U.S. patent application number 12/926302 was filed with the patent office on 2011-07-28 for medical composition for inhibiting the growth of cancer stem cell.
This patent application is currently assigned to Chen-Yu Lee. Invention is credited to Hsiu-Mei Hsieh, Chen-Yu Lee.
Application Number | 20110183015 12/926302 |
Document ID | / |
Family ID | 44309144 |
Filed Date | 2011-07-28 |
United States Patent
Application |
20110183015 |
Kind Code |
A1 |
Hsieh; Hsiu-Mei ; et
al. |
July 28, 2011 |
Medical composition for inhibiting the growth of cancer stem
cell
Abstract
A medical composition for inhibiting the growth of cancer stem
cells is disclosed. The medical composition is prepared by mixing
herbal medicines with water or alcohol, followed by heating and
extraction to obtain a filtrate. One of the medical compositions
according to the present invention includes: Coptis chinensis,
Scutellaria baicalensis, Phellodendron amourense, Gardenia
jasminoides, Radix Glycyrrhizae, and Atractylodes japonica.
Inventors: |
Hsieh; Hsiu-Mei; (Taipei
City, TW) ; Lee; Chen-Yu; (Taipei City, TW) |
Assignee: |
Lee; Chen-Yu
Taipei City
TW
National Taiwan Normal University
Taipei City
TW
|
Family ID: |
44309144 |
Appl. No.: |
12/926302 |
Filed: |
November 9, 2010 |
Current U.S.
Class: |
424/741 ;
424/773 |
Current CPC
Class: |
A61K 36/756 20130101;
A61K 36/484 20130101; A61K 36/756 20130101; A61K 36/744 20130101;
A61K 36/284 20130101; A61K 36/71 20130101; A61K 36/232 20130101;
A61K 36/65 20130101; A61P 35/00 20180101; A61K 36/539 20130101;
A61K 36/718 20130101; A61K 36/744 20130101; A61K 36/718 20130101;
A61K 36/804 20130101; A61K 2300/00 20130101; A61K 2300/00 20130101;
A61K 2300/00 20130101; A61K 2300/00 20130101; A61K 2300/00
20130101; A61K 36/232 20130101; A61K 36/804 20130101; A61K 36/284
20130101; A61K 36/539 20130101; A61K 2300/00 20130101; A61K 2300/00
20130101; A61K 2300/00 20130101; A61K 2300/00 20130101; A61K
2300/00 20130101; A61K 36/484 20130101; A61K 36/71 20130101; A61K
36/65 20130101 |
Class at
Publication: |
424/741 ;
424/773 |
International
Class: |
A61K 36/756 20060101
A61K036/756; A61K 36/718 20060101 A61K036/718; A61P 35/00 20060101
A61P035/00 |
Foreign Application Data
Date |
Code |
Application Number |
Jan 22, 2010 |
TW |
099101774 |
Claims
1. A medical composition for inhibiting the growth of cancer stem
cells, comprising: an extract provided by mixing Coptis chinensis,
Scutellaria baicalensis, Phellodendron amourense, Gardenia
jasminoides, Radix Glycyrrhizae, and Atractylodes japonica with a
water-containing solution or an alcohol-containing solution,
followed by heating and extraction.
2. The medical composition as claimed in claim 1, wherein the
Coptis chinensis ranges from 3 to 5 weight parts, the Scutellaria
baicalensis ranges from 3 to 5 weight parts, the Phellodendron
amourense ranges from 3 to 5 weight parts, the Gardenia jasminoides
ranges from 3 to 5 weight parts, the Radix Glycyrrhizae ranges from
3 to 5 weight parts, and the Atractylodes japonica ranges from 3 to
5 weight parts.
3. The medical composition as claimed in claim 1, wherein the
alcohol-containing solution contains alcohol in 20-40%.
4. The medical composition as claimed in claim 1, wherein the
extract is prepared by heating up to 70.degree. C. or more and
extraction with the water-containing or alcohol-containing
solution.
5. The medical composition as claimed in claim 1, wherein the
extract is prepared by heating up to 70.degree. C. or more and
extraction with the water-containing or alcohol-containing solution
for at least 60 minutes.
6. A medical composition for inhibiting the growth of cancer stem
cells, comprising: an extract provided by mixing Coptis chinensis,
Rhizoma cimicifugae, Angelica sinensis, Rhizome of rehamnnia, and
Cortex Moutan Radicis with a water-containing solution or an
alcohol-containing solution, followed by heating and
extraction.
7. The medical composition as claimed in claim 6, wherein the
Coptis chinensis ranges from 3 to 5 weight parts, the Rhizoma
cimicifugae ranges from 3 to 5 weight parts, the Angelica sinensis
ranges from 3 to 5 weight parts, the Rhizome of rehamnnia ranges
from 3 to 5 weight parts, and the Cortex Moutan Radicis ranges from
3 to 5 weight parts.
8. The medical composition as claimed in claim 6, wherein the
alcohol-containing solution contains alcohol in 20-40%.
9. The medical composition as claimed in claim 6, wherein the
extract is prepared by heating up to 70.degree. C. or more and
extraction with the water-containing or alcohol-containing
solution.
10. The medical composition as claimed in claim 6, wherein the
extract is prepared by heating up to 70.degree. C. or more and
extraction with the water-containing or alcohol-containing solution
for at least 60 minutes.
Description
BACKGROUND OF THE INVENTION
[0001] 1. Field of the Invention
[0002] The present invention relates to a medical composition for
inhibiting the growth of cancer stem cells, which is capable of
inhibiting the growth of cancer stem cells in addition to common
cancer cells.
[0003] 2. Description of Related Art
[0004] Cancer has always been one of ten leading causes of death
but has increased in its perniciousness to become the first cause
of death for 27 years. The main factor causing cancer is that cells
become abnormal and keep dividing to form more cells, resulting in
cancer.
[0005] Western medicine therapy for treatment of cancer, such as
surgery, radiation therapy, chemotherapy, hormone therapy and
biopharmaceutical therapy, is notorious for the distressing
side-effects on patients. In view of those difficulties, more and
more people opt for the less-radical Chinese medicine therapy.
However, for both Chinese and Western medicine therapy, the
commercial anticancer drugs can inhibit only growth of cancer
cells, i.e., not cancer stem cells.
[0006] Cancer stem cells present in tumors are not large in amount,
but they are highly resistant to drugs and can generate more cancer
cells through cell division resulting in tumors. The aforementioned
fact is a thorny problem in treating cancer. The Western
chemotherapeutic drugs nowadays can kill only common cancer cells,
and cannot efficiently inhibit growth of cancer stem cells. That
is, a great amount of standard cancer treatment known in
biomedicine has no influence on cancer stem cells. Thereby, to
efficiently inhibit growth of cancer stem cells is inevitably
advantageous to cancer treatment.
[0007] In general, Chinese herbal medicine is considered more
moderate and more acceptable than Western chemical forms on the
market. Although some clinical treatment results suggest that parts
of medical compositions for treating cancer cells are indeed
capable of controlling cancer, the efficiency of these medical
compositions in inhibiting cancer cells as well as cancer stem
cells has not been confirmed.
[0008] Accordingly, to develop a medical composition that has the
confirmed efficiency in inhibiting cancer cells, particularly the
growth of precursor cells (i.e. cancer stem cells), is helpful to
cancer treatment.
SUMMARY OF THE INVENTION
[0009] The object of the present invention is to provide a medical
composition capable of inhibiting growth of cancer stem cells.
Also, the medical composition according to the present invention
can inhibit growth of common cancer cells.
[0010] To achieve the object, the present invention provides a
medical composition for inhibiting the growth of cancer stem cells
including: an extract provided by mixing Coptis chinensis,
Scutellaria baicalensis, Phellodendron amourense, Gardenia
jasminoides, Radix Glycyrrhizae, and Atractylodes japonica with a
water-containing solution or an alcohol-containing solution,
followed by heating and extraction.
[0011] In the medical composition, Coptis chinensis ranges from 3
to 5 weight parts, Scutellaria baicalensis ranges from 3 to 5
weight parts, Phellodendron amourense ranges from 3 to 5 weight
parts, Gardenia jasminoides ranges from 3 to 5 weight parts, Radix
Glycyrrhizae ranges from 3 to 5 weight parts, and Atractylodes
japonica ranges from 3 to 5 weight parts.
[0012] In addition, the present invention further provides a
medical composition for inhibiting the growth of cancer stem cells
including: an extract provided by mixing Coptis chinensis, Rhizoma
cimicifugae, Angelica sinensis, Rhizome of rehamnnia, and Cortex
Moutan Radicis with a water-containing solution or an
alcohol-containing solution, followed by heating and
extraction.
[0013] In the medical composition, Coptis chinensis ranges from 3
to 5 weight parts, Rhizoma cimicifugae ranges from 3 to 5 weight
parts, Angelica sinensis ranges from 3 to 5 weight parts, Rhizome
of rehamnnia ranges from 3 to 5 weight parts, and Cortex Moutan
Radicis ranges from 3 to 5 weight parts.
[0014] The above-mentioned medical compositions are prepared by
heating and extraction with a water-containing or
alcohol-containing solution. In the case of using an
alcohol-containing solution, preferably, the alcohol-containing
solution contains alcohol in 20-40%. In addition, during
extraction, the solution containing the Chinese herb materials may
be heated up to 70.degree. C. or more. Preferably, the solution
containing the Chinese herb materials is heated up to 70.degree. C.
or more and subjected to extraction for at least 60 minutes.
Accordingly, the medical composition according to the present
invention can be obtained.
BRIEF DESCRIPTION OF THE DRAWINGS
[0015] FIG. 1 is a statistical chart for showing the cell survival
rates of lung cancer cells after treatment with various dosages of
the medical composition according to Example 1 of the present
invention;
[0016] FIG. 2 is a statistical chart for showing the cell survival
rates of lung cancer cells and normal cells after treatment with
the medical composition according to Example 1 of the present
invention;
[0017] FIG. 3 is a chart for showing the correlation between
treatment time and the lung cancer cell survival rate after
treatment with the dose for 50% inhibition (ID50) of the medical
composition according to Example 1;
[0018] FIG. 4 is a statistical chart for showing the percentage of
lung cancer cells in each cycle stage after treatment with the
medical composition according to Example 1 of the present
invention;
[0019] FIG. 5 is a statistical chart for showing the percentage of
lung cancer cells in the G0 stage after treatment with the medical
composition according to Example 1 of the present invention;
[0020] FIG. 6 is a statistical chart for showing the fold change of
cancer stem cells after treatment with the medical composition
according to Example 1 of the present invention;
[0021] FIG. 7 is a statistical chart for showing the cell survival
rates of lung cancer cells after treatment with various dosages of
the medical composition according to Example 2 of the present
invention;
[0022] FIG. 8 is a statistical chart for showing the cell survival
rates of lung cancer cells and normal cells after treatment with
the medical composition according to Example 2 of the present
invention;
[0023] FIG. 9 is a chart for showing the correlation between
treatment time and the lung cancer cell survival rate after
treatment with the dose for 50% inhibition (ID50) of the medical
composition according to Example 2;
[0024] FIG. 10 is a statistical chart for showing the percentage of
lung cancer cells in each cycle stage after treatment with the
medical composition according to Example 2 of the present
invention;
[0025] FIG. 11 is a statistical chart for showing the percentage of
lung cancer cells in the G0 stage after treatment with the medical
composition according to Example 2 of the present invention;
and
[0026] FIG. 12 is a statistical chart for showing the fold change
of cancer stem cells after treatment with the medical composition
according to Example 2 of the present invention.
DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENT
[0027] In the following examples according to the present
invention, A549 cancer cell survival is characterized after
treatment with the medical compositions according to the present
invention. Then, the arresting stage of cell cycle by use of these
medical compositions according to the present invention is
identified by flow cytometric analysis. In addition, the cell
apoptosis resulted from the medical compositions according to the
present invention is studied through cell staining. Moreover, the
efficiency of the medical compositions according to the present
invention killing cancer stem cells is evaluated by double
fluorescence staining and flow cyometric analyses.
Example 1
[0028] Coptis chinensis (10 g), Scutellaria baicalensis (10 g),
Phellodendron amourense (10 g), Gardenia jasminoides (10 g), Radix
Glycyrrhizae (10 g), and Atractylodes japonica (10 g) are cut into
slices and mixed with water to perform heating and extraction to
obtain an extract. Herein, the heating is performed for 90 minutes
at 70.degree. C. Accordingly, the resulting extract is the medical
composition according to the present example.
Test Example 1
Cell Survival Rate Test
[0029] A549 lung cancer cell survival rate is characterized through
MTT assay after treatment with 5 .mu.l, 10 .mu.l and 50 .mu.l of
the medical composition according to Example 1 for 72 hours,
respectively. The results are shown in FIG. 1. Herein, the
horizontal axis represents a control group and various dosages of
medical compositions, and the vertical axis represents the
absorption of cells at 570 nm, which depends on the cell survival
rate.
[0030] FIG. 1 shows that the increased dosage of the medical
composition causes the reduction of A549 cancer cell survival rate
after treatment for 72 hours. In addition, it can be inferred from
FIG. 1 that the dose for 50% inhibition (ID50) of the medical
composition according to Example 1 on A549 cells is 20 .mu.l.
Test Example 2
Cell Survival Rate Test
[0031] After A549 lung cancer cells and MRC-5 normal cells are
treated with 20 .mu.l of the medical composition according to
Example 1 for 72 hours, respectively, the cell survival rates
thereof are characterized by MTT assay. The results are shown in
FIGS. 2 and 3. FIG. 2 is a statistical chart showing the survival
rates of the lung cancer cells and the normal cells after treatment
with the medical composition according to Example 1. FIG. 3 is a
chart showing the correlation between treatment time and the lung
cancer cell survival rate after treatment with the dose for 50%
inhibition (ID50) of the medical composition according to Example
1.
[0032] FIG. 2 suggests that the A549 lung cancer cell survival rate
is significantly reduced with no decrease of the MRC-5 cell
survival rate after treatment with the dose for 50% inhibition
(ID50) of the medical composition according to Example 1 for 72
hours. Thereby, the medical composition according to Example 1 can
significantly inhibit the growth of cancer cells but not the growth
of normal cells.
[0033] FIG. 3 shows that the survival rate of A549 lung cancer
cells is significantly reduced after treatment with ID50 of the
medical composition according to Example 1 for 24, 48 and 72 hours
in comparison with the A549 lung cancer cells with no treatment
with the medical composition according to Example 1. Meanwhile, the
variation of the cell survival rate increases with the increase of
time.
Test Example 3
Arresting Stage of Cell Cycle Test
[0034] A549 lung cancer cells are PI stained after treatment with
20 .mu.l (ID50) of the medical composition according to Example 1
for 72 hours. Subsequently, the DNA content of the lung cancer
cells is detected by flow cytometric analysis to determine cell
cycle distribution of lung cancer cells. The quantitative
statistical results are shown in FIG. 4. Herein, the G0/G1, S and
G2/M on the horizontal axis represent various cell cycle stages,
respectively, and the vertical axis represents the percentage of
cells in each cycle stage.
[0035] FIG. 4 suggests that the cell percentage in the G0/G1 stage
significantly increases in the lung cancer cells treated with the
medical composition according to Example 1 for 24, 48 and 72 hours,
in comparison with the lung cancer cells without treatment with the
medical composition according to Example 1. Thereby, it is
confirmed that the medical composition according to Example 1
causes A549 lung cancer cells to be arrested in the G0/G1
stage.
Test Example 4
Arresting Stage of Cell Cycle Test
[0036] A549 lung cancer cells are double stained with PI and Ki67
antibody and observed by flow cytometric analysis to determine cell
percentage of lung cancer cells in G0 stage after treatment with 20
.mu.l (ID50) of the medical composition according to Example 1 for
72 hours.
[0037] FIG. 5 suggests that the cell percentage in the G0 stage
significantly increased in the lung cancer cells treated with the
medical composition according to Example 1, in comparison with the
lung cancer cells without treatment with the medical composition
according to Example 1. Thereby, it is confirmed that more A549
cells can leave the cell cycle and stay in the quiescent G0 stage
after treatment with the medical composition according to Example
1.
Test Example 5
Cancer Stem Cell Ratio Test
[0038] A549 lung cancer cells are stained with Hoechst33342 in the
presence or absence of reserpine after treatment with 20 .mu.l
(ID50) of the medical composition according to Example 1 for 72
hours. The ratio of side population (SP) cells (i.e. cancer stem
cells) is evaluated by flow cytometric analysis. Reserpine is used
to inhibit ABCG2-mediated Hoechst33342 dye efflux. Accordingly, SP
cancer stem cells in which ABCG2 is highly expressed can be
identified by comparing the presence/absence of reserpine.
[0039] The quantitative statistical results are shown in FIG. 6.
Herein, the horizontal axis represents a fold change, i.e. the
ratio of the measured value in the presence of reserpine to that in
its absence. FIG. 6 suggests that the fold change can be reduced to
about 0.1 after treatment with the medical composition according to
Example 1. That is, the SP cancer stem cell mass is lower and the
ratio of the cancer stem cells is significantly reduced.
[0040] Thereby, it is confirmed that the medical composition
according to Example 1 can inhibit the growth of cancer stem
cells.
Example 2
[0041] Coptis chinensis (10 g), Rhizoma cumicifugae (10 g),
Angelica sinensis (10 g), Rhizome of rehamnnia (10 g) and Cortex
Moutan Radicis (10 g) are cut into slices and mixed with water to
perform heating and extraction to obtain an extract. Herein, the
heating is performed for 90 minutes at 70.degree. C. Accordingly,
the resulting extract is the medical composition according to the
present example.
Test Example 6
Cell Survival Rate Test
[0042] The test method according to the present test example is the
same as that of Test Example 1, except that the medical composition
according to Example 1 is replaced by that according to Example 2.
The results are shown in FIG. 7.
[0043] FIG. 7 shows that the increased dosage of the medical
composition causes the reduction of A549 cancer cell survival rate
after treatment for 72 hours. In addition, it can be inferred from
FIG. 7 that the dose for 50% inhibition (ID50) of the medical
composition according to Example 2 on A549 cells is 11 .mu.l.
Test Example 7
Cell Survival Rate Test
[0044] The test method according to the present test example is the
same as that of Test Example 2, except that 20 .mu.l of the medical
composition according to Example 1 is replaced by 11 .mu.l of the
medical composition according to Example 2. The results are shown
in FIGS. 8 and 9.
[0045] FIG. 8 suggests that the A549 lung cancer cell survival rate
is significantly reduced with no decrease of the MRC-5 cell
survival rate after treatment with the dose for 50% inhibition
(ID50) of the medical composition according to Example 2 for 72
hours.
[0046] FIG. 9 shows that the survival rate of A549 lung cancer
cells is significantly reduced after treatment with ID50 of the
medical composition according to Example 2 for 24, 48 and 72 hours
in comparison with the A549 lung cancer cells with no treatment
with the medical composition according to Example 2. Meanwhile, the
variation of the cell survival rate increases with the increase of
time.
Test Example 8
Arresting Stage of Cell Cycle Test
[0047] The test method according to the present test example is the
same as that of Test Example 3, except that 20 .mu.l of the medical
composition according to Example 1 is replaced by 11 .mu.l of the
medical composition according to Example 2. The quantitative
statistical results are shown in FIG. 10.
[0048] FIG. 10 suggests that the cell percentage in the G0/G1 stage
significantly increases regarding the lung cancer cells treated
with the medical composition according to Example 2 for 24, 48 and
72 hours, in comparison with the lung cancer cells of the control
group without treatment with the medical composition according to
Example 2. Thereby, it is confirmed that the medical composition
according to Example 2 causes A549 lung cancer cells to be arrested
in the G0/G1 stage.
Test Example 9
Arresting Stage of Cell Cycle Test
[0049] The test method according to the present test example is the
same as that of Test Example 4, except that 20 .mu.l of the medical
composition according to Example 1 is replaced by 11 .mu.A of the
medical composition according to Example 2. The quantitative
statistical results are shown in FIG. 11.
[0050] FIG. 11 suggests that the cell percentage in the G0 stage
significantly increases regarding the lung cancer cells treated
with the medical composition according to Example 2, in comparison
with the lung cancer cells without treatment with the medical
composition according to Example 2. Thereby, it is confirmed that
more A549 cells can leave the cell cycle and stay in the quiescent
G0 stage after treatment with the medical composition according to
Example 2.
Test Example 10
Cancer Stem Cell Ratio Test
[0051] The test method according to the present test example is the
same as that of Test Example 5, except that 20 .mu.l of the medical
composition according to Example 1 is replaced by 11 .mu.l of the
medical composition according to Example 2. The quantitative
statistical results are shown in FIG. 12.
[0052] FIG. 12 suggests that the fold change can be reduced to
about 0.2 after treatment with the medical composition according to
Example 2. That is, the SP cancer stem cell mass is lower and the
ratio of the cancer stem cells is significantly reduced. Thereby,
it is confirmed that the medical composition according to Example 2
can inhibit the growth of cancer stem cells.
[0053] From the results of Test Examples 1 to 10, it can be
confirmed that the medical composition according to the present
invention can inhibit the growth of cancer cells as well as cancer
stem cells.
[0054] Although the present invention has been explained in
relation to its preferred embodiment, it is to be understood that
many other possible modifications and variations can be made
without departing from the scope of the invention as hereinafter
claimed.
* * * * *