WO2007026035A2

WO2007026035A2 - Use of a mycosporin-type amino acid (porphyra 334) as an antioxidant

Info

Publication number: WO2007026035A2
Application number: PCT/ES2006/000488
Authority: WO
Inventors: Francisca De La Coba Luque; José AGUILERA ARJONA; Félix LÓPEZ FIGUEROA
Original assignee: Universidad De Málaga
Priority date: 2005-08-31
Filing date: 2006-08-28
Publication date: 2007-03-08
Also published as: ES2301434A1; ES2301434B1; WO2007026035A3; ES2301433A1; ES2301437A1; ES2301293A1; ES2301436B1; ES2301293B1; ES2301435A1; ES2301435B1; ES2301436A1; ES2301437B1

Abstract

The invention relates to a mycosporin-type amino acid (porphyra 334) which can be used as an antioxidant and which is suitable for use in the biotechnological industry. The invention outlines the potential use of a mycosporin (MAA)-type amino acid as an antioxidant substance, specifically porphyra 334 isolated from red alga Porphyra leucostica, and to the possible use thereof in pharmaceutical, nutraceutical or functional food preparations, among others, for the prevention of oxidative stress.

Description

MCOSPORINE TYPE AMINO ACID (PORPHYRA 334) AS AN ANTIOXIDANT

TECHNICAL SECTOR The present invention is part of the biotechnology sector and describes the potential use as antioxidant substances of certain secondary metabolites called mycosporin-type amino acids (MAAs) isolated from red algae and marine lichens in addition to their possible application in pharmaceutical preparations, nutraceuticals, functional foods for the prevention of oxidative stress.

PREVIOUS TECHNIQUE

Ultraviolet radiation is one of biological factors that limit the survival, physiology and growth of many organisms. Some of the multiple harmful effects of UV radiation include the alteration of DNA and protein molecules, inactivation of enzymes and the formation of free radicals, which attack cell membranes and other target molecules altering their functionality. All aerobic organisms have a wide variety of both enzymatic and non-enzymatic antioxidant defense systems that cooperatively coordinate and protect the body from the risks of oxidative stress. Among them, the enzymatic activities of superoxide dismutase (SOD), glutathione peroxidase (GPX) and catalase (CAT) stand out; in addition to ascorbic acid (vitamin C), α-tocopherol (vitamin E), glutathione (GSH), β-carotene, vitamin A, flavonoids and phenolic acids among others.

Free radical means any chemical species that contains one or more missing electrons in its external orbitals so that a compound can become a free radical by capturing or losing an electron. Although free radicals of a very different nature exist, it is the species that derive from the oxygen molecule (ROS) the most abundant in aerobic organisms, highlighting products of the rupture or excitation of O _{2 such} as singlet oxygen ¹ O ₂ and species of oxygen that are partially reduced such as the bidroxyl radical (OH ^» ), superoxide anions (O ₂ ^" ) and hydrogen peroxide (H ₂ O ₂ ). These unstable molecules travel through the body taking electrons, thus recovering their electrochemical stability. it is very dangerous because to achieve this they attack stable molecules.Once the free radical has managed to take the electron it needs to match its free electron, the other molecule becomes a free radical, thus initiating a destructive cycle for our cells . Free radicals give rise to important alterations in molecules such as

DNA, lipids and proteins, seriously altering the cycle and cellular functionality. The DNA can suffer loss of bases as well as the rupture of one or both strands of the genetic material, alterations that can result in irreversible mutations. Many proteins are capable of absorbing a large amount of oxidation without apparently affecting their function. However, there is no doubt that the consequences of alterations in some functions, for example, the reception and transmission of signals, the transport of ions, the duplication and repair of DNA, the responses to stress conditions and energy metabolism, transcription and translation can be critical for the cell. The damage caused by OH ^» and ¹ O ₂ are irreversible and generally mark the proteins for degradation. Cell membranes can also be seriously damaged in oxidative stress because phospholipids that have fatty acids with several double bonds are very susceptible to oxidation due to the loss of a hydrogen (allyl). Once the carbon radical has been generated in a fatty acid, it reacts with molecular oxygen forming a peroxyl radical. The peroxyl radical can take an allylic hydrogen to another methylene whereby the reaction is propagated. Hydroperoxides, which are stable compounds, if they come into contact with transition metal ions will produce more free radicals that will initiate and propagate other chain reactions. Thus, the membranes are seriously damaged and therefore their functionality is altered. Free radicals are associated with a wide range of pathologies and diseases such as Alzheimer's or Parkinson's and conditions related to sun exposure such as the appearance of cataracts, photoaging, inflammatory episodes and neoplasms. They are also responsible for the oxidation of food fats, which is the most important form of deterioration after alterations caused by microorganisms. With oxidation, stale odors and flavors appear, the color and texture are altered, and the nutritional value decreases when some vitamins and polyunsaturated fatty acids are lost. In addition, products formed in oxidation can become harmful to health.

Mycosporin-type amino acids consist of a cyclohexenone or cyclohexenimine ring, conjugated with a nitrogenous substituent of an amino acid or its amino alcohol that acts as a chromophore allowing the absorption of certain shortwave radiation. The metabolites that are isolated in fungi have an absorption range between 310 and 320 nm and have cyclohexenone rings exclusively, being known by the name of mycosporins in reference to their origin. In contrast, metabolites that are isolated from marine organisms and algae contain cyclohexenimine rings, with maximum absorptions between 310 and 360 nm and they are known as mycosporin-like amino acids ό

MAAs. Still, mycosporine-glycine and mycosporine taurine are aminocyclohexenones isolated from marine organisms. There are currently 13 different mycosporins described in fungi and 23 MAAs in marine organisms. They are small molecules, with molecular weights around 330 Da and have high photostability. They behave like amphoteric molecules, similar to amino acids, so that they have positive and negative charges on the same molecule. They show physicochemical characteristics of ionic compounds, for example, high effusion point and high water solubility.

There are many functions that have been attributed to these molecules in the organism: from organic osmolyte in Chlorogloeopsis cyanobacterial communities, photosynthetic accessory pigments or precursors of these, to determining molecules in reproductive processes of some species of fish, however it is the photoprotective role against UV radiation the most accepted and documented since apparently they act partially protecting cellular components and physiological processes. A number of studies have evaluated these types of molecules for their activity as topical photoprotectors, vehiculating natural extracts with a high percentage of MAAs and seeing their SPF and their photoprotective potential in plant cells, human keratinocytes, etc. (US Patent 6787147; WO 02/39974; WO 03/020236). Works have also been published that refer to the antioxidant properties of extracts obtained from algae and corals. Dunlap and Yamamoto in 1995 (Comp. Biochem. Physiol. 112: 105-114) pointed to a possible antioxidant activity of mycosporine-glycine by in vitro assays of lipid peroxidation (phosphatidylcholine method) from extracts of marine organisms containing MAAs, while other iminoMAAs such as porphyra 334, shinorine, palythine, asterine 330 and palythinol were oxidatively robust and did not participate in oxidation-reduction reactions. However, as indicated, these tests were carried out with algal extracts that, in addition to MAAs, contained a high percentage of other cellular components such as polysaccharides, enzymes, etc., so it is not possible to firmly affirm the antioxidant activity of mycosporine-glycine (or M-gly) based on that work.

Nakayama et al. In 1999 (J. Am. OiI Chem. Soc, 76: 649-653) isolated a new mycosporin-like amino acid from the red seaweed Porphyra yezoensis called usujilene, already identified in Palmaria palmata but not in any species of Porphyra. Their results indicated that such MAA showed antioxidant activity against linoleic acid autoxidation (Thiobarbituric acid and ferric thiocyanate methods), donating certain hydrogens to LOO- lipid radicals and giving rise to resonance stabilized MAA molecules as well as ex-tocopherol.

Suh et al. In 2003 (Photochem. Photobiol. 78: 109-113) also suggest the antioxidant function of M-gly, but probably acting together with other active MAAs. M-gly, among others, could play an important role by participating in the elimination of O ₂ ^" generated by endogenous photosynthesizer systems.

Yakovleva et al. In 2004 (Comp. Biochem. PhysioL, 139: 721-739 examined the importance of M-gly as a functional antioxidant against thermal stress in two corals, Platygyra ryukyuensis and Stylophora pistillata, based on the correlation between grade of susceptibility and endogenous concentration of M-gly.

Also the patent US2004228875 refers to the antioxidant properties of extracts obtained from algae of the genus Porphyra, although without concluding on the possible role played by MAAs.

More recent publications analyze the antioxidant properties of extracts obtained from algae but without specifying the participation of MAAs (Yuan et al., 2005, Food Chem. Toxicol., 43: 1073-1081; Kuda et al., 2005, J. Food Composition Anal, 18: 625-633).

It can be concluded, therefore, that the antioxidant role of iminoMAAs as such, that is, purified or isolated in a high degree of purity, is not well known, nor is their behavior at the level of water-soluble free radical sequestration.

An antioxidant is defined as a substance that in low concentrations compared to an oxidizable substrate, delays or prevents its oxidation. The present invention describes the potentiality of the porphyra 334 MAA isolated from Porphyra leucosticta as a radical scavenger and lipid peroxidation inhibitor. In the tests carried out, the new antioxidant is compared with another known antioxidant, α-tocopherol. The described compound could be used in therapeutic applications, and in non-medical applications for the stabilization of compounds susceptible to oxidative deterioration, in the preservation of food or related products, and in nutritional, nutraceutical, functional or parapharmaceutical supplements for their antioxidant properties for prevent oxidative stress. MAAs are found naturally in isolated organisms in the order of mg / g PS, highlighting porphyra 334 found in the Porphyra leucosticta algae in the order of 3-6 mg / gPS. As we know, seaweed has been used as human food since ancient times, especially in Eastern countries and whose culture is expanding worldwide. Porphyra (whose vulgar name in Japan is nori) is one of the most seaweed important used as human food. In recent years, Porphyra has been listed in Japanese fisheries statistics as the third catch in order of importance and has a high content of valuable edible proteins so that this study could give added value to certain types of natural foods.

DISCLOSURE OF THE INVENTION

The present invention presents an isolated compound of Porphyra leucosticta with the following structure and useful as an antioxidant and free radical sequestrant.

Porphyra 334

Compounds of the mycosporin type amino acid type have been purified in the aqueous phase starting from Porphyra leucosticta. The compounds have been detected and characterized by HPLC. A UV-visible detector (photodiode detector 996) was used, which measured the absorbency for each sample between 290 and 400 nm. Once the chromatogram was extracted at 330 nm, the peaks were identified by co-chromatography according to their spectra and retention times, compared to MAAs standards.

Preparative scale extraction was performed by dissolving 60-80 g (PF) of biological material in 1 liter of methanol to 20% v / v and incubated in a thermostatic bath at 45 ⁰ C for

2 hours. The extract is then centrifuged at 14000rpm for 15 min and rotavaporated at

45 ⁰ C to remove part of the methanol from the sample.

The purification was performed in three consecutive steps in which chromatographic absorption techniques are combined by the application of active carbon, precipitation of polysaccharides by adding 100% methanol to the sample and final separation by ion exchange chromatography. Finally, aqueous solutions of MAA were obtained in a high degree of purity at concentrations of the order of mM.

Antioxidant capacity of mycosporin-like amino acids To measure the activity as a water-soluble radical sequestrant, the ABTS peroxidase method has been used, which allows to determine the total antioxidant activity (TAA) of a sample understood as a parameter that allows quantifying the capacity of a sample, natural or processed, of sequestering free radicals present in an aqueous solution.

Porphyra 334 isolated from Porphyra leucosticta has no significant antioxidant activity at any pH tested as an inhibitor of water-soluble free radical production (ABTS ⁺ I

Capacity as a lipid peroxidation inhibitor

Porphyra 334 isolated from Porphyra leucosticta was studied as an inhibitor of lipid peroxidation in vitro using the β-carotene bleaching technique. The method of decolorization of β-carotene is widely used to determine the antioxidant capacity of various substances in lipophilic medium, most of them extracted from fruits, vegetables and other products intended for food consumption in order to determine their greater or lesser degree of self-preservation in a natural state. In this test, α-tocopherol (α-TOC) was used as a positive control.

Porphyra 334 isolated from Porphyra leucosticta shows moderate antioxidant activity at the level of lipid peroxidation inhibition. It is thus constituted as an antioxidant of moderate activity in vitro.

Abduction of superoxide radicals

The protocol that was carried out was based on Marklund & Marklund (1974, Eur. J.

Biochem., 47: 469-474) with some modifications. Dose-response relationships for the MAAs under study were determined at different concentrations.

Porphyra 334 isolated from Porphyra leucosticta at concentrations of 1 mM 50% inhibits the oxidation kinetics of pyrogallol. By acting as an antioxidant and free radical scavenger, this compound, in extracts or preparations containing it, could be used in pharmaceutical preparations or formulations for the prevention and therapeutic treatment of diseases or conditions related to free radicals, in parapharmacy products, in functional foods, nutritional supplements and nutraceutical preparations, and in the food industry as an antioxidant potential (additive).

BRIEF DESCRIPTION OF THE DRAWINGS Figure 1. Area (%) of eluted peaks and concentrations expressed in mg g ^"1 PS of different MAAs present in methanolic extracts of algae Porphyra leucosticta, Gymnogongrus devoniensis, Gelidium sesquipedale and lichen Lichina pygmaea. The presence of one type is observed of MAA majority in each organism (> 66%) together with other minor MAAs and traces of unidentified substances.

Figure 2. Chromatograms of aqueous extracts of MAAs eluted from the column loaded with DOWEX resin.

Figure 3. Table. Dose (μM) - antioxidant activity response (%) of porphyra 334 isolated from Porphyra leucosticta with respect to 10 μM of α-tocopherol by the method of decolorization of β-carotene. The values represent the mean values and standard deviation of 3 experiments. Porphyra 334 isolated from Porphyra leucosticta is a good antioxidant at a concentration of 100-200 μM.

Figure 4. Sequestration capacity of superoxide radicals generated by the porphyra 334 pyrogallol method isolated from Porphyra leucosticta. The mean and standard deviation of three experiments are represented.

WAY (S) OF CARRYING OUT THE INVENTION

Preparative scale purification of porphyra 334 from the red seaweed Porphyra leucosticta

Compounds of the mycosporin type amino acid type have been purified in the aqueous phase starting from Porphyra leucosticta. The compounds have been detected and characterized by HPLC (Waters 600). The column used for the separation of the MAAs in the HPLC was a C ₈ (Sphereclone ™, Phenomenex, Aschaffenburg, Germany), packed with porous silica microparticles of 5 mm in diameter with a derivatized surface with an aliphatic chain of 8 carbon atoms (octadecyl silane). Its size was 250 x 4.6 mm. A pre-column (Phenomenex, Aschaffenburg, Germany) related to the column used was used. The mobile phase used was 2.5% methanol (v / v, HPLC quality) plus 0.1% acetic acid (v / v) isocratically pumped at a flow rate of 0.5 ml min ^"1. A UV detector was used. visible (photodiode detector 996), which measured the absorbance for each sample between 290 and 400 nm. Figure 1 shows the percentages in area of the chromatographed peaks of different algal extracts, some identified as MAAs and others unknown. Purification objective was to isolate the major MAA in Porphyra leucosticta in the aqueous phase, in addition to removing traces and other types of unidentified compounds.

Once the chromatogram was extracted at 330 nm, the peaks were identified by chromatography according to their spectra and retention times, compared with MAA standards, provided by Professor Dr. UIf Karsten (University of Rostock, Germany) extracted from different organisms marine: Mastocarpus stellatus (shinorine), Porphyra yezoensis (porphyra-334), Bostrychia scorpioides (palythine), the eyes of the coral trout Plectropomus leopardus (asterine 330) and the lichen Lichina pygmaea collected in France (M- glycine) The chromatograms of the extracts collected after passing through the DOWEX50 ion exchange column are shown in Figure 2. As can be seen, in the case of the extract from the P. leucosticta algae, initially with the presence of 4 different MAAs, subsequently only two of them appear, porphyra 334 and shinorine, so similar in structure, functional groups and size that their separation was really difficult. The ratio between shinorine and porphyra 334 remained constant throughout the purification process (1: 8.5 approx.).

Preparative scale extraction was performed by dissolving 60-80 g (PF) of biological material in 1 liter of methanol to 20% v / v and incubated in a thermostatic bath at 45 ⁰ C for 2 hours. Extract at 14,000 rpm for 15 min and then centrifuged at rotary evaporation 45 was ⁰ C to remove part of the methanol in the sample.

The purification is carried out in three consecutive steps in which chromatographic absorption techniques are combined by the application of active carbon, precipitation of polysaccharides by adding 100% methanol to the sample and final separation by ion exchange chromatography (Dowex 50 W x 8 resin -100). For the elution of the mycosporine-type amino acid porphyra 334, double-distilled water was used as eluent, with a slightly alkaline pH (7.2). Finally, aqueous solutions of MAA were obtained in a high degree of purity at concentrations of the order of mM.

Antioxidant capacity at the level of ABTS water-soluble radical sequestration

To measure the activity as sequestrants of water-soluble radicals, the ABTS peroxidase method has been used, which allows to determine the total antioxidant activity (TAA) of a sample understood as a parameter that allows quantifying the capacity of a sample, natural or processed, of sequestering free radicals present in an aqueous solution. This parameter is aimed at giving information on the antioxidant activity that a specific sample can present, regardless of the partial activities that each of its components may present or the synergism effects that could be established.

2,2'- Azino -bis- (3- ethyl-benzothiazoline-6- sulfonic acid) or ABTS is a compound that has high chemical stability, high water solubility and maximum absorption in the UVA band at 342 nm . This compound in the presence of H ₂ O ₂ and peroxidases enzymes derives to a metastable radical (ABTS ⁺ ) with a characteristic absorption spectrum and different from ABTS, presenting maximum absorption in the UV spectral region and visible at 413, 645, 727 and 811 nm. ABTS is a product that has great stability over a wide pH range, showing the same absorbance spectrum at pH 4 and pH 8.5. Likewise, the formation of the ABTS ⁺ radical is also carried out in that pH range but the enzymatic activity of the peroxidase itself is dependent on the pH of the reaction medium so that when the activity is alkalized the activity decreases, thus increasing the period of delay or "lag time". The activity of our enzyme could be adjusted to an exponential curve so that it is maximum at pH 4.5 and ceases to be active at pH higher than 10. Our assays will run at pH 6-8.5 so that we ensure the activity of the enzyme.

The quantification of the free radical sequestration capacity of a sample is carried out by decolorization tests in which the formation of ABTS ⁺ results in a characteristic coloration that will decrease proportionally to the amount of substances capable of trapping these. radicals added to the reaction volume. This loss of color can be measured by kinetic monitoring of loss of absorbency at 413 nm (wavelength that does not interfere with other molecules) over a minute using HRP as peroxidase and ascorbic acid (L-ASC) as a negative control. . The reaction medium is composed of 50 mM phosphate buffer pH 6, 7.5, 8, 2 mM H ₂ O ₂ , 2 mM ABTS, 0.25 μM HRP enzyme and sample at increasing concentrations.

The calculation of TAA is established according to the relationship between the slopes (Abs / min) of enzymatic tests in which the course of the reaction is estimated in the absence of antioxidants (positive control), and in the presence of different concentrations of substances with possible activity antioxidant Thus, the slope of the control kinetics would correspond to a TAA of zero percent, based on this the percentage of inhibition of the other curves.

Capacity as a lipid peroxidation inhibitor

Lipid peroxidation is a well-established mechanism of cell damage in plants and animals, as well as food spoilage (thickening). This process leads to the production of lipid peroxides and degradation aldehydes that leads to loss of cell membrane function and integrity. Porphyra 334 isolated from Porphyra leucosticta was studied as an inhibitor of lipid peroxidation in vitro using the β-carotene bleaching technique.

The method of decolorization of β-carotene is widely used to determine the antioxidant capacity of various substances in lipophilic medium, most of them extracted from fruits, vegetables and other products intended for food consumption in order to determine their greater or lesser degree of self-preservation in a natural state. It is a spectrophotometric method that measures the inhibition caused by an antioxidant on the discoloration of β-carotene in an aqueous system emulsified with Tween 20 and linoleic acid.

Linoleic acid self-oxidizes at a high rate in the presence of specially activated hydrogen atoms. Β-carotene, a precursor to vitamin A, is also known as a lipophilic antioxidant that prevents lipid peroxidation in membranes by sequestering singlet oxygen molecules and peroxyl lipid radicals. The β-carotene, when it is in the presence of linoleic acid, yields electrons by delaying the initiation stage of the linoleic acid self-oxidation process as well as limiting the propagation phase of the damage by simultaneously eliminating formed peroxidic radicals. If we add a new substance with possible antioxidant capacity to the reaction medium containing linoleic acid and β-carotene., this new substance will tend to oxidize it preferentially to β-carotene, competing with it for the sequestration of these radicals.

Β-carotene has a maximum absorption at 470 nm. This maximum varies when the molecule oxidizes since it loses double bonds and the structure of the molecule's chromophore is altered, thus losing its characteristic orange color and can be detected spectrophotometrically. The absorbency of the reaction medium will remain unchanged over time in the presence of antioxidant substances, with a drop in the absorbency of the sample when measured in the absence of antioxidants. Thus, the measurement of the antioxidant capacity of a substance will be inversely proportional to the slope drop of the curve that describes the oxidation of β-carotene (measured at the wavelength of

470 nm).

In this test, α-tocopherol (α-TOC) was used as a positive control.

The activity of the solution was evaluated according to the degree of discoloration of β-carotene, applying the formula proposed by Hidalgo et al. (1994, Phytochemistry, 37: 1585-1587) with some modifications:

AA = [P sample - P control / P Pattern - P control] ^• 100

P refers to the slopes of the obtained fading curves (Abs / time).

To do this, we adjust the part of the kinetic curve using linear regression that describes a linear behavior. The correlation coefficients for each replica of each sample were all greater than 0.98.

Abduction of superoxide radicals

The superoxides (O ₂ ^' ) radicals are mediators of autooxidation reactions of some compounds. Most of the time these oxidized compounds are characterized by having a characteristic absorption spectrum quantifiable by spectrophotometry. Pyrogallol (1,2,3-benzenotriol) is a substance that rapidly oxidizes in the presence of oxygen, especially in alkaline solutions. At pH 7.9 the SOD inhibits 99% of the reaction indicating a practically total participation of the superoxide anion O ₂ ^" in the reaction. The oxidized pyrogallol has a maximum absorption at 420 nm so that the ability of MAAs to sequester superoxide radicals was measured as loss of absorbency of spectrophotometrically monitored kinetic assays (Shimadzu UV 1603) during one minute of reaction.The protocol that was carried out was based on Marklund & Marklund (1974, Eur. J. Biochem., 47: 469-474 ) with some modifications.The reaction mixture contained 0.4 mM pyrogallol and MAA at different concentrations in 50 raM phosphate buffer at pH 8.2, containing 1 mM diethylenetriaminepentaacetic acid in a final volume of ImI incubation. The temperature was stable at 20 ± 1 ⁰ C. The positive control was the kinetic curve of generation of oxidized pyrogallol radicals in the absence of antioxidants to compare them with different concentrations of SOD as a known antioxidant. Dose-response relationships for the MAAs under study were determined at different concentrations. The sequestration capacity of superoxide radicals of the purified extracts was evaluated according to the following formula:

AA = 100 - [P sample - 100 / P control]

P refers to the slopes of the kinetic oxidation curves of pyrogallol (Abs

/ weather).

Claims

1. Purified extract of the mycosporin-type amino acid porφhyra-334 extracted from the red algae Porphyra leucosticta Ellis et Soland characterized by not presenting significant antioxidant activity as an inhibitor of the production of water-soluble free radicals in the pH range 6 - 8.5, for presenting a moderate antioxidant activity at the level of inhibition of lipid peroxidation, presenting at an concentration of 10 μM an activity close to 11% of which corresponds to a similar concentration of α-tocopherol; and for presenting significant antioxidant activity at the level of sequestration of superoxide radicals at a concentration equal to or greater than 200 μM, inhibiting approximately 46% the oxidation kinetics of pyrogallol when the concentration of the purified extract is 1 mM.

2. Use of the amino acid extract type mycosporin porphyra 334 extracted from the red algae Porphyra leucosticta according to claim 1 for the preparation of a product for the prevention and therapeutic treatment of coronary heart disease and atherosclerosis.

3. Use of the amino acid extract type mycosporin porphyra 334 extracted from red algae

Porphyra leucosticta according to claim 1 for the preparation of a product for the prevention of carcinogenic processes.

4. Use of the amino acid extract type mycosporin porphyra 334 extracted from the red algae Porphyra leucosticta according to claim 1 for the preparation of a product for the prevention and therapeutic treatment of Parkinson's and Alzheimer's.

5. Use of the porphyra 334 mycosporin-type amino acid extract extracted from the red seaweed Porphyra leucosticta according to claim 1 for the preparation of a product for the prevention of cataracts.

6. Use of the amino acid extract type mycosporin porphyra 334 extracted from the red algae Porphyra leucosticta according to claim 1 for the preparation of a product for the improvement in the performance of mountain sports practices and work performed at height in hypoxic conditions.

7. Use of the porphyra 334 mycosporin-type amino acid extract extracted from the red seaweed Porphyra leucosticta according to claim 1 for the preparation of a product for the prevention and treatment of depressive moods.

8. Use of the porphyra 334 mycosporin-type amino acid extract extracted from the red seaweed Porphyra leucosticta according to claim 1 for the preparation of a product for the prevention and therapeutic treatment of actinic erythema, photocarcinogenesis and photoaging.

9. Use of the porphyra 334 mycosporin-type amino acid extract extracted from the red seaweed Porphyra leucosticta according to claim 1 as an antioxidant or additive potential in the preparation of products in the food industry such as nutraceutical preparations or functional foods.

10. Use of the amino acid extract type mycosporin porphyra 334 extracted from red algae

Porphyra leucosticta according to claim 1 for the prevention of oxidation (deterioration) in cosmetic and pharmaceutical products.

11. Use of the amino acid extract type mycosporin porphyra 334 extracted from the red algae Porphyra leucosticta according to claim 1 for the preparation of parapharmacy products, pharmaceutical products, cosmetic products, nutraceutical preparations or functional foods for the therapeutic treatment of diseases and conditions related to the free radicals listed in claims 2, 3, 4, 5, 6, 7, 8.

12. Use of the porphyra 334 mycosporin-type amino acid extract extracted from the red seaweed Porphyra leucosticta according to the preceding claim for the preparation of parapharmacy products, pharmaceutical products or cosmetic products of topical application for the prevention of radical-related diseases and conditions free ones listed in claim 8.