition (mixed
agent) is about 1 to 400 mg, and preferably about 10 to 300 mg, per
square meter of body surface area of a patient. On the other hand,
the amount of gemcitabine hydrochloride of component (b) is about
50 to 1300 mg, and preferably about 200 to 1000 mg, per square
meter of body surface area of a patient.
In addition, in the case of the combination drug, the amount of the
cisplatin coordination compound of component (a) as cisplatin is
about 10 to 400 mg, and preferably about 30 to 300 mg, per square
meter of body surface area of a patient. On the other hand, the
amount of gemcitabine hydrochloride of component (b) is about 100
to 1300 mg, and preferably about 400 to 1000 mg, per square meter
of body surface area of a patient.
Although not limited thereto, the pharmaceutical composition (mixed
agent) is preferably administered about once every 3 days to about
once every 8 weeks.
In the case of administration of the combination drug, the
cisplatin coordination compound of component (a) and the
gemcitabine hydrochloride of component (b) are administered without
allowing for the passage of time there between or after mixing in
the case of simultaneous administration. In the case of
administering at different times (consecutively), the component (a)
and the component (b) can be administered by repeating cycles
consisting of either first administering the component (a) or the
component (b) followed by administering the other component 1 day
to 2 weeks later (or in other words, by alternating
administration). In addition, the component (a) and the component
(b) can also be administered by repeating cycles consisting of
administering component (a) or component (b) 2 to 5 times at
intervals of 3 days to 2 weeks followed by administering the other
component. At this time, the other component may also be
administered by repeating cycles consisting of administering 2 to 5
times at intervals of 3 days to 2 weeks. Furthermore, in either
case, a period of 3 days to 5 weeks can be provided between cycles,
and a washout period can be provided by observing patient
status.
EXAMPLES
Although the following provides a detailed explanation of the
present invention through examples thereof, these examples do not
limit the scope of the present invention.
COMPARATIVE EXAMPLE 1
In Vitro Cell Growth Inhibitory Effect on Human Prostate Cancer
PC-3 Cells
As the cisplatin (which may also be abbreviated as CDDP), a CDDP
injection solution (Randa.RTM. Injection, Nippon Kayaku, CDDP
concentration: 0.5 mg/mL) was used. Gemcitabine hydrochloride
(Gemzar.RTM.) was purchased from Eli Lilly Japan. Human prostate
cancer PC-3 cells were purchased from the Japan Health Sciences
Foundation Research Resources Bank.
The cell growth inhibitory activities of CDDP, gemcitabine
hydrochloride and combinations of both were evaluated in the manner
described below in accordance with the WST method using the PC-3
cells. Approximately 5000 cells were seeded into each well of a
96-well plate followed by the addition of RPMI1640(Gibco.TM.,
Invitrogen) and 10% FBS (Fetal Bovine Serum, BioWest) to a total of
90 .mu.L. Thereafter, drug diluted three-fold consecutively with
medium (10 .mu.L, or 20 .mu.L but only in the case of combinations)
was added followed by the addition of 10 .mu.L of medium as
necessary to correct to a liquid volume of 110 .mu.L, followed by
culturing for 72 hours at 37.degree. C. under atmosphere of 5%
CO.sub.2. Subsequently, WST reagent (Dojindo Laboratories) was
added (10 .mu.L) followed by continuing culturing for about 72
hours at 37.degree. C. under atmosphere of 5% CO.sub.2. The
absorbance at 450 nm (Abs450) of each well was measured, and cell
growth rate (% cell growth) was measured based on the equation
indicated below.
.times..times..times..times..times..times..times..times..times..times..ti-
mes..times..times..times..times..times..times..times..times..times..times.-
.times..times..times..times..times..times..times..times..times..times.
##EQU00001##
When investigating the effect of the combination of CDDP and
gemcitabine hydrochloride, four predetermined levels of CDDP
concentration were constructed. At the predetermined CDDP
concentration, curves of gemcitabine hydrochloride concentration
vs. cell growth rate with varying the concentration of gemcitabine
hydrochloride are shown in FIG. 1. Although the effect obtained in
the presence of a CDDP concentration of 0.2 .mu.g/mL was similar to
that in the absence of CDDP, in comparisons using the same
concentration of gemcitabine hydrochloride, cell growth rate
decreased with increases in CDDP concentration when CDDP was
present at concentrations of 0.6 .mu.g/mL or more. GI.sub.50 values
(concentrations at which cell growth is inhibited by 50%) were
determined for each of the gemcitabine hydrochloride concentration
vs. growth rate curves shown in FIG. 1 and plotted in FIG. 2. Since
points were present in a region closer to the origin than lines
connecting GI.sub.50 values in the case of using either of the
drugs alone (corresponding to the x-intercept and y-intercept), the
combination of CDDP and gemcitabine hydrochloride was suggested to
demonstrate synergistic effects.
COMPARATIVE EXAMPLE 2
In Vitro Cell Growth Inhibitory Effect on CDDP-Resistant Human Lung
Cancer MOR/CPR Cells
MOR/CPR cells were obtained from the European Collection of Cell
Cultures through Dainippon Sumitomo Pharma Co., Ltd. The cell
growth inhibitory activities of CDDP, gemcitabine hydrochloride and
combinations of both were evaluated in the same manner as
Comparative Example 1 with the exception of changing the cells to
MOR/CPR cells.
In order to investigate the effect of combining CDDP and
gemcitabine hydrochloride, the concentration of CDDP was set to
four predetermined levels, and the concentration of gemcitabine
hydrochloride was varied while setting the predetermined
concentration of CDDP to one of those four levels, and curves of
the concentration of gemcitabine hydrochloride vs. cell growth rate
at those times are shown in FIG. 3. When comparing at the same
concentration of gemcitabine hydrochloride, cell growth rates
decreased as the concentration of CDDP present increased. GI.sub.50
values were determined for each of the gemcitabine hydrochloride
concentration vs. growth rate curves shown in FIG. 3 and plotted in
FIG. 4. The GI.sub.50 value in the case of gemcitabine
hydrochloride alone was estimated to be about 10 .mu.g/mL. It was
suggested from FIG. 4 that CDDP and gemcitabine hydrochloride act
synergistically against MOR/CPR cells as well.
Example 1
Preparation of Cisplatin Coordination Compound
The block copolymer used to prepare the cisplatin coordination
compound had the structure indicated below in which R.sub.1
represents a methyl group, m represents an integer of 272 as an
average value, .DELTA. represents --OCH.sub.2CH.sub.2CH.sub.2NH--,
n represents an integer of 40 as an average value, R.sub.3
represents a hydrogen atom, and all R.sub.2 represent Na.
##STR00003##
A cisplatin coordination compound was prepared using the
aforementioned block copolymer in compliance with the method
described in WO 02/26241.
Example 2
In Vitro Cell Growth Inhibitory Effect on CDDP-Resistant Human Lung
Cancer MOR/CPR Cells
The cisplatin coordination compound obtained in Example 1 was
prepared as a mannitol solution having a final concentration of 5%
so as to contain 2.5 mg/mL in terms of CDDP. The cell growth
inhibitory activities of gemcitabine hydrochloride and a
combination of the two were evaluated in the same manner as
Comparative Example 1 using MOR/CPR cells. Furthermore, the
concentrations or dosages of the cis-platin coordination compound
are all indicated as the concentrations or dosages in terms of
CDDP.
In order to investigate the effect of combining the cisplatin
coordination compound and gemcitabine hydrochloride, the
concentration of the cisplatin coordination compound was set to
four levels, and the concentration of gemcitabine hydrochloride was
changed while setting the concentration of the cisplatin
coordination compound to one of those four levels, and curves of
the concentration of gemcitabine hydrochloride vs. cell growth rate
at those times are shown in FIG. 5. When comparing at the same
concentration of gemcitabine hydrochloride, cell growth rates
decreased as the concentration of cisplatin coordination compound
present increased. GI.sub.50 values were determined for each of the
gemcitabine hydrochloride concentration vs. growth rate curves
shown in FIG. 5 and plotted in FIG. 6. The GI.sub.50 value in the
case of gemcitabine hydrochloride alone was estimated to be about
10 .mu.g/mL. It was suggested from FIG. 6 that the combination of
the cisplatin coordination compound and gemcitabine hydrochloride
acts synergistically.
Example 3
Pharmacological Efficacy Test Using Human Prostate Cancer PC-3
Cells
PC-3 cells were cultured at 37.degree. C. under atmosphere of 5%
CO.sub.2 using RPMI1640+10% FBS medium, and after allowing to
proliferate to the number of cells required for transplant, the
PC-3 cells were suspended in 50 .mu.L of physiological saline and
inoculated subcutaneously into the backs of male nude mice (Balb
nu/nu, Charles River Japan) at 2.times.10.sup.6 cells/mouse. The
nude mice were subsequently housed for 14 days, and administration
of drug was started when tumor volume reached 38.+-.1.3 mm.sup.3
(average.+-.SE). The dosage schedule was administering into a
caudal vein 3 times at a 4-day interval, and time courses of tumor
volume and body weight were measured in the 9 groups (n=7)
indicated below.
(1) control (untreated); (2) cisplatin coordination compound at 5
mg/kg (2/3 of MTD)+gemcitabine hydrochloride at 33 mg/kg; (3)
cisplatin coordination compound at 5 mg/kg+gemcitabine
hydrochloride at 50 mg/kg; (4) cisplatin coordination compound at 5
mg/kg+gemcitabine hydrochloride at 75 mg/kg; (5) cisplatin
coordination compound at 5 mg/kg; (6) CDDP at 3.3 mg/kg (2/3 of
MTD); (7) CDDP at 3.3 mg/kg+gemcitabine hydrochloride at 50 mg/kg;
(8) gemcitabine hydrochloride at 50 mg/kg; and, (9) gemcitabine
hydrochloride at 75 mg/kg (the dose of the cisplatin coordination
compounds is expressed as a dose equivalent to CDDP).
Tumor volume was calculated based on the following equation by
measuring the tumor long axis (a mm) and short axis (b mm) with an
electronic caliper (Mitutoyo). Tumor volume
(mm.sup.3)=a.times.b.sup.2/2
The combined amounts of gemcitabine hydrochloride were varied to
33, 50 (1/2 of MTD) and 75 mg/kg with the dosage of the cisplatin
coordination compound fixed at 5 mg/kg (2/3 of MTD). The time
courses of tumor volume after the start of administration of the
specimens are shown in FIG. 7, while the changes in body weight are
shown in FIG. 8. In comparison with administration of the cisplatin
coordination compound alone, tumor growth inhibitory effects
increased as the combined amount of gemcitabine hydrochloride
increased. Namely, antitumor effects in terms of T/C values changed
from 0.4 to 0.5 (NC-6004 alone) to 0.2 to 0.3, 0.1 to 0.2 and 0.1
or less when the combined amount of gemcitabine hydrochloride was
33, 50 and 75 mg/kg, respectively. Similarly, during administration
at a dosage equal to 2/3 of MTD, tumor volume changed at T/C values
of 0.5 to 0.6 for CDDP alone (3.3 mg/kg), and decreased to roughly
0.2 to 0.3 during combined administration of gemcitabine
hydrochloride at 50 mg/kg. On the other hand, the T/C value was
about 0.3 to 0.5 in the case of administration of gemcitabine
hydrochloride alone. On the basis of these results, the combined
use of the cisplatin coordination compound or CDDP and gemcitabine
hydrochloride was observed to enhance tumor growth inhibitory
effects, and the degree of that enhancement tended to be greater
for combined use with the cisplatin coordination compound.
On the other hand, with respect to body weight loss, which is an
indicator of adverse side effects, body weight loss was a maximum
of 7.0% in the groups that combined the use of the cisplatin
coordination compound during concomitant administration of
gemcitabine hydrochloride at 50 mg/kg. On the other hand, a maximum
weight loss of 14.1% was observed during combined use of CDDP and
gemcitabine hydrochloride.
Example 4
Pharmacological Efficacy Test Using Cisplatin-Resistant Human Lung
Cancer MOR/CPR Cells
MOR/CPR cells were cultured at 37.degree. C. under atmosphere of 5%
CO.sub.2 using RPMI1640+10% FBS medium, and allowed to proliferate
to the number of cells required for transplant. However, CDDP was
added to the medium to a final CDDP concentration of 1 .mu.g/mL at
the rate of once every two rounds of subculturing to maintain CDDP
resistance. The MOR/CPR cells were suspended in 50 .mu.L of
physiological saline and inoculated subcutaneously into the back of
male nude mice (Balb nu/nu) at 2.times.10.sup.6 cells/mouse. The
nude mice were subsequently housed for 9 days, and administration
of drug was started when tumor volume reached 87.+-.3.3 mm.sup.3
(average.+-.SE). The dosage schedule was administering into a
caudal vein 4 times at a 4-day interval, and tumor volume and body
weight were measured three times a week in the 6 groups (n=7)
indicated below. Tumor volume was measured and calculated in the
same manner as Example 3.
(1) control (untreated); (2) cisplatin coordination compound at 5
mg/kg (2/3 of MTD); (3) cisplatin coordination compound at 5
mg/kg+gemcitabine hydrochloride at 75 mg/kg (3/4 of MTD); (4) CDDP
at 3.3 mg/kg (2/3 of MTD); (5) CDDP at 3.3 mg/kgH+gemcitabine
hydrochloride at 75 mg/kg; and (6) gemcitabine hydrochloride at 75
mg/kg (The dose of the cisplatin coordination compounds is
expressed as a dose equivalent to CDDP).
The dosage of the cisplatin coordination compound was fixed at 5
mg/kg (2/3 of MTD), the combined amount of gemcitabine
hydrochloride was fixed at 75 mg/kg (3/4 of MTD), and this was
compared with other combinations and individual drugs alone. The
time courses of tumor volume after the start of administration of
the specimens are shown in FIG. 9, while the changes in body weight
are shown in FIG. 10. FIG. 9 shows clearly that the combination of
the cisplatin coordination compound and gemcitabine hydrochloride
were more effective compared with other combinations or each drug
alone, with a constant T/C value from 0.1 to 0.2. The T/C values
for the combination of CDDP and gemcitabine hydrochloride were
about 0.3, as shown in FIG. 10. The fourth administration was
canceled since maximum body weight loss of about 22% was observed
following the third administration, and subsequent evaluations were
continued based on a total of three administrations. Weight loss
was less than 10% for all administration groups other than this
combination. On the basis of these results, the combined use of the
cisplatin coordination compound and gemcitabine hydrochloride was
demonstrated to be superior to the combination of CDDP and
gemcitabine hydrochloride against cisplatin-resistant human lung
cancer MOR/CPR in terms of both tumor growth inhibitory effects and
adverse side effects.
Example 5
In Vitro Cell Growth Inhibitory Effect on Human Pancreatic Cancer
BxPC-3 Cells
BxPC-3 cells were acquired from the European Collection of Cell
Cultures (ECACC) through Dainippon Sumitomo Pharma Co., Ltd. The
cell growth inhibitory activities of the cisplatin coordination
compound obtained in Example 1, gemcitabine hydrochloride and
combinations of the two were evaluated in the same manner as
Comparative Example 1 with the exception of changing the cells to
BxPC-3 cells. Furthermore, the concentrations of the cisplatin
coordination compound are all indicated as the concentrations in
terms of CDDP as previously described.
In order to investigate the effect of combining the cisplatin
coordination compound and gemcitabine hydrochloride, the
concentration of the cisplatin coordination compound was set to
four levels, and the concentration of gemcitabine hydrochloride was
changed while setting the concentration of the cisplatin
coordination compound to one of those four levels, and curves of
the concentration of gemcitabine hydrochloride vs. cell growth rate
at those times are shown in FIG. 11. When comparing at the same
concentration of gemcitabine hydrochloride, cell growth rates
decreased as the concentration of cisplatin coordination compound
present increased. GI.sub.50 values were determined for each of the
gemcitabine hydrochloride concentration vs. growth rate curves
shown in FIG. 11 and plotted in FIG. 12. It was suggested from FIG.
12 that the combination of the cisplatin coordination compound and
gemcitabine hydrochloride acts synergistically against BcPC-3 cells
as well.
Example 6
In Vitro Cell Growth Inhibitory Effect on Human Breast Cancer
MDA-MB-231 Cells
MDA-MB-231 cells were acquired from the European Collection of Cell
Cultures (ECACC) through Dainippon Sumitomo Pharma Co., Ltd. The
cell growth inhibitory activities of the cisplatin coordination
compound obtained in Example 1, gemcitabine hydrochloride and
combinations of the two were evaluated in the same manner as
Comparative Example 1 with the exception of changing the cells to
MDA-MB-231 cells. Furthermore, the concentrations of the cisplatin
coordination compound are all indicated as the concentrations in
terms of CDDP as previously described.
In order to investigate the effect of combining the cisplatin
coordination compound and gemcitabine hydrochloride, the
concentration of the cisplatin coordination compound was set to
four levels, and the concentration of gemcitabine hydrochloride was
changed while setting the concentration of the cisplatin
coordination compound to one of those four levels, and curves of
the concentration of gemcitabine hydrochloride vs. cell growth rate
at those times are shown in FIG. 13. When comparing at the same
concentration of gemcitabine hydrochloride, cell growth rates
decreased as the concentration of cisplatin coordination compound
present increased. GI.sub.50 values were determined for each of the
gemcitabine hydrochloride concentration vs. growth rate curves
shown in FIG. 13 and plotted in FIG. 14. It was suggested from FIG.
14 that the combination of the cisplatin coordination compound and
gemcitabine hydrochloride acts synergistically against MDA-MB-231
cells as well.
Example 7
In Vitro Cell Growth Inhibitory Effect on Human Colorectal Cancer
LS174T Cells
LS174T cells were acquired from the European Collection of Cell
Cultures (ECACC) through Dainippon Sumitomo Pharma Co., Ltd. The
cell growth inhibitory activities of the cisplatin coordination
compound obtained in Example 1, gemcitabine hydrochloride and
combinations of the two were evaluated in the same manner as
Comparative Example 1 with the exception of changing the cells to
LS174T cells. Furthermore, the concentrations of the cisplatin
coordination compound are all indicated as the concentrations in
terms of CDDP as previously described.
In order to investigate the effect of combining the cisplatin
coordination compound and gemcitabine hydrochloride, the
concentration of the cisplatin coordination compound was set to
four levels, and the concentration of gemcitabine hydrochloride was
changed while setting the concentration of the cisplatin
coordination compound to one of those four levels, and curves of
the concentration of gemcitabine hydrochloride vs. cell growth rate
at those times are shown in FIG. 15. When comparing at the same
concentration of gemcitabine hydrochloride, cell growth rates
decreased as the concentration of cisplatin coordination compound
present increased. GI.sub.50 values were determined for each of the
gemcitabine hydrochloride concentration vs. growth rate curves
shown in FIG. 15 and plotted in FIG. 16. It was suggested from FIG.
16 that the combination of the cisplatin coordination compound and
gemcitabine hydrochloride acts synergistically against LS174T cells
as well.
* * * * *