(12) Patent Application Publication (10) Pub. No.: US 2008/0095869 A1 Archibald Et Al

Home , Sulforaphane

US 2008.0095869A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2008/0095869 A1 Archibald et al. (43) Pub. Date: Apr. 24, 2008

(54) ANTICANCER TREATMENT (30) Foreign Application Priority Data (76) Inventors: David Archibald, West Perth (AU): Aug. 19, 2004 (AU)...... 20049047OO Dorothy Morre, West Lafayette, IN (US); D. James Morre, West Lafayette, Publication Classification IN (US) (51) Int. Cl. Correspondence Address: A6IR 36/00 (2006.01) NATH & ASSOCATES A6II 3 L/65 (2006.01) 112 South West Street A6IP 35/00 (2006.01) Alexandria, VA 22314 (US) (52) U.S. Cl...... 424/760; 514/622 (21) Appl. No.: 11/660,277 (57) ABSTRACT (22) PCT Filed: Aug. 17, 2005 A method of inhibiting tNOX in a living entity which 9 includes administering to the entity, wherein the entity has (86). PCT No.: PCT/AUOS/O1245 cancer cells that express tNOX, a therapeutically active amount of a combination of botanicals selected from the S 371(c)(1), groups consisting of cruciferous vegetables and Capsicum (2), (4) Date: Jan. 4, 2008 plants. Patent Application Publication Apr. 24, 2008 Sheet 1 of 15 US 2008/0095869 A1

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Fig.14 US 2008/0095869 A1 Apr. 24, 2008

ANTICANCER TREATMENT 0012. There is a strong trend towards the use of naturally derived compounds for the effective treatment of cancer. FIELD OF THE INVENTION Patients are more likely to be accepting of a drug that they know has been derived from natural sources such as plants 0001. The present invention relates to compositions and as mere is the belief that the side effects of such drugs will method of treatment providing improved inhibition of not be severe as those that would be considered unnatural. tNOX. 0013 Additionally, when the main source of the active 0002. In particular, the present invention relates to com component is available from natural sources this can then positions and method of treatments that selectively inhibit lead to the final cancer therapy or drug to be significantly tNOX and thus inhibit the growth of cancerous cells. lower in price, and thus more available. DESCRIPTION OF THE PRIOR ART 0014 Compounds derived from Capsicum plants have been used as an anaesthetic (U.S. Pat. No. 4.313,958 and 0003 Cancer is a cellular phenomenon of uncontrolled U.S. Pat. No. 4,493,848). Capsicum compounds have also growth. Normal cells in a mature animal divide in a con been combined with other analgesic compounds, such as trolled manner. Cancer-specific cells arise by abnormal and non-steroidal anti-inflammatory drugs (NSAID) (U.S. Pat. unregulated growth, which can eventually destroy Surround No. 4,812,446) or opioids (U.S. Pat. No. 4,599,342). The ing body tissue. In many instances, cancer may also spread combination of these different classes of analgesic com to other parts of the body in a process called metastasis. pounds produced synergistic effects, in that the combination 0004. In 2003 US mortality statistics, cancer is respon produced greater analgesic effects than either compound sible for 23% of all deaths, with the risk to men of devel alone. oping cancer being 1 in 2 and women 1 in 3. 0.015 U.S. Pat. No. 5,665,378 describe a transdermal 0005 Modern therapies in the treatment of cancer therapeutic composition, administered in patch form, com involve the use either chemotherapy or radiation therapy or prising capsaicin, NSAID and pamabrom. The NSAIDs used Surgery in an attempt to remove the cancer. include diflunisal, fenoprofen, ibuprofen, indomethacin, 0006 Chemotherapy involves the use of various complex meclofenamate, naproxen etc. drugs, many of which are synthesized in a laboratory. Such 0016 Capsicum-based compounds have also been used drugs are often given in combination with other compounds in other compositions for treating ailments such as arthritis, with the aim of disrupting the growth cycle of the cancer strains, bruises and sprains on the outside of the patient, cells. mainly in patch form but there are also a number of creams 0007 Many of these drugs have significant side effects on an aerosols for topical application. humans including hair loss, vomiting, nausea, and reduced 0017. In most instances, along with the active Capsicum white blood cell count, which can lead to an increased compound, there is, by necessity, a second ingredient that in possibility of the patient receiving a secondary infection. Some way reduces the skin irritation caused by the capsaicin. This is due to the fact that many existing drug cancer Usually this is a skin anaesthetic or a compound, which treatments affect normal cells. The presence of these detri binds to the capsaicin. mental side effects can lead to lack of efficacy due to lack of patient compliance with the drug-taking regime. 0018 From the above, the main focus of compositions and methods of treatment involving Capsicum-based com 0008 Additionally, synthetic drugs are often difficult to pounds has been relieving the effects of topical diseases. produce, requiring significant investments in both time and financial resources that are ultimately passed on the patient. 0019 Cruciferous vegetables, such as cauliflower, cab The availability and cost of cancer treatments is an impor bage, and kale contain Sulforaphane, which is an isothiocy tant factor in the patient deciding to proceed with a specific anate that is a known antioxidant. Sulforaphane and other chemotherapy. isothiocyanates are believed to be responsible for the low 0009. A major extant problem of human health is the ered risk of cancer that is associated with the consumption need for inexpensive, safe and effective methods of cancer of broccoli and other cruciferous vegetables. prevention and treatment (Cooper, Elements of Human 0020. However, the use of such isothiocyanates is merely Cancer, Jones and Bartlett, Boston, 1992). seen as being mildly prophylactic. 0010. The use of natural plant extracts in treating differ ent diseases in known. By way of example only, JP OBJECT OF THE INVENTION 10-236968 discloses the use of extracts of paradicsom 0021. It is an object of the present invention to provide a paprika to inhibit cancer cells in a concentration-dependent method of treating cancer. manner. However, in order to achieve this it is first necessary to extract the appropriate compounds from the plant using 0022. More specifically, it is an object of the present organic solvents such as acetone and hexane. invention to provide a synergistic combination, composition 0011. In U.S. Pat. No. 5,830,887 there is a disclosure of and a method of treatment of living entitles each of which is a method for treating cancer using compositions enriched useful in providing in a different way than hitherto inhibition with natural phyto-Oestrogens or analogues thereof that are of tNOX. selected from Genistein, Daidzein, Formononetin and Bio 0023 More specifically, it is an object of the present chanin A. Such phyto-Oestrogens are available from Soya invention to provide for a method of treating cancer by hypocotyl and red clover. increasing the activity of Sulforaphane. US 2008/0095869 A1 Apr. 24, 2008

0024. Another object of the invention is to overcome, or 0042. In preference, the composition includes a pharma at least Substantially ameliorate, the disadvantages and ceutically acceptable carrier. shortcomings of the prior art. 0043. In yet a further form of the invention there is 0025. Other objects of the invention and advantages of disclosed a method of treating cancer in a patient in need of the present invention will become apparent from the fol cancer therapy comprising administering to said patient by lowing description, taken in connection with the accompa ingestion an anti-cancer effective amount of a combination nying figures, wherein, by way of illustration and example, of a purified capsaicinoid and Sulforaphane in a physiologi an embodiment of the present invention is disclosed. cally acceptable formulation. 0044) In preference, the capsaicinoid is derived from the SUMMARY OF THE INVENTION powdered fruits of a Capsicum annum cultivar and/or its 0026. According to the present invention, which follow constituents. ing Statement is not intended to be necessarily the only or indeed the broadest form of this, there is provided a method 0045. In preference, the sulforaphane originates from of inhibiting tNOX in a living entity which includes admin lyophilised broccoli sprouts. istering to the entity, wherein the entity has cancer cells that 0046) The term “pharmaceutically acceptable carrier is express thOX, a therapeutically active amount of a combi intended to mean, but not limited to, a non-toxic solid, nation of botanicals selected from the groups consisting of semisolid or liquid filler, diluents, encapsulating material or cruciferous vegetables and Capsicum plants. formulation auxiliary of any type. 0027) In preference the cruciferous vegetable is broccoli. 0047. In a further form, the invention can be said to reside 0028. In preference, said Capsicum plants are derived in a method of treatment of a living entity to inhibit from the Capsicum annum species. replication of cancer cells within that entity where the entity is of a type that has a life-sustaining process and where a 0029. In preference, finely powdered fruits of the Cap tumour will express tNOX uniquely in contradiction to any sicum plants are used. expression from normal or non-cancer cells, the method including the steps of introducing into the entity so as to be 0030. In preference, said fruits contain Capsicum vanil effectively active within the entity over at least a substantial loids. time together, therapeutic materials which are an extract of 0031. In preference, said vanilloids are capsaicin and/or a en cruciferous vegetable (including a Substantial quantity Vanillylamine. of Sulforaphane) and Capsicum or an extract of Capsicum, in which there is a synergistic effect that leads to the 0032. In preference, said broccoli contains sulforaphane. improvement in the effect of the cruciferous vegetable 0033. In a further form of the invention there is provided eXtract. a method of treating cancer in a patient in need of cancer 0048. In preference the extract of Capsicum is a vanil therapy comprising administering to said patient by inges loid-containing Capsicum preparation. tion an anticancer effective amount of a composition includ ing a product of at least two plants selected from the group 0049. In a further form the invention can be said to reside of Capsicum plants and cruciferous vegetables. in a therapeutic material for the treatment of tumours in living entities which material (whether as a mixture or 0034. In preference, the product of the Capsicum plant is cooperatively packaged or administered or sold together) is finely powdered dried fruit. 100 units by weight of broccoli extract and from 1-10 units 0035) In preference, the cruciferous vegetable is broccoli. by weight of Capsicum extract. 0036). In preference, the product of the broccoli is 0050. In preference, said Capsicum extracts are derived selected from the group of finely ground broccoli sprouts, from the Capsicum annum species. commercially available broccoli sprouts, and a solution of 0051. In preference, said Capsicum extract are finely broccoli sprout extract. powdered fruits of the Capsicum plant. 0037. In preference, the solution of broccoli sprout 0052. In preference, said Capsicum extracts contain Cap extract is an aqueous extract. sicum Vanilloids. 0038. In preference, the method of treating cancer 0053. In preference, said vanilloids are capsaicin and/or involves introducing into the mammal in combination at Vanillylamine. least the two said extracts to an extent that they are active to provide synergistic activation and at least over time there 0054. In preference, said broccoli extract contains sul will be effected by these materials in combination an inhi foraphane. bition of tNOX activity of the cancer cell. 0055. In a further form of the invention this can be said 0039. In preference, said anticancer effective amount by to reside in a botanical Supplement consisting of lyophilised weight of dried extract of broccoli as compared to the dried broccoli sprouts combined with powdered chillies (Capsi Capsicum annum fruits is between 10:1 and 100:1. cum annum species) in ratios of weight between 10:1 and In preference, said broccoli is broccoli sprouts. 100:1 whereby tumour cell division inhibitory activities of 0040 the broccoli sprouts on both the tNOX and cell culture 0041) In preference, the broccoli sprouts are lyophilised. assays are enhanced synergistically. US 2008/0095869 A1 Apr. 24, 2008

0056. A unique plasma membrane NADH oxidase mammary carcinoma) cells in culture. Synergy of inhibition (NOX), a unique cell surface protein with hydroquinone is observed for sulforaphane and the vanilloids in both (NADH) oxidase and protein disulfide-thiol interchange systems. Specifically, a claim is made for a novel botanical activities that is responsive to hormone and growth factors Supplement consisting of lyophilised broccoli sprouts com has been identified. Further, a hormone-insensitive and bined with powdered chillies (Capsicum annum species) in drug-responsive form of NOX designated thOX, which is ratios between 10:1 and 100:1 where activities of the broc specific to cancer cells has been reported. coli sprouts on both the tNOX and cell culture assays are enhanced 2- to 5-fold by the combination compared to 0057. Because the NOX protein is located at the external broccoli sprouts or chilli powders alone when compared at plasma membrane Surface and is not transmembrane, a the same relative concentrations. functional role as an NADH oxidase is not considered likely. While the oxidation of NADH provides a basis for a convenient method to assay the activity, the ultimate elec BRIEF DESCRIPTION OF THE DRAWINGS tron physiological donor is most probably hydroquinones 0063 FIG. 1. Inhibition of NOX activity (fully oxidized) with specific activities for hydroquinone oxidation greater from the HeLa cell surface by sulforaphane. than or equal to that of NADH oxidation and/or protein thiol-disulfide interchange. 0064 FIG. 2. Inhibition of NOX activity (no H202) from the HeLa cell surface by sulforaphane. 0.058 CNOX was originally defined as a drug-indifferent constitutive NADH oxidase activity associated with the 0065 FIG. 3. Inhibition of NOX activity (fully oxidized) plasma membrane of non-transformed cells that was the of 4T1 mouse mammary cells by Sulforaphane. normal counterpart to tNOX. Indeed, a 36 kD protein 0.066 FIG. 4. Sulforaphane does not inhibit NOX activity isolated from rat liver and from plants has NOX activity that of human mammary (non-cancer) epithelia which lack is unresponsive to thOX inhibitors. tNOX. 0059 While cancer cells exhibit both drug-responsive 0067 FIG. 5. Sulforaphane does not inhibit NOX activity and hormone and growth factor-indifferent (tnOX) as well of plasma membranes isolated from dark-grown hypocotyls as drug inhibited and hormone and growth factor dependent of soybean which lack tNOX. (CNOX) activities, non-transformed cells exhibit only the drug-indifferent, hormone- and drug-responsive CNOX. 0068 FIG. 6. Effect of sulforaphane on growth of HeLa Among the first descriptions of so-called constitutive or and human mammary carcinoma (BT-20) cells in culture at CNOX activity of non-transformed cells and tissues was 48 and 72 h of treatment. where the activity of rat liver plasma membranes was 0069 FIG. 7. A-D. Effect of sequential additions of stimulated by the growth factor, diferric transferrin. Subse sulforaphane, capsaicin and EGCg on NOX activity from quent work demonstrated that the observed NADH oxida the HeLa cell surface added in the order given from left to tion was catalysed by a unique enzyme exhibiting respon right. siveness to several hormones and growth factors. Unlike 0070 FIG. 8. Effect of sequential additions of sul mitochondrial oxidases, the hormone-stimulated NADH foraphane, vanillylamine and EGCg on NOX activity from oxidase activity of rat liver plasma membranes is not inhib the HeLa cell surface added in the order given from left to ited by cyanide. The enzyme also was distinguished from right. other oxidase activities by its response to several common oxidoreductase inhibitors, e.g., catalase, azide and chloro 0071 FIG. 9. Inhibition of NOX activity from the HeLa quine, as well as to various detergents e.g., sodium cholate, cell surface by broccoli extract alone (A) and in combination Triton X-100 and CHAPS. Like tNOX of cancer cells, with various pepper powders. CNOX is a unique membrane-associated protein that is 0072 FIG. 10. Survival of HeLa and 4T1 cells after 72 h capable of oxidizing NADH but has an activity which is of treatment with different dilutions of broccoli extract modulated by hormones and growth factors. alone. 0060. There remains a need for treatment of cancer that 0.073 FIG. 11. Survival of HeLa (A and C) and 4T1 (B does not have the adverse effects generally caused by the and D) cells comparing two different sources of pepper non-selective of conventional chemotherapeutic agents. (Capsicum annum) powder; A, B, Ancho; C, D. Piquin. The 0061 Inhibition of thOX, an extracellular membrane optimum ratio for combination is one part pepper powder to associated protein, by the above-mentioned combination of 25 parts lyophilised broccoli sprouts. Capsicum and broccoli products, results in the selective 0074 FIG. 12. As in FIG. 11 except a mixture of two inhibition of cancer cell growth and ultimately, apoptosis. pepper powders. The optimum ratio again is 1 part pepper 0062) What is now provided is a way or method of powder to 25 parts lyophilised broccoli sprouts. treating cancer that was until now unknown. The method enhances the activity, to a previously unknown level, of 0075 FIG. 13. NADH oxidase activity comparing differ Sulforaphane, a major anticancer ingredient of broccoli, by ent ratios of lyophilised broccoli sprouts and pepper pow combination with Capsicum Vanilloids such as capsaicin and ders. The optimum ratio for inhibition was 1 part pepper vanillylamine. Both the sulforaphane and the Capsicum powder to 25 parts lyophilised broccoli sprouts. Vanilloids target the cancer-associated and growth-related 0076 FIG. 14. Survival of LnCap (human prostate can ECTO-NOX protein tNOX, Efficacy evaluations are based cer) cells in culture and response to extract of lyophilised on inhibition oftNOX activity of human cervical carcinoma broccoli sprouts (BSL) with and without pepper powder (HeLa) cells and of growth of HeLa and 4T1 (mouse (PP) in a 25:1 ratio. G=guiallijo. A=ancho. US 2008/0095869 A1 Apr. 24, 2008

DETAILED DESCRIPTION OF THE but required addition of a mixture of pepper powders in the INVENTION 25:1 ratio to achieve a strong inhibitory response (FIG. 14). 0077. This invention has as its basis the discovery of a Growth of Cells cell surface NADH oxidase activity with utility as a screen 0084. HeLa (ATCC CCL-2) human cervical adenocarci ing method for potential anticancer agents. noma cells were cultured in minimal essential medium 0078. Among the more potent NOX inhibitors are cap (Eagle), with 2 mM L-glutamine and Earle's balanced salt saicin (8-methyl-N-Vanillyl-6-moneamide), the pungent Solution adjusted to contain 1.5 g/L Sodium bicarbonate, 0.2 principle of chilli peppers and EGCg((-)-epigallocatechin mM non-essential amino adds, 1.0 mMSodium pyruvate and gallate), the principal tea catechin. In this application, we supplemented with 10% bovine calf serum (heat-inacti describe compositions consisting of powdered fruits of vated) plus 50 mg/L gentamycin Sulfate (Sigma). Capsicum annum cultivars and/or its constituents plus lyo philised broccoli sprouts and/or their constituents with 0085. The 4T1 mammary cancer cell line arose from a potential utility in the treatment and/or prevention of cancer. BALB/c C3H mouse (Miller et al., 1987). The 4T1 cells were grown in DME-10, Dulbecco's modified Eagle's 0079 L-Sulforaphane (sulforaphane), an isothiocyanate medium supplemented with 5% foetal calf serum, 5% new prevalent in broccoli that blocks initiation of cancer caused born calf serum, 1 mM mixed non-essential amino acids, 2 by chemicals, was shown to be a potent inhibitor of the mM L-glutamine, penicillin (100U mL) and streptomycin tNOX cancer target. Activity for L-Sulforaphane with an (100 mg mL). EC, of about 1 mM was shown fortNOX from HeLa (FIGS. 1 and 2) and for tNOX of 4T1 mouse mammary cells (FIG. Preparation of HeLa Cells and Cell-Free Extracts 3). CNOX of non-cancer MCF-10A human mammary epi 0086. HeLa S cells (grown in suspension) were collected thelia was unaffected by L-Sulforaphane (FIG. 4) as was the by centrifugation and shipped frozen by a commercial CNOX activity of soybean plasma membranes (FIG. 5). supplier (Cellex Biosciences, Minneapolis, Minn.) In 0.1 m L-Sulforaphane inhibited the growth of HeLa and human Sodium acetate, pH 5, in a ratio of 1 mL packed cell Volume mammary cancer (BT-20) cells with an ECs of between 0.1 to 1 mL acetate. The cells were thawed at room temperature, and 1 mM (FIG. 6). The margin of safety, however, with resuspended and incubated at 37° C. for 1 h to release the growth of cells was less than a factor of 10 with non-cancer protein. The cells were removed by centrifugation at 37000 MCF-10A cells being inhibited to nearly the same extent as g for 60 min and the cell-free supernatants were refrozen and the cancer cells. stored in 1 mL samples at -70° C. 0080 When sulforaphane was combined with other tNOX inhibitors from natural sources, capsaicin, vanilly Spectrophotometric Assay of NADH Oxidase lamine and EGCg, a beneficial response was seen with 0087 NADH oxidase activity was determined as the capsaicin (FIG. 7) and with vanillylamine (FIG. 8) either disappearance of NADH measured at 340 nm in a reaction preceding (FIG. 7B, FIG. 8B) or following (FIG. 7A, FIG. mixture containing 25 mM Tris-Mes buffer (pH 7.2), 1 mM BA) the addition of Sulforaphane, EGCg was antagonistic in KCN to inhibit low levels of mitochondrial oxidase activity, all combinations (FIG. 7C-F, FIG. 8C-F) as was green tea. and 150 (J-M NADH at 37° C. with temperature control 0081 For development of a botanical based on these (t0.5) and stirring (14). Activity was measured using paired observations, lyophilised broccoli sprouts were used. Dark Hitachi U3210 spectrophotometers. Assays were initiated by grown seedlings from organic broccoli seeds (Sun Organic addition of NADH. With plasma membranes and whole Farm, San Marcos, Calif.) germinated at 25°C. and 4 days cells, assays were for 1 min and were repeated on the same old were harvested, frozen and lyophilised directly. Finely sample every 1.5 min for the time indicated. A millimolar ground and sieved sprouts or a commercial preparation of extinction coefficient of 6.22 was used to determine specific sprouts (Arizona Health Foods) were cold water extracted activity. Proteins were estimated by the bicinchoninic acid overnight prior to assay (FIG. 9). Standard broccoli extracts method with bovine serum albumin as standard. 1.25 mg/ml were diluted 1:10, 1:20 or 1:50 and added to Growth. Measurements HeLa or 4T1 cells grown in a 96 well format at a further dilution of 1:100. The ECs for inhibition of growth of HeLa 0088 Growth was determined using a 96-well plate assay and 4T1 cells by the broccoli extract was 1:500 (final as described by Lin et al. (17). HeLa (5x10') or CHO (10) dilution) (FIG. 10). cells were distributed into each plate well (Costar tissue culture plate). The cells were grown at 37° C. for 24 h after 0082 Combination of broccoli extract with finely pow which the substances to be evaluated were added followed dered Capsicum annum fruits of different varieties (pepper by incubation for an additional 48 or 72 h as indicated. powders) enhanced the cell killing of the broccoli prepara Medium was removed and the cells were washed with tion by a factor of approximately 5 (FIG. 11). Powders phosphate-buffered saline and then fixed by addition of 100 mixed from two different peppers were more active than mL 2.5% (v/v) glutaraldehyde for 0.5 h followed by a single sources (FIG. 12). The optimum ratio for combination distilled water wash. The cells were stained with 100 mL 1% of lyophilised broccoli sprouts and pepper powder was aqueous crystal violet for 0.5 h, washed exhaustively with determined to be 25 parts lyophilised broccoli sprouts to 1 distilled water followed by 200 mL33% (v/v) acetic acid for part pepper power (FIG. 12). Similar results were obtained 5 min. The absorbance was determined at 580 nm using an for inhibition of NADH oxidase activity from the HeLa cell automated plate reader. Growth was determined according surface (FIG. 13). to the formula (b-c)x100/(a-c) where a=absorbance of 0.083 Growth of LnCap (human prostate cancer) cells in cells in medium without treatment b=cells in medium with culture was inhibited by lyophilized broccoli sprout extract treatment and c=medium alone (background). US 2008/0095869 A1 Apr. 24, 2008

0089. From these results we have established that in sera 50. The method of claim 49, wherein the broccoli contains results indicate significant synergistic effects resulting from Sulforaphane. use of the two indicated materials together and that these 51. A method of treating cancer in a patient in need of results will indicate equivalent beneficial enhancement in cancer therapy comprising administering to said patient an V1V3. synergistic anti-cancer effective amount of a composition including a product of at least a plant selected from the In Vivo Trial Results group of Capsicum plants and an aqueous extract of crucif 0090 Three patents with elevated PSA (prostate specific erous vegetables. antigen) levels were provided with the combination of 52. The method of claim 51, wherein product of the broccoli extract with finely powdered Capsicum annum Capsicum plant is finely powdered dried fruit. fruits of different varieties (pepper powders) as described 53. The method of claim 52, wherein the fruits contain previously. Each patient ingested 700 mg of the combination Capsicum Vanilloids. of broccoli extract with finely powdered pepper powders 54. The method of claim 53, wherein the vanilloids are every four hours. selected from the group consisting of capSafoin and Vanil lylamine. 0091. In each case there was a marked response to the 55. The method of claim 54, wherein the cruciferous PSA levels, showing a reduction in the overall rate of vegetable is broccoli. increase of PSA levels or levelling out of PSA levels. 56. The method of claim 55, wherein the product of the 0092. One patient, prior to treatment, had a PSA level that broccoli is selected from the group of finely ground broccoli was increasing at 13.3% over an 8 month period. During sprouts, commercially available broccoli sprouts, and a treatment with the combination of broccoli extract with solution of broccoli sprout extract. finely powdered pepper powders, the rise on PSA was 57. The method of claim 56, wherein the solution of reduced to only an increase of 2.78%. broccoli sprout extract is an aqueous extract. 0093. Another patient had PSA levels that were recoded 58. The method of claim 57, wherein the method of at rising by 5 units per day prior to the distraction of the treating cancer involves introducing into the mammal in combination of broccoli extract with finely powdered pepper combination at least the two said extracts to an extent that powders. During the 14 days that the patient was ingesting they are active to provide synergistic activation and at least over time there will be effected by these materials iri the combination, their PSA levels leveled off and started to combination an inhibition of thOX activity of the cancer decline slightly. After completion of the trial, the patients cell. PSA levels resumed rising at 6 units per day. 59. The method of claim 58, wherein the anti-cancer 0094. The purpose of this description is to illustrate the effective amount by weight of extract of broccoli as com invention and not limit it. pared to the dried Capsicum annum fruits is between 10:1 0.095 Although the invention has been hearing shown and 100:1. and described in one is conceived to be the most practical 60. The method of claim 59, wherein the anti-cancer and preferred embodiment, it is recognized that departures effective amount by weight of extract of broccoli as com can be made within the scope of the invention, which is not pared to the dried Capsicum annum fruits is 25:1. to be limited to the details described herein but it is to be 61. The method of claim 60, wherein the broccoli extract accorded the full scope of the appended claims so as to is broccoli sprouts. 62. The method of claim 61, wherein the broccoli sprouts embrace any and all equivalent methods. are lyophilised. 1-40. (canceled) 63. The method of claim 62, wherein the composition 41. A method of inhibiting tNOX in a living entity having includes a pharmaceutically acceptable carrier. cancer cells that express thOX, which includes administer 64. A method of treating cancer in a patient in need of ing to the entity a synergistic therapeutically effective cancer therapy comprising administering to the patient an amount of a combination of Capsicum plants and aqueous synergistic anti-cancer effective amount of a combination of extract from cruciferous vegetables. a purified capsaicinoid and Sulforaphane in a physiologically 42. The method of claim 1, wherein the cruciferous acceptable formulation. vegetable is broccoli. 65. The method of claim 61, wherein the capsaicinoid is 43. The method of claim 42, wherein the broccoli contains derived from the powdered fruits of a Capsicum annum Sulforaphane. cultivar and/or its constituents. 44. The method of claim 43, wherein the ration of broccoli 66. The method of claim 62, wherein the sulforaphane extract to Capsicum plants is between 10:1 and 100:1 wiw, originates from lyophilised broccoli sprouts. 45. The method of claim 44, wherein the ration of broccoli 67. A composition for treating cancer in a living entity, extract to Capsicum plants is between 25:1 wherein the cancer is a type having cancer cells that express 46. The method of claim 45, wherein the Capsicum plants tNOX, wherein the composition includes a synergistic thera are derived from the Capsicum annum species. peutically active amount of a combination of Capsicum 47. The method of claim 46, wherein finely powdered plants and an aqueous extract of cruciferous vegetables. fruits of the Capsicum plants are used 68. The composition of claim 67, wherein the cruciferous 48. The method of claim 47, wherein the fruits contain vegetable is broccoli. Capsicum Vanilloids. 69. The composition of claim 68, wherein the Capsicum 49. The method of claim 48, wherein the vanilloids are plants are derived from the Capsicum annum species. selected from the group consisting of capsaicin and Vanil 70. The composition of claim 69, wherein the fruits of the lylamine. Capsicum plants are finely powdered. US 2008/0095869 A1 Apr. 24, 2008

71. The composition of claim 70, wherein the fruits 79. The method of claim 78, wherein the broccoli contains contain Capsicum Vanilloids. Sulforaphane. 72. The composition of claim 71, wherein the vanilloids 80. The method of claim 79, wherein the Capsicum plants are selected from the group consisting of capsaicin, Vanil are derived from the Capsicum annum species. lylamine. 81. The method of claim 80, wherein finely powdered 73. The composition of claim 72, wherein the broccoli fruits of the Capsicum plants are used contains Sulforaphane. 82. The method of claim 81, wherein the fruits contain 74. The composition of claim 73, wherein the broccoli is Capsicum Vanilloids. lyophilised broccoli sprouts. 83. The method of claim 82, wherein the vanilloids are 75. The composition of claim 74, wherein the lyophilised selected from the group consisting of capsaicin and Vanil broccoli sprouts combined with powdered chillies (Capsi lylamine. cum annum species) in ratios of weight between 10:1 and 84. A composition for treating cancer in a living entity, 100:1 wherein the cancer is a type having cancer cells that express 76. The composition of claim 75, wherein the lyophilised tNOX, wherein the composition includes a synergistic thera broccoli sprouts combined with powdered chillies (Capsi peutically active amount of a combination of capsaicin and cum annum species) in ratios of weight 25:1. Sulforaphane. 77. A method of inhibiting the division of cancer cells in a living entity, wherein the cancer is a type having cancer 85. The composition of claim 84, wherein the ratio of cells that express thOX, the method including administering capsaicin to sulforaphane is between 10:1 and 100:1 w/w. to the living entity a synergistic combination of Capsicum 86. The composition of claim 85, wherein the capsaicin is plants and an aqueous extract of cruciferous vegetables So as Sourced from Capsicum plants. to provide a prophylactic treatment of cancer by inhibiting 87. The composition of claim 86, wherein the sul the expression of tNOX within the cancer cells. foraphane is sourced from cruciferous vegetables. 78. The method of claim 77, wherein the cruciferous vegetable is broccoli. k k k k k