Supramolecular phospholipids–polyphenolics interactions: The PHYTOSOME ® strategy to improve the...

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Review

Supramolecular phospholipids–polyphenolics interactions: ThePHYTOSOME® strategy to improve the bioavailability of phytochemicals

Ajay Semalty a,⁎, Mona Semalty a, Mohan Singh Maniyari Rawat b, Federico Franceschi c,⁎a Department of Pharmaceutical Sciences, H.N.B. Garhwal University Srinagar (Garhwal), Indiab Department of Chemistry, H.N.B. Garhwal University Srinagar (Garhwal), Indiac Research and Development Laboratories, Indena S.p.A. (Settala), Italy

a r t i c l e i n f o a b s t r a c t

Article history:Received 20 October 2009Accepted in revised form 3 November 2009Available online 14 November 2009

The poor and/or erratic oral bioavailability of polyphenolics can be improved using thePHYTOSOME®1 delivery system, a strategy that enhances the rate and the extent of solubilizationinto aqueous intestinal fluids and the capacity to cross biomembranes. Phospholipids show affinityfor polyphenolics, and form supramolecular adducts having a definite stoichiometry. This articlereviews the preparation and characterization of PHYTOSOME® complexes and their activity invarious medicinal (cardiovascular, anti-inflammatory, hepatoprotective, anticancer) and cosmetic(skin aging) realms of application.

© 2009 Elsevier B.V. All rights reserved.

Keywords:FlavonoidBioavailabilityPHYTOSOME®

Herbal drugsPhospholipid complex

Contents

1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3072. Physical and chemical properties . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3073. Methods of preparation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3084. Characterization . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 308

4.1. Spectroscopy . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3084.2. Thermal gravimetric analysis (TGA)/differential scanning calorimetry (DSC) . . . . . . . . . . . . . . . . . . . . . 308

5. Biological profile of PHYTOSOME® adducts . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3095.1. Cardiovascular properties . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3095.2. Anti-inflammatory properties . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3095.3. Anti-aging properties . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3105.4. Hepatoprotective properties . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3105.5. Anticancer properties . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3125.6. Weight management. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 313

6. Conclusions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 313Acknowledgments . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 313References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 313

Fitoterapia 81 (2010) 306–314

⁎ Corresponding authors. Semalty is to be contacted at Department of Pharmaceutical Sciences, PB No.-32, H.N.B. Garhwal University Srinagar (Garhwal)-246174,India. Franceschi, Indena S.p.A., via Don Minzoni 6, 20090 Settala (MI), Italy.

E-mail addresses: [email protected] (A. Semalty), [email protected] (F. Franceschi).1 PHYTOSOME® is a registered trademark of Indena S.p.A. Milan, Italy.

0367-326X/$ – see front matter © 2009 Elsevier B.V. All rights reserved.doi:10.1016/j.fitote.2009.11.001

Contents lists available at ScienceDirect

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1. Introduction

For good bioavailability, natural products must have a goodbalance between hydrophilicity (for dissolving into the gastro-intestinal fluids) and lipophilicity (to cross lipidic biomem-branes). Many phytoconstituents like polyphenolics have goodwater solubility, but are, nevertheless, poorly absorbed [1]because of their large size, incompatiblewith a process of passivediffusion and/or their poor miscibility with oils and other lipids.As a result, the ability of flavonoids to cross the lipid-rich outermembrane of small intestine enterocytes is severely limited.

Water-soluble phytoconstituents (mainly polyphenolics)can be converted into a lipid-compatible molecular complexknown as PHYTOSOME®. A PHYTOSOME® is generally morebioavailable than a simple herbal extract due to its enhancedcapacity to cross the lipid-rich biomembranes and reachcirculation [2–5]. Phospholipids are small lipid moleculeswhere glycerol is bonded to two fatty acids, with the thirdhydroxyl, normally one of the two primary methylenes,bearing a phosphate group [6]. Phospholipids from soy,mainly phosphatidylcholine, are lipophilic substances andreadily complex polyphenolics. In this context, phosphatidyl-choline, the major molecular building block of cell mem-branes and a compoundmiscible in bothwater and in oil/lipidenvironments, is well absorbed orally, and has the potentialto act as a chaperon for polyphenolics, accompanying themthrough biological membranes [7].

PHYTOSOME® complexes were developed at Indena(Milan, Italy) in the late Eighties. Many popular standardizedherbal extracts [e. g. Ginkgo biloba L., grape (Vitis vinifera L.)seeds, milk thistle (Silybum marianum (L.) Gaertn), green tea(Camellia sinensis (L.) O. Kuntze), ginseng (Panax ginseng C.A.Meyer), licorice (Glycyrrhiza glabra L.), horse chestnut(Aesculus hippocastanum L.), Centella asiatica (L.) Urban,olive (Olea europea L.), Terminalia sericea Burch. Ex. DC, Amnivisnaga (L.) Lam, turmeric (Curcuma longa L.) and hawthorn(Crataegus spp.)] are currently commercially available in thePHYTOSOME® form (see Table 1). Flavonoids and terpenoidsfrom herbal extracts undergo different absorption pathways

when bound to phospholipids, and this article reviews variousaspects and the latest trends of PHYTOSOME® research,highlighting recent advances in their therapeutic potential.

2. Physical and chemical properties

The first PHYTOSOME® generation was prepared bycombining selected polyphenols or polyphenol extractswith phospholipids in non-polar solvents [2], but, morerecently, a newer PHYTOSOME® generation was developedusing hydro-ethanolic solvents. Products obtained in this waycomply with current food specifications [8,9], and expand thePHYTOSOME®' potential from the pharma/cosmetic field tothe health-food one.

A PHYTOSOME® is an amphiphilic substance with adefinite melting point, generally soluble in nonpolar solvents(where its hydrophilic moiety is not), and moderately solublein fats. The low solubility in aqueous media makes theformation of stable emulsions and creams possible (Fig. 1),improving the biopharmaceutical properties of both highlylipid insoluble and poorly water-soluble phytoconstituents.

The PHYTOSOME® formulation increases the absorption ofactive ingredients when topically applied on the skin [10–19],and improves systemic bioavailability when administeredorally [20–24]. In water medium, a PHYTOSOME® will assumea micellar shape, forming a liposome-like structure. Funda-mental differences exit, however, between a PHYTOSOME® anda liposome. In liposomes, the active principles are dissolved inthe central part of the cavity, with no possibility of molecularinteraction between the surrounding lipid and a hydrophilicsubstance. On the contrary, the PHYTOSOME® complex cansomewhat be compared to an integral part of the lipidmembrane (Fig. 2), where the polar functionalities of thelipophilic guest interact via hydrogen bonds with the polarhead of a phospholipid (i.e. phosphate andammoniumgroups),forming a unique arrangement that can be evidenced byspectroscopy [10,11,18,25–28].

Thus, IR andmulti-nuclear spectroscopic studies showthat aPHYTOSOME® is not a mechanical mixture of two constituents,

Table 1Available PHYTOSOME® complexes on the market. PHYTOSOME® and all other trademarks are owned by Indena S.p.A. Milan, Italy.

Trade name Phytoconstituent complexed with phospholipid Indication

Escin ß-sitosterol Phytosome® Escin ß-sitosterol from horse chestnut fruit Anti-oedemaSiliphos® Silybin from milk thistle seed Hepatocyte protectionSilymarin Phytosome® Silymarin from milk thistle seed AntihepatotoxicMeriva™ Curcuminoids from turmeric rhizomeVirtiva® Ginkgoflavonglucosides, ginkgolides, bilobalide from Ginkgo biloba leaf VasokineticGinkgoselect® Phytosome® Ginkgoflavonglucosides, ginkgolides, bilobalide from Ginkgo biloba leaf VasokineticGinselect® Phytosome® Ginsenosides from Panax ginseng rhizome Skin elasticity improver, adaptogenicLeucoselect® Phytosome® Polyphenols from grape seed Antioxidant, capillarotropicCentella Phytosome® Triterpenes from Centella asiatica leaf Cicatrizing, trophodermic18 ß-glycyrrhetinic acid Phytosome® 18 ß-glycyrrhetinic acid from licorice rhizome SoothingCrataegus Phytosome® Vitexin-2″-O-rhamnoside from Hawthorn flower AntioxidantGinkgo biloba Dimeric FlavonoidsPhytosome®

Dimeric flavonoids from Ginkgo biloba leaf Lipolytic, vasokinetic

Ginkgo biloba Terpenes Phytosome® Ginkgolides and bilobalide from Ginkgo biloba leaf SoothingSericoside Phytosome® Sericoside from Terminalia sericea bark root Anti-wrinklesGreenselect® Phytosome® Polyphenols from green tea leaf Prevention of free radical-mediated tissue

damages and weight managementVisnadex® Visnadin from Amni visnaga umbel VasokineticPA2 Phytosome® Proanthocyanidin A2 from horse chestnut bark Anti-wrinkles, UV protectant

307A. Semalty et al. / Fitoterapia 81 (2010) 306–314


but a specific complex between a hydrophilic guest and alipophilic host characterized by specific dipolar interactions, inaccordance with the spectroscopic differences between aPHYTOSOME and amechanical mixture of its two constituents.

3. Methods of preparation

A PHYTOSOME® is prepared by complexing a polyphenolicphytoconstituent or mixture with a phospholipid. Dependingon the product, mass ratios in the 1:1.5–1:4 are observed.Various preparation methods for a PHYTOSOME® [3,8,9,12–16,19,29–33] and the resulting complex can be different

depending on the protocol employed. Thus, the phospholipidcomplex of curcumin [34] prepared in an aprotic solvent [35]showed remarkable differences from the one prepared in aprotic solvent [8], and three different silybin–phospholipidcomplexes have been reported. Two of themwere prepared inaprotic solvent [Silipide (IdB 1016) [30], a pharma-gradePHYTOSOME® which has been extensively characterized [27],and Siliphos®], while a third silybin–phospholipid complexwas obtained in protic solvents [32]. Different phospholipidsafford different complexes, as found for G. biloba extracts,whose PHYTOSOME® extract with phosphatidylserine com-plex is known as Virtiva®, while the standard phosphatidyl-choline complex is known as Ginkgoselect® PHYTOSOME®)[31].

4. Characterization

4.1. Spectroscopy

Complexation and molecular interactions between phy-toconstituents and phosphatidylcholine in solution havebeen studied by 1H-NMR [2,8,10,25–27], 13C-NMR[2,8,10,27], 31P-NMR, [8–10] as well as by IR spectroscopy[18]. Complex formation is associated with changes ofchemical shift and line broadening of some characteristicsignals in NMR spectra and with appearance of new bands inIR spectra.

4.2. Thermal gravimetric analysis (TGA)/differential scanningcalorimetry (DSC)

Detection and measurement of thermal effects such asfusion, solid–solid transitions, glass transitions, loss ofsolvent, and decomposition, can be used to characterize asolid PHYTOSOME®.

The TG/DT analyses of two complexes and themechanicalmixtures of phytoconstituent and phospholipid componentsare reported in Fig. 3 [36]. The DT profile of SilymarinPHYTOSOME® (green solid line in Fig. 3a) shows anendothermic peak with a maximum at about 173 °C, cor-responding to a ca. 3% weight loss, and a less intense andbroader endothermic signal with a maximum at about148 °C. The TG profile (blue solid line in Fig. 3a) of somePHYTOSOME® adducts shows an initial loss on drying below100 °C (about 4%), and a massive weight loss starting atabout 175 °C due to degradation. The TG/DT profiles of thephytoconstituent/phospholipid mechanical mixture(dashed lines) show two broad endothermic peaks, thefirst onewith amaximum at about 146 °C and correspondingto a weight loss of ca. 2%, and the second one at about 167 °Cwith a loss of weight of ca. 3% followed by themassiveweightloss expected for degradation. Fig. 3b shows the TG/DTprofile of Ginkgoselect® PHYTOSOME® and the one of aGinkgoselect®/phospholipid mechanical mixture. The DTprofile of the PHYTOSOME® (green solid line in Fig. 3b)shows only an endothermic peak with a maximum at about179 °C, linked to aweight loss of about 2%. Conversely, the TGprofile (blue solid line) shows an initial loss on drying below100 °C (about 2%) and amassive weight loss starting at about180 °C due to degradation.

Fig. 1. Electron-microscopic examination of lyophilized microdispersion ofGinselect® PHYTOSOME® 0.1% in water. Magnification 1×4000.

Fig. 2. Major difference between liposome and PHYTOSOME®.

308 A. Semalty et al. / Fitoterapia 81 (2010) 306–314


The TG/DT profiles of the mechanical mixture ofGinkgoselect® and a phospholipid (dashed lines) showstwo broad endothermic peaks: the first with a maximum atabout 135 °C simultaneous with a loss of weight of about 3%,and the second with a maximum at about 171 °C with a lossof weight of about 2% followed by a massive weight due todegradation.

In both cases, phytoconstituent–phospholipid molecularinteractions in the complex lead to a radically differentthermal profile and behavior of the PHYTOSOME® complexcompared to its corresponding mechanical mixture.

5. Biological profile of PHYTOSOME® adducts

5.1. Cardiovascular properties

Preliminary studies have shown that thePHYTOSOME®ofG.biloba L. and that of grape seed extracts are more potent thantheir non-complexed forms. The efficacy of Ginkgoselect®

PHYTOSOME® to treat peripheral vascular disease (e.g.,Raynaud's disease and intermittent claudication) was foundto be 30–60% higher than that of Ginkgoselect® [37]. Ginkgo-select® PHYTOSOME® was also recently validated as acardioprotective agent [38]. The two G. biloba L. PHYTOSOME®

complexes showed different cognitive effects, with thephosphatidylserine complex (Virtiva®) being active atlower doses than the standard phosphatidylcholine complex(Ginkgoselect® PHYTOSOME®) [31,39].

The activity of Leucoselect® PHYTOSOME® to decreaselow-density lipoprotein susceptibility to oxidation andoxidative stress damage in a group of heavy smokers wasalso investigated, with positive results [40].

5.2. Anti-inflammatory properties

PHYTOSOME® complexes show better anti-inflammatoryactivity than their uncomplexed herbal extracts. Croton oil-induced dermatitis was used, as an inflammation model

Fig. 3. a: TG curves (blue) and DT profiles (green) of Silymarin PHYTOSOME® (solid line) and silymarin/phospholipid mechanical mixture (dashed line). b: TGcurves (blue) and DT profiles (green) of Ginkgoselect® PHYTOSOME® (solid line) and Ginkgoselect®/phospholipid mechanical mixture (dashed line).



involving both tissue and vascular damage, to evaluate theanti-inflammatory activity of PHYTOSOME® complexes inanimals [18]. The results obtained with glycyrrhetinic acid, apowerful anti-inflammatory and anti-allergic triterpenoid,and with silymarin, a free radical scavenger flavanolignanendowed with anti-phlogistic activity, were discussed. Theproducts were applied topically as microdispersion in water,and the reduction of edema was used as the end point.Phospholipids are per se devoid of anti-inflammatory activity,but it was found that their complexation with glycyrrhetinicacid prolonged, its activity. Thus, the anti-edematous activityof 18-β-glycyrrhetinic acid gradually waned with time andwas almost completely terminated after 24 h, a time at whichits phosphatidyl complex still induced an 80% reduction inedema. The silymarin complex showed greater reduction ofedema (76% reduction in 6 h) as compared to that with thefree form (33% even after 12 h) in the croton oil test in mice(Fig. 4). Therefore, the PHYTOSOME® was about two-foldmore effective than the free phytoconstituent. In anotherstudy, the 18-β-glycyrrhetinic acid complex exhibited potentactivity against the erythema secondary to UVA radiation[28].

5.3. Anti-aging properties

PHYTOSOME® as a delivery system offers interestingapplications and opens new opportunities for the use ofactive ingredients in the cosmetic field. The G. bilobaPHYTOSOME® was investigated for the treatment of skinaging connected to superficial capillary blood flow circula-tion. Extracts from G. biloba are used orally to improveperipheral circulation [41], and their phospholipid complexeswere found to improve skin microcirculation after topicalapplication. Activation of microcirculation meliorated skinaging associated to dystrophic alteration of the epidermis anddermis, and the regressive abiotrophic pannicular disease oflower limbs and breast associated to venous stasis and/orchronic venous insufficiency. The role of PHYTOSOME® in

functional cosmetics was reviewed [25], and the use ofSilymarin PHYTOSOME® on aging skin was also reported[18,42].

5.4. Hepatoprotective properties

Most studies on PHYTOSOME® are focused on Silybummarianum Gaertner and its liver-protectant flavolignans. Thefruit of milk thistle (Silybum marianum) contains flavonoidsshowing hepatoprotective properties [43,44]. Silymarin hasbeen shown to be effective in the treatment of liver diseasesof various kinds, including hepatitis, cirrhosis, fatty infiltra-tion of the liver (chemical and alcohol induced fatty liver),and inflammation of the bile duct [45–48]. The antioxidantproperties of silymarin substantially boost the liver resistanceto toxic insults [49]. Silymarin has a complex composition,based on three patterns of covalent interaction between itsflavonol and lignan constituents (Fig. 5). Silybin, in turn themixture of two diastereomers, is the major and more potentconstituent of silymarin [43]. Silybin protects the liver byconserving glutathione in the parenchymal cells [49], whilephosphatidylcholine helps repair and replace cell membranes[7]. These constituents act synergistically, sparing liver cellsfrom destruction. While silybin was found active in manymodels of human disease, clinical translation of these resultswas hampered by its extremely poor oral bioavailability. In apharmacokinetic study [32], the oral bioavailability of silybinwas increased remarkably when administered as silybin–phospholipid complex. The increased lipophilicity of thesilybin–phospholipid complex, have been claimed to explainthe improved bioavailability [7].

AFB1 (aflatoxin B1, a fungal compound produced byvarious strains of Aspergillus flavus and Aspergillus para-siticus) causes aflatoxicosis in chickens, and is characterizedby mortality, listlessness, anorexia, decreased growth rates,negative feed conversions, fatty liver, decreased egg pro-duction, poor pigmentation, and increased susceptibility toother diseases. For these reasons, intensive research hasbeen carried out to develop cost-effective and safe methodsto reduce the effects of AFB1. Remarkably, SilymarinPHYTOSOME® showed better antihepatotoxic activity thansilymarin alone, and could provide protection against thetoxic effects of aflatoxin B1 on safety and quality perfor-mance of broiler chickens [50].

A series of studies on silymarin administered as SilymarinPHYTOSOME® found that the complex could protect the fetusfrom maternally ingested EtOH [51]. In similar studies, it wasreported that Silymarin PHYTOSOME® shows a better fetopro-tectant activity against ethanol-induced behavioral deficitsthan uncomplexed silymarin on fetuses of both sexes [52]. Inthe first study, gammaglutamyl transpeptidase (GGTP) activitywas determined for liver and brain tissue from both the fetusand the dams on day 21 of pregnancy, while the offspringweretested at age 90 days on the social recognition task and at75 days on the radial armmaze in the later study. Female EtOH-exposed offsprings performed more poorly on the radial armmaze than did female EtOH/Silymarin PHYTOSOME® offspringsand that of controls. In a similar way, male EtOH-exposedoffspringswere less able to form socialmemories than themaleEtOH/Silymarin PHYTOSOME® offsprings and the male off-spring control.

Fig. 4. Reduction of edematous response to croton oil-induced by silymarin andits complex with distearoylphosphatidylcholine at different times (dose0.5 µmol/ear).



Silymarin PHYTOSOME®, was also found to have a muchhigher and specific activity and a longer lasting activity thanany single silymarin component in terms of oedema,inhibition of myeloperoxidase activity, antioxidant and freeradical scavenging properties [42].

When silybin was administered as Silipide (a pharma-grade silybin–phospholipid complex) at a dose of 200 mg perkg body weight, plasma concentration of silybin remainedhigh 70 h after the oral dosing, while, after 25 h, silybin barelyrose above the detection threshold [21]. In another study onrats, silybin given as Silipide at 200 mg/kg was detectable inthe plasma within minutes, peaked after 1 h, and its plasmalevels remained elevated for over 6 h. Silipide was also shownto rapidly reach the liver, cross the liver cells, and appear inthe bile within 2 h. The amount of silybin reaching the bilefrom PHYTOSOME® dosing was at least 6.5 times higher thanthat from non-complexed silybin (13% versus 2%, over 24 h)[22]. In general, Silipide was found to show higher pharma-cological potency in animal models of hepatic injury thanuncomplexed silybin, and the oral bioavailability and organdistribution profile of Silipide was much better than that ofsilybin from silymarin [53]. The pharmacokinetics of Silipidein healthy human subjects showed that complexation withphosphatidylcholine improved the oral bioavailability ofsilybin 4.6 fold compared with silymarin, probably because

of a facilitated passage across the gastrointestinal mucosa[20].

A pilot study on the liver protective effect of Silipide inchronic active hepatitis (CAH) found that Silipide improvedliver function tests related to hepatocellular necrosis and/orincreased membrane permeability in patients affected byCAH [54].

The iron chelating properties of Silipide were alsoinvestigated during a 12 week study on 37 patients withchronic hepatitis C and Batts–Ludwig fibrosis. Treatment withsilybin–phosphatidylcholine complex was clearly associatedwith a reduction in iron stores in patients with advancedfibrosis [55]. Hepatitis C infection was also improved by acombination of Silipide and vitamin E (SPV) [56].

In another study, the role of SPV complex for thetreatment of non alcoholic fatty liver disease (NAFLD) wasinvestigated [57]. Significant correlation among indexes offibrosis, body mass, insulinemia, plasma levels of transform-ing growth factor-β, tumor necrosis factor-α, degree ofsteatosis and γ-glutamyl transpeptidase was observed.Reduction in ultrasonographic scores for liver steatosis andimprovement of liver enzyme levels, hyperinsulinemia andindexes of liver fibrosis suggest that SPV complex could beused as a complementary approach to the treatment ofpatients with chronic liver damage.

Fig. 5. Flavolignans of Silybum marianum and hydrogen bond interactions in phosphatidylcholine complex of silybin.



The addition of increasing concentrations of Silipide toisolated rat hepatocytes caused a dose-dependent inhibitionof lipid peroxidation induced by cumene hydroperoxide,concluding that Silipide acted as potentially useful protectiveagent against free radical mediated toxic liver injury [58].Prompted by these observations, a phase II randomized openclinical trial to evaluate the dose response relationship ofSilipide in patients with viral or alcoholic hepatitis wascarried out, finding a dose-dependent improvement [59]. In apilot study on eight patients with chronic active hepatitis Band/or C, treatment with Silipide, at 240 mg silybin, for twomonths, the liver enzymes alanine aminotransferase (ALT)and aminotransferase (AST)were significantly reduced, whileglutamyltranspeptidase (GGT) and malondialdehyde (MDA)levels, a byproduct of lipid peroxidation were not statisticallyimproved [60].

Healthy volunteers (total number not disclosed) receiving360 mg Silipide three times daily for three weeks did notshow any adverse effect [61], while treatment of 232 patientswith liver disorder for up to four months with either 240 or360 mg Silipide daily showed an excellent tolerability. Minoradverse effects (nausea, heartburn, dyspepsia, transientheadache) were reported only in 12 patients (5.2% of thetotal studied), compared with 8.2% of patients who receiveduncomplexed silybin and 5.1% of patients treated with theplacebo. In this study, Silipide produced no clinically relevantblood changes, and its efficacy was evaluated by measuringserum levels of aspartate aminotransferase (AST), alanineaminotransferase (ALT), and gamma-glutamyltranspeptidase(GGT). Besides antioxidant activity, Silipide has also theability to scavenge ethanol-derived free radicals damage, andhas therefore potential to prevent liver damage from alcoholabuse [62]. Silipide was also able to scavenge free radicalsgenerated by CCl4 or INH [63].

Silipide was also investigated in rodents in various models ofliver damage. After oral administration, Silipide exhibited asignificant and dose related protective effect against hepatotox-icity inducedbyCCl4, acetaminophen, ethanol and galactosamine[64]. A controlled clinical study on the effect of Silipide in chronicpersistent hepatitis was carried out [65]. The drug treatmentsavailable for this condition have limited efficacy, do not work atall for many patients, and have major adverse effects. In thisstudy, patients were randomized to receive either 240 mgSilipide (n=31) or placebo (n=34) twice daily for threemonths. The Silipide group showed significant lowering of bothserum ALT and AST, while in the placebo group these markersworsened. The Silipide treatment was well tolerated, with feweradverse events than for the placebo group, and no patientdiscontinued the trial due to adverse effects.

A quercetin–phospholipid complex was also developed,and investigated in a murine model of liver injury (carbontetrachloride-induced damage) [23]. In this study, treatmentwith free quercetin (20 mg/kg) as well as quercetin–phospholipid complexes (10 mg/kg and 20 mg/kg) showedsignificant (pb0.05) decrease in thiobarbituric acid reactivesubstance (TBARS) levels in liver homogenate when com-pared to CCl4-treated animals. It was also observed that lowerdoses of uncomplexed quercetin (10 mg/kg) failed to producesignificant results. Quercetin at 20 mg/kg in phospholipidcomplex gave better results than free quercetin (20 mg/kg)restoring the normal enzyme levels (Fig. 6).

The phospholipid complexes of naringenin [66] andcurcumin [33] were also investigated. The phospholipidcomplex of naringenin produced better antioxidant activitythan the free compound with a prolonged duration of action,while the curcumin–phospholipid complex was active oncarbon tetrachloride-induced acute liver damage in rats, withsignificantly higher aqueous and n-octanol solubility andhigher antioxidant activity at all doses investigated.

5.5. Anticancer properties

Over the past few years, silymarin and silybin havereceived a growing attention for properties unrelated toliver protection, like anticancer, chemopreventive, hypocho-lesterolemic, cardioprotective, neuroactive and neuroprotec-tive activities [67]. The anti-radical potential and thecytoprotective activity may be responsible for the use ofsilymarin as a chemoprotective and anticancer agent. Due totheir chemopreventive activity, silybin and silymarin caninhibit the carcinogenic activity of many chemicals [68], andhave the potential to be successfully used in the adjuvanttherapy of cancer. In this context, silybin might act essentiallyas an antioxidant, protecting tissues against the oxidativestress generated by chemotherapeutics, preventing at thesame time hepatotoxicity, a major side-effect of manychemotherapy treatment. A phase I and pharmacokineticstudy of silybin-PHYTOSOME® in prostate cancer patientswas carried out [69]. The study concluded that oral silybin-PHYTOSOME® at a daily dose of 13 g seems well tolerated inpatients with advanced prostate cancer, and this dosage wasrecommended for the phase II study [69].

Silybin and its phospholipid complex IdB1016 have beenshown to have antitumor activity and to potentiate the effectof cisplatin [70], and repeated administration of IdB1016 tomice bearing human ovarian cancer xenografts significantlyinhibited tumor growth [71].

Turmeric is probably the most thoroughly researcheddietary supplement from the standpoint of potential value incancer management [72,73]. Compared bioavailabilities ofpure curcumin, turmeric active principle, and Meriva™(curcumin-PHYTOSOME®) has been evaluated in rats, show-ing that Meriva™ is five times more bioavailable than the

Fig. 6. Effect of quercetin–phospholipid complex on TBARS. Values are mean±SEM (n=6); *pb0.05 [significant with respect to control (CCl4-treatedgroup)], complex denotes quercetin–phospholipid complex.



parent compound [24]. Recent studies have confirm that theinteraction between curcumin and lipids layers plays afundamental role in the wide bioactivity of this compound[74], and Meriva™ thus appears to be a favorite candidate toenhance biological action of curcumin through a higherbioavailability.

5.6. Weight management

Results of a clinical trial investigating the role of Green-Select® PHYTOSOME® in the treatment of obesity wererecently published [75]. The study showed the efficacy ofgreen tea PHYTOSOME® to potentiate weight loss in dietingobese subjects.

6. Conclusions

A PHYTOSOME® is a complex between polar polypheno-lics and dietary phospholipids that shows definite physico-chemical and spectroscopic features. PHYTOSOME®

complexes were first investigated for cosmetic applications,but mounting evidence of potential for drug delivery has beencumulated over the past two decades, with beneficial activityin the realms of cardiovascular, anti-inflammatory, hepato-protective and anticancer applications. While the exactnature of the supramolecular interactions between specificphospholipids and polyphenolics still awaits clarification, theburgeoning literature on the biomedical applications of thePHYTOSOME® bodes well for the elucidation of the structuralbases of these remarkable interactions as well as for theirtranslation in biomedical research.

Acknowledgments

A. Semalty, M. Semalty and M.S.M. Rawat acknowledgethe grant provided by the Department of Science andTechnology, Govt. of India for the research work and arealso thankful to LIPOID GmbH Germany for providing the giftsample of phosphatidylcholine for the research work. Theyalso thankfully acknowledge UGC-DAE Consortium for Sci-entific Research, Indore (M.P.) for the provided facilities.

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