U.S. patent application number 14/844027 was filed with the patent office on 2015-12-31 for cholestanol derivative for combined use.
This patent application is currently assigned to NATIONAL UNIVERSITY CORPORATION GUNMA UNIVERSITY. The applicant listed for this patent is NATIONAL UNIVERSITY CORPORATION GUNMA UNIVERSITY, OTSUKA PHARMACEUTICAL CO., LTD.. Invention is credited to Takayuki Asao, Toyo Nishimura, Shin YAZAWA.
Application Number | 20150374730 14/844027 |
Document ID | / |
Family ID | 41055800 |
Filed Date | 2015-12-31 |
United States Patent
Application |
20150374730 |
Kind Code |
A1 |
YAZAWA; Shin ; et
al. |
December 31, 2015 |
CHOLESTANOL DERIVATIVE FOR COMBINED USE
Abstract
The invention provides a cancer chemotherapeutic agent which has
fewer side effects and excellent efficacy. The cancer
chemotherapeutic agent of the invention includes a cholestanol
derivative represented by formula (1): ##STR00001## (wherein G
represents GlcNAc-Gal-, GlcNAc-Gal-Glc-, Fuc-Gal-, Gal-Glc-, Gal-,
or GlcNAc-) or a cyclodextrin inclusion compound thereof, and an
anti-cancer agent.
Inventors: |
YAZAWA; Shin; (Osaka-shi,
JP) ; Nishimura; Toyo; (Osaka-shi, JP) ; Asao;
Takayuki; (Maebashi-shi, JP) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
NATIONAL UNIVERSITY CORPORATION GUNMA UNIVERSITY
OTSUKA PHARMACEUTICAL CO., LTD. |
Maebashi-shi
TOKYO |
|
JP
JP |
|
|
Assignee: |
NATIONAL UNIVERSITY CORPORATION
GUNMA UNIVERSITY
Maebashi-shi
JP
OTSUKA PHARMACEUTICAL CO., LTD.
TOKYO
JP
|
Family ID: |
41055800 |
Appl. No.: |
14/844027 |
Filed: |
September 3, 2015 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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14193512 |
Feb 28, 2014 |
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14844027 |
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13705304 |
Dec 5, 2012 |
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14193512 |
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13235877 |
Sep 19, 2011 |
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13705304 |
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12553355 |
Sep 3, 2009 |
8088744 |
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13235877 |
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PCT/JP2009/000985 |
Mar 4, 2009 |
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12553355 |
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Current U.S.
Class: |
424/450 ;
424/617; 514/26 |
Current CPC
Class: |
A61K 31/337 20130101;
A61K 31/282 20130101; A61K 31/282 20130101; A61K 31/664 20130101;
A61K 31/7072 20130101; A61K 31/724 20130101; A61K 31/7072 20130101;
C07J 17/005 20130101; A61K 31/4745 20130101; A61K 2300/00 20130101;
A61K 33/24 20130101; A61K 31/704 20130101; A61K 9/127 20130101;
A61P 43/00 20180101; A61K 33/24 20130101; A61K 2300/00 20130101;
A61K 2300/00 20130101; A61P 35/00 20180101; A61K 2300/00 20130101;
A61K 45/06 20130101; A61K 31/337 20130101; A61K 31/513 20130101;
A61K 31/704 20130101; A61K 2300/00 20130101 |
International
Class: |
A61K 31/704 20060101
A61K031/704; A61K 31/282 20060101 A61K031/282; A61K 9/127 20060101
A61K009/127; A61K 31/337 20060101 A61K031/337; A61K 31/4745
20060101 A61K031/4745; A61K 31/664 20060101 A61K031/664; A61K 33/24
20060101 A61K033/24; A61K 31/513 20060101 A61K031/513 |
Foreign Application Data
Date |
Code |
Application Number |
Mar 6, 2008 |
JP |
2008-055284 |
Claims
1. A cancer chemotherapeutic agent comprising, in combination, a
cholestanol derivative represented by formula (1): ##STR00005##
(wherein G represents GlcNAc-Gal , or GlcNAc or a cyclodextrin
inclusion compound thereof, and an anti-cancer agent selected from
the group consisting of Paclitaxel, Docetaxcel, 5-fluorouracil,
Cisplatin, Oxaliplatin, Pemetrexed, Cyclophosphamide, and
Irinotecan.
2-10. (canceled)
11. The cancer chemotherapeutic agent of claim 1, which is a
compounding agent.
12. The cancer chemotherapeutic agent of claim 11, wherein the
cholestanol derivative in the compounding agent is a liposomal
formulation.
13. The cancer chemotherapeutic agent of claim 1, which is in the
form of a kit comprising a drug comprising a cholestanol derivative
and a drug comprising an anti-cancer agent.
14. The cancer chemotherapeutic agent of claim 13, wherein the drug
comprising a cholestanol derivative is a liposomal formulation.
15. A method of providing chemotherapy to a cancer patient in need
thereof, the method comprising administering to the patient a
cholestanol derivative represented by formula (1): ##STR00006##
wherein G represents GlcNAc-Galor GlcNAc- or a cyclodextrin
inclusion compound thereof, and an anti-cancer agent selected from
the group consisting of Paclitaxel, Docetaxcel, 5-fluorouracil,
Cisplatin, Oxaliplatin, Pemetrexed, Cyclophosphamide, and
Irinotecan.
16. The method of claim 15, wherein the cholestanol derivative and
the anti-cancer agent are administered simultaneously.
17. The method of claim 15, wherein the cholestanol derivative and
the anti-cancer agent are administered separately.
18. The method of claim 15, wherein the cholestanol derivative and
the anti-cancer agent are administered intermittently.
Description
BACKGROUND OF THE INVENTION
Field of the Invention
[0001] The present invention relates to a chemotherapeutic agent
for cancer (hereinafter referred to as a "cancer chemotherapeutic
agent") and, more particularly, to a cancer chemotherapeutic agent
employing a cholestanol derivative and an anti-cancer agent in
combination.
BACKGROUND ART
[0002] A variety of anti-cancer agents used in chemotherapy for
cancer, which is one mode of cancer therapy, have hitherto been
developed and classified based on structure, action mechanism, etc.
However, the efficacy of such an anti-cancer agent employed as a
single agent is unsatisfactory. Instead, multi-drug therapy
employing a plurality of anti-cancer agents has been predominantly
carried out in recent years from the viewpoint of suppressing
adverse side effects, and the effecacy of multi-drug therapy has
been recognized.
[0003] Under such circumstances, both of the development of new
anti-cancer combination chemotherapy, which has fewer adverse side
effect and higher efficacy than conventional chemotherapies, and
the development of new chemotherapeutic agents for use in the
chemotherapy are desired.
[0004] Meanwhile, a cholestanol derivative, in which. a sugar chain
such as GlcNAc-Gal-, GlcNAc-Gal-Glc-, Fuc-Gal-, Gal-Glc-, Gal-, or
GlcNAc- is bonding to cholestanol (the compound that the double
bond in the B ring of the cholesterol is saturated), were
previously found to have excellent anti-tumor activity.
JP-A-2000-191685, JP-A-1999-60592, WO 2005/007172 (pamphlet), and
WO 2007/026869 (pamphlet) disclose the effects of such cholestanol
derivatives.
[0005] However, no cases have been reported in which the
aforementioned cholestanol derivatives and another anti-cancer
agent are employed in combination.
SUMMARY OF THE INVENTION
[0006] Thus, the present invention is directed to provision of a
cancer chemotherapeutic agent which has fewer side effects and
excellent efficacy.
[0007] In view of the foregoing, the present inventors have carried
out extensive studies, and have found that a remarkably potentiated
anti-cancer effect can be attained through employment, in
combination, of a cholestanol derivative represented by formula (1)
or a cyclodextrin inclusion compound thereof and a known
chemotherapeutic agent (anti-cancer agent), and thus the combined
use of these pharmaceutical agents in cancer chemotherapy is very
useful.
[0008] Accordingly, the present invention is directed to the
following (1) to (10). [0009] (1) A cancer chemotherapeutic agent
comprising, in combination, a cholestanol derivative represented by
formula (1):
##STR00002##
[0009] (wherein G represents GlcNAc-Gal-, GlcNAc-Gal-Glc-,
Fuc-Gal-, Gal-Glc-, Gal-, or GlcNAc-) or a cyclodextrin inclusion
compound thereof, and an anti-cancer agent. [0010] (2) A cancer
chemotherapeutic agent according to (1) above, wherein, in formula
(1), G is GlcNAc-Gal- or GlcNAc-. [0011] (3) A cancer
chemotherapeutic agent according to (1) or (2) above, wherein the
anti-cancer agent is one or more species selected from the group
consisting of a taxane anti-cancer agent, a platinum complex
anti-cancer agent, a pemetrexed compound, and fluorouracil. [0012]
(4) A cancer chemotherapeutic agent according to (3) above, wherein
the anti-cancer agent is one or more species selected from the
group consisting of Paclitaxel, Docetaxcel, Pemetrexed, 5-FU,
Cisplatin, Oxaliplatin, Cyclophosphamide, and Irinotecan. [0013]
(5) A cancer chemotherapeutic agent according to any of (1) to (4)
above, which is a compounding agent. [0014] (6) A cancer
chemotherapeutic agent according to any of (1) to (4) above, which
is in the form of a kit including a drug containing a cholestanol
derivative and a drug containing an anti-cancer agent. [0015] (7) A
cancer chemotherapeutic agent according to (6) above, wherein the
drug containing a cholestanol derivative is a liposomal
formulation. [0016] (8) Use, in combination, of a cholestanol
derivative represented by formula (1):
##STR00003##
[0016] (wherein G represents GlcNAc-Gal-, GlcNAc-Gal-Glc-,
Fuc-Gal-, Gal-Glc-, Gal-, or GlcNAc-) or a cyclodextrin inclusion
compound thereof and an anti-cancer agent, for producing a cancer
chemotherapeutic agent. [0017] (9) A cancer chemotherapy,
characterized by comprising administering, in combination, a
cholestanol derivative represented by formula (1):
##STR00004##
[0017] (wherein G represents GlcNAc-Gal-, GlcNAc-Gal-Glc-,
Fuc-Gal-, Gal-Glc-, Gal-, or GlcNAc-) or a cyclodextrin inclusion
compound thereof and an anti-cancer agent, to a patient in need
thereof. [0018] (10) A cancer chemotherapy according to (9) above,
wherein the cholestanol derivative or a cyclodextrin inclusion
compound thereof and the anti-cancer agent are administered to a
patient in need thereof simultaneously, or separately at
intervals.
[0019] Through employment of the cancer chemotherapeutic agent and
the cancer chemotherapy according to the present invention,
prevention and treatment of cancer can be realized with safety and
higher efficacy.
BRIEF DESCRIPTION OF THE DRAWINGS
[0020] FIG. 1 is a graph showing the cell proliferation inhibitory
effects of CDDP, GC-CD, and CDDP+GC-CD on colon 26 cells;
[0021] FIG. 2 is a graph showing the cell proliferation inhibitory
effects of CDDP, GC-CD, and CDDP+GC-CD on MKN45 cells, NCIH226
cells, or colo201 cells;
[0022] FIG. 3 is a graph showing the cell proliferation inhibitory
effects of CDDP, GGC-CD, and CDDP+GGC-CD on colon26 cells, MKN45
cells, NCIH226 cells, or colo201 cells;
[0023] FIG. 4 is a graph showing the anti-tumor effect of single
administration of CDDP, GC-CD, and CDDP+GC-CD against peritoneal
dissemination caused by colon26 cells intraperitoneally inoculated
in mice;
[0024] FIG. 5 is a graph showing the anti-tumor effect of multiple
administration of CDDP, GC-CD, and CDDP+GC-CD against peritoneal
dissemination caused by colon26 cells intraperitoneally inoculated
in mice;
[0025] FIG. 6 is a graph showing the anti-tumor effect of single,
delayed administration of CDDP, GC-CD, and CDDP+GC-CD against
peritoneal dissemination caused by colon26 cells intraperitoneally
inoculated in mice, after confirmation of peritoneal dissemination
on the mesothelium of mice;
[0026] FIG. 7 is a graph showing the survival rate of mice to which
colon26 cells were intraperitoneally inoculated, upon single
administration of CDDP, GC-CD, or CDDP+GC-CD (single administration
of CDDP and double administration of GC-CD, respectively);
[0027] FIG. 8 is a graph showing the effect of suppressing or
reducing the tumor growth to which colon26 cells were
subcutaneously inoculated in mice, upon single administration of
CDDP, GGC-CD, or CDDP+GGC-CD;
[0028] FIG. 9 is a graph showing the effect of inhibiting
metastatis of colon26 cells to the lung, upon single administration
of CDDP, GC-CD, GGC-CD, CDDP+GC-CD, and CDDP+GGC-CD;
[0029] FIG. 10-A is a graph showing the cell proliferation
inhibitory effects of an anti-cancer agent (5-FU, PTX, DTX, or
CPT), GC-CD, and the anti-cancer agent+GC-CD on colon 26 cells;
and
[0030] FIG. 10-B is a graph showing the cell proliferation
inhibitory effects of an anti-cancer agent (L-OHP or CPA), GC-CD,
and the anti-cancer agent+GC-CD on colon 26 cells.
DETAILED DESCRIPTION OF PREFERRED EMBODIMENTS
[0031] The specific cholestanol derivatives represented by formula
(1) and employed in the present invention are all known
compounds.
[0032] Among the cholestanol derivatives which are represented by
formula (1) and in which G is GlcNAc-Gal-, G is preferably
GlcNA.beta.1,3-Gal.beta.- or GlcNAc.beta.1,4-Gal.beta.-. Among the
cholestanol derivatives (1) in which G is GlcNAc-Gal-Glc-, G is
preferably GlcNAc.beta.1,3-Gal.beta.1,4-Glc-. Among the cholestanol
derivatives (1) in which G is Fuc-Gal-, G is preferably
Fuc.alpha.1,3Gal-. Among the cholestanol derivatives (1) in which G
is Gal-Glc-, G is preferably Gal.beta.1,4Glc.beta.-. Among the
cholestanol derivatives (1) in which G is Gal-, G is preferably
Gal.beta.-. Among the cholestanol derivatives (1) in which G is
GlcNAc-, G is preferably GlcNAc.beta.-.
[0033] Of these, species in which G is GlcNAc-Gal- and GlcNAc- are
more preferred, with those in which G is GlcNAc.beta.1,4-Gal.beta.-
and GlcNAc.beta.- being still more preferred.
[0034] The aforementioned cholestanol derivatives may be produced
through a method, for example, disclosed in the aforementioned
Patent Documents or a similar method.
[0035] The cholestanol derivative represented by (1) readily forms
an inclusion complex with a cyclodextrin or a derivative thereof.
Thus, the cholestanol derivative employed in the present invention
may be a cyclodextrin inclusion compound thereof. In formation of
such inclusion compounds, the size of the guest molecule to be
included, Van der Waals interaction between the guest molecule and
cyclodextrin, and hydrogen bond between the hydroxyl groups of
cyclodextrin and the guest molecule must be taken into
consideration. Therefore, insoluble guest compounds do not always
form the corresponding inclusion compounds. However, the
cholestanol derivative of the present invention can form good
inclusion complexes with cyclodextrin.
[0036] Examples of the cyclodextrin forming the cyclodextrin
inclusion compound of the present invention include cyclodextrins
such as .alpha.-cyclodextrin, .beta.-cyclodextrin, and
.gamma.-cyclodextrin; and cyclodextrion derivatives such as
methyl-.beta.-cyclodextrin, 2-hydroxypropyl-.beta.-cyclodextrin,
monoacetyl-.beta.-cyclodextrin, and
2-hydroxypropyl-.gamma.-cyclodextrin. Of these,
2-hydroxypropyl-.beta.-cyclodextrin is preferred for obtaining
improved solubility.
[0037] The cyclodextrin inclusion compound may be prepared through,
for example, the following procedure: an aqueous solution of a
cyclodextrin or a derivative thereof having an appropriate
concentration (e.g., 20 to 40%) is prepared, and the cholestanol
derivative of the present invention is added to the aqueous
solution, followed by stirring of the resultant mixture.
[0038] No particular limitation is imposed on the concentration of
the solution of the cholestanol derivative (1), so long as the
cholestanol derivative can form an inclusion compound with
cyclodextrin. Generally, the concentration is about 1 to about 50
mass %, preferably about 10 to about 30 mass %.
[0039] The thus-produced cyclodextrin inclusion compound is highly
water-soluble and, therefore, effectively exhibits the effect of
the guest in vivo. Another advantage of the cyclodextrin inclusion
compound is to ensure consistent in vitro test results.
[0040] Alternatively, the cholestanol derivative (1) may be
prepared into a liposomal formulation, whereby the cholestanol
derivative can be more effectively delivered to the action
expression site. Another advantage of the cyclodextrin inclusion
compound is to ensure consistent in vitro test results.
[0041] Preferably, the liposomal formulation includes the
cholestanol derivative of the present invention, a membrane
component, and an aliphatic or aromatic amine.
[0042] The cholestanol derivative content in the liposomal
formulation is preferably 0.3 to 2.0 mol, more preferably 0.8 to
1.5 mol, with respect to 1 mol of the membrane component.
[0043] The membrane component may be a phospholipid. Specific
examples of preferably employed phospholipids include natural and
synthetic phospholipids such as phosphatidylcholine,
phosphatidylethanolamine, phosphatidylserine, phosphatidylinositol,
and phosphatidic acid; mixtures thereof; and modified natural
phospholipids such as aqueous lecithin. Examples of more preferred
species include phosphatidylcholine
(1.alpha.-dipalmitoylphosphatidylcholine (DPPC)).
[0044] The aliphatic or aromatic amine is employed mainly for
positively charging the surface of lipid membrane. Examples of such
amines include aliphatic amines such as stearylamine and
oleylamine; and aromatic amines such as fluorenethylamine. Among
them, stearlylamine is particularly preferably employed.
[0045] Preferably, the amine is contained in an amount of 0.04 to
0.15 mol, more preferably 0.1 to 0.15 mol, with respect to 1 mol of
membrane component (phospholipid).
[0046] In addition to the aforementioned components, if required,
the liposome may further contain a membrane structure stabilizer
such as cholesterol, fatty acid, diacetyl phosphate, etc.
[0047] The aqueous solution employing for dispersing the membrane
component is preferably water, physiological saline, buffer,
aqueous sugar solution, or a mixture thereof. Either an organic or
an inorganic buffer may be used, so long as the buffer has a
buffering action in the vicinity of the hydrogen ion concentration
of body fluid. Examples of such buffers include a phosphate
buffer.
[0048] No particular limitation is imposed on the method of
preparing the liposomal formulation, and generally employed methods
may be selected. Examples of the employable method include methods
disclosed in JP-A-1982-82310, JP-A-1985-12127, JP-A-1985-58915,
JP-A-1989-117824, JP-A-1989-167218, JP-A-1992-29925, and
JP-A-1997-87168; a method disclosed in Methods of Biochemical
Analysis (1988) 33, p. 337; or a method disclosed in "Liposome"
(published by Nankodo).
[0049] No particular limitation is imposed on the anti-cancer agent
which is used in combination with the cholestanol derivative
represented by formula (1) or a cyclodextrin inclusion compound
thereof, and known cancer chemotherapeutic agents may be used.
Standard therapeutic agents which have been established with
respect to the cancer of therapy target are preferably
employed.
[0050] Specific examples include alkylating agents such as
Cyclophosphamide, Ifosfamide, Melphalan (L-PAM), Busulfan, and
Carboquione; metabolism antagonists such as 6-Mercaptopurine
(6-MP), Methotrexate (MTX), 5-Fluorouracil (5-FU), Tegafur,
Enocitabine (BHAC), and pemetrexed compounds (Pemetrexed , Alimta,
MTA), etc.; carcinostatic antibiotics such as Actinomycin D,
Daunorubicin, Bleomycin, Peplomycin, Mitomycin C, Aclarubicin, and
Neocarzinostatin (NCS); plant alkaloids such as Vincristine,
Vindesine, Vinblastine, taxane anti-cancer agents (Taxotere
(Docetaxel) and Taxol (Paclitaxel, TXL), etc.), and Irinotecan
(CPT-11); and platinum compounds such as Cisplatin (CDDP), and
Carboplatin, Oxaliplatin (L-OHP). These anti-cancer agents may be
used singly or in combination of two or more species.
[0051] As shown in the Examples described hereinbelow, when the
cholestanol derivative represented by formula (1) or a cyclodextrin
inclusion compound thereof is used in combination with an
anti-cancer agent, proliferation of cancer cells of various types
are strongly suppressed, as compared with the case of
administration of only each agent. Therefore, this combined
chemotherapy can drastically enhance therapeutic efficacy and
mitigation of adverse side effects, and a pharmaceutical product
containing these ingredients is a useful cancer chemotherapeutic
agent.
[0052] No particular limitation is imposed on the cancer which can
be effectively treated by administering the cancer chemotherapeutic
agent of the present invention. Examples of the target cancer
include malignant tumors such as gastric cancer, large bowel
cancer, pancreatic cancer, uterus cancer, ovaria cancer, lung
cancer, gallbladder cancer, esophageal cancer, liver cancer, breast
cancer, mesothelioma, and prostatic cancer.
[0053] The form of the cancer chemotherapeutic agent of the present
invention may be a compounding agent in which the aforementioned
ingredients are mixed at an appropriate ratio, each at an effective
amount, to form a single dosage form (single-formulation type), or
may be a kit that consists of the respective dosage form of the
aforementioned ingredients, each of which is formed independently
including each effective amount, and that enables the dosage forms
to be administered simultaneously or separately at intervals
(double-formulation type).
[0054] Similar to general pharmaceutical formulations, no
particular limitation is imposed on the dosage form of the
above-described formulation, and the form may be any of the solid
form such as tablet, liquid form such as injection, dry powder
dissolving before use, etc.
[0055] No particular limitation is imposed on the administration
route of the formulation, and an appropriate route may be
determined depending on the dosage form of the agents. For example,
an injection solution may be administered intravenously,
intramuscularly, subcutaneously, intradermally, or
interperitoneally, and a solid form may be administered orally or
enterally.
[0056] The formulation may be prepared through a known method in
the art. All pharmaceutically acceptable carriers (excipients or
diluents such as a filler, a bulking agent, and a binder) generally
employed in the art may also be employed.
[0057] For example, a peroral solid form may be prepared through
mixing the drug ingredients of the present invention with a
excipient, and with an optional binder, disintegrant, lubricant,
colorant, flavoring agent, deodorant, etc., and forming the mixture
into tablets, coated-tablets, granules, powder, capsules, etc.
through a method known in the art. These additive may be those
generally employed in the art. Examples of the excipient include
lactose, sucrose, sodium chloride, glucose, starch, calcium
carbonate, kaolin, microcrystalline cellulose, and silicic acid.
Examples of the binder include water, ethanol, propanol, simple
syrup, liquid glucose, liquid starch, liquid gelatin, carboxymethyl
cellulose, hydroxypropyl cellulose, hydroxypropyl starch, methyl
cellulose, ethyl cellulose, shellac, calcium phosphate, and
polyvinylpyrrolidone. Examples of the disintegrant include dry
starch, sodium alginate, agar powder, sodium hydrogencarbonate,
calcium carbonate, sodium lauryl sulfate, monoglyceryl stearate,
and lactose. Examples of the lubricant include purified talc,
stearate salts, borax, and polyethylene glycol. Examples of the
flavoring agent include sucrose, orange peel, citric acid, and
tartaric acid.
[0058] An oral liquid formulation may be prepared by mixing the
drug ingredients of the present invention with a flavoring agent,
buffer, stabilizer, deodorant, etc., and forming the mixture into
internal liquid agent, syrup, elixir, etc. through a method known
in the art. The flavoring agent employed in the preparation may be
any of the aforementioned members. Examples of the buffer include
sodium citrate. Examples of the stabilizer include traganth, gum
arabic, and gelatin.
[0059] Injection solutions may be prepared by mixing the drug
ingredients of the present invention with additives such as a
pH-regulator, buffer, stabilizer, tonicity agent, and local
anesthetic agent, etc., and forming the mixture through a method
known in the art, to thereby provide subcutaneous, intramuscular,
and intravenous injection liquids. Examples of the pH-regulator and
buffer include sodium citrate, EDTA, thioglycolic acid, and
thiolactic acid. Examples of the local anesthetic include procaine
hydrochloride and lidocaine hydrochloride. Examples of the tonicity
agent include sodium chloride and glucose.
[0060] Suppositories may be prepared by mixing the drug ingredients
of the present invention with a carrier for formulation known in
the art such as polyethylene glycol, lanolin, cacao butter, and
fatty acid triglyceride, and with an optional surfactant such as
Tween (registered trademark), and forming the mixture into
suppositories through a method known in the art.
[0061] Ointments may be prepared by mixing the drug ingredients of
the present invention with optional additives generally employed in
the art such as a base, stabilizer, moisturizer, and preservatives,
and forming the mixture into ointments through a method known in
the art. Examples of the base include liquid paraffin, white
petrolatum, white beeswax, octyldodecyl alcohol, and paraffin.
Examples of the preservative include methyl p-hydroxybenzoate,
ethyl p-hydroxybenzoate, and propyl p-hydroxybenzoate.
[0062] Cataplasms may be prepared by applying the aforementioned
ointment, gel, cream, paste, etc. to a generally employed support
through a routine method. Examples of appropriate supports include
woven and nonwoven fabric made of cotton, staple fiber, or chemical
fiber, and film and foamed sheet made of soft vinyl chloride,
polyethylene, polyurethane, etc.
[0063] Generally, the formulation is preferably prepared so as to
have a cholestanol derivative content and an anti-cancer agent
content of 0.0001 to 80 wt. % (as effective ingredient).
[0064] When the cancer chemotherapeutic agent of the present
invention is provided as a kit, the kit may be designed to pack
independently the respective dosage form including separately the
cholestanol derivative represented by formula (1) or a cyclodextrin
inclusion compound thereof and an anti-cancer agent, each of which
have been prepared in the above manner, and to be used each
pharmaceutical formulation taken separately from the corresponding
respective package before use. Alternatively, each pharmaceutical
formulation may be held in a package suitable for each time of
combined administration.
[0065] The dose of the cancer chemotherapeutic agent of the present
invention varies depending on the body weight, age, sex, symptoms
of a patient in need thereof, route and frequency of administration
to a patient in need thereof, etc. Generally, for example, the
daily dose for an adult is about 0.1 to 30 mg/kg as the cholestanol
derivative (1), preferably 3 to 10 mg/kg. The dose of the
anti-cancer agent may fall within a range established with respect
to the agent, or may be lower than that range.
[0066] No particular limitation is imposed on the frequency of
administration, and the agent may be administered once or several
times a day. Single administration a day is preferred. When the kit
is used, each of the formulation including separated drug
ingredients may be administered simultaneously or
intermittently.
EXAMPLES
[0067] The present invention will next be described in more detail
by way of examples, which should not be construed as limiting the
invention thereto.
Example 1
Effect of Drug Addition on Inhibition of Cancer Cell
Proliferation
[0068] Colon26 cells (derived from mouse colon cancer) were
inoculated to a 96-well plate (1.times.10.sup.4 cells/50 .mu.L, 10%
FCS-RPMI medium/well), and incubated at 37.degree. C. for 16 hours.
To each well, cisplatin (abbreviated as "CDDP") and/or a
cyclodextrin inclusion compound (abbreviated as "GC-CD") of a
cholestanol derivative in which G in formula (1) is GlcNAc.beta.-
(abbreviated as "GC") was added (multi-fold dilution by
FCS(-)-medium: final concentration: .ltoreq.500 .mu.m, 50 .mu.L),
followed by incubation at 37.degree. C. for two days. GC-CD was
prepared in accordance with a method disclosed in Example 1(2) in
WO 2007/026869. Specifically, a 40% aqueous solution of
hydroxypropyl-.beta.-cyclodextrin was prepared, and GC was added to
the solution, followed by mixing with stirring (80.degree. C. for
30 minutes), to thereby prepare GC-CD.
[0069] As a control, wells to which only FCS(-)-medium had been
added were employed. Viable count was performed by means of a cell
counting kit (product of Dojin).
[0070] Cell proliferation inhibition (CPI) rate (%) was calculated
by the following equation. FIG. 1 shows the results.
CPI rate ( % ) = ( 1 - treated cells OD 450 - 650 untreated cells
OD 450 - 650 ) .times. 100 ##EQU00001##
Example 2
Effect of Inhibition of Proliferation of Various Cancer Cells
[0071] The procedure of Example 1 was repeated, except that colon26
cells were changed to MKN45 (derived from human gastric cancer),
NCIH226 (derived from human lung cancer), and Colo201 (derived from
human colon cancer). CPI rate (%) was determined in a similar
manner. FIG. 2 shows the results.
[0072] In Example 2, a cyclodextrin inclusion compound (abbreviated
as "GGC-CD") of a cholestanol derivative in which G in formula (1)
is GlcNAc.beta.1,4-Gal.beta.- (abbreviated as "GGC") was also used.
GGC-CD was produced in a manner similar to the method as the
aforementioned GC-CD production method, except that the cholestanol
compound was changed to GGC. CPI rate with respect to the cancer
cells was determined. FIG. 3 shows the results.
Example 3
Effect of Drug Addition on Inhibition of Cancer Cell Proliferation
In Vivo
[0073] In the following Examples, Balb/c mice (6-weeks old, female)
were employed as test animals.
[0074] (1) Colon26 cells (1.times.10.sup.4 cells/mouse) were
intraperitoneally inoculated to the mice (day 0). On the following
day after inoculation (day 1), CDDP and/or GC-CD was adjusted with
physiological saline (Otsuka normal saline) to a concentration of
interest, and CDDP, GC-CD, or CDDP+GC-CD (500 .mu.L) was
intraperitoneally administered to the mice, followed by breeding.
On day 19, mice were dissected, and the weight of the mesentery and
the greater omentum was measured. To the control group, only
physiological saline (500 .mu.L) was administered (n=10; 10
mice/group).
[0075] FIG. 4 shows the results.
[0076] (2) Colon26 cells (1.times.10.sup.4 cells/mouse) were
intraperitoneally inoculated to the mice (day 0). On day 1, day 2,
day 3, day 6, day 7, and day 8, CDDP and/or GC-CD was adjusted with
physiological saline (Otsuka normal saline) to a concentration of
interest, and CDDP, GC-CD, or CDDP+GC-CD (500 .mu.L) was
intraperitoneally administered to the mice, followed by breeding.
On day 21, mice were dissected, and the weight of the mesentery and
the greater omentum was measured. To the control group, only
physiological saline (500 .mu.L) was administered (n=10; 10
mice/group).
[0077] FIG. 5 shows the results.
[0078] (3) Colon26 cells (1.times.10.sup.4 cells/mouse) were
intraperitoneally inoculated to the mice (day 0). On day 7, CDDP
and/or GC-CD was adjusted with physiological saline (Otsuka normal
saline) to a concentration of interest, and CDDP, GC-CD, or
CDDP+GC-CD (500 .mu.L) was intraperitoneally administered to the
mice, followed by breeding. On day 18, mice were dissected, and the
weight of the mesentery and the greater omentum was measured. To
the control group, only physiological saline (500 .mu.L) was
administered (n=10; 10 mice/group).
[0079] FIG. 6 shows the results.
Example 4
Anti-Tumor Effect by Drug Addition
[0080] Balb/c mice (6 weeks old, female) were employed as test
animals. Colon26 cells (1.times.10.sup.4 cells/mouse) were
intraperitoneally inoculated to the mice (day 0). On day 2 and/or
day 3, CDDP and/or GC-CD was adjusted with physiological saline
(Otsuka normal saline) to a concentration of interest, and CDDP
(once, on day 2), GC-CD (twice, on day 2 and 3), or CDDP (once, on
day2)+GC-CD (twice, day 2 and 3) (500 .mu.L) was intraperitoneally
administered to the mice, followed by breeding. The survival
duration (days) was counted to day 43. To the control group, only
physiological saline (500 .mu.L) was administered (n=10; 10
mice/group).
[0081] FIG. 7 shows the results.
Example 5
Anti-Tumor Effect by Drug Addition
[0082] Balb/c mice (6 weeks old, female) were employed as test
animals. Colon26 cells (5.times.10.sup.4 cells/mouse) were
subcutaneously inoculated to the mice (day 0). After confirmation
that the tumor size reached about 4 mm (day 7 to 10 after
inoculation), CDDP and/or GGC-CD was adjusted with physiological
saline (Otsuka normal saline) to a concentration of interest, and
CDDP, GGC-CD, or CDDP+GGC-CD (200 .mu.L) was administered to the
mice through the tail vein, followed by breeding. Time-dependent
change in tumor size was monitored to day 21, and the corresponding
tumor volume was determined. To the control group, only
physiological saline (200 .mu.L) was administered (n=7; 7
mice/group).
[0083] FIG. 8 shows the results.
Example 6
Cancer Metastasis Inhibitory Effect by Drug Addition
[0084] Balb/c mice (6 weeks old, female) were employed as test
animals. Colon26 cells (5.times.10.sup.4 cells/mouse) were
intraperitoneally inoculated to the mice (day 0). Immediately after
inoculation, CDDP and/or GC-CD or GGC-CD was adjusted with
physiological saline (Otsuka normal saline) to a concentration of
interest, and CDDP, GC-CD (or GGC-CD), or CDDP+GC-CD (or GGC-CD)
(200 .mu.L) was administered to the mice through the caudal vein,
followed by breeding. On day 14, mice were dissected, and the tumor
nodes in the lungs were counted. To the control group, no substance
was administered (n=10; 10 mice/group).
[0085] FIG. 9 shows the results.
Example 7
Effect of Drug Addition on Inhibition of Cancer Cell
Proliferation
[0086] Colon26 cells (derived from mouse colon cancer) were
inoculated to a 96-well plate (1.times.10.sup.4 cells/50 .mu.L, 10%
FCS-RPMI medium/well), and incubated at 37.degree. C. for 16 hours.
To each well, well-known anti-cancers agent (Oxaliplatin
(abbreviated as "L-OHP"), Fluorouracil (5-FU), Paclitaxel (TXL;
abbreviated as "PTX"), Docetaxel (TXT; abbreviated as "DTX"),
Irinotecan (CPT-11; abbreviated as "CPT"), or Cyclophosphamide
(abbreviated as "CPA") and/or a cyclodextrin inclusion compound
(abbreviated as "GC-CD") of a cholestanol derivative in which G in
formula (1) is GlcNAc.beta.- (abbreviated as "GC") was added
(multi-fold dilution by FCS(-)-medium: final concentration:
.ltoreq.500 .mu.M, 50 .mu.L), followed by incubation at 37.degree.
C. for two days. GC-CD was prepared in accordance with a method
disclosed in Example 1(2) in WO 2007/026869. Specifically, a 40%
aqueous solution of hydroxypropyl-.beta.-cyclodextrin was prepared,
and GC was added to the solution, followed by mixing with stirring
(80.degree. C. for 30 minutes), to thereby prepare GC-CD.
[0087] As a control, wells to which only FCS(-)-medium had been
added were employed. Viable count was performed by means of a cell
counting kit (product of Dojin).
[0088] Cell proliferation inhibition (CPI) rate (%) was calculated
by the following equation. FIG. 10 (FIGS. 10-A and 10-B) shows the
results.
CPI rate ( % ) = ( 1 - treated cells OD 450 - 650 untreated cells
OD 450 - 650 ) .times. 100 ##EQU00002##
[0089] As described hereinabove, through employment, in
combination, of the cholestanol derivative of the present invention
or a cyclodextrin inclusion compound thereof and an anti-cancer
agent, proliferation of various cancer cells is strongly inhibited,
and synergistic effect and/or effect of potentiating anti-tumor
action of a known anti-cancer agent can be obtained.
* * * * *