US 20150079208A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2015/0079208A1 Bates et al. (43) Pub. Date: Mar. 19, 2015

(54) PROCESSES FOR EXTRACTING Publication Classification FROM SOLIDS AND COMPOSITIONS AND (51) Int. Cl. METHODS OF USE THEREOF A61E36/85 (2006.01) (52) U.S. Cl. (71) Applicant: POM Wonderful LLC, Los Angeles, CPC ...... A61K 36/185 (2013.01) CA (US) USPC ...... 424/776 (72) Inventors: Byron Bates, Bakersfield, CA (US); (57) ABSTRACT Erich Fritz, Westlake Village, CA (US); Processes for producing an extract containing phytochemi Yair Henig, Beverly Hills, CA (US); cals from are disclosed. The processes gener Harley Liker, Beverly Hills, CA (US) ally comprise providing pomegranate solids, such as the peri carp, inner membrane and seeds; creating a mixture Assignee: POM Wonderful LLC, Los Angeles, comprising the pomegranate Solids in an aqueous solution; (73) adding enzymes to the mixture in an amount Sufficient to at CA (US) least partially degrade the pomegranate solids; heating the mixture to a temperature that permits the maximum rate of (21) Appl. No.: 14/174,613 catalysis of the enzyme; maintaining the temperature of the heated mixture for a time sufficient to allow at least partial (22) Filed: Feb. 6, 2014 degradation of the pomegranate solids; and removing residual insoluble solid materials from the mixture. Compositions containing the extract may be used as a food product, bever Related U.S. Application Data age, pharmaceutical preparations, nutritional Supplements, (63) Continuation of application No. 12/564.878, filed on Vitamin Supplements, food additives, and food Supplements. Sep. 22, 2009, now Pat. No. 8,658,220, which is a The compositions may also be used for preventing or ame continuation of application No. 1 1/137,248, filed on liorating disease conditions by administering an effective May 24, 2005, now Pat. No. 7,611,738. amount of the composition to a subject in need thereof. US 2015/0079208 A1 Mar. 19, 2015

PROCESSES FOR EXTRACTING 110° F. to 160°F. Enzymes are added to the mixture in an PHYTOCHEMICALS FROM POMEGRANATE amount Sufficient to at least partially degrade the pomegran SOLIDS AND COMPOSITIONS AND ate Solids and liberate phytochemicals from the plant tissues METHODS OF USE THEREOF and/or cells. Once liberated, the phytochemicals may react and/or polymerize to create new compounds CROSS REFERENCE TO RELATED or reaction products. The residual insoluble solid materials APPLICATIONS are removed from the mixture to provide an extract containing 0001. This application is a Continuation of prior filed and phytochemicals. co-pending U.S. patent application Ser. No. 1 1/137,248, filed 0011. In another preferred embodiment, extracts contain May 24, 2005, of the same name, which is hereby incorpo ing phytochemicals from a pomegranate are provided. Such extracts are characterized by a significantly higher total rated by reference in its entirety. content, particularly of the high molecular weight BACKGROUND OF THE INVENTION polyphenol (e.g., ), than is found in pomegranate juice. Such extracts may be obtained from the methods dis 0002 1. Field of the Invention closed herein. 0003. The present invention relates generally to pome 0012. In a further preferred embodiment, food products granate extracts, and more particularly, to methods for obtain and beverages are provided comprising the extract containing ing and using extracts from pomegranate solids and compo phytochemicals from a pomegranate. sitions comprising pomegranate extracts. 0013. In yet a further preferred embodiment, composi 0004 2. Description of the Related Art tions comprising the extract containing phytochemicals from 0005. It is well-known that fruits and vegetables are an a pomegranate are provided. Such compositions may be in essential part of a healthy diet. Chief, among the reasons, is form of tablets, Suspensions, implants, solutions, emulsions, that fruits and vegetables are rich Sources of important phy capsules, powders, syrups, liquid compositions, ointments, tochemicals, which provide essential nutrients and enhance lotions, creams, pastes, and gels. Such compositions may also the body’s ability to prevent and fight disease. There is a beinform of pharmaceutical preparations, nutritional Supple multitude of phytochemicals, in unique combinations, in dif ments, vitamin Supplements, food additives, and food Supple ferent fruits and vegetables, and each functions differently in mentS. the body: as anti-oxidants, as anti-allergenic, as anti-carcino 0014. In a further preferred embodiment, compositions genic, as anti-inflammatory, as anti-viral, and/or anti-prolif containing the extract and the pomegranate juice are pro erative. vided. The combination of the extract and pomegranate juice 0006. The pomegranate has recently been acclaimed for not only produces a composition having a higher total its health benefits and for its disease-fighting antioxidant polyphenol content, as compared to the pomegranate juice potential. Antioxidants are important because they are alone, but it also provides the broad spectrum of the different believed to protect the body against free radicals, the harmful which predominate the pomegranate juice and molecules that can cause heart disease, premature aging, eXtract. Alzheimer's disease, blindness, and a variety of cancers. 0015. In another preferred embodiment, methods are pro 0007 Studies have shown that pomegranate juice has vided for preventing or ameliorating disease conditions in a more polyphenol antioxidants than any other drink, Such as Subject by administering to the Subject an effective amount of red wine, green tea, blueberry juice, cranberry juice and the composition Suitable for use as a pharmaceutical or nutri orange juice. Currently, the two common ways of consuming tional preparation. Such disease conditions include polyphe pomegranates are by eating the fleshy arils of the pomegran nolmediated diseases and cancer. Examples of polyphenol ate and by drinking the juice obtained from the arils. mediated diseases include circulatory disorders such as 0008. There are many kinds of antioxidants, some pro hypertension and coronary artery disease, erectile dysfunc duced by the body and others derived from the foods we eat. tion, lung disorders such as asthma, cancers of various types, When the body's natural antioxidant defenses are lowered, or inflammatory conditions, certain liver conditions, diabetes, greater amounts of free radicals are being produced, the body mood disorders, eye disorders such as cataracts, weak eye becomes more dependent upon food sources of antioxidants. sight due to aging, macular degeneration, and other age related disorders, such as Alzheimer's disease and dementia. BRIEF SUMMARY OF THE PREFERRED 0016. In yet another preferred embodiment, methods are EMBODIMENTS provided for modulating the growth and progression of can 0009 Methods are provided for producing an extract con cerous cells, the methods comprising selecting a Subject hav taining phytochemicals from pomegranate Solids. The pome ing cancerous cell growth and administering to the Subject an granate Solids are anyone or more of the group consisting of effective amount of the composition containing the extract. the pericarp, inner membrane and seeds. The extract pro 0017. In yet a further preferred embodiment, methods are duced differs from commercially-available pomegranate provided for preventing or slowing increases in the Prostate juice in that the extract is substantially derived from the Specific Antigen (PSA) levels in a subject having prostate pomegranate Solids, whereas pomegranate juice is Substan cancer. The method comprises selecting a Subject having tially derived from the Sweet, fleshy arils. prostate cancer and administering to the Subject an effective 0010. In one preferred embodiment, the method includes amount of the composition containing the extract. the following steps. Anyone or a combination of the pericarp, 0018. Other objects, features and advantages of the inner membrane and seeds are selected and a mixture is present invention will become apparent to those skilled in the formed comprising the pomegranate solids and an aqueous art from the following detailed description. It is to be under solution. The mixture is then heated to about 60°F. to 210°F., stood, however, that the detailed description and specific preferably of about 85° F. to 185° F. and optimally of about examples, while indicating preferred embodiments of the US 2015/0079208 A1 Mar. 19, 2015 present invention, are given by way of illustration and not extract, these lower molecular weight polyphenols comprise limitation. Many changes and modifications within the scope a higher proportion of the total polyphenol content in pome of the present invention may be made without departing from granate juice (approximately 50%) than in the extract. the spirit thereof, and the invention includes all such modifi 0027. Accordingly, methods are provided for producing cations and equivalents thereof. an extract containing phytochemicals from pomegranate Sol ids. The extract produced from the methods disclosed herein DETAILED DESCRIPTION OF THE PREFERRED differ from the commercially-available pomegranate juice in EMBODIMENTS that the extract is substantially derived from the pomegranate 0019. As used herein, the term “phytochemicals' refers Solids, whereas pomegranate juice is Substantially derived collectively to compounds which are naturally-occurring in from the Sweet, fleshy arils that Surround the pomegranate the pomegranate and to reaction products and metabolites of seed. The extract is characterized as containing polyphenols these compounds, which are considered to have a beneficial and, particularly, high molecular weight polyphenols, such as effect on the human health. Examples of such phytochemicals punicalagin. include, but are not limited to polyphenols, estrogens and 0028. In one preferred embodiment, the method com phytoestrogens. prises providing anyone or a combination of pomegranate 0020. As used herein, the term “polyphenols' refers gen Solids selected from the group consisting of the pericarp, erally to a family of naturally-occurring compounds in the inner membrane and seeds and creating a mixture comprising pomegranate and includes phenols and polyphenols. Phenols the pomegranate solids in an aqueous solution. In a preferred are a class of chemical compounds consisting of a single embodiment, the mixture of the pomegranate solids is created phenol unit in their structure. Although similar to alcohols, by adding water in an amount that is about 20-80% w/v, and phenols have unique properties including relatively higher more preferably about 50% w/v. of the pomegranate solids. acidities due to the aromatic ring tightly coupled to the oxy The mixture is preferably crushed or milled to create a rough gen and a relatively loose bond between the oxygen and the grind of pomegranate solids dispersed in the aqueous solu hydrogen. Examples of phenolic compounds within this tion. group include elagic acid and . Polyphenols are a 0029. The mixture is then heated to a temperature of about group of compounds, characterized by the presence of more 60° F to 210° F., preferably of about 85° F. to 185° F and than one phenolic group. Polyphenols include (e.g., optimally of about 110° F to 160° F. The temperature to and gallotannins), (e.g., anthocya which the mixture is heated depends upon the selection of nins and isoflavones) and stilbenes (e.g., resveratrol). enzymes, or combination of enzymes, that is added to the 0021. As used herein, the term “pomegranate juice' refers mixture. Preferably, the mixture is heated to a temperature to the juice that is substantially obtained from the arils of the that permits the maximum catalysis of the enzyme or combi pomegranate. nation of enzymes. 0022. As used herein, the term “pomegranate solids' 0030 Alternatively, enzymes may be added before the refers to anyone or a combination of the pericarp, the inner mixture is heated. Thus, the order of the steps of heating the membrane and seeds of a pomegranate. mixture and adding the enzymes is not critical, so long as the 0023. It has been surprisingly discovered that extracts mixture is heated to a temperature that permits the enzymes to obtained from the pomegranate Solids, in accordance with the at least partially degrade the pomegranate Solids and liberate methods disclosed herein, have a substantially higher total phytochemicals from the plant tissues and/or cells. Once lib polyphenol content than is found in the juice from the pome erated, the phytochemicals may react and/or polymerize to granate arils. This is particularly true with respect to the create new phytochemical compounds or reaction products. higher molecular weight polyphenols and, in particular, puni 0031 Enzymes suitable for use in accordance with this calagin. embodiment include those which are capable of at least par 0024 Punicalagin is a powerful antioxidant, protecting tially degrading the plant tissue or cells to liberate the phy cardiovascular function and accurate cellular replication. tochemicals from the pomegranate Solids. Such enzymes Thus, punicalagin is responsible, in part, for the high antioxi include anyone or a combination of pectinase, cellulase, dant activity of the extract. While the antioxidant and other hemicellulase, amylase, arabanase, and other hydrolyzing beneficial health effects of the extract are due to the presence enzymes, to name a few. The enzymes added to the mixture of polyphenols, the presence of other phytochemical com may be naturally-occurring or synthetic. They may be derived pounds in the extract, or the synergistic effect of these phy from any one or a combination of Sources, such as animal, tochemicals, may also be responsible for the anti-oxidant and plant, fungal, and bacterial sources. The amount of the other beneficial health effects of the extract. enzyme or combination of enzymes added to the mixture 0025. In addition to punicalagin, other high molecular depends on the temperature of the mixture and the amount of weight polyphenols have been characterized in the extract of pomegranate solids present in the mixture. pomegranate solids. These high molecular weight polyphe 0032. After enzymes are added, the mixture is maintained nols include elagitannin and other hydrolysable tannins, at a temperature for a time sufficient to allow at least partial Such as punicacortein A, punicalin, , and gallo degradation of the pomegranate solids. The temperature and tanin dimmers and trimers. length of time required depends on the type of enzymes added 0026. Moreover, a large number of anthocyanins have to the mixture, the rate of enzyme catalysis and the amount of been characterized in the extract of the pomegranate Solids. the pomegranate solids contained in the mixture. Examples of the anthocyanins include pelargonidin 3-gluco 0033. Thus, in one preferred embodiment, a combination side, cyaniding 3-glucoside, delphinidin 3-glucoside, pelar of pectinase, cellulase and hemicellulase enzymes are added gonidin 3,5-diglucoside, cyaniding 3,5-diglucoside, and del to the mixture, which is heated to a temperature of about 60° phinidin 3,5-diglucoside. Although these anthocyanins have F. to 210° F., preferably about 110° F. to 160° F., and opti been characterized in both the pomegranate juice and the mally of about 120° F. The mixture is maintained at these US 2015/0079208 A1 Mar. 19, 2015

temperatures, preferably with agitation or stirring, for about talc, magnesium Stearate, calcium Stearate, colloidal silica or 45-195 minutes, preferably for about 45-75 minutes, and Stearic acid, and binders such as polyvinylpyrrolidone, cellu optimally for about 60 minutes. lose derivatives, carboxymethyl cellulose, hydroxylpropyl 0034. After the enzymes have at least partially degraded cellulose, hydroxypropylmethyl cellulose, methyl cellulose the pomegranate Solids, the residual insoluble solid materials or gelatin. Conventional procedures for preparing Such com are removed from the mixture. Optionally, a clarification positions in appropriate, dosage forms of the extract may be agent, such as bentonite, may be added before the step of utilized. Such compositions may be administered orally or removing the residual insoluble materials from the mixture. parenterally employing liquid form preparations containing The removal of residual insoluble materials from the mixture the extract. may be accomplished by filtration, centrifugation, chromato 0041. The compositions may be administered orally, in graphic techniques, and other techniques. Filtration tech appropriate dosage units of the extract in a pharmaceutically niques suitable for the practice of the methods disclosed acceptable carrier or excipient. Thus, the compositions may herein include micro-filtration at a molecular weight cut-off be formulated into Solid or liquid preparations, such as cap of at least 1,000 Da, preferably of about 4,500 Da, and opti Sules, pills, tablets, powders, solutions, Suspension, or emul mally of about 5,500 Da. sions and may be prepared according to methods known in the 0035. The resulting liquid extract may be concentrated in art for the manufacture of such compositions. The solid unit an evaporator under vacuum to about 50-90 Brix (BX), pref dosage forms may be in form of a hard or soft shelled gelatin erably to about 60-80 Bx, and optimally to about 70 Bx, and capsule containing the extract and a suitable carrier or excipi pasteurized at a temperature and for a length of time Sufficient ent. to kill microorganisms that could cause disease, spoilage or 0042. The composition may also be administered undesired fermentation. In one preferred embodiment, the parenterally as injectable dosages in a physiologically accept extract may be pasteurized at a temperature of about 140° able carrier. Parenteral administration may be subcutaneous, F-280° F., preferably of about 195° F-240° F., and optimally intravenous, intramuscular, or interperitoneally. of about 205 F. The pasteurization may also denature the 0043. The effective amount of a composition is the amount remaining enzymes that were added to the mixture. or dosage unit of the extract sufficient to achieve the intended 0036. In another preferred embodiment, extracts contain beneficial health results. Accordingly, the effective amount of ing phytochemicals from a pomegranate are provided. Such the composition to be administered depends on consider extracts are characterized by a significantly higher total ations such as the dosage unit employed, the mode of admin polyphenol content, particularly of the high molecular weight istration, the period of treatment, the age, sex and weight of polyphenol (e.g., punicalagin), than is found in pomegranate the person treated and the nature and extent of the condition juice. Such extracts may be obtained from the methods dis treated. The effective amount can readily be determined closed herein. In a further preferred embodiment, extracts based upon standard techniques known to evaluate whether containing phytochemicals, polyphenols, punicalagin, puni the intended effect of the composition has been achieved, by calin, elagic acid, and metabolite thereof are provided. standard toxicity tests and by standard pharmacological 0037. In yet another preferred embodiment, food products assayS. and beverages are provided comprising the extract containing 0044. In a further preferred embodiment, compositions phytochemicals from a pomegranate. For example, due to the containing the extract and the pomegranate juice are pro significantly higher total polyphenol content in the extract, an vided. The combination of the extract and pomegranate juice 8 oz. sports beverage containing 0.33 oz of the extract may be not only produces a composition having a higher total formulated to deliver the same total polyphenols as a 20 oz polyphenol content, as compared to the pomegranate juice single-strength pomegranate juice. The polyphenol content of alone, but it also provides the broad spectrum of the different pomegranate juice is approximately about 1 to 2.25 mg/mL polyphenols which predominate the pomegranate juice and and the amount of polyphenols present in 20 OZ of juice is extract, for example the lower molecular weight polyphenols approximately 567 to 1.256 mg. In contrast, the extract may (e.g., anthocyanins) which is present in greater quantities in contain a polyphenol content of about 60 to 120 mg/mL, the pomegranate juice and the higher molecular weight depending on the method employed. Thus only 0.33 oz of the polyphenols (e.g., punicalagin, punicalin, elagic acid glyco 70 BX extract would be needed to provide the equivalent sides, polyphenols, and other amount of polyphenols in 20 oz of the juice. hydrolysable tannins, such as punicacortein A, punicalin, 0038. In a further preferred embodiment, compositions pedunculagin, and gallotanin dimmers and trimers). comprising the extract containing phytochemicals from a 0045. In yet a further preferred embodiment, methods are pomegranate are provided. The compositions may be formu provided for ameliorating disease conditions in a Subject by lated in the form of tablets, Suspensions, implants, Solutions, administering to the Subject an effective amount of the com emulsions, capsules, powders, syrups, liquid compositions, position Suitable for use as a pharmaceutical or nutritional ointments, lotions, creams, pastes, gels, and the like. preparation. Such disease conditions include polyphenolme 0039. The compositions may also be prepared in forms diated diseases and cancer. Suitable for use as pharmaceutical preparations, nutritional 0046 Polyphenols and countless other phytochemicals in Supplements, vitamin Supplements, food Supplements, and the extract are necessary for the various organs and tissues food additives. As such, the compositions may optionally and for the proper functioning of the human body. Accord include a Suitable carrier or excipient. ingly, many disease conditions may be prevented or amelio 0040 Suitable carriers or excipients are inert ingredients rated by the administration of polyphenols to patients with and include, by way of example, fillers, e.g. Sugars such as polyphenolmediated diseases. These polyphenol-mediated lactose, or Sucrose, Sugar alcohols such as mannitol, diseases include circulatory disorders such as hypertension Sorbitol or xylitol, starch Such as wheat, corn or potato starch, and coronary artery disease, erectile dysfunction, lung disor modified Starch or Sodium starch glycolate, lubricants such as ders such as asthma, cancers of various types, inflammatory US 2015/0079208 A1 Mar. 19, 2015

conditions, certain liver conditions, diabetes, mood disorders, 0055. The pomegranate solids were then transferred to eye disorders such as cataracts, weak eyesight due to aging, three Reitz, Mills with 3/8-inch screens. The material was macular degeneration, and other age-related disorders. Such milled to a fine puree and heated to approximately 125° F. as Alzheimer's disease and dementia. This step, coupled with the following enzyme addition, 0047 Thus, in one preferred embodiment, methods are assisted in breaking down the colloidal structure of the provided for formulating a composition Suitable for use as a remaining pomegranate solids, thereby releasing the remain pharmaceutical or nutritional preparation for improving the ing soluble Solids. health of a Subject comprising obtaining an extract containing 0056. The mixture was heated to a temperature of about 125° F. for two hours. Three enzymes were added to the phytochemicals from a pomegranate and admixing an effec mixture: pectinase (Rohapect(R) DA6L), cellulase/pectinase tive amount of the extract with a suitable carrier or excipient. (Rohapect(R) CL), and hemi-cellulase/pectinase (Rohapect(R) In another preferred embodiment, methods are provided for B1L). These enzymes were used to liberate the remaining treating a polyphenol-mediated condition in a Subject com pomegranate soluble Solids, such as Sugars, minerals, antho prising selecting a subject having a polyphenol-mediated cyanins, and remaining polyphenols. condition and administering to the Subject an effective 0057 The mixture was then pumped from the extraction amount of the composition comprising the extract. plant to the primary processing plant where it was held in the 0048. In yet another preferred embodiment, methods are mash treatment tanks for approximately one hour. After one provided for modulating the growth and progression of can hour, 50-100 pounds of bentonite in a 125 gallon water slurry, cerous cells, the methods comprising selecting a subject hav per 8,000 gallons of the mixture, was added for protein ing cancerous cell growth and administering to the Subject an removal. The treated mixture was then passed through a West effective amount of the composition containing the extract. phalia 755 Decanter for removal of solids. The residual 0049. In yet a further embodiment, methods are provided insoluble material was discharged as waste. for preventing or slowing increases in the Prostate Specific 0058. The remaining liquid extract was processed in a Antigen (PSA) levels in a Subject having prostate cancer. The Schmidt evaporator. In this step, the extract was stripped and method comprises selecting a Subject having prostate cancer rectified. In addition, the liquid extract was pre-concentrated and administering to the Subject an effective amount of the and then pasteurized to 205 F. for 45 seconds. The liquid composition containing the extract. extract then exited the evaporator and was filtered on Koch 0050 Prostate cancer is the most commonly detected can Micro-Filtration membranes at a 4,500 Da molecular weight cer in men in the U.S., affecting approximately 1 out of every cut-off for liquid extract soluble solids. 6 men. It is the second leading cause of cancer death among 0059. The liquid extract then re-entered the evaporator for men in the U.S. Laboratory testing can assist with screening, final concentration. Initial heat on this step was about 185 diagnosis, staging, prognosis, detection of residual or recur 195° F. At about 70 Bx, the liquid extract was cooled to less rent disease, and therapeutic monitoring. The primary test than about 45° F. and pumped to the concentrate batching used for these purposes is a PSA test. room where it was blended and standardized. 0051. The PSA test was approved by the FDA in 1986 to help detect prostate cancer. A number of prostate problems Example 2 can be identified by testing and monitoring the levels of PSA circulating in the bloodstream. The level of PSA in the blood Comparison of Polyphenol Content in Extracts of stream may be elevated by an process that leads to an increase Pomegranate Solids and in Pomegranate Juice in the number of cells making PSA or to a breakdown of the 0060. The concentrations of punicalagin, punicalin, normal barriers in the prostate that prevent much PSA from ellagic acid glycosides, and ellagic acid polyphenols in the getting into the bloodstream. pomegranate juice and the pomegranate extract were ana 0052 Increases in levels of PSA in the blood following lyzed and compared in a University study. treatment for localized prostate cancer with Surgery or radia 0061 All samples (50 mL injection volume) were filtered tion often indicates the presence of residual cancer and the (0.22 mm) and analyzed on a Novapak (Waters Corp.) C-18 eventual development of metastatic cancer. Moreover, PSA column, 150x3.9 mm i.d., 5 mm. The mobile phase, solvent A doubling times are correlated with diagnostic tumor stage and (2% CH3COOH/H20) and solvent B (2% aqueous grade CH3COOH/CH3OH) was used under linear gradient condi 0053. The following examples further illustrate the tions starting with 99% solvent A in solvent B to 40% solvent embodiments disclosed herein. These examples are provided A in solvent B over 40 minutes, hold time, 5 minutes with a only for purpose of illustrating the preferred embodiments of flow rate of 1 mL/min. All compounds were detected at 254 the invention and do not limit the invention in any manner. nm, and at 378 nm () and 366 (ellagic acid) for quantification. Table 1 shows a side-by-side comparison of Example 1 the concentration of the polyphenols punicalagins, punicalin, ellagic acid glycosides, and elagic acid in the pomegranate Production of Liquid Extract from Pomegranate extract and the pomegranate juice. Solids 0054 The starting material for the production of the TABLE 1 extract is the pomegranate solids, which generally comprise Pomegranate Extract Pomegranate Juice the pericarp, the inner membrane and seeds of the pomegran Concentration Concentration ate. The pomegranate solids were obtained and collected after Compound Name (mg/ml) (mg/ml) the primary juice from the arils had been substantially Punicalagin (f isomer) 4.79 O.O2 expelled or otherwise removed from the pomegranate by Punicalagin (C. isomer) 21.80 O.15 pressing, crushing, or other methods known to the art for Punicalin 3.62 NA extracting pomegranate juice. US 2015/0079208 A1 Mar. 19, 2015

TABLE 1-continued Example 4 Pomegranate Extract Pomegranate Juice Preventing or Slowing Increases in the PSA Levels Concentration Concentration of Patients with Prostate Cancer Compound Name (mg/ml) (mg/ml) Ellagic Acid 1965 O.33 0067. While both pomegranate juice and pomegranate Glycosides Solid extract contain various types of the anti-oxidant Ellagic Acid 18 O.74 polyphenols, pomegranate solid extract contains a higher total polyphenol content than the pomegranate juice. Accord Total 6786 1.24 ingly, to the extent that the administration of 8 oz of pome granate juice to patients with prostate cancer has been dem onstrated to increase the PSA doubling time in patients with 0062 Although other polyphenols are present in both the prostate cancer, the administration of pomegranate solid pomegranate extract and juice, and this example highlights extract also achieves at least the same, and preferably an the unexpected and Surprising results in that significantly improved, effect. higher concentrations of polyphenols, particularly of puni 0068 Accordingly, methods and compositions are pro calagin, are present in the pomegranate extract than in the vided for preventing or slowing increases in the Prostate pomegranate juice. Table 1 shows a total punicalagin (for Specific Antigen (PSA) levels in a patient. The methods com both C. and B isomers) concentration for the pomegranate prise selecting a Subject having prostate cancer and adminis extract that is over 26-fold greater than for the pomegranate tering to the Subject an effective amount of a composition Ju1ce. containing the extract containing phytochemicals from a pomegranate solid. Example 3 0069. In preferred one embodiment, the composition may be inform of a liquid comprising the extract and pomegranate juice. The total polyphenol content provided by the liquid Effects of Pomegranate Juice in Men with Rising may be varied by the changing the amount of the pomegranate PSA Following Surgery or Radiation for Prostrate extract and pomegranate juice contained in the liquid. Table 2 Cancer provides examples of the formulations of the liquid compo sition and the total polyphenol content in the formulations 0063. The positive and significant beneficial effects of relative to the total polyphenol content in standard pomegran pomegranate juice on Prostrate Specific Antigen (PSA) ate juice. parameters have been demonstrated in a clinical trial in patients with recurrent prostrate cancer, coupled with corre TABLE 2 sponding laboratory effects on prostrate cancer in in vitro cell Extract Pomegranate growth and apoptosis. Juice Liquid Pomegranate Pomegranate 0064. To determine the clinical effects of pomegranate Composition Extract (oz) Juice (OZ) juice on patients with prostate cancer, a clinical trial was Formulation 1 O.13 O.8 performed. A two-year, single center, phase II, Simon two 2x polyphenol content Formulation 2 O.26 O.8 stage clinical trial for men with rising PSA after surgery or 3 x polyphenol content radiotherapy was designed based on a 20% response rate, an Formulation 3 O.39 O.8 alpha of 5%, and 90% power. Eligible patients had a detect 4 x polyphenol content Formulation 4 O.S2 O.8 able PSA greater than 0.2 ng/ml and less than 5 ng/ml, and a 5 x polyphenol content Gleason score of 7 or less. Serial PSA measurements deter mined a baseline PSA doubling time. 0070 For purposes of this embodiment, the effective 0065 Patients were treated with 8 oz of pomegranate juice amount of the extract that is administered to the patient is at by mouth daily (Wonderful variety, equivalent to 1.5 mmol of least 0.13 oz (or an equivalent unit or measurement) of the total polyphenols per day) until disease progression. Clinical extract administered daily, whether the extract is provided endpoints included safety, effect on serum PSA, and explor alone or in a composition. Because the administration of 8 oz. atory laboratory studies. Patients were followed in 3-month of pomegranate juice was found to be effective in slowing the intervals for serum PSA, and blood and urine were collected rising PSA levels in patients with prostate cancer, the admin for laboratory studies. istration of at least 0.13 oz of the pomegranate solid extract is 0066. The study was fully accrued to 48 participants in two believed to achieve the same, if not improved, results. The stages after efficacy criteria were met. There were no serious dosage of the extract may be increased by administering a adverse events reported and the treatment was well tolerated. greater dosage or increasing the frequency at which the No patients developed metastatic disease on study. Mean extract is administered. PSA doubling time significantly increased with treatment, 0071. In addition to the liquid compositions containing the from a mean of 14 to 26 months (p<0.048). The slope of the extract, the extract may also be administered in a solid form, mean log PSA decreased from 0.08 to 0.04 on treatment Such as pharmaceutical or nutritional preparation that com (p<0.019). In vitro assays using pre- and post-treatment prises the extract and a pharmaceutically acceptable carrier or patient serum on the growth of LNCaP showed decreased cell excipient. proliferation and increased apoptosis (p<0.07). Pomegranate 0072 The invention described and claimed herein is not to polyphenols were detected in the urine of all participants by be limited in scope by the specific embodiments herein dis liquid chromatography mass spectrometry (LC-MS). closed, since these embodiments are intended as illustrations US 2015/0079208 A1 Mar. 19, 2015 of several aspects of the invention. Any equivalent embodi heating said mixture to a temperature that permits enzyme ments are intended to be within the scope of this invention. catalysis of said one or more pomegranate solids; Indeed, various modifications of the invention in addition to maintaining said mixture at a temperature and time suffi those shown and described herein will become apparent to cient to allow at least partial degradation of said one or those skilled in the art from the foregoing description. Such more pomegranate solids; modifications are also intended to fall within the scope of the removing residual insoluble Solid materials from said mix appended claims. ture; and What is claimed: wherein a resulting extract contains polyphenols of a level 1. A method of modulating growth and progression of greater than that which occurs in pomegranate juice cancerous cells comprising: made from arils of a pomegranate. Selecting a Subject having cancerous cell growth; 5. The method of claim 4, wherein said one or more pome administering to said Subject a composition comprising an granate solids are selected from the group consisting of peri effective amount of a pomegranate extract and pome carp, inner membrane and seeds from a pomegranate fruit. granate juice, whereby said pomegranate extract is pro 6. The method of claim 4, wherein said one or more pome duced by a process comprising: granate solids comprise a remainder of a whole pomegranate providing one or more pomegranate Solids; fruit following extraction of substantially all liquid from arils creating a mixture comprising said one or more pomegran of said whole pomegranate fruit. ate Solids in an aqueous solution; 7. A method of increasing Prostate Specific Antigen dou adding enzymes to said mixture in an amount Sufficient to bling time in a subject having prostate cancer, the method at least partially degrade said one or more pomegranate comprising: Solids; heating said mixture to a temperature that permits enzyme selecting a Subject having prostate cancer, catalysis of said one or more pomegranate Solids; administering to said Subject a composition comprising an maintaining said mixture at a temperature and time Suffi effective amount of a pomegranate extract and pome cient to allow at least partial degradation of said one or granate juice, whereby said pomegranate extract is pro more pomegranate solids; duced by a process comprising: removing residual insoluble Solid materials from said mix providing one or more pomegranate Solids; ture; and, creating a mixture comprising said one or more pomegran wherein a resulting extract contains polyphenols above a ate Solids in an aqueous solution; level found in said pomegranate juice made from arils. adding enzymes to said mixture in an amount Sufficient to 2. The method of claim 1, wherein said one or more pome at least partially degrade said pomegranate solids; granate Solids are selected from the group consisting of peri heating said mixture to a temperature that permits enzyme carp, inner membrane and seeds from a pomegranate fruit. catalysis of said one or more pomegranate solids; 3. The method of claim 1, wherein said one or more pome maintaining said mixture at a temperature and time suffi granate Solids comprises a remainder of a whole pomegranate cient to allow at least partial degradation of said one or fruit following extraction of substantially all liquid from arils more pomegranate solids; of said whole pomegranate fruit. removing residual insoluble Solid materials from said mix 4. A method of increasing Prostate Specific Antigen dou bling time in a Subject having prostate cancer comprising: ture; and, Selecting a Subject having prostate cancer, wherein a resulting extract contains polyphenols of a level administering to said Subject a composition comprising an greater than that which occurs in pomegranate juice effective amount of a pomegranate extract, whereby said made from arils of a pomegranate. pomegranate extract is produced by a process compris 8. The method of claim 7, wherein said one or more pome ing: granate solids are selected from the group consisting of peri obtaining one or more pomegranate solids; carp, inner membrane and seeds from a pomegranate fruit. creating a mixture comprising said one or more pomegran 9. The method of claim 7, wherein said one or more pome ate Solids in an aqueous solution; granate solids comprise a remainder of a whole pomegranate adding enzymes to said mixture in an amount Sufficient to fruit following extraction of substantially all liquid from arils at least partially degrade said one or more pomegranate of said whole pomegranate fruit. Solids; k k k k k