U.S. patent application number 10/942723 was filed with the patent office on 2005-02-10 for composition and method for treatment of neoplastic diseases associated with elevated matrix metalloproteinase activities using catechin compounds.
Invention is credited to Ivanov, Vadim, Netke, Shrirang, Niedzwiecki, Aleksandra, Rath, Matthias, Roomi, Waheed M..
Application Number | 20050032715 10/942723 |
Document ID | / |
Family ID | 21916454 |
Filed Date | 2005-02-10 |
United States Patent
Application |
20050032715 |
Kind Code |
A1 |
Netke, Shrirang ; et
al. |
February 10, 2005 |
Composition and method for treatment of neoplastic diseases
associated with elevated matrix metalloproteinase activities using
catechin compounds
Abstract
The present invention relates to a composition comprising a
catechin compound, ascorbic acid, proline and lysine. The present
invention also relates to a method for treating neoplastic disease
using a composition comprising a catechin compound, ascorbic acid,
proline and lysine.
Inventors: |
Netke, Shrirang; (San Bruno,
CA) ; Ivanov, Vadim; (Castro Valley, CA) ;
Roomi, Waheed M.; (Sunnyvale, CA) ; Niedzwiecki,
Aleksandra; (San Jose, CA) ; Rath, Matthias;
(Almelo, NL) |
Correspondence
Address: |
KENYON & KENYON
ONE BROADWAY
NEW YORK
NY
10004
US
|
Family ID: |
21916454 |
Appl. No.: |
10/942723 |
Filed: |
September 15, 2004 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
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10942723 |
Sep 15, 2004 |
|
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10041427 |
Jan 8, 2002 |
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Current U.S.
Class: |
514/27 ; 514/251;
514/423; 514/456; 514/564 |
Current CPC
Class: |
A61P 25/00 20180101;
A61K 31/375 20130101; A61K 31/525 20130101; A61P 31/10 20180101;
A61P 31/22 20180101; Y10S 514/824 20130101; A61K 31/355 20130101;
A61K 33/04 20130101; A61P 37/08 20180101; A61P 9/10 20180101; A61P
11/00 20180101; A61P 35/04 20180101; A61P 29/00 20180101; A61P 1/02
20180101; A61K 31/198 20130101; A61P 25/28 20180101; A61P 19/02
20180101; A61P 31/18 20180101; A61K 31/401 20130101; A61K 31/401
20130101; A61P 31/16 20180101; A61P 35/00 20180101; A61P 31/20
20180101; A61K 2300/00 20130101; A61K 2300/00 20130101; A61K 33/04
20130101; A61K 2300/00 20130101; A61P 17/00 20180101; A61K 31/525
20130101; A61P 43/00 20180101; A61P 1/16 20180101; A61P 31/04
20180101; A61P 9/04 20180101; A61K 31/385 20130101; A61P 19/10
20180101; A61P 37/02 20180101 |
Class at
Publication: |
514/027 ;
514/423; 514/456; 514/251; 514/564 |
International
Class: |
A61K 031/7048; A61K
031/525; A61K 031/401; A61K 031/198; A61K 031/353 |
Claims
What is claimed is:
1. A composition of biochemical substances comprising a catechin
compound, an anti-oxidant, proline and lysine that is effective in
inhibiting a matrix-metalloproteinase.
2. The composition according to claim 1, wherein the catechin
compound is selected from the group consisting of epicatechins,
epigallocatechin, epicatechin gallate, and epigallocatechin
gallate.
3. The composition according to claim 1, wherein the catechin
compound is epigallocatechin gallate.
4. The composition according to claim 1, wherein the anti-oxidant
is selected from the group consisting of ascorbic acid,
tocopherols, tocotrienols, carotinoids, glutathione, alpha-lipoic
acid, ubiquinols, bioflavonoids, and carnitine.
5. The composition according to claim 1, wherein the anti-oxidant
is ascorbic acid.
6. The composition according to claim 5, wherein the ascorbic acid
is selected from the group consisting of ascorbate precursors,
ascorbate metabolites and an ascorbate salt.
7. The composition according to claim 1, wherein the anti-oxidant
further comprises a folic acid.
8. The composition according to claim 7, wherein the folic acid is
folate.
9. The composition according to claim 1, wherein the composition
further comprises a selenium.
10. The composition according to claim 9, wherein the selenium is
selected from the group consisting of selinite and methyl
selinate.
11. The composition according to claim 1, wherein the
matrix-metalloproteinase is selected from the group consisting of
matrix-metalloproteinase 1, matrix-metalloproteinase 2,
matrix-metalloproteinase 3, matrix-metalloproteinase 4,
matrix-metalloproteinase 5, matrix-metalloproteinase 6,
matrix-metalloproteinase 7, matrix-metalloproteinase 8,
matrix-metalloproteinase 9, matrix-metalloproteinase 10,
matrix-metalloproteinase 11, matrix-metalloproteinase 12, and
matrix-metalloproteinase 14.
12. The composition according to claim 1, wherein the
matrix-metalloproteinases is matrix-metalloproteinase 2.
13. The composition according to claim 1, wherein the
matrix-metalloproteinase is matrix-metalloproteinase 9.
14. A method of treating human diseases associated with an elevated
matrix-metalloproteinase activity by administering to a patient in
need a composition of biochemical substances comprising a catechin
compound, an anti-oxidant, proline and lysine that are effective in
inhibiting a matrix-metalloproteinase.
15. The method according to claim 14, wherein the catechin compound
is selected from the group consisting of epicatechins,
epigallocatechin, epicatechin gallate, and epigallocatechin
gallate.
16. The method according to claim 14, wherein the catechin compound
is epigallocatechin gallate.
17. The method according to claim 14, wherein the anti-oxidant is
selected from the group consisting of ascorbic acid, tocopherols,
tocotrienols, carotinoids, glutathione, alpha-lipoic acid,
ubiquinols, bioflavonoids, and carnitine.
18. The method according to claim 14, wherein the anti-oxidant is
ascorbic acid.
19. The method according to claim 14, wherein the ascorbic acid
further comprises an ascorbate selected from the group consisting
of ascorbate precursors, ascorbate metabolites and ascorbate
salts.
20. The method according to claim 14, wherein the anti-oxidant
further comprises a folic acid.
21. The method according to claim 20, wherein the folic acid is
folate.
22. The method according to claim 14, wherein the composition
further comprises a selenium.
23. The method according to claim 14, wherein the selenium is
selected from the group consisting of selinite and methyl
selinate.
24. The method according to claim 14, wherein the human disease is
selected from the group consisting of neoplastic diseases,
inflammatory diseases, infectious diseases, cardiovascular
diseases, degenerative diseases, neurological diseases and
autoimmune diseases.
25. A method of treating neoplastic diseases related to an
excessive degradation of extracellular matrix comprising
administering an effective amount of a composition comprising a
catechin compound, an anti-oxidant, proline and lysine.
26. The method according to claim 25, wherein the catechin is
selected from the group consisting of epicatechins,
epigallocatechin, epicatechin gallate, and epigallocatechin
gallate.
27. The method according to claim 25, wherein the catechin is
epigallocatechin gallate.
28. The method according to claim 25, wherein the anti-oxidant is
selected from the group consisting of ascorbic acid, tocopherols,
tocotrienols, carotinoids, glutathione, alpha-lipoic acid,
ubiquinols, bioflavonoids, and carnitine.
29. The method according to claim 25, wherein the anti-oxidant is
ascorbic acid.
30. The method according to claim 25, wherein the ascorbic acid
further comprises ascorbate precursors, ascorbate metabolites and
an ascorbate salt.
31. The method according to claim 25, wherein the anti-oxidant
further comprises a folic acid.
32. The method according to claim 31, wherein the folic acid is
folate.
33. The method according to claim 25, wherein the composition
further comprises a selenium.
34. The method according to claim 33, wherein the selenium is
selected from the group consisting of selinite and methyl
selinate.
35. A pharmacological preparation according to claim 1.
36. A pharmaceutical preparation/s according to claim 1, where the
pharmacological preparation is used for oral administration,
parental administration or topical application.
Description
FILED OF THE INVENTION
[0001] The present invention relates to the use of catechin
compounds in combination with other dietary constituents in
inhibiting matrix-metalloproteinases. More particularly, the
present invention relates to the use of a composition comprising
catechin, ascorbic acid, lysine and proline in treating neoplastic
diseases.
BACKGROUND OF THE INVENTION
[0002] Polyphenolic compounds, also known as catechins, are present
in green tea and have been suggested to provide protection against
variety of illnesses including cancer (Mukhtar H., Ahmed N. Am. J.
Clin. Nutr. 71:1698S-1702S (2000)). Sadzuka et al. showed that oral
administration of green tea enhanced the tumor-inhibitoy effects of
doxorubicin in mice.
[0003] The anti-cancer activity of catechins may relate to their
effects on several factors involved in proliferation of cancer
cells and their metastasis. Catechins are known to cause cell cycle
arrest in human carcinoma cells (Ahmad N., Feyes D. K., Nieminen A.
L., Agarwal R., Mukhtar H. J. Natl. Cancer Inst. 89: 1881-1886
(1997)). Polyphenolic fraction from green tea is shown to protect
against inflammation and cytokines induced by tumors.
[0004] Polyphenolic compounds present as 30% dry weight in green
tea. They include flavanols, flavandiols, flavonoids, and phenolic
acids. Flavanols are the most abundant among the polyphenols in
green tea and are commonly known as catechins. There are four major
catechins in green tea: 1) (-)-epicatechin, 2)
(-)-epicatechin-3-gallate, 3) (-)-epigallocatechin, and 4)
(-)-epigallocatechin-3-gallate (EGCG). Among the catechins, EGCG is
the major polyphenolic constitutents present in green tea.
[0005] EGCG is a potent anti-oxidant compound (J. Cell. Biochem.
265:236-257 (1996)) and may attribute to the anti-cancer activity
of green tea. Catechin compounds were reported to exercise its
anti-metastatic activity by preventing the angiogenesis process
(Cao Y., Cao R. Nature 398:381 (1999)). EGCG has also been shown to
interfere with the activity of urokinase (u-plasminogen activator),
one of the most frequently expressed enzymes in human cancers
(Jankun J., Selman S. H., Swiercz R., Skrzypczak J. E. Nature:
387-567 (1997)).
[0006] However, it is established that the bioavailability of
polyphenols in humans is extremely low (Chen L., Lee M. J., Yang C.
S. Drug Metab. Dispos. 25: 1045-1050 (1997); Yang C. S., Chen L.,
Lee M. J., Balentine D. A., Kuo M. C., Schantz S. Cancer Epidemol.
Biomark. Prev. 7: 351-35 (1998); Bell J. R., Donovan J. L., Wong
R., Waterhouse H., German J. B., Walzem R. L., Kasim K. Am. J.
Clin. Nutr. 71:103-108 (2000); Sherry Chow H. H., Cai Y., Alberts
D. S., Hakim I., Dorr R., Shahi F., Crowell J. A., Yang S. C., Hara
H. Cancer Epidemol. Biomark. Prev. 10: 53-58 (2001)). The
references cited are hereby incorporated by reference by its
entireties. The low tissue concentration greatly reduces the
therapeutic value of polyphenols including EGCG. There is a
constant need in finding a better composition containing
polyphenols that is effective in the treatment of neoplastic
diseases. We surprisingly found a composition comprising catechins,
ascorbic acid, proline, and lysine that can exert a potent
anti-proliferative and anti-metastatic activity against neoplastic
diseases.
SUMMARY OF THE INVENTION
[0007] The present invention relates to a composition of
biochemical substances comprising a catechin, an anti-oxidant,
proline and lysine that are effective in treating human
diseases.
[0008] The present invention relates to a composition of
biochemical substances comprising a catechin, an anti-oxidant,
proline and lysine that are effective in inhibiting a
matrix-metalloproteinase.
[0009] The present invention relates to a method of treating
neoplastic diseases related to excessive degradation of
extracellular matrix comprising administering an effective amount
of a composition comprising a catechin compound, an anti-oxidant,
proline and lysine.
BRIEF DESCRIPTION OF THE DRAWINGS
[0010] FIG. 1 depicts the inhibitory effects of EGCG on cell
proliferation of breast cancer cells (MDA-MB 231).
[0011] FIG. 2 depicts the synergistic inhibitory effects of the
combination of EGCG, ascorbic acid, proline, and lysine on cell
proliferation of breast cancer cells (MDA-MB 231).
[0012] FIG. 3 depicts the synergistic inhibitory effects of the
combination of EGCG, ascorbic acid, proline, and lysine on cell
proliferation of colon cancer cells (HCT116).
[0013] FIG. 4 depicts that EGCG (20 .mu.g/ml) inhibits the Matrigel
invasion by breast cancer cells by about 25%. A combination of
ascorbic acid, proline and lysine inhibits about 65%. A combination
of ascorbic acid, proline and lysine with EGCG (20 .mu.g/ml)
completely inhibits (about 100%) the Matrigel invasion.
[0014] FIG. 5 depicts the combination of ascorbic acid, proline and
lysine with the EGCG (20 .mu.g/ml) synergistically inhibits to 100%
of Matrigel invasion by melanoma cells (A2058).
[0015] FIG. 6 depicts a zymogram showing EGCG decreases the
activity of MMP2 secreted by breast cancer cells.
[0016] FIG. 7 depicts the normal morphology of melanoma cells after
the Matrigel invasion assay.
[0017] FIG. 8 depicts the changes induced by the combination of
ascorbic acid, proline and lysine in the morphology of the melanoma
cells.
[0018] FIG. 9 depicts the apoptotic effects of the combination of
ascorbic acid, proline and lysine with EGCG.
DETAILED DESCRIPTION OF THE INVENTION
[0019] In another embodiment, the present invention relates to a
composition comprising catechins present in tea extracts, red wine
in combination with other dietary constituents, for synergistic
effects, against neoplastic diseases and a variety of other
illnesses. The dietary constituents covered by this application
include, but are not limited to those detailed in this
application.
[0020] Catechin compounds may be used in combination with other
anti-oxidants such as vitamin E, glutathione, with other
flavinoids, with facilitatory agents like folic acid and with
metals like selenium which are known to suitably modify the
activity of matrix-metalloproteinases, will enable us to reduce the
effective concentration at which EGCG can manifest its anti-tumor
activity.
[0021] In another embodiment, the present invention relates to a
composition comprising catechins that is effective in reducing
transformation of normal body cells into cancerous cells.
[0022] In another embodiment, the present invention relates to a
composition comprising catechins that is effective in preventing
cell proliferation of cancerous cells and in reducing synthesis,
secretion and/or activity of various matrix-metalloproteinases that
digest extra-cellular matrix (ECM).
[0023] In another embodiment, the present invention relates to a
method of preventing and treating neoplastic conditions by
administering such a composition comprising catechins, ascorbic
acid, proline and lysine orally or by topical application.
[0024] The present invention is described in further detail with
reference to the following examples, with no limitation of the
invention implied.
[0025] General Experimental Conditions
[0026] (a) Cell lines
[0027] The following cancer cell lines obtained from ATCC were used
in the studies:
[0028] (i) Human Breast Cancer Cells MDA-MB 231
[0029] (ii) Human Colon Cancer Cells HCT 116
[0030] (iii) Human Melanoma Cells A 2058
[0031] (b) Cell Proliferation Studies
[0032] To study the effects of catechins and dietary composition on
cell proliferation of human cancer cells, various human cancer cell
lines were cultured in 24 well plates using the culture conditions
specified by ATCC (supplier of cell lines). The cells were
generally incubated for 3 to 4 days (before coalescence was
reached). The total number of cells in a culture well was
determined by staining the cells with vital stain (MTT) followed by
the determination of OD of the stain solution. MTT only stain the
dead cells and the amount of stain uptake correlates with the
number of dead cells in the culture. Percent inhibition was
calculated by comparing the OD of the treatment groups with the OD
of the control groups.
[0033] (c) Matrigel Invasion Studies
[0034] The Matrigel Invasion studies were conducted using Matrigel
(Becton Dickinson) inserts in compatible 24 well plates. This assay
is a reliable assay for evaluating cancer metastasis.
[0035] Human fibroblast cells were seeded and grown in the 24-well
plates using culture media containing .about.10% serum. When the
fibroblasts reached coalescence, the culture media with serum was
withdrawn and replaced with fresh media without serum. A
combination of catechin compounds plus dietary composition were
added to the media without serum and human cancer cells were seeded
on the upper surface of the Matrigel inserts.
[0036] After 18 hours, the media were withdrawn. Some media were
saved for zymogram studies. The cells on the upper surface of the
inserts were gently scrubbed away with cotton swab. The cells that
had penetrated the Matrigel membrane and had migrated into the
lower surface of the Matrigel were stained with Quick Stain and
were counted under a microscope.
[0037] (d) Zymogram Studies
[0038] The media (25-30 .mu.l) from Matrigel Invasion studies was
applied to Novex zymogram gels (Invitrogen). The gel plates were
developed and stained as recommended by the manufacturer. The
matrix-metalloproteinases (MMPs) bands were identified on the basis
of their known molecular weights.
[0039] (e) Morphological Studies
[0040] The morphology of human cancer cells that had migrated into
the lower surface of the Matrigel membrane were stained with Quick
Stain and were photographed under a microscope (100.times.).
EXAMPLE 1
[0041] Inhibitory Effects of Cpigallocatechin Gallate (EGCG) on
Cell Proliferation of Human Breast Cancer Cells (MDA MB 231)
[0042] In these studies, 5.times.10.sup.4 breast cancer cells
(MDAMB 231) were seeded in each of the wells of 24-well plate.
Control group refers to breast cancer cells that were grown in
Liebovitz's media supplemented with 10% fetal bovine serum (FBS).
Treatment group refers to breast cancer cells that were grown in
Liebovitz's media supplemented with 10% fetal bovine serum (FBS)
plus either 0, 10, 20, 50, 100 or 200 mg/ml of EGCG. Plates were
incubated in ambient air (without supplemental CO.sub.2) for a
period of 4 days.
[0043] At the end of the period, the culture media were withdrawn
and the cells in each well were stained with MTT. Excess MTT stain
was washed off. The MTT stained cancer cells were dissolved in 1 ml
DMSO solution. The optical density (OD) of the solution was
determined for each well. The OD for the well was directly
proportional to the number of dead cells. The OD of the MTT stained
cancer cells that were previously cultured in the absence of EGCGT
was used as a reference and was considered as 100. Percent
inhibition was calculated by using the formula: % Inhibition=(OD of
Reference -OD of the Test Treatment)/OD of Reference .times.
100%.
[0044] EGCG at 20, 50, 100 and 200 .mu.g/ml caused 3, 34, 66 and
70% inhibition of cell proliferation of human cancer cells
respectively (FIG. 1). EGCG at 10 ug/ml did not inhibit the cell
proliferation.
EXAMPLE 2
[0045] Inhibitory Effect of A Combination of Ascorbic acid, Proline
and Lysine with Various Concentrations of EGCG on Cell
Proliferation of Human Breast Cancer Cells (MDA MB 231)
[0046] The general procedure of these studies remains to be the
same as in Example 1.
[0047] In these studies, basal culture media were supplemented with
the followings:
[0048] i) ascorbic acid (100 .mu.M)+proline (140 .mu.M);
[0049] ii) ascorbic acid (100 .mu.M)+proline (140 .mu.M)+Lysine
(400 .mu.M);
[0050] iii) ascorbic acid (100 .mu.M)+proline (140 .mu.M)+Lysine
(400 .mu.M) plus 20 mg/ml EGCG;
[0051] iv) ascorbic acid (100 .mu.M)+proline (140 .mu.M)+Lysine
(400 .mu.M) plus 50 mg/ml EGCG; or
[0052] v) ascorbic acid (100 .mu.M)+proline (140 .mu.M)+Lysine (400
.mu.M) plus 100 mg/ml EGCG.
[0053] Ascorbic acid+proline did not cause any inhibition of cell
proliferation. Ascorbic acid+proline+lysine inhibited cell
proliferation by about 14% (FIG. 2). A combination of ascorbic
acid+proline+lysine plus 20 .mu.g/ml of EGCG caused 20% inhibition
(FIG. 2). 20 .mu.g/ml of EGCG alone caused only 3% inhibition
(Example 1). Thus, a combination of ascorbic acid, proline and
lysine act synergistically with EGCG to inhibit cancer cell
proliferation.
EXAMPLE 3
[0054] Inhibitory Effects of a Combination of Ascorbic acid,
Proline and Lysine with Various Levels of EGCG on Cell
Proliferation of Human Colon Cancer Cells (HCT116)
[0055] In this study, human colon cancer cells were grown in
McCoy's 5A medium with 10% fetal bovine serum in 5% C0.sub.2
atmosphere. The general procedure and the treatment investigated
were the same as used in Example 2.
[0056] A combination of ascorbic acid, proline and lysine with EGCG
synergistically increased the inhibitory effects on cell
proliferation from 0% to 31% at 20 ug/ml EGCG and to about 95% at
50 .mu.g/ml EGCG (FIG. 3).
[0057] Inhibitory Effects of EGCG and a Combination of Ascorbic
Acid, Proline and Lysine on Invasion of Matrigel by Cancer
Cells
EXAMPLE 4
[0058] Inhibitory Effects of Graded Levels of EGCG and Combination
of Ascorbic Acid, Proline and Lysine with Various Levels of EGCG on
Invasion Through Matrigel by Breast Cancer Cells (MDA MB 231)
[0059] The general procedure for Matrigel Invasion Assay has been
described above. In this assay, human breast cancer cells
(5.times.10.sup.4) were seeded on each insert. Various supplements
were added to Leibovitz's media. The plates were incubated in an
incubator in ambient air without supplemental CO.sub.2.
[0060] A composition comprising 20 or 50 .mu.g/ml EGCG in the media
inhibited the invasion by the breast cancer cells by about 26% and
100% respectively. While ascorbic acid (100 .mu.M)+proline (140
.mu.M)+lysine (400 .mu.M) in the media caused 65% inhibition, a
combination of ascorbic acid (100 .mu.M)+proline (140 .mu.M)+lysine
(400 .mu.M) plus 20 .mu.g/ml of EGCG completely inhibited (100%
inhibition) of cancer cell invasion (FIG. 4).
EXAMPLE 5
[0061] Inhibitory Effects of Graded Levels of EGCG and Combination
of Ascorbic Acid, Proline and Lysine with Various Levels of EGCG on
Invasion Through Matrigel by Human Melanoma Cells (A2058)
[0062] The general procedure for Matrigel Invasion assay has been
described above. Human melanoma cells (A2058) (5.times.10.sup.4)
were seeded on each insert. Various supplements were added to DMEM.
The plates were incubated in an incubator under 5% CO.sub.2
atmosphere.
[0063] While a combination of ascorbic acid (100 .mu.M)+proline
(140 .mu.M)+lysine (400 .mu.M) caused only 13% inhibition, a
combination of ascorbic acid (100 .mu.M)+proline (140 .mu.M)+lysine
(400 .mu.M) plus 20 .mu.g/ml EGCG completely prevented the invasion
of melanoma cells through the Matrigel (FIG. 5).
[0064] Zymogram Studies
EXAMPLE 6
[0065] Effects of Graded Levels of EGCG on MMP2 Production by Human
Breast Cancer Cells (MDA MB231)
[0066] The media from various treatments in the Matrigel Invasion
assay (Example 4) were applied to Novex Zymogram Gel (Invitrogen).
The plates were developed and stained as recommended by the
manufacturer. The matrix metalloproteinases (MMPs) bands were
identified on the basis of their known molecular weights (FIG.
6).
[0067] Zymogram of the culture media from the Matrigel Invasion
Assays indicated that 20 ug/ml EGCG in the media reduced the
production of MMP2 and completely inhibited the production of MMP9
(FIG. 6). At concentrations of 50 .mu.g/ml and 100 ug/ml of EGCG,
the activities of both MMP2 and MMP9 were completely inhibited
(FIG. 6).
[0068] Cell Morphology
EXAMPLE 7
[0069] Effects of EGCG and a Combination of Ascorbic Acid, Proline
and Lysine on the Cell Morphology of Human Melanoma Cells
(A2058)
[0070] The micrographs of the cancer cells in basal media as they
migrated through the Matrigel are shown FIG. 7. Inclusion of the
combination of ascorbic acid (100 .mu.M)+proline (140 .mu.M)+lysine
(400 .mu.M) in the media altered the morphology of the cells (FIG.
8). The distension of the cells with distinct enlargement of
nucleus was evident. Addition of 20 ug/ml of EGCG to the
combination of ascorbic acid (100 .mu.M)+proline (140 .mu.M)+lysine
(400 .mu.M) in the media caused extensive apoptotic changes (FIG.
9).
[0071] These findings described in the examples 1-7 indicate that a
strong synergistic effect exerted by EGCG, when EGCG was used with
a combination of ascorbic acid+ proline+lysine. Therefore, these
studies show a surprising synergistic effect of a combination of
EGCG and ascorbic acid+proline+lysine makes it possible to take
full advantage of anti-proliferative and anti-metastasis activity
of EGCG at a comparatively low level of its tissue
concentration.
[0072] Hence, the present findings are of immense importance as
they can bring effective level of catechins closer to those, which
can be achieved, in the tissues.
[0073] It has been suggested that the proliferation of cancer cells
and up-regulation of their enzymes are caused by increased
concentration of reactive oxygen species (ROS). In this situation,
use of combinations of various biological antioxidants such as
tocopherols, carotinoids, along with other facilitating agents like
ubiquinols, biflavonoides, lipoic acid, carnitine will provide a
more potent synergistic mixture for treatment of the
above-mentioned maladies.
[0074] The present invention provides a surprising observation that
a combination comprising catechin compounds would exert synergistic
activity and thereby make it possible to achieve very efficient
anti-cancer activity at lower levels of tissue catechins. The above
findings open the possibility of using various constituents in
combination of different constituents at effective levels for the
prevention and treatment of neoplastic diseases.
[0075] One skill in the art will appreciate that proline and lysine
are not merely limited to proline and lysine. The scope of the
present invention is intended to cover lysine derivatives and its
precursors, proline derivatives and its precursors.
[0076] One skill in the art will appreciate that the anti-oxidant,
ascorbic acid, should cover the derivatives and precursors of
ascorbic acid.
[0077] Other biological anti-oxidants include tocopherols and
related compounds, trans-retinoic acid and related compounds,
carotinoids and related compounds, glutathione and related
compounds, ubiquinols and related compounds, folates and related
compounds, bioflavonoids and related compounds as well as compounds
of selenium.
[0078] Clinical Applications
[0079] The invention focuses on the preventive and therapeutic use
of a catechin in combination with an anti-oxidant, proline and
lysine. The combined use of a catechin with an anti-oxidant,
proline and lysine increases the efficiency of the catechin
compound in treating human diseases.
[0080] Human diseases include but are not limited to neoplastic
diseases, inflammatory conditions, infectious diseases,
cardiovascular diseases and other pathological conditions involving
degradation of extra-cellular matrix. Such conditions include
abnormal angiogenesis, pathological intravasation, rheumatoid and
osteoarthritis, atherosclerosis, dilated cardiomyopathy, emphysema
and other chronic conditions.
[0081] The present invention provides a method of treating and
preventing human diseases involving degradation of extra-cellular
matrix such as: i) neoplastic diseases; ii) inflammatory conditions
(including but not limited to allergies, emphysema, rheumatoid
arthritis, osteoarthritis, periodontitis, neurodermatitis); iii)
infectious diseases (including but not limited to viral infections
such as common cold, influenza, hepatitis, herpes, HIV; bacterial
infections such as pneumonia, tuberculosis, meningitis, gonorrhea,
syphilis, and or fungal diseases; iv) cardiovascular diseases
(including but not limited to atherosclerosis, cardiomyopathy,
restonosis after angioplasty); v) degenerative diseases (including
but not limited to osteoporosis and arthritis); vi) neurological
disorders (including but not limited to Alzheimer Disease, multiple
sclerosis); and vii) autoimmune diseases (including but not limited
to arthritis) by administering effective amounts of compositions
described in this application.
* * * * *