Curcumin, a polyphenolic antioxidant derived from a dietary spice, exhibits anticancer activity in rodents and in humans. Its efficacy appears to be related to induction of glutathione S-transferase enzymes, inhibition of prostaglandin E 2 (PGE 2 ) production, or suppression of oxidative DNA adduct (M 1 G) formation. We designed a dose-escalation study to explore the pharmacology of curcumin in humans. Fifteen patients with advanced colorectal cancer refractory to standard chemotherapies consumed capsules compatible with curcumin doses between 0.45 and 3.6 g daily for up to 4 months. Levels of curcumin and its metabolites in plasma, urine, and feces were analyzed by highpressure liquid chromatography and mass spectrometry. Three biomarkers of the potential activity of curcumin were translated from preclinical models and measured in patient blood leukocytes: glutathione S-transferase activity, levels of M 1 G, and PGE 2 production induced ex vivo. Dose-limiting toxicity was not observed. Curcumin and its glucuronide and sulfate metabolites were detected in plasma in the 10 nmol/L range and in urine. A daily dose of 3.6 g curcumin engendered 62% and 57% decreases in inducible PGE 2 production in blood samples taken 1 hour after dose on days 1 and 29, respectively, of treatment compared with levels observed immediately predose (P < 0.05). A daily oral dose of 3.6 g of curcumin is advocated for Phase II evaluation in the prevention or treatment of cancers outside the gastrointestinal tract. PGE 2 production in blood and target tissue may indicate biological activity. Levels of curcumin and its metabolites in the urine can be used to assess general compliance.

Purpose: Curcumin, a major constituent of the spice turmeric, suppresses expression of the enzyme cyclooxygenase 2 (Cox-2) and has cancer chemopreventive properties in rodents. It possesses poor systemic availability. We explored whether formulation with phosphatidylcholine increases the oral bioavailability or affects the metabolite profile of curcumin. Methods

Epidemiological and preclinical evidence suggests that polyphenolic phytochemicals exemplified by epigallocatechin gallate from tea, curcumin from curry and soya isoflavones possess cancer chemopreventive properties. Whilst such naturally occurring polyphenols have been the subject of numerous mechanistic studies in cells, information on their clinical properties, which might help assess their promise as human cancer chemopreventive agents, is scarce. Therefore, we present a review of pilot studies and trials with a cancer chemoprevention-related rationale, in which either healthy individuals or patients with premalignant conditions or cancer received polyphenolic phytochemicals. The review identifies trial design elements specifically applicable to polyphenolic phytochemicals. The available evidence for tea polyphenols tentatively supports their advancement into phase III clinical intervention trials aimed at the prevention of progression of prostate intraepithelial neoplasia, leukoplakia or premalignant cervical disease. In the case of curcumin and soya isoflavones more studies in premalignacies seem appropriate to optimise the nature and design of suitable phase III trials. The abundance of flavonoids and related polyphenols in the plant kingdom makes it possible that several hitherto uncharacterised agents with chemopreventive efficacy are still to be identified, which may constitute attractive alternatives to currently used chemopreventive drugs. ' 2006 Wiley-Liss, Inc.Key words: cancer chemoprevention; clinical trial; curcumin; flavonoids; tea polyphenolsIn recent years there has been increasing interest in the potential cancer chemopreventive properties of diet-derived agents. This interest has been elicited by epidemiological research linking variations in geographical distribution of cancer incidence to intake of specific diets, and it has been supported by convincing evidence of chemopreventive efficacy of specific diet constituents in rodent models of carcinogenesis. The ultimate proof of efficacy of a putative cancer chemopreventive agent is provided by long-term phase III clinical intervention studies involving large numbers of individuals. Such studies are complex and expensive to conduct. Only relatively few dietary constituents have undergone, or are currently undergoing, phase III cancer chemoprevention studies. Prominent among them are folate, 1 b-carotene plus vitamins A and E, 2,3 calcium plus vitamin D 4 and selenium plus vitamin E. 5 Polyphenolic pytochemicals such as epigallocatechin gallate (EGCG) from tea, the flavonoids quercetin and genistein from onions and soya, respectively, curcumin in curry spice and resveratrol from red grapes (for structures see Fig. 1) constitute a class of diet constituents with notable efficacy in preclinical models of carcinogenesis, including those of the colorectum, breast and prostate. 6 To our knowledge, none of these species have to date been the subject of phase III clinical trials. A prominent feature rendering polyphenolic phytochemicals worthy of s...

We describe a reversed-phase HPLC method that uses gradient elution and UV detection (325 nm) to determine levels of resveratrol and identify 6 major conjugated metabolites in the plasma and urine of human volunteers after administration of a single oral dose of 1 g. Waters Atlantis C 18 3μ served as the stationary phase. The gradient was formed using ammonium acetate and methanol, containing 2% propan-2-ol. Detection is linear between 5 ng/mL and 500 ng/mL in plasma (5-1000 ng/mL in urine). The coefficient of variation for intra-and inter-day variation is <10%. The average recovery of resveratrol from plasma and urine is 58 ± 3%. The data presented in this report demonstrate a rapid, sensitive and accurate method for the analysis of resveratrol and its metabolites in human plasma and urine for pharmacokinetic studies.

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Anthocyanins are dietary flavonoids, which can prevent carcinogen-induced colorectal cancer in rats. Here, the hypotheses were tested that Mirtoselect, an anthocyanin mixture from bilberry, or isolated cyanidin-3-glucoside (C3G), the most abundant anthocyanin in diet, interfere with intestinal adenoma formation in the Apc Min mouse, a genetic model of human familial adenomatous polyposis, and that consumption of C3G or Mirtoselect generates measurable levels of anthocyanins in the murine biophase. Apc Min mice ingested C3G or Mirtoselect at 0.03, 0.1 or 0.3% in the diet for 12 weeks, and intestinal adenomas were counted. Plasma, urine and intestinal mucosa were analyzed for presence of anthocyanins by high-pressure liquid chromatography with detection by UV spectrophotometry (520 nm) or tandem mass spectrometry (multiple reaction monitoring). Ingestion of either C3G or Mirtoselect reduced adenoma load dose-dependently. At the highest doses of C3G and Mirtoselect adenoma numbers were decreased by 45% (p < 0.001) or 30% (p < 0.05), respectively, compared to controls. Anthocyanins were found at the analytical detection limit in the plasma and at quantifiable levels in the intestinal mucosa and urine. Anthocyanin glucuronide and methyl metabolites were identified in intestine and urine. Total anthocyanin levels in mice on C3G or Mirtoselect were 43 ng and 8.1 lg/g tissue, respectively, in the intestinal mucosa, and 7.2 and 12.3 lg/ml in the urine. The efficacy of C3G and Mirtoselect in the Apc Min mouse renders the further development of anthocyanins as potential human colorectal cancer chemopreventive agents worthwhile. ' 2006 Wiley-Liss, Inc. Key words: colorectal cancer chemoprevention; anthocyanin; analysisThere is an increasing interest in the role of dietary constituents in the prevention of cancer. Examples of dietary components, which have been implicated as cancer chemopreventive agents, are flavonoids such as genistein from soya, quercetin from onions or apigenin from leafy vegetables.1 Dietary intake of these flavonoids is estimated to be within the range of 20-25 mg/day. Anthocyanins are flavonoids responsible for the bright blue and red colors of many fruits and berries. They are glycosides of anthocyanidins, mainly cyanidin, delphinidin, malvidin, pelargonidin, peonidin and petunidin, conjugated with sugars, including glucose, galactose and arabinose (for structures see Fig. 1). The daily intake of anthocyanins in the US is in excess of 200 mg/day, amounts far superior to those of genistein or quercetin.3 Anthocyanins and anthocyanidins have demonstrated antiproliferative activity in some cancer cell lines with IC 50 values for growth inhibition in the range of 100-800 and 18-200 lM, respectively.4,5 Anthocyanins-and anthocyanin-rich fruit extracts decreased aberrant crypt foci in rats exposed to azoxymethane 6 and reduced colonic adenocarcinoma burden in rats that had received azoxymethane, 1,2-dimethylhydrazine or 2-amino-1-methyl-6-phenylimidazo [4,5-b] pyridine.6-8 The Apc Min (multiple intestin...

C3G and its metabolites were recovered from murine tissues which may be targets for cancer chemopreventive intervention. Anthocyanin levels achieved in the gi mucosa, prostate and the kidneys were of an order of magnitude consistent with pharmacological activity.

Running Title: Measurement of anandamide in human plasma Abbreviations: AEA, N-arachidonoylethanolamine (anandamide); AEA-d8, octo-deuterated anandamide; LOD, limit of detection; LOQ, limit of quantification; RSD, relative standard deviation; UPLC, ultra performance liquid chromatography, HPLC, high performance liquid chromatography; MS/MS, tandem mass spectrometry Keywords: anandamide, endocannabinoid, liquid chromatography, mass spectrometry, plasmaThe authors affirm that they have no conflicts of interest.