DECAFFEINATED MATE EXTRACTS AND THE USE THEREOF

Abstract: There are disclosed decaffeinated Maté extracts, the use thereof as antioxidant and radical scavengers, the pharmaceutical, cosmetic and dietary/alimentary compositions containing the extracts.

Description: MATE DECAFFEINATED EXTRACTS AND THE USE THEREOF The present invention relates to decaffeinated Mate extracts, characterized in caffeic acid derivatives and standardized in polyphenol content, to their use as antioxidantε and radical scavengers in the preparation of medicaments, dietary supplements, cosmetics, and to the pharmaceutical, cosmetic, alimentary/dietary compositions containing them. It is well known from epidemiological and clinical studies [1] that the intake of antioxidants/radical scavengers through the diet or their administration through pharmaceutical and nutritional products is of great benefit in wellnesε, health maintenance, and in the prevention of chronic and degenerative diseases associated to oxidative damage (atherosclerosis, cancer, coronary heart disease, rheumatoid arthritis, Alzheimer's and Parkinson's diseases and aging), since a free radical overload is massively involved in their development [2-7]. In the cosmetic field, compounds with antiradicalar activity able to enhance the natural defenses of the skin, hold the greatest promise for the development of photoprotective agents, since free radicals generated on epidermal skin layer by ultraviolet exposure, are recognized as potent inducers of premature skin aging and skin cancer, in particular of malignant melanoma [8]. Mate tea is one of the most consumed stimulating beverages in several South American countries, including Uruguay, Paraguay, Argentina and Brazil, prepared as a hot infusion of the caffeine-rich leaves of Ilex Paraguayenεis St. Hill (Aquifoliaceae ) and of other Ilex species. As occurs for other widely used stimulating beverages like coffee and tea, its use is linked to the presence of caffeine, responsible for the stimulating effects on central nervous system and the heart. While the use of Mate extracts in cosmetic or pharmaceutical products since the presence of xanthines is known [9,10], the use of decaffeinated extracts from Mate as natural antioxidants/radical scavengers is not documented. Although in two publications [11,12] has been reported the antioxidant action of water infusions of Mate leaves on the Cu^÷-induced peroxidation of isolated low density lipoproteins , and in other two patents Mate extracts have been proposed as antioxidant/radical scavengers [13,14], surprisingly it has been found that the decaffeinated extracts of the invention, enriched in well characterized polyphenols and standardized in polyphenols/chlorogenic acid content, possess a significantly greater radical scavenging activity against all free radicals involved in the onset (εuperoxide anion O^-, hydroxyl radical HO"; preventive antioxidants ) and the propagation (peroxyl radical LOO"; chain-breaking antioxidantε) of the peroxidation procesεes. As a first object, the invention provides decaffeinated extracts from plants of Ilex species. Preferably, the extracts of the invention are obtained from plants of Ilex Paraguayensis species, and they contain no more than 0,1% by weight of caffeine. Such extracts have been prepared according to the following procedure: Leaves of Il ex Paraguayenεiε dried and finely minced were mixed with boiling water (50 mg/ml) under mechanical stirring and cooled; the extracts were centrifuged at 10,000 x g for 10 min and the supernatant exhaustively extracted with chlorinated solvents, preferably ethylene chloride, to eliminate caffeine, theobro ine and other not phenolic substances. The aqueous residue were concentrated in vacuo to drynesε to obtain a pale-yellow powder (yield = 23%), the decaffeinated aqueous extract of Mate. The total or partial decaffeination process of Mate, in analogy to coffee or tea, could be done by several other techniques, as liquid-solid extraction, liquid-liquid extraction, chromatographic separation, supercritical or εubcritical extraction (CO2, etc.), and so on. Alternatively decaffeination could be carried out directly on the Mate leaves before the extraction of the final polyphenol-rich extract. The HPLC profile of the decaffeinated aqueous extract of Mate (Fig.l) obtained according to the above process, indicates the presence of at least 7 peaks with absorption maximum at 330 n , typical of chlorogenic, caffeoyl derivatives [15] and flavonoids: all the components of the polyphenol fraction were characterized by LC-MS analysis. In Fig. 4 is shown a typical mass chro atogram (in the bottom box the corresponding UV profile at 330 nm), in which 7 main peaks are detectable, and in Fig. 5-8 the negative ion mass spectra of the single components are shown. Peaks 1-3 show a molecular ion [M-H]- at m/z 353 (M.W. 354) and fragmentation patterns with ions at m/z 179 [Caffeic acid-H] _ and m/z 191 [quinic acid - H] ~ consistent with the structures of caffeoylquinic acid derivatives [16], peak 2 being chlorogenic acid (3-0- caffeoyl quinic acid), as determined by comparison with the authentic standard. Peaks 1 and 3 were identified as isomers of chlorogenic acid (i.e. isochlorogenic acid = 5-0-caffeoyl quinic or neochlorogenic acid, pεeudochlorogenic acid), but in the abεence of εtandards, not commercially available, no definitive structure asεignement can be done for peaks 1 and 3. Peaks 4-6 show molecular anion at m/z 515 (M.W. 516), and similar ragmentation pattern, with the diagnostic peak at m/z 353 due to losε of a caffeoyl reεidue (162 μ) [M - Caff - H] ~ and the ions at m/z 179 and 191, tipical of dicaffeoyl eεters of quinic acid [16]. On the basis of molecular weight and the fragmentation patterns we have detected and identified in this polyphenol fraction three iεomeric dicaffeoyl esterε (i.e. 3 , 4-O-dicaffeoyl , 4 , 5-O-dicaffeoyl , 3,5-0- dicaffeoyl or 1 , 5-dicaffeoyl esters of quinic acid). By contraεt, peak 7 shows a molecular anion [M-H] ~ at m/z 609 (M.W. 610) and a fragmentation pattern typical of glycosyl derivatives (losεes of terminal sugarε ) with ions at m/z 463 [M-H-Rha] ~ and m/z 301 [M-H-Rha-Glu] _ , this last corresponding to the deprotonated molecular ion of quercetin. On the basis of the fragmentation pattern and by comparison with the LC-MS profile of the reference standard, to peak 7 was asεigned the εtructure of rutin ( quercetin-3-rutinoεide ) . The total polyphenol content of the decaffeinated aqueouε extract of Mate obtained according to the deεcribed procedure waε 50 ± 0.8 % (w/w), determined by the Folin-Ciocalteau method [17]; the content in chlorogenic acid, determined by HPLC analyεis (Fig. la, peak 2, R.T. 21.86 min) was 5.1 ± 0.2% and assuming the same extinction coefficient for chlorogenic acid isomerε, the total content of chlorogenic acids waε 17.7 ± 0.5 % (8.2 % peak 1 Fig. la, R.T. 13.3; 4.4 % peak 3 Fig. la, R.T. 23.4); the content in rutin (peak 4, R.T. 33.68) waε 1.96 ± 0.3 %. In most caεes the caffeine content of the preparations was below 0.1%. The extracts of the invention have been examined for their antilipoperoxidant and radical scavenging activities in different experimental models like cell- free, membranous and cellular systems. The antilipoperoxidant (chain breaking) activity resulted much greater than that of a conventional Mate infusion, both in terms of IC 5Q value (8.1 vs 12.5 μg/ml ) , and of concentration which giveε total inhibition of the peroxidation process (20 vε 37.5 μg/ml) [11,12]. The overall antioxidant activity of the decaffeinated extracts, determined as quenching ability of the stable radical DPPH (see below), resulted to be even greater than that of a conventional Mate infusion (ICcg = 4 μg/ml vs 6.2 μg/ml). The radical scavenging activity of the extracts of the invention is due to the presence of low molecular weight caffeic acid derivatives, chlorogenic acid and other congeners, which have been fully characterized by means of HPLC and LC-MS . However, the teεtε have demonstrated that chlorogenic acid iε leεε active than the Mate extract as radical εcavenging agent, indicating a cooperative antioxidant interaction between the polyphenol componentε of the decaffeinated Mate extract (εee table 1). The decaffeinated extracts of Mate can be used alone or in combination with other natural extracts and/or different εubεtanceε , εuch aε hydrophilic/lipophilic antioxidantε, natural/synthetic flavors in foodε, beverage , dietary, coεmetic and pharmaceutical productε; in addition they can be formulated with εuitable vehicleε, excipientε or additives, according to the techniques conventionally used in the pharmaceutical, cosmetic and alimentary fieldε. The decaffeinated extracts standardized in their polyphenol contents can be both in liquid, liquid viscouε, or solid (powder, granules, etc.) physical forms. They can also be in a powder form obtained by a spray drying proceεε on suitable supporting agents like sugars, maltodextrines , gumε , etc. A further object of the present invention relates to pharmaceutical, cosmetic, alimentary/dietary compositions containing the extracts of the invention, which are, in pharmaceutical forms such as tablets, capsules, sachets, syrups, suppoεitorieε , vialε and oint entε, in coεmetic forms such aε emulεionε, gels, lotions or powders and dietary forms εuch aε foods, beverages and dietary products, are specifically suited as antioxidants and radical scavengers, in the prevention/treatment of free radical-mediated pathologies, such aε atherosclerosis, cancer, cardiovascular diεeases, rheumatoid arthritis, neurological disorders (Alzheimer's and Parkinson' diseaseε), aged-related cataract, infective diεeaεes and in the maintenance of the immune function, as photoprotectants to prevent the accelerated aging of the skin and the phootoxidative degeneration of the εkin and the hair, aε dietary εupplementε for the prevention of the phyεical aging, and for the maintenance of the physical performance under εtrenuouε exercise, for profesεional athletε. In particular, in the prevention/tretament of all the free-radical mediated phyεiopathological εituations, the product of the invention can be advantageously combined with other endogenous antioxidants, such as carotenoids, vitamin E, ascorbic acid, glutathione and sulphurated aminoacidε, in order to preserve and maintain the integrity of the physiological antioxidant pool through a maximization of the antioxidant effect due to synergiεtic interaction. Brief description of the figures Fig.l - HPLC profile of the decaffeinated aqueouε extract of Mate. Chromatographic conditionε: reverse phase column LC-18; mobile phase: linear gradient of 100% A (0.0127 N H 3 P0 4 /CH 3 CN/Ethylacetate 96.5/3/0.5) to 30% B (CHnCN) in 30 min; flow rate: 1 ml/min; detection: UV-DAD 330 nm. Seven peaks are detectable with absorbption maximum at 330 nm, typical of chlorogenic acid, caffeoyl derivatives [15] and flavonoids. Fig.2a - HPLC profile (detection 270 nm) of the methylene chloride extract, containing caffeine (R.T. 18.63 min) and theobro ine (R.T. 8.28 min). Fig.2b - HPLC profile (detection 270 nm) of the decaffeinated aqueous extract of Mate (see the absence of caffeine at 18.63 min and of theobromine at 8.28 min ) . Fig.3 - HPLC chromatograms (detection 330 nm) of the decaffeinated aqueouε extract of Mate before (a) and after reaction with DPPH (b = 0.0625 mM; c = 0.125 nM; d = 0.25 M; e = 0.5 M ) . Fig.4 - LC-MS profile of the decaffeinated aqueouε extract of Mate (bottom box UV tracing). Experimental conditions: Finnigan LCQ mass spectrometer; reverεe phaεe column LC-18; mobile phaεe : linear gradient of 100% A (95% ammonium formiate / 5% CH3CN, pH 4.5 with HCOOH) to 30% B (95% CH3CN / 5% ammonium formiate, pH 4.5 with HCOOH) in 30 min; flow rate: 1 ml/min; negative ion detection. Peak 1 = [M-H]- m/z 353 chlorogenic acid iεomer; peak 2 = [M-H]- m/z 353 chlorogenic acid; peak 3 = [M- H]- m/z 353 chlorogenic acid iεomer; peakε 4, 5, 6 = [M- H]- m/z 515 dicaffeoyl eεterε of quinic acid; peak 7 = [M-H]- m/z 609 rutin. Fig.5 - Negative ion ElectroSpray (ES) mass spectrum of peak 1. Fig.6 - Negative ion ElectroSpray (ES) masε spectra of peaks 2 and 3. Fig.7 - Negative ion ES mass spectra of peakε 4 and 5. Fig.8 - Negative ion ES εpectra of peaks 6 and 7 Fig.9 - Time-course of conjugated dienes in the propagation and breakdown phaεeε of lipid peroxidation in phosphatidylcholine liposomeε: chain-breaking antioxidant activity of decaffeinated aqueouε extract of Mate. Valueε are the meanε ± S.D. of 5 determinations. Fig.10 - Protective effect of decaffeinated aqueous extract of Mate on UVB-induced hemolyεiε. Red blood cellε (0.2% εuεpenεionε) were exposed to UVB radiation for 210 min in the absence and in presence of different concentrations of decaffeinated aqueous extract of Mate; percentage hemolysiε waε turbidimetrically determined at 710 nm at 30 min intervalε. Valueε are the meanε ± S.D. of 5 determinationε . The following exampleε illustrate the invention in more details. Example 1 Antioxidant and radical scavenging activity aεsays of decaffeinated Mate extracts a ) 2, 2-Dyphenyl -l -pi crylhydrazyl (DPPH) radical The activity of the extract in quenching the stable free radical DPPH through hydrogen transferring reaction, has been tested by monitoring the loεε in abεorbance at 517 nm of a methanol solution of DPPH [18] in the presence and the absence of increasing concentrations of Mate extract. The resultε, εummarized in Table I, indicate that Mate extract ( IC 5Q = 4.2 μg/ml), although equipotent to chlorogenic acid ( ICC Q = 4.34), is significantly more effective than ascorbic acid or vitamin E (ΪC50 = 19 μg/ml) in quenching DPPH radical. All the components detected by HPLC analysiε (Fig.l), contribute to the DPPH quenching ability of the extract, εince after reaction with increaεing concentrationε of DPPH (0.625, 0.125, 0.25, 0.5 mM), a progreεsive disappearance of peaks 1-7 is observed (Fig.3). b) Superoxide anion The decaffeinated aqueouε extract of Mate dose- dependently inhibit the rate of reduction of nitrotetrazolium blue chloride (NBT) by superoxide anion generated in the phenazine methosulfate/NADH syεtem [18], with a minimal effective concentration of 5 μg/ml and an ICC Q value of 15 μg/ml; chlorogenic acid waε leεs potent in entrapping εuperoxide radicals, with an ICc 0 value of 24 μg/ml (Table I). The bimolecular rate constant for the reaction of εuperoxide radicals with Mate extract, determined by kinetic competition studies [18] gives a K value of 1.2 x 10 5 M -1 cm -1 , more favourable than that of chlorogenic acid ( K = 0.5 x 10^ M -1 cm -1 ) . c) Hydroxyl radical Using the deoxyriboεe assay [19], it has been demonεtrated that the decaffeinated aqueouε Mate extract inhibit the HO°-induced deoxyribose degradation, as indicated by the progreεsive decrease in thiobarbituric acid-malondialdehyde (TBA-MDA) chromogen formation in the presence of increasing concentration of the extract. The rate constant of reaction of Mate extract with HO° ( K) , calculated according to Aruo a [19] using a K value for deoxyribose of 3.1 x 10 9 M -1 ε-^ , waε 4.5 ± 0.3 x 10_ M -l s -l. Chlorogenic acid waε leεε effective than Mate extract, with a l value of 2.1 ± 0.2 x 10 9 M -1 s -1 (Table I ) . d) Antilipoperoxidant activi ty in phoεphatidylcholine liposomes The chain-breaking antioxidant activity of the decaffeinated aqueouε extract of Mate waε determined in a well eεtabliεhed model of HO°-induced peroxidation (by ultraεoundε expoεure ) of phoεphatidylcholine liposomes, by monitoring the formation of conjugated hydroperoxideε during the propagatory phaεe of the peroxidation process [20-22]. Mate extract markedly and doεe-dependently inhibited the increaεe in conjugated dieneε (6 hourε after induction), by 17% at 1 μg/ml, 36% at 5 μg/ml, 66% at 10 μg/ml and complete inhibition waε obεerved at 20 μg/ml (IC jj g = 8 -l μg/ml); chlorogenic acid waε lesε effective aε chain-breacking antioxidant, with an ICC Q value of 18.4 μg/ml (Table I). In addition, as εhown in Fig. 9, in the εampleε protected with Mate extract, the plateau of conjugated dienes was reached later than the control (t = 6h), with a lag time of 28 hours at 10 μg/ml (t = 34 h); at the highest concentration the conjugated dienes were far from the plateau even after 50 h. e ) UVB-induced lyεis of rat erythrocyteε (RBC) Finally we evaluated in a cellular model (0.2% erythrocyte suεpensionε) the protective effect of the decaffeinated aqueouε extract of Mate on the peroxidative εtreεs induced by UVB exposure (0.4-4 J/cm^) [23], and monitoring the time-course of hemolysiε ( turbidimetric determination at 710 nm) at 30 min intervalε. The protective effect of Mate extract (1-20 μg/ml) on UVB-induced he olyεis was compared to that of chlorogenic acid (1-20 μg/ml). Aε shown in Fig. 10, hemolysiε in the controls starts after 60 min irradiation and plateaued between 90 and 120 min (71.2 ± 4.8 % and 95.5 ± 5.2% hemolysiε). The decaffeinated aqueouε extract of Mate significantly and dose-dependently delay the onset of the UVB-induced hemolysis, by 30 min at 10 μg/ml and by 60 min at 20 μg/ml; the protective effect starts from low concentrations, since at 1 μg/ml % hemolysis at 90 min irradiation was only 56.6 ± 2.8 %, 33.0 ± 3.2 % at 2.5 μg/ml and 18.2 ± 2.6 % at 5 μg/ml (IC 5Q determined at 90 min = 2.35 μg/ml). Chlorogenic acid at the same concentrations (data not shown) was slightly leεε effective than decaf einated aqueouε extract of Mate, with an ICC Q value (90 min irradiation) of 3.2 μg/ml. Table I - Radical scavenging activity of decaffeinated aqueous extract of Mate EXPERIMENTAL Ma έ extract Chlorogenic acid MODEL DDPH radical IC-50 (μg/ml) = 4.2 ICββ (μg ml) = 4.3 Superoxide anion ICso (μg/ml) = 15.0 ICso (μg/ml) = 24.2 (NBT reduction) = 1.2 x 10 5 -ι _ -1 K = 0.5 x 10° M _1 s _ -1 Hydroxyl radical (deoxyribose assay) K = 4.5 x 1Q 9 Wr 1 s 1 K = 2. x 0 9 M- 1 s- 1 Peroxyl radical ICβo (μg ml) = 8.1 ICβo (μg/ml) = 18.4 (chain-breaking antioxidant) Example 2 1000 ml of diet beverage contain: Acesulfame K 0 .09 g Aspartame 0. .09 g - Cyclamate 0. .20 g Decaffeinated aqueous extract of Mate 1. .00 g White grape juice 4 fold 5. .00 g Citric acid 1, .80 g Apple Flavor 1, .00 g - Citron flavor 0. .20 g Water q. . ε REFERENCES 1. Frei B. Natural Antioxidantε in Human Heal th and Diεeaεe . Academic Preεε, Inc., San Diego (CA) 1994. 2. Croεε C.E. Oxygen radicals and human diseaεe. Ann . In tern . Med . , 107, 526-45 (1987). 3. Ameε B.N., Shigenaga M.K., Hagen T.M. Oxidants, antioxidantε and the degenerative diseaεeε of aging. Proc. Natl . Acad . Sci . , 90, 7915-22 (1993). 4. Halliwell B., Gutteridge J.M.C. ed . Free Radi calε in Biology and Medi cine, 2nd edition, Clarendon Preεε, Oxford (1989). 5. Sieε H. ed. Oxidati ve εtreεε : Oxidan tε and Antioxi dantε . Academic Preεε, London (1991). 6. Packer L. Oxidative stress and the antioxidant vitamins C and E in functional medicine. In " Oxidati ve Proceεεeε and An tioxidantε " (Paoletti R. , Samuelsson B., Catapano A., Poli A., Rinetti M. eds . ) Raven Preεs, New York, 1994, pp. 153-64. 7. Cadenaε E. and Packer L. edε. Handbook of antioxidant , Marcel Dekker Inc. (1996). 8. Fuchs J. and Packer L. Photooxidative streεε in the εkin. In " Oxida tive εtreεε Oxidantε and An ti oxidantε . Academic Preεε, London (1991), pp 559-83. 9. Honerlagen H.J. and Bolardt S.M. Maεse, welche mεbeεondere fur die Einkapεelung in ein festes Umhullungεmaterial geeignet iεt, und Verfahren zur Herεtellung dieser Maεεe. EP 0 496 705 Al (1992). 10. Voβ H. , Deppe M., Deppe R. Koε etiεcheε Mittel mit anticellulitischer Wirkung. DE 44 01 308 Al (1995). 11. Gugliucci A. and Stahl A.J.C. Low density lipoprotein oxidation is inhibited by extracts of Il ex Paraguari enεiε , Bi ochem . Mol ec . Bi ol . In t . , 35, 47-56 (1995). 12. Gugliucci A. Antioxidant effects of Ilex paraguayensis : induction of decreased oxidability of human LDL in vivo. Bi ochem . Bi ophyε . Reε . Commun . , 224, 338-44 (1996). 13. Kuroεe S., Matεu ura T. Extracting flavonoidε and catechinε from Mate tea leaveε for use as antioxidants and active agents in cosmeticε. Braz.Pedido, PI BR 93 03,217. 14. Hibino M., Nakamura I., Sakanaka S. Extraction of oxygen radical εcavengerε from Ilex paraguayensis. Jpn. Kokai Tokkyo Koho JP 06,135,848 [94,135,848]. 15. Clifford M.N. and Ramirez-Martinez J.R. Chlorogenic acids and purine alkaloids contents of Mate (Ilex Paraguayensiε ) leaf and beverage. Food Chem . , 35, 13-21 (1990). 16. Maffei Facino R., Carini M., Aldini G., Marinello C. Direct characterization of caffeoyl esters with antihyaluronidaεe activity in crude extractε from Echinacea anguεtifolia rootε by faεt Atom Bombardment Mass Spectromatry . II Farmaco, 48, 1447-61 (1993). 17. Singleton S.L., Ross J.A. Colorimetry of total phenolics with phosphomolybdic-phoεphotungstic acid reagents. Am . J. Enol . Vi ti c , 16, 144-58 (1965). 18. Maffei Facino R., Carini M., Saibene L. Scavenging of free radicals by tenoxicam: a participating mechanism in the antirheumatic/antiinflammatory efficacy of the drug. Arch . Pharm . Pharm . Med . Chem . , 329 , 457 - 63 ( 1996 ) . 19. Aruoma 1.0. Deoxyribose asεay for detecting hydroxyl radicalε. Meth . Enzy ol . , 233, 57-66 (1994) . 20. Maffei Facino R., Carini M., Aldini G., Bombardelli E., Morazzoni P., Morelli R. Free radical scavenging and anti-enzyme activities of procyanidines from Vi tiε vinifera . A mechanism for their capillary protective action. Arznei . Forεch . /Drug Reε . , 44, 592-601 (1994). 21. Maffei Facino R., Carini M., Aldini G., Saibene L., Morelli R. Differential inhibition of superoxide, hydroxyl and peroxyl radicals by nimeεulide and its main metabolite 4-hydroxynimesulide . Arzneim. Forεch . /Drug Reε . , 45, 1102-9 (1995). 22. Maffei Facino R., Carini M., Aldini G., Berti F., Rosεoni G., Bombardelli E., Morazzoni P. Procyanidines from Vi tiε vinifera seedε protect rabbit hearth from iεche ia/reperfuεion injury: antioxidant intervention and/or iron/copper εequeεtering ability. Planta Med. , 62, 495-502 (1996) . 23. Maffei Facino R., Carini M., Aldini G. , Calloni M.T., Bombardelli E., Morazzoni, P. Sparing effect of procyanidinε from Vitiε vinifera L. on vitamin E: in vitro εtudies . Planta Med. , 64 (1998 In preε ) .

Claims: CLAIMS 1. Decaffeinated extractε from plants of Ilex specieε. 2. Extractε according to claim 1, containing no more than 0.1% by weight of caffeine. 3. Extractε according to claimε 1-2, obtained from plantε of the Ilex Paraguayenεiε specieε. 4. Decaffeinated extracts according to claimε 1-3 with a chromatographic profile basically comparable to that reported in Fig.l 5. Decaffeinated extracts according to claims 1-3 with a LC-MS profile baεically comparable to that reported in Fig. 4. 6. Process for the preparation of polyphenol-rich decaffeinated extracts which comprises: treatment of dried and finely minced leaves of Ilex Paraguayenεiε with boiling water, preceded or followed by extraction of caffeine, and, finally, drying of the aqueouε reεidue. 7. Proceεε according to claim 6, wherein the extraction of caffeine iε carried out with chlorinated εolventε . 8. Proceεε according to claim 7, wherein εaid εolvent iε methylene chloride. 9. Procesε according to claim 6, wherein the extraction of caffeine iε carried out with liquid-εolid extraction, liquid-liquid extraction, chromatographic separation, εupercritical or εubcritical extraction. 10. Polyphenol-rich decaffeinated extracts obtainable by the process of claims 6-9. 11. Use of the extracts of the claimε 1-5 and 10 aε antioxidant and radical εcavenging agents. 12. Use of the extracts according to claim 11 in liquid, liquid-viscous or solid forms. 13. Use of the extracts of claims 1-5 and 10, for the preparation of medicaments for the prevention and the treatment of atheroεclerosiε , cancer, cardiovascular diseaεes, rheumatoid arthritis, Alzheimer'ε disease, Parkinson's diεeaεe and other age-related chronic degenerative pathologieε. 14. Uεe of the extractε of claims 1-5 and 10 for the preparation of coεmeticε for the prevention of premature εkin aging and the photooxidative degeneration of skin and of the hair. 15. Uεe of the extractε of claimε 1-5 and 10 for the preparation of dieraty εupplementε for the prevention of phyεical aging, and for the maintenance of the physical performance under strenuous exerciεe, for profeεsional athletε. 16. Pharmaceutical, coεmetic or dietary/alimentary compositions containing the extracts of claims 1-5 and 10 alone or in asεociation with other natural extractε, hydrophilic and lipophilic antioxidantε, ingredientε or flavorε, formulated with εuitable vehicleε, excipientε or additives. 17. Compositions according to claim 16, in form of tablets, capsuleε , εachetε, εyrupε, εuppositories , vialε and ointments, emulsionε, gelε, lotionε or powderε and dietary εupplementε, foods, beverages.

Inventors: CARINI MARINA, MAFFEI FACINO ROBERTO, MARIANI MARCO, CARINI, MARINA, MAFFEI FACINO, ROBERTO, MARIANI, MARCO
Applicants: CARINI MARINA, MAFFEI FACINO ROBERTO, MARIANI MARCO, UNIVERSAL FLAVORS S R L, CARINI, MARINA, MAFFEI FACINO, ROBERTO, MARIANI, MARCO, UNIVERSAL FLAVORS S.R.L.
Application year: 1998
Main classification: A23L1
IPC codes: A23F3, A23L1, A23L3, A61K8, A61K36, A61P9, A61P25, A61P29, A61P35, A61Q17, A61Q19, C09K15