CAM Conference

ZYFLAMEND, AN HERBAL COX-2 INHIBITOR WITH IN VITRO ANTI-PROSTATE CANCER

ACTIVITY

D.L. Bemis, K.A. Kozakowski, A.G. Anastasiadis, B.C. Stisser, L. Salomon, A.E.

Katz, Department of Urology, Columbia University, New York, USA.

Purpose: Elevated expression of COX-2, the inducible cyclooxygenase isoform, has been observed in a variety of tumors including prostate cancer and several clinical trials are currently evaluating COX-2 inhibitors for prostate cancer.

We are studying a unique preparation (Zyflamend) comprised of ten different standardized herbal extracts (rosemary, tumeric, ginger, holy basil, green tea, hu zhang, Chinese goldthread, barberry, oregano, and scutallaria baicalensis) for COX-2 and growth inhibitory activities in the LNCaP human prostate cancer cell line.

Methods: Zyflamend was provided by the manufacturer, New Chapter, Inc. (Brattleboro, VT). A series of dilutions was prepared in DMSO and the dilutions were added to LNCaP growth medium so that all doses tested had equivalent (0.1%) DMSO levels.

Cell growth curves were done by counting cells at 24, 48, and 72 hr and were compared to control cells treated at the same times with 0.1% DMSO alone. Apoptosis in these cultures was evaluated by Western blot analysis of PARP cleavage and measurement of caspase-3 activity using a colorimetric substrate assay.

Effects on purified COX-2 enzyme activity was measured using a colorimetric assay (Cayman Chemicals, Ann Arbor), and effects on COX-2 protein expression was determined via Western blot analysis of protein extracts from treated cells.

These activities were also compared to the effects of purified curcumin (dissolved in DMSO) at levels equivalent to those that would be found in Zyflamend, at similar doses.

Results: LNCaP cell growth was suppressed by Zyflamend extracts and by 72 hrs there was a significant reduction (78%) in cell number in Zyflamend- treated cultures (7.9×104 vs 3.53×105 cells / well, p , 0.01) compared to controls. PARP cleavage fragments were evident and caspase-3 activity was upregulated 2-fold by 72 hr treatment, demonstrating a potential apoptotic effect of this substance that was not found in controls.

COX-2 activity was also significantly decreased in the presence of the herbal extract, while equivalent doses of curcumin alone did not inhibit COX-2 in the assay utilized. COX-2 protein expression in LNCaP cells was not affected by the agent.

Conclusions: Our results suggest that Zyflamend, an herbal extract and COX-2 inhibitor, strongly suppresses LNCaP cells and induces apoptosis.

These effects appear to be more potent that those observed with curcumin alone. We are pursuing further in vitro and in vivo studies to determine if Zyflamend could be an effective prostate cancer chemopreventative supplement.

Presented at the December 13, 2003 meeting of the Society of Urologic Oncology at the National Institutes of Health in Bethesda, Maryland.


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