Ulva australis extract with antioxidant activity and cell proliferation activity

Abstract

The present invention relates to an ulva australis extract with antioxidant activity and cell proliferation activity for use as a drug to prevent and / or treat neurodegenerative diseases of the retina, especially those associated with oxidative stress or loss of photoreceptors, such as age-related macular degeneration, glaucoma, inherited retinal dystrophies, diabetic retinopathy and retinitis pigmentosa.

Description

[0001] Ulva australis extract with antioxidant activity and cell proliferative activity

[0002] DESCRIPTION

[0003] TECHNICAL FIELD

[0004] The present invention falls within the field of biomedicine, specifically providing a natural source of antioxidant compounds that also have the ability to enhance cell proliferation of the photoreceptors of the retina, and therefore its use is proposed for the treatment of neurodegenerative diseases, especially those that affect the retina.

[0005] PRIOR ART

[0006] A common characteristic of all neurodegenerative diseases is the presence of a high degree of oxidative stress, as previously described in various neurodegenerative diseases, such as Alzheimer's disease or Parkinson's disease, among others.

[0007] Oxidative stress also plays a fundamental role in neurodegenerative diseases of the retina, such as age-related macular degeneration (AMD), glaucoma, or hereditary retinal dystrophies.

[0008] Age-related macular degeneration (AMD) is a complex and progressive chronic neurodegenerative disease characterized by drusenoid deposits in the retina, lipofuscin deposits, loss of retinal pigment epithelium cells, and choroidal neovascularization.

[0009] Glaucoma is an optic neuropathy characterized by the progressive degeneration of retinal ganglion cells (RGCs). Many modern therapeutic approaches have focused on reducing elevated intraocular pressure. Elevated intraocular pressure in the pathogenesis of glaucoma has been shown to increase endogenous reactive oxygen species within the trabecular meshwork. It has been postulated that this increased production of reactive oxygen species creates an imbalance between pro-oxidant and antioxidant capacity and is a crucial factor in early cell injury.

[0010] Inherited retinal dystrophies are characterized by the progressive degeneration of the retina, the retinal pigment epithelium (RPE), and the choriocapillaris, the vascular source of nutrients and oxygen. Retinal degeneration involves inflammation, oxidative stress, and cell death.

[0011] Other neurodegenerative diseases of the retina that, like those already mentioned, also involve the loss of retinal cells are diabetic retinopathy (DR) or retinitis pigmentosa (RP).

[0012] Diabetic retinopathy (DR) is a progressive chronic complication of type 1 or type 2 diabetes mellitus characterized by retinal neurodegeneration in the context of chronic diabetes. It is a microvascular complication of diabetes resulting from capillary damage, exhibiting a loss of penocytes and endothelial cells due to accelerated apoptosis. The reduction in the number of penocytes leads to various symptoms, including the presence of ghost cells, an increased number of acellularly occluded capillaries, the development of microaneurysms, and thickening of the capillary basement membrane.

[0013] Retinitis pigmentosa (RP) is a group of genetic eye disorders characterized by the progressive degeneration of the photoreceptor cells in the retina, leading to vision loss. Symptoms include difficulty seeing at night and decreased peripheral vision (side vision and upper or lower visual field). As peripheral vision worsens, individuals may experience tunnel vision.

[0014] The retina is one of the most metabolically active tissues and is exposed to high levels of light and oxygenation, an environment that fosters a high degree of oxidative stress. Photoreceptors are specialized light-sensitive neurons located in the outer retina of vertebrates and are classified into two subtypes: cones and rods. Photoreceptors are among the largest consumers of oxygen in the central nervous system, mainly due to the large accumulation of mitochondria in the ellipsoid. Because of the relationship between oxidative stress and the progression of neurodegenerative retinal diseases, the use of antioxidants is postulated as a possible treatment to prevent the progression of these diseases [1]. The administration of a wide variety of antioxidant substances, such as N-acetylcysteine, amide, thioredoxin, saffron, minocycline, melatonin, curcumin, quercetin, lutein (L), zeaxanthin (Z), catechins, and resveratrol,Dietary antioxidants, Fructus lycii, alpha-lipoic acid, (Z)-7,40-Dimethox¡-6- hydroxy¡-aurone-4-O-_-glucop¡ranóside¡de (DHAG), multi-target iron chelators, creatine, sulforaphane, NOS inhibitors; SigmaI R(+)-Pentazocine (PTZ) Ligand; Norbixin (bixin extracted from Bixa Orellana), (3R)-5,6,7-trihydroxy-3-isopropyl-3-methylisochroman-1-one, Glycyrrhizic acid / Glycyrrhizin; Tudca, Rasagiline, Norgestrel / Progesterone, Proinsulin, Anthocyanin Cyanidin-3-glucoside (C3G) Cranberry Extract, 4-Phenylbutyric Acid, Celastrol, Salvia miltiorrhiza Bunge, Vitamin A and / or E. has been tested in preclinical models and clinical trials [2, 3, 4].

[0015] Although compounds such as tauroursodeoxycholic acid (TlDCA) or natural compounds from saffron exist with antioxidant properties and that increase the proliferation of retinal cells, these are expensive and not always environmentally friendly. [5]

[0016] Algae of various species have been shown to be a reservoir of antioxidant compounds. In fact, compounds obtained from green, red, and brown algae have proven to be an effective therapy for preventing and treating neurodegenerative diseases. In the case of green algae, numerous studies have demonstrated their protective capacity against neurodegenerative diseases. Different extracts of Caulerpa racemosa [6], containing phenolic compounds and flavonoids, have been shown to increase cell viability in the SH-SY5Y cell line (a human neuroblastoma cell line used as an in vitro model of neuronal function and differentiation). Extracts of other species, such as Codium fragrans [7], have shown the ability to inhibit the cyclooxygenase-2 enzyme and reduce reactive nitrogen species and factors related to molecular stress.

[0017] Regarding the genus U. llva, methanol and methanol / water extracts of U. fasciata have shown antifungal and antibacterial activity [8], and the ethanol extract antioxidant activity in vitro [9]. For its part, the methanolic extract of U. reticulata has shown the ability to inhibit acetylcholinesterase, involved in the progression of some neurodegenerative diseases

[0010] , and the extract of U. lactuca has shown healing properties of bone tissue

[0011] . Sulfated polysaccharides of U. pertusa have shown a reduction of superoxide radicals in in vitro studies

[0012] .

[0018] However, the antioxidant potential of these extracts has not been tested in retinal cells, which are subjected to much greater oxidative stress.

[0019] Ulva australis Areschoug, 1854, or U. australis, is a species of the genus Ulva that is very abundant in the Mediterranean, especially in environments with high nutrient levels. Although the antioxidant and chelating capacity of U. australis extracts on various metals (iron and copper) has been demonstrated

[0013] , there are no studies on the antioxidant capacity of U. australis extracts in cell cultures or biological tissues. Specifically, no studies have been conducted on the antioxidant capacity of extracts of this alga on retinal cells.

[0020] DESCRIPTION OF THE INVENTION

[0021] The use of U. australis algae extract is proposed to protect retinal cells from oxidative damage and increase their cell proliferation, an effect not described in any of the other algal extracts with antioxidant effects mentioned.

[0022] The present invention demonstrates the antioxidant capacity of the U. australis extract on cells of the 661W cell line, an immortalized retinal cell line derived from murine cone photoreceptors. The ability of said extract to promote cell proliferation is also demonstrated.

[0023] The extract obtained from U. australis is an extract with natural antioxidant capacity, environmentally friendly, highly available and easy to obtain.

[0024] Thanks to its antioxidant capacity, proven in in vitro cell cultures, the use of U. australis extract helps reduce the effects of neurodegenerative diseases associated with oxidative stress, especially degenerative retinal diseases. This effect is enhanced by its ability to promote cell proliferation in retinal cells, thus facilitating cell regeneration in this organ.

[0025] Therefore, the U. australis extract described in the present invention helps to prevent, delay the progression and treat various neurodegenerative diseases that involve oxidative stress, specifically degenerative diseases of the retina, acting on the one hand by preventing cell death of cones and rods, and on the other hand by promoting their proliferation.

[0026] Therefore, a first aspect of the invention describes an extract of Ulva australis for use as a medicine.

[0027] The extract obtained from U. australis is a deep green color. Organic solvents, such as methanol or ethyl acetate, are preferably used for its preparation, instead of the aqueous solvents commonly used in the prior art. Preferably, the extract is obtained by a cooking-maceration method. Cooking allows for the extraction of compounds with a medium yield, while maceration allows for the extraction of heat-sensitive compounds. Other extraction techniques, such as Soxhlet extraction, damage heat-sensitive molecules because they require temperatures above 60°C. Therefore, the present method allows for a higher yield, a greater number of heat-sensitive molecules, and greater molecular integrity.

[0028] Regardless of the solvent used for its preparation, methanol or ethyl acetate, the extract of the invention is composed of various types of lipids, phenols, and flavonoids. The phenol content ranges from 35 to 50 pg gallic acid equivalents / mg dry weight of extract, and the flavonoid content ranges from 40 to 65 pg quercetin equivalents / mg dry weight of extract. The lipids, phenols, and flavonoids of the extract are preferably dissolved in dimethyl sulfoxide (DMSO).

[0029] A second aspect of the invention describes the composition comprising the Ulva australis extract for use as a medicine.

[0030] In a preferred embodiment of this aspect of the invention, the composition of the invention is a pharmaceutical composition. As used herein, the term "pharmaceutical composition" refers to any substance used for the diagnosis, prevention, relief, treatment, or cure of a disease in humans or animals. The pharmaceutical composition may comprise a single composition or separate compositions. The pharmaceutical composition of the invention may be used alone or in combination with other pharmaceutical compositions, preferably comprising another compound useful in the treatment of neurodegenerative diseases, more preferably neurodegenerative diseases of the retina, in which case it may also comprise a single composition or separate compositions. In one particular embodiment, the pharmaceutical composition of the invention further comprises a pharmaceutically acceptable carrier or excipient.

[0031] The term "pharmaceutically acceptable excipient" refers to a substance that aids in the absorption of the pharmaceutical composition comprising the composition of the invention, stabilizes said pharmaceutical composition, or assists in its manufacture by providing consistency, shape, flavor, or any other specific functional characteristic. Thus, excipients may have the function of binding the ingredients together, such as starches, sugars, or cellulose; a sweetening function; a coloring function; a protective function, such as isolating it from air and / or moisture; a filler function for a tablet, capsule, or any other form of formulation, such as dibasic calcium phosphate; or a disintegrating function to facilitate the dissolution of the components and their absorption, without excluding other types of excipients not mentioned in this paragraph.A "pharmaceutically acceptable carrier" (or "pharmacologically acceptable") refers to any substance, or combination of substances, known in the pharmaceutical industry, used in the manufacture of pharmaceutical dosage forms and includes, among others, solids, liquids, solvents, or surfactants. The carrier may be an inert substance or have an action similar to any of the compounds of the present invention, serving to facilitate the incorporation of the drug, as well as other compounds, allowing for improved dosage and administration, or providing consistency and form to the pharmaceutical composition. When the pharmaceutical form is liquid, the carrier is the diluent. The term "pharmacologically acceptable" refers to the fact that the compound in question is permitted and evaluated to be harmless to the organisms to which it is administered.

[0032] The pharmaceutical composition of the invention can be administered via any route of administration, and as such, said composition shall be formulated in the pharmaceutical form appropriate to the chosen route of administration. Thus, the pharmaceutical composition of the invention can be administered orally, nasally, ocularly, topically, intradermally, intracranially, intravenously, or intraperitoneally. Preferably, it is administered ocularly.

[0033] The composition of the invention may include an effective amount of Ulva australis extract. The term "effective amount" as used herein refers to an amount sufficient to effectively treat neurodegenerative diseases, more preferably neurodegenerative diseases of the retina. The effective amount may be appropriately selected by a person skilled in the art based on the individual being treated, and factors such as age, health condition, duration of administration, route of administration, excretion rate, and others may be involved, including the use of a drug in combination with or concurrently with the pharmaceutical composition, and other factors known in the field of medicine.Therapy is considered "personalized" when the compound administered to the individual to treat a disease is specially adapted to the genotypic and phenotypic characteristics of the individual to be treated, thus avoiding wasting time with ineffective therapies.

[0034] A third aspect of the invention describes the extract of Ulva australis and compositions comprising it for use as a medicament for the prevention and / or treatment of neurodegenerative diseases, preferably neurodegenerative diseases of the retina, and more preferably neurodegenerative diseases of the retina that involve oxidative stress and / or photoreceptor loss. These degenerative diseases of the retina include, but are not limited to, age-related macular degeneration, glaucoma, hereditary retinal dystrophies, diabetic retinopathy, and retinitis pigmentosa.

[0035] The terms "prevent" and "prevention" refer to avoiding totally or partially, or minimizing the adverse effects or the severity thereof, of a disease or condition, in this case, neurodegenerative diseases, and more preferably, neurodegenerative diseases of the retina.

[0036] The terms “treat” and “treatment,” as used here, refer to avoiding, lessening, or completely eliminating the effects of a disease or condition, in this case, neurodegenerative diseases, and more preferably, neurodegenerative diseases of the retina.

[0037] A fourth aspect of the invention describes the use of U. australis extract as an antioxidant and as a cell proliferation enhancer in in vitro cell cultures, especially in nerve cell cultures or neurons, and more specifically, photoreceptors.

[0038] A final aspect of the invention relates to a kit comprising the extract of U. australis for use in the treatment and / or prevention of neurodegenerative diseases, preferably neurodegenerative diseases of the retina.

[0039] The term "patient," "individual," or "subject," as used herein, refers to a mammal and includes, but is not limited to, domestic and farm animals, primates, and humans, e.g., humans, non-human primates, cows, horses, pigs, sheep, goats, dogs, cats, or rodents such as rats and mice. In a preferred embodiment, the subject is a human being.

[0040] Unless otherwise defined, all technical and scientific terms used herein have the same meaning commonly understood by a person skilled in the art to which this invention pertains. Similar or equivalent methods and materials to those described herein may be used in the practice of the present invention. Throughout the description and claims, the word "comprises" and its variations are not intended to exclude other technical features, additives, components, or steps. Other objects, advantages, and features of the invention will be evident to those skilled in the art upon examination of the description or may be discovered through the practice of the invention. The following examples and drawings are provided for illustrative purposes and are not intended to limit the scope of the present invention.

[0041] BRIEF DESCRIPTION OF THE FIGURES Figure 1: Antioxidant effect. Viability of cells from the 661W line subjected to oxidative stress by incubation with 0.1 mM NSP, maintained for 24 h, after incubation with A) methanol extract and B) ethyl acetate extract at the different concentrations indicated and evaluated by the XTT assay. Cell viability in the absence of NSP is shown as a control. C: control. NSP: 0.1 mM sodium nitroprusside. The concentrations of each extract are expressed in pg / ml. The letters indicate statistically significant differences. Mean value of the cell viability assay (XTT) in % with respect to the control, undamaged cells ± SD (n = 8 wells in each condition). * p <0.05, ANOVA and post-hoc test (TukeyHSD). The letters a, b, and c mark conditions where there are no significant differences between the results obtained.All of them show significant differences compared to the other conditions not marked with that specific letter.

[0042] Figure 2: Cell proliferation. Cell viability of cell line 661 W after incubation with A) methanol extract and B) ethyl acetate extract at the indicated concentrations. C: control. Extract concentrations are expressed in pg / ml. Letters indicate statistically significant differences. Mean cell viability assay (XTT) value in % with respect to the control, undamaged cells ± SD (n = 8 wells in each condition). * p < 0.05, ANOVA and post-hoc test (TukeyHSD). Letters a, b, and c mark conditions where there are no significant differences between the results obtained. All of them show significant differences with the other conditions not marked with that specific letter.

[0043] DETAILED EXPOSURE OF MODES OF REALIZATION

[0044] Obtaining the extract of U. australis

[0045] To prepare the U. australis extract, the algal biomass is preferably cleaned several times with reverse osmosis water to remove epiphytes and salts. It is then freeze-dried for 48 hours. Once freeze-dried, it is ground with liquid nitrogen and extracted in methanol or ethyl acetate in a 1:10 ratio (dry weight / volume). The mixture is then boiled at 40 °C for 2 hours with stirring. Afterward, it is macerated for 48 hours at 4 °C with continuous stirring. Following maceration, the mixture is filtered through a Whatman No. 1 filter. The resulting extract is filtered through a 0.22 µm pore diameter filter. The solvent is evaporated using a rotary evaporator or Speedvac at 35 °C until a solid residue is obtained. Finally, the solid residue is resuspended in DMSO to achieve a final concentration of 100 mg of dry residue / mL.

[0046] The lipid composition of the methanol extract, analyzed by HPLC mass-mass spectrometry, in decreasing order of proportion, is: diacylglycerol trimethylhomoserin (22.70%), triacylglycerol (17.46%), fatty acids (11.79%), lyso-diacylglycerol trimethylhomoserin (8.73%), ceramide alpha-hydroxylated fatty acid dihydrosphingosine (5.24%), ceramide alpha-hydroxylated fatty acid phytosphingosine (4.80%), ceramide non-hydroxylated fatty acid dihydrosphingosine (3.93%), diacylglycerol (3.93%), hydroxylated fatty acid (3.05%), sulfoquinovosyl diacylglycerol (3.05%), monogalactosyldiacylglycerol (2.62%), ceramide alpha-hydroxylated fatty acid sphingosine (2.18%), ceramide non-hydroxylated fatty acid-phytosphingosine (2.18%), digalactosyldiacylglycerol (1.74%), lyso-phosphatidylglycerol (1.31%), phosphatidylglycerol (1.31%), sulfoquinovosyl diacylglycerol (1.31%), ceramide non-hydroxylated fatty acid-sphingosine (0.87%), phosphatidylinositol (0.87%), lyso-phosphatidylinositol (0.43%), phosphatidylglycerol (0.43%),

[0047] The lipid composition of the ethyl acetate extract, analyzed by HPLC mass-mass spectrometry, in decreasing order of proportion, is: diacylglycerol trimethylhomoserine (21.05%), triacylglycerol (17.54%), fatty acids (15.79%), ceramide alpha-hydroxylated fatty acid-dihydrosphingosine (5.84%), ceramide non-hydroxylated fatty acid-dihydrosphingosine (5.26%), ceramide alpha-hydroxylated fatty acid-phytosphingosine (5.26%), diacylglycerol (4.09%), monogalactosyldiacylglycerol (4.09%), lyso-diacylglycerol trimethylhomoserine (3.50%), hydroxylated fatty acid (3.50%), ceramide alpha-hydroxylated fatty acid-sphingosine (2.92%), phosphatidylinositol (2.92%). digalactosyldiacylglycerol (2.34%), fatty acid acyl esters (1.75%), ceramide non-hydroxylated fatty acid-sphingosine (1.75%), phosphatidylglycerol (0.58%), acylated glucosylacylglycerol (0.58%), sulfoquinovosyl diacylglycerol (0.58%) and monogalactosyldiacylglycerol homologue (0.58%).

[0048] Effects of U. australis extract on the retina-derived 661W cell line

[0049] The antioxidant effects of the extract and its protective effect against retinal degeneration were studied in the 661W cell line, derived from photoreceptors. The 661W cell line was obtained from retinal tumors of transgenic mice expressing the SV40 T antigen under the control of the IRBP gene promoter (PBR-3). It is a homogeneous cell line that expresses several markers of cone-type photoreceptors: opsins, transducin, and X-arrestin. A wealth of information is available on the pathways activated in these cells in response to various apoptotic stimuli, and they have been successfully used as a tool in research associated with retinal dystrophies.

[0050] The 661W cell line was cultured in DMEM-glutamax medium supplemented with 10% heat-inactivated fetal bovine serum (FBS) at 37 °C in a humidified atmosphere of 5% CO2. To assess the antioxidant effect of methanol and ethyl acetate extracts of U. australis, the 661W cell line was subjected to damage by the oxidizing agent sodium nitroprusside (SNP), and cell viability was measured in the presence and absence of the extracts. Cell cultures were pre-incubated for 2 h with different concentrations of the extracts (50–800 µg / ml). After the pre-incubation period, the treated cells were exposed to a toxic stimulus (100 µM sodium nitroprusside), and cell survival was measured after 24 h of incubation with the toxic stimulus.

[0051] To assess the ability of methanol and ethyl acetate extracts of U. australis to increase cell proliferation, the 661W cell line was incubated for 2 h with different concentrations of the extracts (50-800 µg / ml). Cell viability was measured after the incubation period.

[0052] In both cases, cell viability was assessed by measuring the reducing capacity of compound XTT (2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-5-[(phenylamino)carbonyl]-2H-tetrazolium hydroxide), which was quantified spectrophotometrically according to the manufacturer's instructions.

[0053] Antioxidant effect

[0054] The methanol extract (Figure 1A) significantly increased neuronal cell survival at a concentration of 400 .g / ml, while the ethyl acetate extract (Figure 1B) did so at a concentration of 50 to 600 .g / ml.

[0055] Cell proliferation

[0056] The methanol extract (Figure 2A) increased cell proliferation in a statistically significant way at a concentration of 100-400 .g / ml, while the ethyl acetate extract (Figure 2B) did so at a concentration of 200 .g / ml.

[0057] It is therefore concluded that the U. australis extract has antioxidant activity in vitro in cell cultures, making its use as an antioxidant in the medical field possible.

[0058] Furthermore, its effect in reducing oxidative damage in the 661W cell line supports its use in preventing and treating neurodegenerative diseases associated with oxidative stress, especially those affecting photoreceptors (specialized neuronal cells), such as neurodegenerative diseases of the retina. Moreover, its cell proliferation-promoting action reinforces the interest in this extract for use in the prevention and treatment of retinal diseases, particularly neurodegenerative diseases involving photoreceptor loss.

[0059] References

[0060] 1. Wang P, Chin EK, Almeida D. Antioxidants for the Treatment of Retinal Disease: Summary of Recent Evidence. Clin Ophthalmol. 2021 Apr 19;15:1621-1628. doi:10.2147 / OPTH.S307009. PMID: 33907376; PMCID: PMC8064715.

[0061] 2. Pinilla I, Maneu V, Campello L, Fernández-Sánchez L, Martinez-Gil N, Kutsyr O, Sánchez- Sáez X, Sánchez-Castillo C, Lax P, Cuenca N. Inherited Retinal Dystrophies: Role of Oxidative Stress and Inflammation in Their Physiopathology and Therapeutic Implications. Antioxidants (Basel). 2022 May 30; 11 (6): 1086. doi: 10.3390 / antioxl 1061086. PMID: 35739983; PMCID: PMC9219848.

[0062] 3. B Doménech E, Marfany G. The Relevance of Oxidative Stress in the Pathogenesis and Therapy of Retinal Dystrophies. Antioxidants (Basel). 2020 Apr 23;9(4):347. doi: 10.3390 / antiox9040347. PMID: 32340220; PMCID: PMC7222416.

[0063] 4. Pinilla I, Maneu V. Oxidative Stress as a Main Contributor of Retinal Degenerative Diseases. Antioxidants (Basel). 2022 Jun 17; 11 (6): 1190. doi: 10.3390 / antiox11061190. PMID: 35740087; PMCID: PMC9229683.

[0064] 5. Fernández-Sánchez L, Lax P, Noailles A, Angulo A, Maneu V, Cuenca N. Natural Compounds from Saffron and Bear Bile Prevent Vision Loss and Retinal Degeneration. Molecules. 2015 Jul 31 ;20(8): 13875-93. doi: 10.3390 / molecules200813875. PMID: 26263962; PMCID: PMC6332441.

[0065] 6. Yang P, Liu DQ, Liang TJ, Li J, Zhang HY, Liu AH, Guo YW, Mao SC. Bioactive constituents from the green alga Caulerpa racemosa. Bioorg Med Chem. 2015 Jan 1 ;23(1):38-45. doi: 10.1016 / j.bmc.2014.11.031. Epub 2014 Nov 27. PMID: 25497963.

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[0067] 9. Rengasamy, Kannan & Amoo, Stephen & Aremu, Adeyemi & Stirk, Wendy & Gruz, Jiri & Subrtová, Michaela & Dolezal, Karel & van Staden, Johannes. (2014). Phenolic profiles, antioxidant capacity, and acetylcholinesterase inhibitory activity of eight South African seaweeds. Journal of Applied Phycology. 27. 10.1007 / s10811-014-0438-8.

[0068] [ PubMed ] [ Cross Ref ] 10. Suganthy N, Karutha Pandian S, Pandima Devi K. Cholinesterase inhibitory effect of Hypnea valentiae and lllva reticulata. Neurosci Lett. 2010 Jan 14;468(3):216-9. doi: 10.1016 / j.neulet.2009.11.001. Epub 2009 Nov 6. PMID:19897016.

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Claims

CLAIMS 1. Ulva australis extract for use in the prevention and / or treatment of neurodegenerative diseases of the retina.

2. Extract of Ulva australis for use according to the preceding claim, characterized in that it is obtained by maceration-cooking.

3. Extract of Ulva australis for use according to the preceding claim, characterized in that the maceration-cooking is carried out in an organic solvent 4. Extract of Ulva australis for use according to the preceding claim, characterized in that the organic solvent is selected from methanol, ethyl acetate, or a mixture of both.

5. Ulva australis extract for use according to any of claims 2 to 4 characterized in being resuspended in DMSO.

6. Composition comprising the extract of Ulva australis for use according to any of claims 1 to 5.

7. Ulva australis extract for use according to any of claims 1 to 5 and composition for use according to claim 6 characterized in that neurodegenerative diseases of the retina are associated with oxidative stress.

8. Extract of Ulva australis for use according to any of claims 1 to 5 and composition for use according to claim 6 characterized in that neurodegenerative diseases of the retina involve loss of photoreceptors.

9. Ulva australis extract for use according to any of claims 1 to 5 and composition for use according to claim 6 characterized in that the degenerative diseases of the retina are selected from the list comprising: age-related macular degeneration, glaucoma, hereditary retinal dystrophies, diabetic retinopathy and retinitis pigmentosa.

10. Non-therapeutic use of an extract of Ulva australis as an enhancer of cell proliferation in vitro.

11. Non-therapeutic use of an extract of Ulva australis according to the preceding claim, characterized in that the extract of Ulva australis is obtained by maceration-cooking.

12. Non-therapeutic use of an extract of Ulva australis according to the preceding claim, characterized in that the maceration-cooking is carried out in an organic solvent 13. Non-therapeutic use of an extract of Ulva australis according to the preceding claim, characterized in that the organic solvent is selected from methanol, ethyl acetate, or a mixture of both.

14. Non-therapeutic use of an extract of Ulva australis according to any of claims 11 to 13 characterized by being resuspended in DMSO.

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