Cosmetic Use of Petroselinic Acid Kosmetische Verwendung Von Petroselinsäure Utilisation Cosmétique De L’Acide Pétrosélinique

(19) &

(11) EP 1 013 178 B2

(12) NEW EUROPEAN PATENT SPECIFICATION After opposition procedure

(45) Date of publication and mention (51) Int Cl.: of the opposition decision: A23L 1/30 (2006.01) 16.09.2009 Bulletin 2009/38

(45) Mention of the grant of the patent: 28.09.2005 Bulletin 2005/39

(21) Application number: 99204230.9

(22) Date of filing: 09.12.1999

(54) Cosmetic use of petroselinic acid Kosmetische Verwendung von Petroselinsäure Utilisation cosmétique de l’acide pétrosélinique

(84) Designated Contracting States: • Powell, Jonathan, Richard, AT BE CH CY DE DK ES FI FR GB GR IE IT LI LU Unilever Res. Colworth MC NL PT SE Sharnbrook Bedford, MK44 1LQ (GB) (30) Priority: 22.12.1998 EP 98310627 • Rawlings, Anthony, Vincent, 22.12.1998 EP 98310626 Unilever Res. Colworth 17.03.1999 EP 99302067 Sharnbrook Bedford, MK44 1LQ (GB) (43) Date of publication of application: • Rogers, Julia, Sarah, 28.06.2000 Bulletin 2000/26 Unilever Res. Colworth Sharnbrook Bedford, (73) Proprietors: MK44 1LQ (GB) • Unilever N.V. • Watkinson, Allan, 3013 AL Rotterdam (NL) Unilever Res. Colworth Designated Contracting States: Sharnbrook Bedford, AT BE CH DE DK ES FI FR GR IT LI LU MC NL PT SE MK44 1LQ (GB) • Unilever PLC • Cain, Frederick, William, London Loders Croklaan B.V. Greater London EC4P 4BQ (GB) 1521 AZ Wormerveer (NL) Designated Contracting States: CY GB IE (74) Representative: Hugot, Alain et al Unilever Patent Group (72) Inventors: Colworth House • Alaluf, Simon, Sharnbrook Unilever Res. Colworth Bedford, MK44 1LQ (GB) Sharnbrook Bedford, MK44 1LQ (GB) (56) References cited: • Hu, Heng-Long, EP- - 0 709 084 EP- - 3 155 842 Unilever Res. Colworth EP-A- 0 777 971 EP-A- 0 888 773 Sharnbrook Bedford, WO- -00//15179 WO- -99//47110 MK44 1LQ (GB) WO-A-95/00136 GB- - 980 556 4.3 • Green, Martin, Richard, US- - 6 365 175 US- - 09/ 150 841 Unilever Res. Colworth US-A- 4 918 063 US-A- 5 605 929 Sharnbrook Bedford, MK44 1LQ (GB) EP 1 013 178 B2

Printed by Jouve, 75001 PARIS (FR) (Cont. next page) EP 1 013 178 B2

• YAGALOFF K A ET AL: "ESSENTIAL FATTY • ELLENBRACHT F. ET AL: ’UNUSUAL FATTY ACIDS ARE ANTAGONISTS OF THE ACIDS IN THE LIPIDS FROM ORGANS AND CELL LEUKOTRIENE B4" PROSTAGLANDINS CULTURES OF PETROSELINUM CRISPUM’ LEUKOTRIENES AND ESSENTIAL FATTY ACIDS, PLANTA vol. 150, 1980, pages 114 - 119 vol. 52, no. 5, 1 January 1995 (1995-01-01), pages • T.P.KRISHNAKANTHA: ’Scavenging of 293-297, XP002050601 superoxide anoins by spice principles’ INDIAN • AFIFI N A ET AL: "SOME PHARMACOLOGICAL JOURNAL OF BIOCHEMISTRY AND BIOPHYSICS ACTIVITIES OF ESSENTIAL OILS OF CERTAIN vol. 30, 30 April 1993, page 133-134 UMBELLIFEROUS FRUITS" VETERINARY • J.LEDERER: ’Selenium, Vitamine et MEDICAL JOURNAL GIZA, vol. 42, no. 3, 1 vieilissement’ LOUVAIN MED. 1991, page 285-300 January 1994 (1994-01-01), pages 85-92, • BERNSTEIN ET AL: ’Differential Expression of XP002050599 the Versican and Decorin....’ LABORATORY • PATENT ABSTRACTS OF JAPAN vol. 011, no. 115 INVESTIGATION vol. 72, no. 6, 1995, page 662-669 (C-415), 10 April 1987 (1987-04-10) & JP 61 257944 • ’USA NEWS’ COSMETIC RESEARCH April 1994, A (NISSHIN FLOUR MILLING CO LTD), 15 • BIRGIT REITER ET AL RESEARCH PAPERS / THE November 1986 (1986-11-15) FATTY ACIDS PROFILES.... no. 11, 1998, page • PATENT ABSTRACTS OF JAPAN vol. 011, no. 003 498-502 (C-395), 7 January 1987 (1987-01-07) & JP 61 • DAMOUR, MARTINI, ROUSSELLE: ’Le 181352 A (HORIUCHI:KK), 14 August 1986 vieillissement cutané’ EDITION MEDIAFLASH (1986-08-14) 1998, • DATABASE WPI Section Ch, Week 199248 Derwent Publications Ltd., London, GB; Class Remarks: D13, AN 1992-396691 XP002120504 & SU 1 706 The file contains technical information submitted after 514 A (BAKINSK SECT MEAT IND RES INST), 23 the application was filed and not included in this January 1992 (1992-01-23) specification • DATABASE WPI Section Ch, Week 199823 Derwent Publications Ltd., London, GB; Class D13, AN 1998-259525 XP002120503 & RU 2 091 033 C (AZNAURYAN M P), 27 September 1997 (1997-09-27)

2 EP 1 013 178 B2

Description

[0001] Petroselinic acid is a well known compound that eg is present in coriander oil in relatively high amounts. Its use in food products is also disclosed in literature. EP 777971 eg discloses that fat compositions can be obtained from 5 fats that have high contents of asymmetric monounsaturated fatty acids, such as petroselinic acid, ie PSA (> 79 wt%) and that have simultaneously low contents of trans acids (< 5 wt%). The fat compositions obtained can be used as fat replacer with a number of benefits such as providing good textural properties to the food, while not raising the LDL- cholesterol levels in the blood serum. When considering example 5, in particular table 3 of this patent application one must conclude that good product performance is only obtained if the fat applied contains about 12 wt% PSA. Products 10 based on fats without PSA perform badly (=control fat 1). Fats with only small amounts PSA are still grainy, while using a fat with a high content of PSA (=fat 4 with about 64 wt% PSA via the coriander oil) has a poor overall appearance. Food products that are indicated are margarines, dressings, confections, spreads, frozen desserts, ice cream, mayon- naise, mustards, cheese, dip sauces, bread, biscuit, dairy products, frying oils, CBE’s, candy, meat, egg products, nut products, vegetable or fruit products, toppings, creams, puddings, cookies, pastries, pies, crackers, cakes, bread rolls 15 and ingredients or premixes herefore. As PSA is the active ingredient that provides the health benefits to the food product there is a big need to be able to use fats with high levels of PSA but that perform well when applied in food products. Ie the appearance of the food products should be good as well. [0002] It is also known that certain fatty acids can be used as anti-inflammatory agent, from e.g. Afifi et al "Some pharmacological activities of essential oils of certain umbelliferous fruits" Vet Med Journal Giza vol 42, No, 3 1 Jan 1994, 20 and also Yagaloff et al "Essential fatty acids are antagonists of the Leukotriene B4 Receptor" Prostaglandins Leukotriened and Essential Fatty acids, vol 52, no. 5, 1 Jan 1995. These publications mention petroselinic acid. [0003] A well recognized consumer problem is formed by the aging of the skin. In recent years the demand for methods to improve the appearance of skin and, in particular, for reducing or preventing the visible signs of wrinkled, aged and/or photodamaged skin has grown enormously. It is known in the art that the levels of collagen, a dermal structural protein 25 and decorin, a structuring proteoglycan in skin are significantly reduced with aged and/or photodamaged skin (Lavker et al J.Inv.Derm.1979 73:79-66, Bernstein et al Lab. Invest. 1995 72:662-669). The reduction in these proteins is asso- ciated with a decrease in the tensile strength of the skin causing wrinkles and laxity. [0004] Extensive research has focused on identifying actives that have anti-aging activity. One of the main current actives is retinoic acid, giving wrinkle effacement and dermal repair through boosting for example by collagen synthesis 30 (e.g. Griffiths et al. N. Eng. J. med. 1993 329:530-535). According to GB 2 181 349 another solution was found in the use of triglycerides derived from long chain polyunsaturated fatty acids in cosmetic or dermatological compositions. These compositions are thus not edible in general because they also will contain non edible components. Moreover the triglycerides derived from these polyunsaturated fatty acids are unstable in particular with respect to oxygen stability [0005] We studied whether we could find a natural component that could be used as anti-aging agent without creating 35 problems of side effects and not having the negative aspects from the known anti-aging compositions. [0006] The study on antiaging activity resulted in our finding that petroselinic acid derivatives (ie free fatty acid, short alkyl ester, short being C1-C4, wax-esters, mono-di- or triglycerides) can be applied as an anti-aging agent that act by boosting the levels of two structural proteins in the dermis of the skin, collagen and decorin. Therefore our invention concerns the use of edible compositions containing petroselinic acid for the preparation of 40 functional food compositions or food supplements, wherein the composition containing petroselinic acid is used as an anti-aging component that boosts decorin levels with a positive impact on skin conditions selected from the group consisting of wrinkling and sagging. [0007] Functional food compositions being defined as food compositions containing at least one component with a health benefit. Food supplements being defined as compositions that are not used as food per se, but that are used as 45 supplement to the daily food intake, in general in the form of encapsulated essential food ingredients. These edible compositions containing petroselinic acid can be applied in many different forms, but we prefer to apply these compositions as a composition comprising 5-99.9 wt% of fat or fat blend. The fats that can be used in the compositions containing the petroselinic acid derivative can be selected from the group consisting of: cocoa butter equivalents, palm oil or fractions thereof, palm kernel oil or fractions thereof, interesterified 50 mixtures of above fats, hardened fats or fractions thereof, liquid oils, selected from sunflower oil, high oleic sunflower oil, soybean oil, rape seed oil, cotton seed oil, safflower oil, high oleic safflower oil, maize oil, MCT oils, hardened liquid oils or fractions thereof and mixtures hereof. These fats are nearly all natural fats (the exception being MCT oils which are synthetic fats based on medium chain fatty acids ie fatty acids with 6-12 carbon atoms). 55 The level of petroselinic acid in the compositions that we can apply for our purposes is 2-80 wt%, preferably 5-40 wt% of petroselinic acid derivative (calculated as petroselinic acid). [0008] As already indicated above we found that the petroselinic acid could be used in different forms. Very good results were obtained by using the petroselinic acid as free fatty acid, or in a form wherein it is bound to a glycerol

3 EP 1 013 178 B2

backbone as in mono-di- or triglycerides. In this latter case the glycerides must contain at least one petroselinic acid moiety. Other forms are short alkyl esters of the petroselinic acid, short meaning having one to eight, preferably 1 to 4 carbon atoms. Also wax esters, ie long chain alcohol esters of petroselinic acid can be applied. Of course also mixtures of above forms can be used. 5 [0009] We also found that the antiaging effect of the petroselinic acid can be increased, even in a synergistic way by using the petroselinic acid derivative in combination with one or more anti-oxidants. Typical examples of useful antioxidants can be selected from the group consisting of natural or synthetic tocopherols, natural polyphenols, in particular as present in tea extracts, BHT, BHA, free radical scavengers and enzymes with anti-oxidant properties. From these anti-oxidants we prefer the tocopherols and the polyphenols as present in tea extracts the most. 10 EXAMPLES

Methodology

15 Procedure For Measuring Procollagen-I and Decorin Synthesis In Human Dermal Fibroblasts

Preparation of Dermal Fibroblast Conditioned Medium

[0010] Primary human foreskin fibroblasts at passage 2 (P2) were seeded into 12-well plates at 10000 cells/cm2 and 20 maintained for 24 hours in an atmosphere of 5% carbon dioxide and 4% oxygen in Dulbeccos Modified Eagles Medium (DMEM) supplemented with 10% foetal calf serum. After this time the cells were washed with serum free DMEM and then incubated in fresh serum free DMEM for a further 60 hours. The fibroblast monolayers were then washed again with serum free DMEM. Test reagents (petroselinic acid, EGCG and gallic acid) and vehicle controls were added to the cells in triplicate in a final volume of 0.4ml/well fresh serum free DMEM and incubated for a further 24 hours. This 25 fibroblast conditioned medium was either analysed immediatedly or snap frozen in liquid nitrogen and stored at -70°C for future analysis. The cells were then counted and data from the dot-blot analysis subsequently standardised to cell number.

Dot Blot Assay for Decorin Protein in Dermal Fibroblast Conditioned Medium 30 [0011] Samples of conditioned medium from dermal fibroblasts treated with vehicle (as a control) or test reagents were supplemented with 20mM dithiothreitol (1:10 dilution of 200mM stock solution) and 0.1% sodium dodecylsulphate (1:100 dilution of 10% stock solution), mixed well and then incubated at 75°C for 2 minutes. A standard for the assay was generated by serial dilution of neat fibroblast conditioned medium from fibroblasts seeded at 10000 cells/cm2 in a 35 175cm2 flask and maintained in serum free DMEM as described above. Assay samples were subsequently applied in triplicate to a prewetted sheet of Immobilon-P transfer membrane using the 96-well Bio-Dot Apparatus from Bio-Rad as described in the manufacturers guidelines. Approximately 200Pl of medium was applied per well. The medium was allowed to filter through the membrane under gravity (30 minutes) after which the membrane was washed twice with PBS (200Pl). These PBS washes were allowed to filter through the membrane under gravity (2x15 minutes). The Bio- 40 Dot apparatus was then attached to a vacuum manifold and a third and final PBS wash carried out under suction. The apparatus was disassembled, the membrane removed and quickly cut as required before being placed in blocking buffer overnight at 4°C. Membranes prepared for decorin analysis were blocked with 3% (w/v) bovine serum albumin (BSA)/ 0.1% (v/v) Tween 20 in phosphate buffered saline (PBS), whilst those for procollagen-I analysis were blocked with 5% (w/v) non fat dried milk powder/ 0.05% Tween 20 in PBS. The following day, the membranes were probed with 1:10000 45 dilution of primary antibodies to human decorin (rabbit polyclonal; Biogenesis) for 2 hours at room temperature. The membranes were subsequently washed with TBS/ 0.05% Tween 20 (3 x 5 minutes) and then incubated with 1:1000 dilution of 125I-conjugated anti-rat or anti-rabbit F(ab’)2 fragments (Amersham) as required for 1 hour at room temperature. Following this the Immobilon strips were again washed with TBS/Tween 20 (3 x 5 minutes) before being allowed to dry in air at room temperature. The dried membranes were wrapped in cellophane and exposed to a Molecular Dynamics 50 storage phosphor screen for 16-18 hours. At the end of this time the exposed screen was scanned by a phosphorimager (Molecular Dynamics Phosphorimager SF) using ImageQuant™ software. Dot intensity was assessed by computer- assisted image analysis using the quantification tools in ImageQuant™, standardised to cell number and the effects of various test reagents on decorin and procollagen-I synthesis were determined relative to a vehicle treated control value of 100 arbitrary units. 55

4 EP 1 013 178 B2

Examples:

Anti-ageing effects - determined by measurement of decorin levels and synergy with antioxidants

5 [0012] Petroselinic acid has been demonstrated to boost decorin levels in fibroblasts, consistent with dermal repair and antiageing activity (Table 1). Antioxidants such as ECGC, gallic acid, diadzein and genistein can also exert this effect (Table 2). In combination, a synergistic boost in decorin levels is observed (Table 3).

Table 1. 10 Boosting of decorin by petroselinic acid. Petroselinic acid (PM) Decorin (% of control) 0100.0 13.6 1137.6 13.3 15 10 152.5 14.2

[0013] The production of decorin by fibroblasts was determined in control cells (no treatment) and petroselinic acid treated cells. The data is represented mean +/- standard deviation as decorin levels as a percentage of the control (no 20 petroselinic acid). Each data point was determined in triplicate. All petroselinic acid values were significantly increased compared to the control.

Table 2. Boosting of decorin by antioxidants 25 Antioxidants Decorin (% of control) EGCG (Pg / ml) 0 100.0 6.4

30 1167.5 11.8 2.5 188.6 9.4 5 258.2 6.1 Gallic acid (Pg / ml)

35 0 100.0 7.0 1 136.9 4.2 2.5 163.4 6.5 5 161.8 6.6

40 [0014] The production of decorin by fibroblasts was determined in control cells (no treatment) and antioxidant treated cells. The data is represented as mean +/- standard deviation for decorin levels as a percentage of the control (no antioxidant). All antioxidant values were significantly increased compared to the control. Each data point was determined in triplicate. 45 Table 3. Synergistic boosting of decorin by petroselinic acid and antioxidants Treatments Decorin (% of control)

50 Control 100.0 1.2 Petroselinic acid (0.01M) 93.7 10.3 EGCG (0.005Pg/ml) 118.4 3.1 Petroselinic acid (0.01M) + EGCG 0.005Pg/ml 140.5 16.7 Control 100.0 3.5 55 Petroselinic acid (0.01M) 91.6 5.1 EGCG (0.05Pg/ml) 111.7 9.9 Petroselinic acid (0.01M) + EGCG 0.05Pg/ml 131.2 9.2

5 EP 1 013 178 B2

(continued) Synergistic boosting of decorin by petroselinic acid and antioxidants Treatments Decorin (% of control) 5 Control 100.0 7.7 Petroselinic acid (0.01M) 102.7 4.5 EGCG (0.5Pg/ml) 120.2 3.7 Petroselinic acid (0.01M) + EGCG 0.5Pg/ml 155.9 8.9

10 Control 100.0 3.3 Petroselinic acid (0.01M) 100.9 8.2 Gallic acid (0.001Pg/ml) 96.6 7.4 Petroselinic acid (0.01M) + Gallic acid 0.001Pg/ml 129.2 13.9 Genistein (1Pm) 101.7 2.3 15 Petroselinic Acid (0.01 PM) 117.3 4.5 Genistein + Petroselinic acid 130.2 9.0 0.1 PM Daidzein 115.4 5.5 0.01 PM Petroselinic acid 104.3 3.0 Daidzein + Petroselinic acid 130.8 4.4 20

[0015] The production of decorin by fibroblasts was determined in control cells (no treatment) and petroselinic acid and/or antioxidant treated cells. The data is represented as mean +/- standard deviation (n=3) for decorin levels as a percentage of the control (no antioxidant). A synergistic effect in boosting decorin levels of both antioxidants and pet- 25 roselinic acid was observed, compared to the control.

Claims

30 1. Cosmetic use of an edible composition containing 2-80 wt% petroselinic acid as an anti-aging component with a positive impact on skin conditions selected from the group consisting of wrinkling and sagging.

2. Use of an edible composition containing petroselinic acid according to claim 1, wherein the petroselinic acid containing composition is used as a composition comprising 5-99.9 wt% of fat or fat blend. 35 3. Use of an edible composition containing petroselinic acid according to claims 1-2, wherein fat in the compositions that are used is selected from the group consisting of: cocoa butter equivalents, palm oil or fractions thereof, palm kernel oil or fractions thereof, interesterified mixtures of above fats, hardened fats or fractions thereof, liquid oils, selected from sunflower oil, high oleic sunflower oil, soybean oil, rape seed oil, cotton seed oil, safflower oil, high 40 oleic safflower oil, maize oil, MCT oils, hardened liquid oils or fractions thereof and mixtures hereof.

4. Use of an edible composition containing petroselinic acid according to claims 1-3, wherein the petroselinic acid containing composition contains 5-40 wt% of petroselinic acid.

45 5. Use of an edible composition containing petroselinic acid according to claims 1-4, wherein the petroselinic acid is applied in the form of free fatty acid, or in the form of mono-, di- or triglyceride with at least one petroselinic acid group, or in the form of wax esters of petroselinic acid or in the form of C1-C4 alkyl esters of petroselinic acid or as mixtures hereof.

50 6. Use of an edible composition containing petroselinic acid according to claims 1-5, wherein the edible composition also contains an anti-oxidant, preferably selected from the group consisting of natural or synthetic tocopherols, natural polyphenols, in particular as present in tea extracts, BHT, BHA, free radical scavengers and enzymes with anti-oxidant properties.

55 Patentansprüche

1. Kosmetische Verwendung einer essbaren Zusammensetzung, welche 2-80 Gew.-% Petroselinsäure als Alterungs-

6 EP 1 013 178 B2

schutzbestandteil enthält, mit einem positiven Einfluss auf Hautzustände, ausgewählt aus der Gruppe bestehend aus Faltenbildung und Durchhängen.

2. Verwendung einer essbaren Zusammensetzung, welche Petroselinsäure enthält, gemäß Anspruch 1, wobei die 5 Petroselinsäure enthaltende Zusammensetzung als Zusammensetzung verwendet wird, welche 5-99,9 Gew.-% Fett oder Fettmischung umfasst.

3. Verwendung einer essbaren Zusammensetzung, welche Petroselinsäure enthält, gemäß Ansprüchen 1-2, wobei das Fett in den Zusammensetzungen, die verwendet werden, ausgewählt wird aus der Gruppe bestehend aus 10 Kakaobutter-Äquivalenten, Palmöl oder Anteilen davon, Palmkernöl oder Anteilen davon, interveresterten Gemi- schen der obigen Fette, gehärteten Fetten oder Anteilen davon, flüssigen Ölen, ausgewählt aus Sonnenblumenöl, High-Oleic-Sonnenblumenöl, Sojaöl, Rapsöl, Baumwollsaatöl, Safloröl, High-Oleic-Safloröl, Maisöl, MCT-Ölen, ge- härteten flüssigen Ölen oder Anteilen davon und Gemischen daraus.

15 4. Verwendung einer essbaren Zusammensetzung, welche Petroselinsäure enthält, gemäß Ansprüchen 1-3, wobei die Petroselinsäure enthaltende Zusammensetzung 5-40 Gew.-% Petroselinsäure enthält.

5. Verwendung einer essbaren Zusammensetzung, welche Petroselinsäure enthält, gemäß Ansprüchen 1-4, wobei die Petroselinsäure in Form von freier Fettsäure oder in Form von Mono-, Di- oder Triglycerid mit mindestens einer 20 Petroselinsäuregruppe oder in Form von Wachsestern von Petroselinsäure oder in Form von C1-C4-Alkylestern von Petroselinsäure oder als Gemische daraus angewendet wird.

6. Verwendung einer essbaren Zusammensetzung, welche Petroselinsäure enthält, gemäß Ansprüchen 1-5, wobei die essbare Zusammensetzung ebenfalls ein Antioxidans enthält, vorzugsweise ausgewählt aus der Gruppe beste- 25 hend aus natürlichen oder synthetischen Tocopherolen, natürlichen Polyphenolen, insbesondere wie sie in Tee- Extrakten vorhanden sind, BHT, BHA, freien Radikalfängern und Enzymen mit Antioxidans-Eigenschaften.

Revendications 30 1. Utilisation cosmétique d’une composition comestible contenant 2-80 % en poids d’acide pétrosélinique comme composant anti-vieillissement avec un impact positif sur l’état de la peau, sélectionné dans le groupe consistant en rides et affaissement.

35 2. Utilisation d’une composition comestible contenant de l’acide pétrosélinique selon la revendication 1, dans laquelle la composition contenant de l’acide pétrosélinique est utilisée comme une composition comprenant 5-99,9 % en poids de graisse ou d’un mélange de graisses.

3. Utilisation d’une composition comestible contenant de l’acide pétrosélinique selon les revendications 1 à 2, dans 40 laquelle la graisse dans les compositions qui sont utilisées est sélectionnée dans le groupe consistant en: équivalents de beurre de coco, huile de palme ou fractions de celle-ci, huile de noyau de palme ou fractions de celle-ci, mélanges interestérifiés des graisses ci-dessus, graisses durcies ou fractions de celles-ci, huiles liquides sélectionnées parmi huile de tournesol, huile de tournesol hautement oléique, huile de soja, huile de graines de colza, huiles de graines de coton, huile de carthame, huile de carthame hautement oléique, huile de maïs, huiles MCT, huiles liquides durcies 45 ou fractions de celles-ci et leurs mélanges.

4. Utilisation d’une composition comestible contenant de l’acide pétrosélinique selon les revendications 1 à 3, dans laquelle la composition contenant l’acide pétrosélinique contient 5-40% d’acide pétrosélinique.

50 5. Utilisation d’une composition comestible contenant de l’acide pétrosélinique selon les revendications 1 à 4, dans laquelle l’acide pétrosélinique est appliqué sous la forme d’acide gras libre, ou sous la forme de mono-, di- ou triglycéride avec au moins un groupe acide pétrosélinique, ou sous la forme d’esters cireux d’acide pétrosélinique ou sous la forme d’alkyl esters en C1-C4 d’acide pétrosélinique ou sous la forme de leurs mélanges.

55 6. Utilisation d’une composition comestible contenant de l’acide pétrosélinique selon les revendications 1 à 5, dans laquelle la composition comestible contient également un antioxydant, sélectionné de préférence dans le groupe consistant en tocophérols naturels ou synthétiques, polyphénols naturels, en particulier ceux présents dans les extraits de thé, BHT, BHA, capteurs de radicaux libres et enzymes avec propriétés antioxydantes.

7 EP 1 013 178 B2

REFERENCES CITED IN THE DESCRIPTION

This list of references cited by the applicant is for the reader’s convenience only. It does not form part of the European patent document. Even though great care has been taken in compiling the references, errors or omissions cannot be excluded and the EPO disclaims all liability in this regard.

Patent documents cited in the description

• EP 777971 A [0001] • GB 2181349 A [0004]

Non-patent literature cited in the description

• Afifi et al. Some pharmacological activities of essen- • Lavker et al. J.Inv.Derm., 1979, vol. 73, 79-66 [0003] tial oils of certain umbelliferous fruits. Vet Med Jour- • Bernstein et al. Lab. Invest., 1995, vol. 72, 662-669 nal Giza, 01 January 1994, vol. 42 (3 [0002] [0003] • Yagaloff et al. Essential fatty acids are antagonists • Griffiths et al. N. Eng. J. med., 1993, vol. 329, of the Leukotriene B4 Receptor. Prostaglandins Leu- 530-535 [0004] kotriened and Essential Fatty acids, 01 January 1995, vol. 52 (5 [0002]