WO2002081027A1

WO2002081027A1 - An antioxidant composition and a cosmetic or pharmaceutical composition

Info

Publication number: WO2002081027A1
Application number: PCT/BR2002/000052
Authority: WO
Inventors: Simone Fanan; Elizabete Fernandes Vicentini Rosin; Luciana Villa Nova Silva; Rita De Cassia Rabadji Zornoff; Jean-Luc Gezstesi; Walter Albuquerque
Original assignee: Natura Cosméticos S.A.
Priority date: 2001-04-10
Filing date: 2002-04-10
Publication date: 2002-10-17
Also published as: PE20021038A1; BR0101449A; AR033112A1

Abstract

The present invention relates to an antioxidant composition comprising from 1X10-7 to 0.02 % of a carotenoid, from 1x10-5 to 1 % of polyphenols, from 0.0002 to 6.5 % of vitamins and from 0.001 to 3 % of helioxine, all the percentages being expressed by weight based on the total weight of the antioxidant composition. In another aspect, the invention refers to a cosmetic or pharmaceutical composition comprising the antioxidant composition defined above. The compositions according to the invention have an improved effect on protecting the skin against damages to the cells and skin caused by free radicals of oxygen.

Description

Title. "AN ANTIOXIDANT COMPOSITION AND A COSMETIC OR PHARMACEUTICAL COMPOSITION".

Field of the Invention

The present invention relates to an antioxidant composition and, more particularly, to its use in cosmetic or pharmaceutical compositions for the purpose of protecting the skin, thus preventing damages to the cell and skin caused by free radicals of oxygen. Background of the Invention

There are many cosmetic, pharmaceutical or nutritional products that contain, in its composition, components with antioxidant action. Such compounds are most used in the field of cosmetics.

Free radicals of oxygen and the reactions triggered by these chemical species occur in a wide diversity of physiological and pathological processes, including aging (HARMAN, D. 1992 - Free radical theory of ag- ing. Mutation Research, 275: 257-266). It is already well established that such chemical species are produced in the aerobic cells from agents in the environment, as well as from normal metabolic processes of the organism, such as cellular respiration. It is also known that the aerobic organisms have efficient defense mechanisms against free radicals of oxygen and that they are characterized by a set of antioxidant enzymes and compounds that maintain the oxidant species at levels below a concentration considered to be toxic. However, when there is an unbalance between the process for producing of said oxidant species and the mechanisms for destroying them, an oxidative stress condition establishes (SIES, H. 1986 - Biochemistry of oxi- dative stress. Angew. Chem. Int. Ed. Engl., 25: 1058-1071 ), in which the oxidative attack of the biomolecules is favored.

The cause relationship observed between the oxidative stress and various pathologies and degenerative processes in the human being has aroused interest for the study and exploitation of molecules capable of inter- fering with the generation of free radicals of oxygen or with the reactions triggered by them, thus acting as antioxidants.

As described in CADENAS, E. (1989). Biochemistry of oxygen toxicity. Ann. Rev. Biochem., 58:9-110, the oxigen-reactive species (EROs) are formed in the aerobic eukaryotic cells by means of successive steps of monoelectronic reductions of oxygen, sequentially generating the superoxide (02) anion radical, the hydrogen peroxide (H2O₂) radical and the hydroxyl radicals (HO^"), as schematized below:

O₂ + e^" → O₂ ^"

2H⁺ + O₂ ^" + e^" → H₂O₂

H₂O₂ + Fe⁺² → Fe⁺³ + HO + HO- (Fenton reaction)

Another ERO, the singlet oxygen (¹O₂), may be produced by photochemical reactions in which transfer of energy from a sensibilizer to the fundamental oxygen takes place, and this sensibilizer may be released by phagocytic cells of the immune system or by enzymatic reactions. As exoge- nous sources, one can cite, for example, some drugs, toxins, cigarette smoke, radiations, thermal shock end pollution. Singlet oxygen is capable of spreading over a long distance, since it has a relatively long half-life (microseconds) in comparison with that of the hydroxyl radical (nanoseconds), and may cause damage to lipids, proteins and DNA. All these damages are being more and more associated with pathologies such as arteriosclerosis, muta- genesis, arthritis and diabetes.

However, among the oxygen-reactive species, the most powerful oxidant is represented by the hydroxyl radical, which is capable of reacting rapidly with virtually every biomolecules, such as lipids, proteins, carbohy- drates and nucleic acids, leading to the development of cellular toxicity (HALLIWELL, B. & GUTTERIDGE, J.M.C. (1981). Formation of thiobarbituric acid reactive substance from deoxyribose in the presence of iron salts. FEBS Letters, 128: 347-352; SIES, H. (1986). Biochemistry of oxidative stress. Angew. Chem. Int. Ed. Engl., 25: 1058-1071). Therefore, substances capable of reacting with the EROs or preventing chains of their radical reactions constitute valuable elements in antioxidant protection of biologic systems. Such substances are generically called free radical scavenges.

Antioxidants such as superoxide dismutase, reductase glutathi- one and glutathione peroxidase, vitamin E, vitamin C and carotenoids are present in the skin to protect it against free radicals (MUIZZUDDIN, N et al., (1999). Effect of antioxidants and free radical scavengers on protection of human skin against UVB, UVA and IR irradiation. Skin Research and Technology, 5: 260-265). Even so, continuous damages to the skin may be generated, since the ability of those compounds already present in the organism to eliminate free radicals is not 100% efficient. Studies have shown that topi- cal application of antioxidants may be effective in decreasing cellular damage caused by external factors, such as excessive exposure of the skin to sunshine (MUIZZUDDIN, N., et al., (1999). Effect of antioxidants and free radicals scavengers on protection of human skin against UVB, UVA and IR irradiation. Skin Research and Technology, 5: 260-265; SHINDO, Y et al (1993). Antioxidant defense mechanisms in murine epidermis and dermis and their responses to ultraviolet light. J. Invest. Dermatol., 100: 260-265).

Compositions that are usually employed as antioxidants comprise a mixture of vitamins, preferably vitamins A and E, or vitamins C and E or else vitamins A, C and E. Brazilian patent document PI 9909273-1 , also filed in the name of the present applicant, describes a process for stabilizing levorotatory ascorbic acid (vitamin C) and cosmetic compositions that contain vitamin C associated with vitamin A and OPC glucospheres. Other studies have also shown that there may be an increase in the ability of scavenging free radicals when a mixture of vitamins is made. For instance, KITAZAWA, M. et al - Interactions between vitamin E homologues and ascorbate free radicals in murine skin homogenates irradiated with ultraviolet light. Photochemistry and Photobiology, 65(2): 355-3657, have shown that vitamin C recovers vitamin E after scavenging free radicals produced by exposure to ultraviolet light. Another document from the prior art that teaches a composition with an antioxidant effect, containing a mixture of vitamins and their derivatives, is US Patent 5,023,235, which teaches an antioxidant system com- prising ascorbic ester stabilized with an aliphatic acid, a complexing agent (formed by ethylenediamine tetracetic acid, diethylenetriamine pentacetic acid, hexadecylamine salicylate, citric acid, dibenzyldithiocarbamate), to- copherol (vitamin E), caffeic acid and its derivatives, N-acetyl-L-cysteine and glutathione.

Other antioxidant compounds already known from the prior art comprise carotenoids and flavonoids. Carotenoids are a group of lipophilic pigments found in photosynthestic complexes of plants and photosynthetic bacteria, fungi and algae. Over 600 different natural carotenoids are known, α,β-carotene, lycopene, β-cryptoxanthine, lutein and zeaxanthine being the one of highest concentration in the human being. Carotenoids that predominate in the human epithelial tissue are β-carotene and lycopene; however, the latter has a great advantage over β-carotene, since it is not consumed during the "capture" of singlet oxygen, is not a precursor of vitamin A and can be used in products that are exposed to light. On the other hand, flavonoids are polyphenols and comprise compounds such as the proantocyanidins known as OPCs, quercetins, leucoantocyanidins and catechins.

Document US 5,804,168 describes a pharmaceutical composition capable of preventing and protecting against skin damages caused by exposure to sunshine, inhibiting the formation of free radicals. An example of the composition described in that document contains at least one flavonoid, vitamin E and carotenoid. The composition preferably comprises a component of cysteine, magnesium, copper and selenium, and it may also contain wild yan, rosemary, gentian root and myrrh gum. On the other hand, document EP 968,709 describes an antioxidant cosmetic composition comprising a synergistic mixture of chrysanthel- lum extract, which is a flavonoid-rich vegetable compound , and at least one compound selected from the group comprising vegetable extracts rich in phenolic compounds, vegetable extracts rich in carotenoids, vegetable ex- tracts rich in tocopherols, an antioxidant of natural or synthetic origin and an enzymatic system.

Document US 5,296,500 teaches a method for regulating wrin- kles and/or atrophies on skins of mammals by treating the skin with an effective and safe amount of N-acetyl-L-cysteine and/or its derivatives. The composition used in the treatment described in the above-mentioned document may also contain other components, such as an antioxidant compound, among others.

In document US 5,709,816, one describes a molecule and its derivatives, developed for preventing and treating damages to the organism caused by singlet oxygen, and a composition that has anti-inflammatory, anti- aging action and that brings about denaturation of proteins, lipoperoxidation and damages to the DNA.

Document US 5,827,880, in turn, presents a metal complex that has an activity similar to that of superoxide, catalase and peroxidase for pharmaceutical administration in the treatment or prevention of diseases associated with damages to cell or tissue caused by free radicals, such as su- peroxide, and a cosmetic formulation containing compounds that capture these compounds.

Another document from the prior art, US 4,797,421 , describes a composition in which protocyanidin dimers and decimers are used as antioxidant compounds. By the teachings known from the prior art there was not yet an antioxidant that could act on every cell compartments and in all layers of the skin.

Therefore, the object of the present invention is to provide an antioxidant composition that has a more effective effect than that of known antioxidants.

Summary of the Invention

The present invention relates to an antioxidant composition comprising from 1x10^"7 to 0.01% of a carotenoid, from 1x10^"5 to 1% of polyphenols, from 0.0002 to 6.5% of vitamins and from 0.001 to 3% of helioxine, all the percentages being expressed by weight based on the total weight of the antioxidant composition.

In another aspect, the invention relates to a cosmetic or pharma- ceutical composition comprising the antioxidant composition defined above. Detailed Description of the Invention

It has now been found that a mixture of antioxidant compounds such as carotenoids, polyphenols and vitamins together with helioxine in de- termined concentration ranges provides elimination of free radicals in different media, either hydrophilic of lipophilic ones, and different types of EROs (HO , ¹O₂). When incorporated in topical cosmetic compositions, for instance, the antioxidant combination of the present invention is effective in protecting all the compartments of the cell and all the layers of the skin. The antioxidant composition of the present invention comprises, as a first component, at least one compound selected from the group of carotenoids. Carotenoids useful for the present invention comprise β-carotene, zeaxanthine, lutein, lycopene, β-cryptoxanthine; preferably, lycopene is used. The carotenoid component is present in the antioxidant composition of the invention in an amount of from 1x10^"7 to 0.01% by weight, more preferably from 5x10^"7 to 0.002% by weight, based on the total antioxidant composition.

The antioxidant combination of the invention also comprises a compound selected from the group of polyphenols such as flavonoids and terpenes. Among the flavonoids, one may use proantocyanidins (OPC), quercetins, leucoantocyanidins and catechins. Preferably, one employs proantocyanidins and more preferably OPCs from grape seed, which are oli- gomers that, when applied to cosmetic compositions, for example, protect the corneous stratum against attack by the hydroxyl radical (HO), peroxyl radical (HOO) and the lipoperoxides (LO)³. In a more particularly advantageous way, the OPCs from grape seed are used enveloped by a glucosphere, which favors its stabilization within the formulation. According to the invention, the polyphenols are used in an amount of from 1x10^"5 to 1% by weight, more preferably from 8x10^"5 to 0.3% by weight, based on the total antioxidant composition. The vitamins present in the antioxidant composition of the invention are advantageously selected from the group comprising tocopherols, retinoids, complex-B vitamins and ascorbic acid and their derivatives, and one preferably uses tocopherols such as α,β and γ tocopherol and tocopherol palmitate acetate. Still more preferably, the vitamin component is tocopherol acetate (Vitamin E), since it has greater cutaneous permeation and protects the cytoplasm membrane against attacks by the hydroxyl radical (HO), su- peroxide anion radical (O -), peroxyl radical (HOO) and the lipoperoxides (LO)². Therefore, the utilization thereof is more advantageous when the antioxidant composition is included in topical cosmetic or pharmaceutical compositions.

The vitamins are used in amounts ranging from 0.0002 to 6.5% by weight, based on the total weight of the antioxidant composition, preferably from 0.001 to 5%.

On the other hand, the helioxine component is present in a range from 0.001 to 3% by weight, based on the total weight of the antioxidant composition. Helioxine useful in the present invention is the sunflower-seed extract (Helianthus annus).

According to preferred embodiments of the invention, in addition to the components mentioned above, the antioxidant composition may also contain other conventional ingredients of this type of formulation such as amino acids, chelating agents, chromophorous agents other than carotenoids such as nucleotides and microelements. Examples of amino acids include glutathione, L-prolihistamine, glucagon, serilglycyltrosinilalanileucine. On the other hand, the chelating agents usable for the objectives of the invention are EDTA, nitrilo-tri-acetate, DEQUEST® and lactoferrine, DEQUEST® being preferably used. The microelements may be selected from the group com- prising manganese, selenium, silicon, copper, cobalt, zinc and molybdenum. Preferably, manganese, selenium and zinc are used. In addition to the latter, various other conventional ingredients of this kind may be added to the antioxidant composition of the invention; these ingredients may easily be defined by any person skilled in the art and do not limit the invention. The antioxidant composition of the present invention, when incorporated in cosmetic or pharmaceutical compositions, provides an improved effect of protecting the skin and in preventing damages to the cells and the skin, as well as in the immune system of the skin, which may be a result of a physiological unbalance between the production and the elimination of oxigen-reactive species caused by internal or external factors to the human organism. Among the internal factors, for example, poor feeding, chemical, physical and emotional stress can be pointed out, and among the external factors, pollution, solar radiation, cigarette smoke, among others. In this way, the antioxidant composition of the present invention may be included, for instance, in products for mouth hygiene, treatment of the hair, treatment of the skin, protection against sun rays, perfume and nutritional supplements.

Cosmetic or pharmaceutical compositions preferably comprise the antioxidant combination in a range of from 0.001 to 10.5% by weight, based on the total weight of the composition.

In experiments for appraising the antioxidant action of the anti- oxidant composition of the invention, control assays were carried out, exposed to the oxidative system Fe⁺³/NTA/H₂H2, in the presence and in the absence of a known hydroxyl radical scavenger, namely manitol (data not shown). The treatment with the oxidative system for 20 minutes, in the absence of manitol, caused extensive degradation of the deoxyribose, as ob- served from the intensive coloration achieved after reaction with TBA, thio- barbituric acid (data not shown). The average value of the absorbency of the control assays in the absence of manitol, at 532nm, was taken as 100% of oxidation. In the presence of a relatively high manitol concentration (50mM = 0.9%), one has observed an intense reduction in the coloration and, conse- quently, in the degradation of deoxyribose.

Figure 1 illustrates a curve obtained in an assay in which deoxyribose (2.8mM) was incubated with the oxidative system Fe⁺³/NTA/H₂θ₂, for 20 min at 37°C in the presence of increasing concentrations of the antioxidant composition of the present invention. At the end, reaction with TBA and detection of the complex formed at 532nm were effected. The results were exposed in oxidation percentage with respect to the average value of the control assays. The curve of Figure 1 shows that there was a significant increase in the inhibition of desoxyribose with the increase in the concentration of said antioxidant composition, and an IC₅₀ value of 0.009% (a value that reduces oxidative degradation of deoxyribose by 50%) was achieved. 93% of inhibi- tion of oxidative degradation at concentration of 1 % of the composition was achieved. At this concentration, the composition contains, respectively, 0.65% of Vitamin E acetate, 0.32% of Helioxine, 0.0026% of Lycomate at 6%, and 0.026% of OPC glucosphere.

In this figure one can see that, at this concentration, a maximum level of inhibition of oxidative degradation of desoxyribose was achieved, and there was no need to increase the concentration of the antioxidants for the purpose of improving protection of the deoxyribose against oxidative degradation caused by free radicals.

For a concentration of 0.03% of the antioxidant composition (containing 0.0194% of vitamin E acetate, 0.0097% of helioxine, 0.000078% of Lycomate at 6% and 0.00078% of OPC Glycospheres), one achieved inhibition of 87.3% in oxidative degradation of deoxyribose caused by the free radicals generated by the system Fe⁺³/NTA/H O₂. Above this concentration, the inhibition curve reached a level of protection against oxidative degrada- tion of deoxyribose at which the maximum inhibition (97%) was achieved at concentration of 3% of the antioxidant composition, which is the objective of the present invention.

Thus, any concentration of the composition described now, in the range of 0.03 - 3%, has good protection of deoxyribose against the free radicals generated specifically by the oxidant system Fe⁺³/NTA/H₂O₂.

The results shown in Figure 1 lead to the conclusion that there is synergism between the tested antioxidants, and low concentrations of these antioxidant may be utilized to achieve a good result in scavenging free radicals. The illustrative examples presented below will serve to describe the present invention in a better way. However, the illustrated data and procedures merely refer to some embodiments of the present invention and should not be taken as limiting the scope of the invention. Example 1 :

The following cosmetic composition was prepared for treating the skin with SPF 15:

*Vitamin and acetate, compounds of the antioxidant combination according to the invention.

Example 2:

A hydrating emulsion was prepared - SPF 15 with the following composition:

Example 3:

A hydrating emulsion was prepared with the following composition:

PHOTONYL® Example 4:

A composition in the form of a gel was prepared - SPF 15, com- prising:

Example 5:

A composition in the form of a gel was prepared for skin treat- ment:

Claims

1. An antioxidant composition characterized by comprising from 1x10^~7 to 0.01% of at least one carotenoid, from 1x10^"5 to 1% of one or more polyphenols, from 0.0002 to 6.5% of one or more vitamins and from 0.001 to 3% of helioxine, all the percentages being expressed by weight, based on the total weight of the antioxidant composition.

2. An antioxidant composition according to claim 1 , characterized in that the carotenoid is present in an amount ranging from 5x10^'7 to 0.002% by weight, based on the total weight of the antioxidant composition.

3. An antioxidant composition according to claim 1 , characterized in that the carotenoid is selected from the group comprising α and β- carotene, zeaxanthine, lutein, lycopene, β-cryptoxanthine.

4. An antioxidant composition according to claim 3, characterized in that the carotenoid is lycopene.

5. An antioxidant composition according to claim 1 , characterized in that the polyphenol is present in an amount raging from 8x10^"5 to 0.3%.

6. An antioxidant composition according to claim 1 , characterized in that the polyphenol is selected from the group comprising flavonoids and terpenes.

7. An antioxidant composition according to claim 6, characterized in that the polyphenol is selected from proantocyanidins (OPC), quercetins, leucoantocyanidins and catechins.

8. An antioxidant composition according to claim 7, characterized in that the polyphenol is proantocyanidin.

9. An antioxidant composition according to claim 1 , characterized in that the vitamins are present in a range from 0.001 to 5%.

10. An antioxidant composition according to claim 1 , characterized in that the vitamins are chosen from the group comprising tocopherols, retinoids, B-complex vitamins and ascorbic acid and derivatives thereof, to- copherols such as α, β and γ tocopherol and tocopherol palmitate acetate being preferably used.

11. An antioxidant composition according to claim 10, character- ized in that the vitamin is tocopherol acetate.

12. An antioxidant composition according to claim 1 , characterized in that it further comprises amino acids, chelating agents, chromopho- rous agents other than carotenoids such as nucleotides and microelements.

13. An antioxidant composition according to claim 1 , characterized in that it comprises from 1x10^"7 to 0.01 % of lycopene, from 1x10^"5 to 1% of glucospheres of grape-seed OPCs, from 0.001 to 5% of vitamin E acetate, and from 0.01 to 3% of helioxine, all the percentages by weight based on the total weight of the oxidant composition.

14. A cosmetic or pharmaceutical composition characterized in that it comprises, as an antioxidant agent, a composition as defined in any one of claims 1 to 13.

15. A cosmetic or pharmaceutical composition according to claim 14, characterized in that the antioxidant agent is present in an amount rang- ing from 0.001 to 10.5% by weight, based on the total weight of the composition.

16. A cosmetic or pharmaceutical composition according to claim 14 or 15, characterized in that it is a composition for topical use.