supplement combinations

Rich66 · 01-12-2010, 12:55 AM

Arch Biochem Biophys. 2008 Aug 15;476(2):107-12. Epub 2008 Feb 7.
Are polyphenols antioxidants or pro-oxidants? What do we learn from cell culture and in vivo studies?

FULL TEXT

Halliwell B.
Department of Biochemistry, National University of Singapore, University Hall, Lee Kong Chian Wing, UHL #05-02G, 21 Lower Kent Ridge Road, Singapore 119077, Singapore. bchbh@nus.edu.sg
Diets rich in polyphenols are epidemiologically associated with lower risk of developing some age-related diseases in humans. This apparent disease-protective effect of polyphenols is often attributed to their powerful antioxidant activities, as established in vitro. However, polyphenols can also exert pro-oxidant activities under certain experimental conditions. Neither pro-oxidant nor anti-oxidant activities have yet been clearly established to occur in vivo in humans, nor are they likely given the limited levels of polyphenols that are achievable in vivo after consumption of foods and beverages rich in them. Other actions of polyphenols may be more important in vivo. Many studies of the biological effects of polyphenols in cell culture have been affected by their ability to oxidise in culture media, and awareness of this problem can avoid erroneous claims.

PMID: 18284912 [PubMed - indexed for MEDLINE]

Altern Med Rev. 2009 Sep;14(3):226-46.
Bioavailability and activity of phytosome complexes from botanical polyphenols: the silymarin, curcumin, green tea, and grape seed extracts.

Kidd PM.
Cell biology, University of California, Berkeley, USA. dockidd@dockidd.com
Plant-derived polyphenols are increasingly receiving attention as dietary supplements for the homeostatic management of inflammation, to support detoxication, and for anticancer, weight loss, and other benefits. Their pro-homeostatic effects on genes, transcription factors, enzymes, and cell signaling pathways are being intensively explored, but the poor bioavailability of some polyphenols likely contributes to poor clinical trial outcomes. This review covers four polyphenol preparations with poor bioavailability and their complexation into phytosomes to bypass this problem. Silybin and the other silymarin flavonolignans from milk thistle conserve tissue glutathione, are liver-protective, and have anticancer potential. Curcumin and its related diphenolic curcuminoids have potent antioxidant, anti-inflammatory, and anti-carcinogenic properties. The green tea flavan-3-ol catechins have antioxidant, anti-inflammatory, cardio- and neuro-protective effects, and anti-carcinogenic benefits, with fat oxidation effects coupled to weight loss. The complex grape seed proanthocyanidin mix (including catechin and epicatechin monomers and oligomers) counters oxidative stress and protects the circulatory system. For each of these preparations, conversion into phytosomes has improved efficacy without compromising safety. The phytosome technology creates intermolecular bonding between individual polyphenol molecules and one or more molecules of the phospholipid, phosphatidylcholine (PC). Molecular imaging suggests that PC molecule(s) enwrap each polyphenol; upon oral intake the amphipathic PC molecules likely usher the polyphenol through the intestinal epithelial cell outer membrane, subsequently accessing the bloodstream. PC itself has proven clinical efficacy that contributes to phytosome in vivo actions. As a molecular delivery vehicle, phytosome technology substantially improves the clinical applicabilities of polyphenols and other poorly absorbed plant medicinals.

PMID: 19803548 [PubMed - indexed for MEDLINE]

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A Specific Combination Of Ascorbic Acid, Lysine, Proline And Epigallocatechin Gallate Inhibits Proliferation And Extracellular Matrix Invasion Of Various Human Cancer Cell Lines
FULL TEXT

Shrirang P Netke, M Waheed Roomi, Vadim Ivanov, Aleksandra Niedzwiecki* and Matthias Rath

Matthias Rath, Inc., Research and Development, Santa Clara, CA 95054
Author for correspondence and reprints. Phone (408) 807-5564; FAX: (408) 986-9403;
EMERGING DRUGS – VOL. II
Copyright © 2003 by
PJD Publications Limited
Research Communications
Westbury, NY 11590-0966 USA

Abstract

We found that a specific combination of ascorbic acid (AA, 100µM), proline (P, 140 µM) and lysine (L, 400 µM) had a significant antiproliferative and antimetastatic effect against some human cancer cell lines, breast (MDA-MB-231), colon (HCT116) and skin (melanoma, A2058). In the presence of this nutrient combination the invasion of the extracellular matrix by human breast cancer cells, melanoma cells and colon cancer cells was inhibited by 50%, 10% and 30% respectively. Addition of epigallocatechin gallate (EGCG) further enhanced this nutritional synergy producing a more pronounced inhibitory effect on both cellular proliferation and matrix invasion. The proliferation of breast cancer cells MDA-MB-231 in the presence of AA, P, L and 20µg/ml of EGCG was reduced to 74% and colon cancer cells HCT116 to 69% compared to the unsupplemented medium. The increase in concentration of EGCG to 50µg/ml did not cause much further reduction in the number of breast cancer cells. However it reduced proliferation of colon cancer cells to 4,6% and melanoma cells to 30% of the control. Matrigel invasion by breast cancer cells and human melanoma cells in the presence of AA, P, L and 20µg/ml of EGCG was stopped completely. At a similar concentration, invasion by colon cancer cells was reduced to 24,9%. However, the expression of matrix metalloproteinases (MMPs) –2 and –9 was not altered by this nutrient combination in melanoma cells as visualized by gelatinase zymography. MMP-2 and MMP-9 were significantly inhibited by EGCG in a dose dependent manner, and L, P, AA have no additional effect. Thus the combination of AA, P, L and EGCG shows great potential for control of cancer using a safe and multi-targeted approach.

Quote:
The antiproliferative activity of the nutrient combinations used in these studies varied with the type of cancer cells. In breast cancer cells, the combination of AA, P and L with EGCG had higher anti-proliferative effects than when these nutrients were used individually

In vivo and in vitro antitumor effect of ascorbic acid, lysine, proline, arginine, and green tea extract on human fibrosarcoma cells HT-1080.Roomi MW et al. Med Oncol. (
2006)
In vivo and in vitro antitumor effect of ascorbic acid, lysine, proline and green tea extract on human melanoma cell line A2058.Roomi MW et al. In Vivo. (
2006)
In vivo antitumor effect of ascorbic acid, lysine, proline and green tea extract on human prostate cancer PC-3 xenografts in nude mice: evaluation of tumor growth and immunohistochemistry.Roomi MW et al. In Vivo. (2005)

Transl Oncol. 2008 Mar;1(1):19-27. Links: Combined resveratrol, quercetin, and catechin treatment reduces breast tumor growth in a nude mouse model.

Schlachterman A, Valle F, Wall KM, Azios NG, Castillo L, Morell L, Washington AV, Cubano LA, Dharmawardhane SF.
Department of Anatomy and Cell Biology, Universidad Central del Caribe School of Medicine, Bayamon, PR 00960, USA.
Grape polyphenols can act as antioxidants, antiangiogenics, and selective estrogen receptor (ER) modifiers and are therefore especially relevant for gynecological cancers such as breast cancer. The major polyphenols of red wine (resveratrol, quercetin, and catechin) have been individually shown to have anticancer properties. However, their combinatorial effect on metastatic breast cancers has not been investigated in vivo. We tested the effect of low dietary concentrations of resveratrol, quercetin, and catechin on breast cancer progression in vitro by analyzing cell proliferation and cell cycle progression. The effects of these compounds on fluorescently tagged breast tumor growth in nude mice were assessed using in situ fluorescence image analysis.; Individual polyphenols at 0.5 microM neither decreased breast cancer cell proliferation nor affected cell cycle progression in vitro. However, a combination of resveratrol, quercetin, and catechin at 0.5, 5, or 20 microM each significantly reduced cell proliferation and blocked cell cycle progression in vitro. Furthermore, using in situ image analysis, we determined that combined dietary polyphenols at 0.5, 5, or 25 mg/kg reduced primary tumor growth of breast cancer xenografts in a nude mouse model. Peak inhibition was observed at 5 mg/kg. These results indicate that grape polyphenols may inhibit breast cancer progression.

Biochem Biophys Res Commun. 2000 Jun 24;273(1):50-3.
Artifacts in cell culture: rapid generation of hydrogen peroxide on addition of (-)-epigallocatechin, (-)-epigallocatechin gallate, (+)-catechin, and quercetin to commonly used cell culture media.

Long LH, Clement MV, Halliwell B.
Department of Biochemistry, Faculty of Medicine, National University of Singapore, 10 Kent Ridge Crescent, 119260, Singapore.
There is considerable current interest in the possible beneficial health effects of quercetin, catechins, epigallocatechins, epigallocatechin gallates, and related phenolic compounds found in teas, wines, and other plant products. As a result, many laboratories are studying the effects of these compounds on cells in culture. The present paper shows that addition of these compounds to commonly used cell culture media leads to generation of substantial amounts of hydrogen peroxide (H(2)O(2)). Dulbecco's modified Eagle medium gives the highest H(2)O(2) level for all the compounds tested, with levels reaching >400 microM within 2 h for addition of 1 mM concentrations of gallic acid, epigallocatechin gallate, and epigallocatechin. Catechin and quercetin produced lower, but still significant, levels of H(2)O(2). McCoy's 5A and RPMI 1640 media also promoted H(2)O(2) production from the above phenolic compounds. This rapid generation of H(2)O(2) could account for some or all of the reported effects of phenolic compounds on cells in culture. Copyright 2000 Academic Press.

PMID: 10873562 [PubMed - indexed for MEDLINE]

Nutr Cancer. 2008;60(3):401-11.Links: Vitamin E analog alpha-TEA, methylseleninic acid, and trans-resveratrol in combination synergistically inhibit human breast cancer cell growth.

Snyder RM, Yu W, Jia L, Sanders BG, Kline K.
Institute for Cell and Molecular Biology, University of Texas at Austin, Austin, Texas 78712-1097, USA.
Alpha-tocopherol ether-linked acetic acid analog [2,5,7,8-tetramethyl-2R-(4R, 8R-12-trimethyltridecyl) chroman-6-yloxyacetic acid (alpha-TEA)] is a novel form of vitamin E effective at killing cancer cells but not normal cells. alpha -TEA alone and together with methylseleninic acid (MSA) and trans-resveratrol (t-RES) were investigated for ability to induce apoptosis, DNA synthesis arrest, and cellular differentiation and inhibit colony formation in human MDA-MB-435-F-L breast cancer cells in culture. The 3 agents alone were effective in inhibiting cell growth by each of the 4 different assays, and 3-way combination treatments synergistically inhibited cell proliferation in each assay in comparison to individual treatments. Furthermore, combinations of alpha -TEA, t-RES, and MSA significantly enhanced levels of apoptosis in human breast (MDA-MB-231, MCF7, and T47D) and prostate (LnCaP, PC-3, and DU-145) cancer cell lines as well as in immortalized but nontumorigenic MCF10A cells but not primary cultures of human mammary epithelial cells. Western immunoblotting confirmed the induction of apoptosis in that the 3 agents induced poly(adenosine diphosphate-ribose) polymerase cleavage, with earlier detection and more complete cleavage seen in the combination treatment. Mechanistic studies showed combination treatments to inhibit cell proliferation via downregulation of cyclin D1 and induce apoptosis via activation of caspases 8 and 9 and downregulation of prosurvival proteins FLIP and survivin. In summary, the combination of alpha-TEA, MSA, and t-RES is more effective than single treatments for inhibiting cell proliferation, inducing cellular differentiation, and inducing cell death by apoptosis in human cancer cells in culture.
PMID: 18444175 [PubMed - indexed for MEDLIN

1: Eur J Gynaecol Oncol. 2001;22(5):347-9.Links: Herbal complex suppresses telomerase activity in chemo-endocrine resistant cancer cell lines.

Lian Z, Fujimoto J, Yokoyama Y, Niwa K, Tamaya T.
Department of Obstetrics and Gynecology, Gifu University School of Medicine, Gifu City, Japan.
A herbal complex consisting of Hoelen, Angelicae radix, Scutellariae radix and Glycyrrhizae radix suppressed cell viability and telomerase activity in hormone-refractory and chemo-resistant cancer cell lines, namely poorly differentiated uterine endometrial cancer cell line AN3 CA, adriamycin-resistant breast cancer cell line MCF7/ADR and cisplatin-resistant ovarian cancer cell line A2780. Furthermore, the herbal complex suppressed the expression of the full length of human telomerase reverse transcriptase (hTERT), which is related to telomerase activity. This indicates that the herbal complex can suppress the tumor growth of chemoendocrine resistant cancers, at least in part via suppression of telomerase activity associated with down-regulated hTERT.
PMID: 11766737

Anticancer Res. 2009 Oct;29(10):3917-24.
The effect of a novel botanical agent TBS-101 on invasive prostate cancer in animal models.

Evans S, Dizeyi N, Abrahamsson PA, Persson J.
Department of Clinical Sciences, Clinical Research Center, Lund University, University Hospital, 205 02, Malmo, Sweden.

Evans S, Dizeyi N, Abrahamsson PA, Persson J.
Department of Clinical Sciences, Clinical Research Center, Lund University, University Hospital, 205 02, Malmo, Sweden.
BACKGROUND: Traditional Botanical Supplement-101 (TBS-101) is a newly developed proprietary botanical agent containing seven standardized botanical extracts, including: Panax ginseng, cranberry, green tea, grape skin, grape seed, Ganoderma lucidum and chamomile. Each of the components has been consumed either in the regular diet or as natural supplement. When used as a single agent, each of these seven botanicals has been implicated in chemoprevention and therapy in various types of cancer. The anticancer effect of TBS-101, with the specific combination of these anti-cancer botanicals for the treatment of prostate cancer (PCa), has not been tested. MATERIALS AND METHODS: The IC(50) and the effect of TBS-101 on the proliferation and apoptosis of PC-3 cells were determined. Tumor xenograft mice were generated by subcutaneously implanting PC-3 cells into mice and tumors were allowed to grow to different sizes before starting the treatment. The effects of TBS-101 on tumor growth were assessed by measuring tumor size and by histological, pathological and immunohistochemical analyses. A basic toxicity study was performed to test the tolerance of the mice to high doses of TBS-101. RESULTS: Treatment of the PC-3 cells with TBS-101 resulted in a dose-dependent inhibition of cell growth, with an IC(50) of 1.4 microg/ml. A concomitant induction of apoptosis in PC-3 cells treated with TBS-101 was also observed. Upon the treatment with TBS-101, all three groups of mice bearing moderate or large tumors showed significant inhibition of tumor growth and invasion. In contrast, control mice treated with vehicle alone had significant tumor growth and lymph node metastasis. In the basic toxicity studies, high doses of TBS-101 exerted no toxicity in healthy or tumor-bearing mice. CONCLUSION: The natural botanical agent TBS-101 has a good safety profile and significant anticancer activities in hormone-refractory PC-3 cells and large aggressive PC-3 tumors in a xenograft mouse model and has great potential for the treatment of aggressive prostate cancer.

PMID: 19846929 [PubMed - indexed for MEDLINE]

Int J Oncol. 2009 Nov;35(5):1183-9.
Fucoidan-Vitamin C complex suppresses tumor invasion through the basement membrane, with scarce injuries to normal or tumor cells, via decreases in oxidative stress and matrix metalloproteinases.

Saitoh Y, Nagai Y, Miwa N.
Cell-Death Control BioTechnology Laboratory, Faculty of Life and Environmental Sciences, Prefectural University of Hiroshima, Shobara, Hiroshima 727-0023, Japan.
Fucoidan (Fucdn) and vitamin C (VC) were saturatedly dissolved in water and lyophilized and thoroughly ethanol-rinsed until no detection for supernatant vitamin C to form the Fucdn-VC (1:0.23 wt/wt) inclusion body (Fucdn-VC-IB). Fucdn-VC-IB increased not only VC-stabilizing at 37 degrees C, but also hydroxyl-radical scavenging as shown by ESR/spin-trap method, more markedly than a mere mixture of Fucdn:VC (1:0.23 wt/wt). Invasion of human fibrosarcoma cells HT-1080 through the basement membrane was repressed by Fucdn-VC-IB of non-cytotoxic concentrations without significant inhibition to human skin dermal fibroblasts DUMS-16 cells. Fucdn-VC-IB suppressed the invasiveness-related gelatinases MMP-2/9, and diminished reactive oxygen species inside the cytoplasm around the nucleus, in HT-1080 cells as shown by electrophoretic zymography and the redox indicator NBT assay, respectively. Thus Fucdn-VC-IB markedly exhibits antioxidant and MMP-suppressing activities and preferentially inhibited tumor invasion without cytotoxicity to normal cells, and is suggested as a potent tumor-invasion suppressor.

PMID: 19787274 [PubMed - indexed for MEDLINE]

Oncol Rep. 2009 Nov;22(5):1077-83.
A traditional Chinese medicine formulation consisting of Rhizoma Corydalis and Rhizoma Curcumae exerts synergistic anti-tumor activity.

Gao JL, He TC, Li YB, Wang YT.
Institute of Chinese Medical Sciences, University of Macau, Macau 999078, PR China.
Synergy analysis of anticancer agents is an important approach to determining the ratio and/or dose of drugs for clinical combination therapy. However, this method is rarely used to evaluate the composition of traditional Chinese medicine formulation. 'Yanhusuo San' (YHSS), which consists of yanhusuo (Rhizoma Corydalis) and Ezhu (Rhizoma Curcumae), has been an archaic Chinese medicine prescription since the Song dynasty (960-1279 AD). We previously demonstrated that either yanhusuo or ezhu has strong anticancer effect. Herein, we sought to determine the possible synergic effect between these two Chinese herbs. We measured the IC50 of each herb extract and both extracts at different ratios of doses by MTT assay. Isobologram and combination index (CI) analyses were used to evaluate the synergistic effect of yanhusuo and ezhu in different fixed ratios. Our results indicated that a combination of two herbal extracts exhibits the strongest anticancer cell proliferation effect at the ratio of 3:2 (ezhu to yanhusuo; referred to as E3Y2). Using Boyden Chamber assay, flow cytometry, and fluorescence microscopy analysis, we found that E3Y2 could markedly reduce the cell invasion ability and induce cytochrome c release rather than single use, but E3Y2 could not influence the cell cycle distribution. When the levels of ERK1/2, p-ERK1/2 and p-Rb were determined by Western blot analysis, we found that the E3Y2 significantly suppresses the level of p-ERK. Thus, our studies provide a plausible molecular basis of the synergistic anti-tumor effect of ezhu and yanhusuo.

PMID: 19787224 [PubMed - indexed for MEDLINE]

Am J Chin Med. 2005;33(3):381-95.
Immunomodulatory activities of Yunzhi and Danshen in post-treatment breast cancer patients.

Wong CK, Bao YX, Wong EL, Leung PC, Fung KP, Lam CW.
Department of Chemical Pathology, The Chinese University of Hong Kong, Prince of Wales Hospital, Shatin, Hong Kong, PR. China.
Breast cancer is the most common cancer among women worldwide. Discomfort and fatigue are usually arisen from anticancer therapy such as surgery, radiotherapy, chemotherapy, hormonal therapy, or combination therapy, because of the suppressed immunological functions. Yunzhi (Coriolus versicolor) can modulate various immunological functions in vitro, in vivo, and in human clinical trials. Danshen (Salvia miltiorrhiza) has been shown to benefit the circulatory system by its vasodilating and anti-dementia activity. The purpose of this clinical trial was to evaluate the immunomodulatory effects of Yunzhi-Danshen capsules in post-treatment breast cancer patients. Eighty-two patients with breast cancer were recruited to take Yunzhi [50 mg/kg body weight, 100% polysaccharopeptide (PSP)] and Danshen (20 mg/kg body weight) capsules every day for a total of 6 months. EDTA blood samples were collected every 2 months for the investigation of immunological functions. Flow cytometry was used to assess the percentages and absolute counts of human lymphocyte subsets in whole blood. Plasma level of soluble interleukin-2 receptor (sIL-2R) was measured by enzyme-linked immunosorbent assay (ELISA). Results showed that the absolute counts of T-helper lymphocytes (CD4+), the ratio of T-helper (CD4+)/T suppressor and cytotoxic lymphocytes (CD8+), and the percentage and the absolute counts of B-lymphocytes were significantly elevated in patients with breast cancer after taking Yunzhi-Danshen capsules, while plasma slL-2R concentration was significantly decreased (all p < 0.05). Therefore, the regular oral consumption of Yunzhi-Danshen capsules could be beneficial for promoting immunological function in post-treatment of breast cancer patients.

Am J Chin Med. 2008;36(4):729-44.
Green tea extract enhances the selective cytotoxic activity of Zizyphus jujuba extracts in HepG2 cells.

Huang X, Kojima-Yuasa A, Xu S, Norikura T, Kennedy DO, Hasuma T, Matsui-Yuasa I.
Department of Food and Human Health Sciences, Graduate School of Human Life Science, Osaka City University, Osaka 558-8585, Japan.
Anticarcinogenic effects attributed to phytochemicals may be based on synergistic, additive, or antagonistic interactions of many compounds. In our previous study, we demonstrated that the chloroform fraction (CHCl(3)-F) from Z. jujuba has anticancer activity in HepG2 cells. In China, many people drink jujuba tea and believe in the synergic effects of jujuba and tea for better health. We therefore investigated the effects of CHCl(3)-F and green tea extract (GTE), and their underlying mechanisms of action in HepG2 cells. Our results showed that GTE enhanced the effect of CHCl(3)-F on cell viability in HepG2 cells, without cytotoxicity in rat hepatocytes, which was used as a normal cell model. Furthermore, combination of CHCl(3)-F and GTE caused an effect on G1 phase arrest but not on apoptosis. Interestingly, the mechanism of the G1 arrest was associated, not with an increase in p27(Kip1) levels and the hypohosphorylation of Rb, which are pathways used by CHCl(3)-F on G1 arrest in HepG2 cells, but with increases in p53 and p21(Waf1/Cip1) levels, and a decrease in cyclin E levels. Collectively, our findings suggest that combination of CHCl(3)-F and GTE produces an enhanced cell growth inhibition effect, and that the resultant G1 arrest was caused via a different mechanism as that of CHCl(3)-F treatment alone.

PMID: 18711770 [PubMed - indexed for MEDLINE]

Green tea with Capsaicin: http://www.newcancerresearch.com/about.htm
Capsibiol-T

Recently concluded preliminary tests at Purdue University using cancer cells in culture have shown that the synergistic formulation of Capsibiol-T inhibits tNOX. When tNOX activity is inhibited, cancer cells fail to grow to the size required for division and undergo programmed cell death (apoptosis). Capsibiol-T® is a unique, synergistic formulation of modified capsaicin and decaffeinated green tea. Before the proprietary process to modify capsaicin for exclusive use in Capsibiol-T® was developed, the dosage required for capsaicin to be effective (its therapeutic dosage) had exceeded the level at which its pungency and neurological discomfort could be tolerated. The amount of modified capsaicin (12.5mg per capsule) in synergy with decaffeinated green tea (637.5mg per capsule) enables Capsibiol-T® to achieve a ten-fold increase in potency. The synergistic combination of Capsibiol-T® is 10 times more potent than either green tea or capsaicin separately. In an effort to provide the greatest benefit, Capsibiol-T® should be taken three times daily, one capsule every eight hours.

Green tea appears to block growth of new blood vessels that tumors require for growth and metastasis.

Green tea catechin EGCg inhibits special enzyme cancer cells secrete to penetrate and colonize tissue.

Green tea enhances the effectiveness of conventional therapies.

EGCg inhibits telomerase, the enzyme that "immortalizes" cancer cells.

Capsaicin inhibits tNOX.

ActivaMune
http://www.activamune.com/

At the University of California at Berkeley, the Chairman of the Nutritional Sciences Department and the Director of the National Institutes of Health Cancer Research Program were studying the anticancer properties of Diindolylmethane (DIM), a naturally occurring compound found in Brassica vegetables (broccoli, cauliflower, cabbage, kale, brussels sprouts), when they made a remarkable discovery: DIM is a potent activator of the immune response system. Activation of the immune system in part explains DIM's anticancer properties, and for the first time, sheds light on its potent antiviral and antibacterial properties.The scientists patented their discovery and ActivaMune was launched as a first-in-class nutritional supplement to enhance the immune system and support multiple organs throughout the body: breast, prostate, cardiovascular, vision, skin and colon health. ActivaMune's unique and patented formula combines multiple nutrients for maximum effectiveness: Diindolylmethane (DIM), Sulforaphane, Selenium, Lycopene, Lutein, Zeaxanthin, Calcium and Vitamins C, D3 & E.

ActivaMune's formula is exclusively licensed from the University of California at Berkeley. The supplement provides phytonutrients equivalent to approximately five pounds of fresh organic uncooked broccoli, tomatoes and spinach per day. All broccoli extract supplements on the market today, without exception, do not have the active ingredients in ActivaMune as their manufacturing process (heat or freeze drying) destroys the enzyme necessary for the production of the key phytonutrients present in this supplement.

Zyflamend thread HERE

Nutr Cancer. 2007;57(1):78-87.
Zyflamend, a polyherbal preparation, inhibits invasion, suppresses osteoclastogenesis, and potentiates apoptosis through down-regulation of NF-kappa B activation and NF-kappa B-regulated gene products.

Sandur SK, Ahn KS, Ichikawa H, Sethi G, Shishodia S, Newman RA, Aggarwal BB.
Cytokine Research Laboratory, Department of Experimental Therapeutics, The University of Texas M. D. Anderson Cancer Center, Houston, Texas 77030, USA.
Zyflamend, a polyherbal preparation, was designed based on constituents that exhibit antiproliferative, antiinflammatory, antioxidant, antiangiogenic, and apoptotic activities through a mechanism that is not well defined. Because the nuclear factor (NF)-kappaB has been shown to regulate proliferation, invasion, and metastasis of tumor cells, we postulated that Zyflamend modulates the activity of NF-kappa B. To test this hypothesis, we examined the effect of this preparation on NF-kappaB and NF-kappaB-regulated gene products. We found that Zyflamend inhibited receptor activator of NF-kappa B ligand-induced osteoclastogenesis, suppressed tumor necrosis factor (TNF)-induced invasion, and potentiated the cytotoxicity induced by TNF and chemotherapeutic agents, all of which are known to require NF-kappa B activation. Zyflamend suppressed NF-kappa B activation induced by both TNF and cigarette smoke condensate. The expression of NF-kappa B-regulated gene products involved in antiapoptosis (inhibitor-of-apoptosis protein 1/2, Bcl-2, Bcl-xL, FADD-like interleukin-1betaconverting enzyme/caspase-8 inhibitory protein, TNF receptor-associated factor-1, and survivin) and angiogenesis (vascular endothelial growth factor, cyclooxygenase-2, intercellular adhesion molecule, and matrix metalloproteinase-9) was also down-regulated by Zyflamend. This correlated with potentiation of cell death induced by TNF and chemotherapeutic agents. Overall, our results indicate that Zyflamend suppresses osteoclastogenesis, inhibits invasion, and potentiates cytotoxicity through down-regulation of NF-kappa B activation and NF-kappa B-regulated gene products.

PMID: 17516865 [PubMed - indexed for MEDLINE]

Curr Med Chem. 2009;16(12):1451-62.
Chemoprotective mechanism of the natural compounds, epigallocatechin-3-O-gallate, quercetin and curcumin against cancer and cardiovascular diseases.

Jagtap S, Meganathan K, Wagh V, Winkler J, Hescheler J, Sachinidis A.
Center of Physiology and Pathophysiology, University of Cologne, Germany.
Cancer and cardiovascular disease (CVD) chemoprevention can be achieved by the use of natural, synthetic, or biologic compounds to reverse, suppress, or prevent the development of diseases. Chemoprevention is a potential anti-cancer approach, which has reduced secondary effects in comparison to classical prophylaxis. Natural compounds such as flavonoids reduce oxidative stress, which is the most likely mechanism in the protective effects of these compounds. Even though the exact mechanisms of action are not well understood another central action mechanism of polyphenolic flavonoids seems to be an induction of apoptosis as demonstrated in numerous cellular systems. Moreover, flavonoids may modulate protein and lipid kinase signaling pathways. Understanding the mechanism of these natural products will contribute to the development of more specific preventive strategies against cancer and CVD. Much of the research in the field is focused on epigallocatechin-3-O-gallate (EGCG), quercetin and curcumin, which were found to have beneficial effects against cancer and CVD. We review the chemoprotective mechanisms through which these natural compounds exert their beneficial effects against cancer and CVDs.

PMID: 19355899 [PubMed - indexed for MEDLINE]

Free Radic Res. 2002 May;36(5):593-9.
Hydrogen peroxide generation in caco-2 cell culture medium by addition of phenolic compounds: effect of ascorbic acid.

FULL TEXT

Roques SC, Landrault N, Teissèdre PL, Laurent C, Besançon P, Rouane JM, Caporiccio B.
Unité Nutrition, Laboratoire Génie Biologique et Sciences des Aliments, Université Montpellier II, France.
Phenolic compounds have recently attracted special attention due to their beneficial health effects; their intestinal absorption and bioavailability need, therefore, to be investigated and Caco-2 cell culture model appeared as a promising tool. We have shown herein that the addition of a grape seed extract (GSE) to Dulbecco's modified Eagle's medium (DMEM) used for Caco-2 cell culture leads to a substantial loss of catechin, epicatechin and B2 and B3 dimers from GSE in the medium after 24 h and to a production of hydrogen peroxide (H2O2). When 1420 microM ascorbic acid is added to the DMEM, such H2O2 production was prevented. This hydrogen peroxide generation substantially involves inorganic salts from the DMEM. We recommend that ascorbic acid be added to circumvent such a risk.

PMID: 12150547 [PubMed - indexed for MEDLINE]

Patent: herbal complex, wubeizi, Lonicera japonica, Astragalus membranaceus, Rehmanniae Radix, Glycyrrhizae Radix and Panax schinseng‏

http://www.freepatentsonline.com/7527812.html

US Patent References:
Adminiculum for antitumor agents
Aburada et al. - September, 1986 - 4613591

Inventors: Hsieh, Chen Fung (Taipei, TW)

Application Number:
11/535483

Publication Date:
05/05/2009

Filing Date:
09/26/2006

View Patent Images:
Download PDF 7527812

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Assignee:
Sheng Foong Pharmaceutical Co., Ltd. (Taipei, TW)

Primary Class:
424/728

Other Classes:
424/773, 424/757, 424/725

International Classes:
A01N65/00; A61K36/258; A61K36/481; A61K36/236; A61K36/355; A61K36/00; A61K36/254; A61K36/48

Foreign References:

CN1220891	June, 1999
CN1421224	June, 2003
CN1470276	January, 2004
JP360025933	February, 1985

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Lian Z, et al., Association of cellular apoptosis with anti-tumor effects of the Chinese herbal complex in endocrine-resistant cancer cell line, 2003;27(2):147-54, Cancer Detect Prev.
Kim OS, et al., Establishment of In Vitro Test System for the Evaluation of the Estrogenic Activities of Natural Products, Sep. 2004;27(9):906-11, Arch Pharm Res.
Loo WT, et al., The inhibitory effect of a herbal formula comprising ginseng and Carthamus tinctorius on breast cancer, Nov. 26, 2004;76(2):191-200, Life Sci.
095136827; Taiwan Office Action; May 2, 2008; Taiwan.

Primary Examiner:
Meller, Michael V.

Assistant Examiner:
Mi, Qiuwen

Attorney, Agent or Firm:
WPAT, PC
King, Anthony

Claims:
What is claimed is:

1. A composition consisting of Wubeizi, Lonicera japonica, Astragalus membranaceus, Rehmanniae Radix, Glycyrrhizae Radix and Panax ginseng, wherein weight ratios of any two components range from 1:10 to 10:1; wherein Wubeizi is a gall produced by parasitic aphids Melaphis chinensis (Bell) Baker on a leave or stem of a plant selected from the group consisting of Rhus chinensis Mill, Rhus potaninii Maxim and Rhus punjabensis Stew. var. sinica (Diels) Rehd. et Wils.; wherein Lonicera japonica is prepared from flower buds of a plant selected from the group consisting of Lonicera japonica and Lonicera confuse, wherein Astragalus membranaceus is prepared from a root of a plant selected from the group consisting of Astragalus membranaceus (Fisch.) Bge. and Astragalus membranaceus Bge. var. mongholicus (Bge.) Hsiao; wherein Rehmanniae Radix is prepared from a root of Rehmannia glutinosa Libosch; wherein Glycyrrhizae Radix is prepared from a whole plant of a plant selected from the group consisting of Glycyrrhiza uralensis Fisch., Glycyrrhiza glabra L., Glycyrrhiza kansuensis Chang et Peng and Glycyrrhiza inflata Batal; and wherein Panax ginseng is prepared from the root of a plant selected from a group consisting of Panax ginseng and Panax pseudoginseng.

2. The composition of claim 1, wherein weight ratios of any two components range from 1:5 to 5:1.

3. The composition of claim 2, wherein weight ratios of any two components range from 1:3 to 3:1.

4. The composition of claim 3, wherein weight ratios of any two components are 1:1.

Description:
FIELD OF THE INVENTION

The present invention relates to a herbal composition for treating cancer, especially for breast cancer.
BACKGROUND OF THE INVENTION

The popularity of traditional herbal medicine being used as complementary medicines or alternative medicines is rapidly increasing in recent years. The current understanding on the molecular mechanisms of immunopotentiating polysaccharides from medicinal herbs have provided important clues on the anti-cancer effects of Chinese herbal medicines [Shao BM, et al., Biochem Biophys Res Commun. 2004 Aug. 6;320(4):1103-11]. Also, it was found that the combination of anticancer drugs with some herbal extracts contributes to the enhancement of clinical outcomes in cancer chemotherapy [Takara K, et al., Biol Pharm Bull. 2005 Jan.; 28(1):138-42]. Chinese herbal medicines have emerged as an effective tool for treating cancer.
In previous publications, several herbal compounds or complexes have been indicated to be candidate drugs for treating breast cancer. For example, the Glycyrrhizae radix extract was suggested to exhibit inhibitory effects on E2-related endometrial carcinogenesis in mice [Niwa K. et al., Jpn J Cancer Res. 1999 Jul.; 90(7):726-32]. A herbal complex consisting of Hoelen, Angelicae radix, Scutellariae radix and Glycyrrhizae radix suppresses the tumor growth of chemoendocrine resistant cancers [Lian Z, et al., Eur J Gynaecol Oncol. 2001; 22(5): 347-9]. Still another herbal complex consisting of Hoelen, Angelicae radix, Scutellariae radix and Glycyrrhizae radix is shown to be a potential candidate for treating endocrine-resistant gynecologic carcinomas [Lian Z, et al., Cancer Detect Prev. 2003;27(2):147-54]. In addition, it is suggested that Puerariae radix and Ginseng radix Rubra extracts have effective estrogenic actions and could be developed as estrogenic supplements [Kim OS, et al., Arch Pharm Res. 2004 Sep.;27(9):906-11]. Furthermore, the complex named Zhu-xiang containing ginseng and Carthamus tinctorius also exhibits the ability to inhibit proliferation in MDA-MB-231 breast cancer cell [Loo WT, et al., Life Sci. 2004 Nov. 26;76(2):191-200]. These herbal medicines have shed some lights on the therapy of breast cancer.
All of the above-mentioned references are herein incorporated by reference in their entireties.
SUMMARY OF THE INVENTION

The present invention relates to a herbal composition, comprising wubeizi, Lonicera japonica, Astragalus membranaceus, Rehmanniae Radix, Glycyrrhizae Radix and Panax schinseng . The weight ratios of any two components range from 1:10 to 10:1.
BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows the inhibitory effects of the composition of the present invention, Taxol and Doxorubicin on cell growth of human cancer cell A498, HCT 116, Hep 3B, LNCaP clone FGC, MCF-7, MKN45, NCI-H226, NPC-TW01 and the human normal renal cell RPTEC.
FIG. 2 shows the morphological effects of the composition of the present invention, Taxol and Doxorubicin on human cancer cell A498, HCT 116, Hep 3B, LNCaP clone FGC, MCF-7, MKN45, NCI-H226, NPC-TW01 and the human normal renal cell RPTEC.
DETAILED DESCRIPTION OF THE INVENTION

The present invention relates to a herbal composition that provides anti-cancer effects, especially for breast cancer.
Herbal Composition
The present invention relates to a composition of herbal medicines comprising Wubeizi, Lonicerajaponica, Astragalus membranaceus, Rehmanniae Radix, Glycyrrhizae Radix and Panax schinseng.
Wubeizi is the gall produced by the parasitic aphids Melaphis chinensis (Bell) Baker on the leaves or stems of a plant selected from the group consisting of Rhus chinensis Mill, Rhus potaninii Maxim and Rhus punjabensis Stew. var. sinica (Diels) Rehd. et Wils.
Lonicera japonica is prepared from the flower buds of a plant selected from the group consisting of Lonicerajaponica and Lonicera confuse.
Astragalus membranaceus is prepared from the roots of a plant selected from the group consisting of Astragalus membranaceus (Fisch.) Bge. and Astragalus membranaceus Bge. var. mongholicus (Bge.) Hsiao.
Rehmanniae Radix is prepared from the roots of Rehmannia glutinosa Libosch.
Glycyrrhizae Radix is prepared from the whole plant of a plant selected from the group consisting of Glycyrrhiza uralensis Fisch., Glycyrrhiza glabra L., Glycyrrhiza kansuensis Chang et Peng and Glycyrrhiza inflata Batal.
Panax schinseng is prepared from the roots of a plant selected from the group consisting of Panax schinseng and Panax pseudoginseng.
The weight ratios of any two components in the composition of the present invention range from 1:10 to 10:1. In a preferred embodiment, the weight ratios of any two components range from 1:7 to 7:1. In a more preferred embodiment, the weight ratios of any two components range from 1:5 to 5:1. In a furthermore preferred embodiment, the weight ratios of any two components range from 1:3 to 3:1. In the most preferred embodiment, the weight ratio of wubeizi, Lonicerajaponica, Astragalus membranaceus, Rehmanniae Radix, Glycyrrhizae Radix and Panax schinseng is 1:1:1:1:1:1.
Anti-cancer Effects
The present composition can induce the apoptosis of the cancer cell in accordance with this invention. The cancer cells include abnormal cancer cells associated with lymphoma, leukemia, plasma cell dyscrasias, multiple myeloma, amylodosis, also as known as hepatocellular cancer, colorectal cancer, renal cancer, breast cancer, prostate cancer, stomach cancer, lung cancer, nasal-pharyngeal cancer, ovarian cancer, bone cancer, gastric cancer, pancreatic cancer, and melanoma. In a preferred embodiment, the present composition can induce apoptosis of the tumor cells of hepatocellular cancer, colorectal cancer, renal cancer, breast cancer, prostate cancer, stomach cancer, lung cancer and nasal-pharyngeal cancer. In the most preferred embodiment, the present composition induces apoptosis of breast cancer cells.
EXAMPLES

The examples below are non-limiting and are merely representative of various aspects and features of the present invention.
Example 1

Preparation of Single-herb Herbal Medicines

A whole herb of medicinal plant was obtained, washed with cold water and dried. The plant materials were then extracted with boiling water. The weight ratio of plant material to water was approximately 1:5 to 1:10. The amount of water used should at least cover the plant material in the extraction vessel. To allow effective extraction of the desired components, samples were boiled for 0.5 to one hour, but not more than 3 hours. Then, the aqueous solution was separated from the plant residues by filtration. After that, the aqueous solution was spray-dried, freeze-dried or absorbed by powdered material of starch. The single-herb herbal medicine was prepared in powdered form.
Example 2

Preparation of the Composition of the Invention

The herb mixture was prepared by mixing six single-herb herbal medicines wubeizi, Lonicera japonica, Astragalus membranaceus, Rehmanniae Radix, Glycyrrhizae Radix and Panax schinseng at a ratio of 1:1:1:1:1:1 by weight. The resulted mixture was the composition of the present invention.
Example 3

Anti-cancer Testing of the Herb Mixtures

Cell Lines
To examine the effects of the present composition SF-01 on inhibition of cancer/tumor cells, various human cell lines were used, including one normal cell line and eight cancer/tumor cell lines:

- 1. RPTEC (Cambrex): Normal renal proxima tubule epithelial cell, Human
- 2. A-498 (ATCC HTB-44): Kidney carcinoma, Human
- 3. HCT 116 (ATCC CCL-247): Colorectal carcinoma, Human
- 4. Hep 3B (ATCC HB-8064): Hepatocellular carcinoma, Human
- 5. LNCap clone FGC (ATCC CRL-1740): Prostate carcinoma, Human
- 6. MCF-7 (ATCC HTB-22): Mammary gland adenocarcinoma, Human
- 7. MKN45 (FDSC JCRB0245): Stomach carcinoma, Human
- 8. NCI-H226 (ATCC CRL-5826): Lung carcinoma, Human
- 9. NPC-TW01 (School of Medicine, NTU): Nasal-pharyngeal carcinoma, Human

All of the cell lines used were negative for mycoplasma test. Among these cell lines, renal proximal tubule epithelial cell (RPTEC), the only normal cell line, were maintained in REGM™ BulletKit at 37° C. in a humidified atmosphere of 5% CO ₂/95% air, in the absence of antibiotics according to the manufacture's recommendation. On the other hand, the eight tumor cell lines were maintained in DulBeccco's modified essential medium (DMEM) supplied with 10% fetal bovine serum at 37° C., in a humidified atmosphere of 5% CO ₂/95% air in the absence of antibiotics.
Reagents
Fetal calf serum was purchased from BioWhittaker(Walkersville, Md.), MTS was obtained from Promega (Madison, Wis.). All of the other chemicals were from Sigma Chemical (St. Louis, Mo.) and were standard analytic grade or higher. SF-01 was dissolved in the complete culture medium at final concentration of 10mg/ml and was filter sterilized. Doxorubicin and Taxol were prepared in DMSO at concentration of 20 mM and were used as the positive controls.
To evaluate the antitumoral activity of SF-01 towards various human cancer cell lines and measure the potential cytotoxicity of SF-01 towards the primary human renal cell lines (RPTEC), an in vitro cytotoxicity assay was carried out. Tumor cells were seeded at a density of 2×10 ³to 8×10 ³cells/well in 96-well plate 16 hours prior to the chemicals treatment. After exposure to different concentrations of SF-01 (5.0 to 0.0016 mg/ml), Doxorubixin (10 μM to 3.2 nM) or Taxol (2 μM to 0.02 nM) for 72 hours, cells were washed, replaced with medium containing 0.4mg/MTS [3-(4,5-dimethythiazol-2-yl)-5-(3-carboxymethosyphenyl)-2-(4 -sulfophenyl)-2H-tetrazolium] and further incubated for 2 hours. To quantify the metabolically viable cells, the conversion of MTS to formazan was measured by absorbance at 490 nm in a 96-well microtiter plate reader. The mock-treated control was used to evaluate the effect of the chemical on cell growth and to determine the concentration of chemical that inhibited 50% of cell growth (IC ₅₀). The percentage of cytostasis was calculated by the formula:
Cytostasis (%)=[1−(B/A)]×100,
where A is the absorbance of the untreated control and B is the absorbance of the treated cell.
To evaluate the effects of the tested drugs on cell growth, the concentration of chemical that caused 50% reduction of the treated cells (IC ₅₀) was calculated from the dose response curve (FIG. 1) for each cell line. The IC ₅₀for both Doxorubicin and Taxol to various human tumor cellines were found between 800 to 11 nM and 11 to 0.003 nM (Table 1) respectively, which was consistent with previously data. Our data showed that the cytotoxicity of SF-01 toward cancer cell line depends on the nature of the tumor cells, showing the selectivity of SF-01. Among all the tumor cell lines tested, MCF-7 was found to be most sensitive to SF-01, as the concentration of SF-01 as low as 0.59 mg/ml was able to cause 50% reduction of the treated cells, demonstrating the efficient anti-tumor activity of SF-01 on breast cancer cells. On the other hand, no noticeable morphological change or cytotoxicity was found when the normal cell line RPTEC was cultured with SF-01 for 72 hours compared to the untreated cells (FIG. 2). In addition, cell growth stimulation was found when RPTEC was cultured in higher concentration of SF-01. The results showed that SF-01 has little or no toxicity to the normal cells and also might promote its growth at higher concentration. This report was finished from Department of Toxicology and Preclinical Sciences, Pharmaceutical R&D Laboratories, Development Center for Biotechnology. No. 103, Lane 169, Kang Ning St. Hsichih city, Taipei county, Taiwan.

TABLE 1

The concentration of SF-01, Taxol and Doxorubicin that
inhibit 50% of cell growth (IC ₅₀) of human cancer cell
A498, HCT 116, Hep 3B, LNCaP clone FGC, MCF-7, MKN45,
NCI-H226, NPC-TW01 and the human normal renal cell RPTEC.
	IC ₅₀
Cell line	SF-01 (mg/mL)	Taxol (uM)	Doxorubicin (uM)

A498	4.59	0.0111	0.782
HCT 116	2.85	<0.0000256	0.033
Hep 3B	1.97	0.0000256	0.116
LNCaP clone FGC	>5	<0.0000256	0.014
MCF-7	0.59	<0.0000256	0.070
MKN45	1.75	0.0000256	0.058
NCI-H226	3.12	<0.00064	0.123
NPC-TW01	1.89	<0.0000256	0.011
RPTEC	>5	0.4059	3.18