ES2428665A1 - Composition for use in the treatment and/or prevention of inflammation, oxidative stress and ocular neovascularization - Google Patents

Abstract

Composition for use in the treatment and/or prevention of inflammation, oxidative stress and ocular neovascularization. The present invention refers to the use of quercetin (QCT) or a composition comprising QCT and at least one other polyphenolic compound, preferably resveratrol (RES), for the treatment and/or prevention of injuries or diseases that occur with inflammation, stress oxidative and/or ocular neovascularization, preferably from the anterior segment, more preferably from the ocular surface and even more preferably from the corneal epithelium. The invention also relates to the use of QCT and said composition as an antimicrobial, antioxidant and anti-inflammatory agent in contact lens cleaning and maintenance solutions.

Description

Composition for use in the treatment and / or prevention of inflammation, oxidative stress and ocular neovascularization

The present invention falls within the field of ophthalmology, specifically within the ophthalmic compositions useful for the treatment and / or prevention of lesions or diseases that occur with inflammation, oxidative stress and / or ocular neovascularization, preferably of the tear system (secretor and excretory), eye attachments (eyelids, including the Meibomian glands) and the anterior segment of the eye, which includes conjunctiva (including mucin-producing cells and glands of the secondary tear system), cornea, sclerocorneal limbus, iris, pupil, crystalline, Zinn zonula, ciliary body, anterior chamber, aqueous humor, posterior chamber; and more preferably of the ocular surface, which comprises the epithelia of the cornea, sclero-corneal and conjunctive limbus, the overlying tear film and the stroma underlying these structures.

STATE OF THE TECHNIQUE

Ocular inflammation is one of the problems with the highest incidence in ophthalmic pathology. The treatment of this condition can be performed specifically, attacking the etiology, unspecifically, reducing inflammatory symptoms, or joining both. In this sense, corticosteroids have been used as drugs of choice in the nonspecific treatment of ocular inflammation, but the adverse effects they present have raised the need to increase the use of other drugs such as non-steroidal anti-inflammatory drugs (NSAIDs) (Thadani SM ., Foster CS., 2004, Pediatr Drugs, 6: 289-301). The anti-inflammatory activity of these latter agents lies in the reversible inhibition of the activity of the enzyme cyclooxygenase (COX) in its COX-1 and COX-2 isoforms, preventing the formation of prostaglandins, substances that mediate inflammation.

The eye inflammation may be due to various causes and have some characteristics or others depending on the area of the eye in which it is triggered. It can be caused, for example, by a surgical intervention, however, this is only one of the causes of the pathology, which must be treated with anti-inflammatory in any of its manifestations to alleviate the symptoms and avoid the serious complications that can lead to and which can worsen the quality of life of patients. These complications depend on the location of the inflammation and can range from the loss of the eyelashes, if it occurs on the eyelids, worsening of the prognosis of eye injuries and even blindness in the case of uveitis or intraocular inflammatory pathology.

Inflammation can occur in all structures of the eye anatomy. Thus, both the eye attachments and the eyeball or optic nerve could be affected, and inflammation of both infectious and non-infectious origin could be found. And so, we will talk about orbital, palpebral, intraocular inflammations (uveitis, also called, depending on the parts it predominantly affects, retinitis, choroiditis, papillitis or vasculitis) or inflammations of the ocular surface (blepharitis, conjunctivitis, keratitis, affections of the sclero-corneal limbo and tear pathology, dry eye syndrome (SOS) included).

It is estimated that around 7% of young people suffer from some phenomenon of ocular inflammation in Spain, while in adulthood the appearance of other diseases that cause this phenomenon, such as SOS, causes its prevalence to increase up to 30%. from 60 years.

SOS is an inflammatory, chronic, multifactorial inflammatory disease that could originate in any of the components of the lacrimal functional unit, formed by the ocular surface, the main and accessory lacrimal glands and the innervation of interconnection between them - afferent branches and efferent - and that, at some point, causes damage to the ocular surface (corneal epithelia, sclero-corneal and conjunctive limbus, overlying tear film and stroma underlying these structures) that are detected with suitable tests.

As for ocular neovascularization, it is directly linked to the stimulation of hypoxia, tissue necrosis and the activation of complex cascades of interaction between cytokines / chemokines (Ex: IL-8) and growth factors stimulating the formation of neo-vessels (VEGF, bFGF, TGF-a, PDGF, IGF). All these events are triggered, more frequently, by intense and / or prolonged and / or recurrent inflammation, regardless of the cause that causes it. For example, an acute and very intense event such as caustication (chemical burn) or a less intense but recurring event (herpetic keratitis) or prolonged (severe SOS) may be the origin of a corneal neovascularization. In recent years, research studies in neovascularization have shown a direct interrelation between cytokines, growth factors, cell adhesion molecules (integrins) and metalloproteinases (MMP). These MMPs play an essential role in tissue invasion by neovases in different corneal diseases (inflammatory, autoimmune, infectious), so that the inhibition of MMPs by natural inhibitors (or TIMPs) and MMP inhibitor drugs, would result in neovascularization inhibition.

One of the cases in which neovascularization may occur is, for example, when extensive destruction of the corneal surface occurs, and especially of the pluripotential stem cells located at the level of the sclero-corneal limbus, which would lead to the loss of the normal corneal re-epithelialization and finally a conjunctivalization or corneal pannus with a neovasal invasion. This process, at present, is defined as Limbic Insufficiency Syndrome (SIL).

Experimental findings (in vitro and in vivo), which allow a better understanding of the molecular biology of neovascularization and scarring, have helped to carry out new medical treatments with drugs that inhibit angiogenic growth factors or MMPs, as well as with the use of new growth factors that inhibit neovascularization ("Pigment Epithelium Derived Factor" or PEDF), and by surgical treatments such as transplantation of limbal cells and amniotic membrane.

With regard to oxidative stress, reactive oxygen species (ROS), which produce adverse effects at the cellular level, are generated by multiple endogenous mechanisms, such as inflammatory reactions or ischemia; and exogenous, such as exposure to ultraviolet light. At the ocular level, it is worth mentioning for example the inflammatory process of the lacrimal functional unit that occurs in the SOS, which has an oxidative process associated with it taking place on the ocular surface.

In reference to the tear film, the lipid layer is formed by cholesterol esters and phospholipids. If we take into account that the lipid layer is in direct contact with the air and the oxidation processes that lipids undergo in the presence of radical molecules, the generation of ROS in the inflammatory oxidative process can degrade fatty acids, increasing the evaporation of the aqueous layer of tear film. This is corroborated by the increase in the concentration of lipid peroxidase on the ocular surface.

In addition to ROS, there is also the formation of oxidizing species related to nitrogen. Nitric oxide and oxidizing species related to nitrogen can play a very important role in SOS. The expression of nitric oxide synthase 2 and 3 (NOS2, NOS3) in the conjunctival epithelium is increased in the SOS compared to the conjunctival epithelium of healthy eyes, which aggravates the symptomatology.

This oxidative process produces a high concentration, and therefore a high activity, of myeloperoxidase (MPO) and xanthine oxidoreductase / xanthine oxidase in the conjunctival epithelium, among others. These enzymes generate radical species that could be involved in the oxidative processes that occur in the SOS. In addition, the concentration of antioxidant enzymes, responsible for neutralizing radical molecules, is decreased in the dry eye, which could further contribute to the oxidative damage produced in the inflammatory process.

On the other hand, polyphenolic compounds, within which flavonoid and non-flavonoid polyphenols are found, are compounds whose main characteristic is the presence of benzyl and hydroxyl groups in their structure and whose antioxidant properties have been extensively studied. Among the flavonoid polyphenolic compounds, the quercetin molecule (QCT), 3,3 ', 4', 5,7-pentahydroxy-2-phenylchromen-4-one stands out. It is the most abundant flavonoid compound, with many potentially beneficial effects on human health (Kelly GS, 2011, Alternative Medicine Review, 16 (2): 172-194). Similarly, within the non-flavonoid polyphenolic compounds, within which are the stilbenoids that are compounds whose main characteristic is the presence of benzyl and hydroxyl groups in their structure, it is worth highlighting a molecule whose importance has increased in recent years also due to its multiple qualities, uses and health benefits (Frémon L., 2000, Life Sciences, 66 (8): 663-673), the resveratrol molecule (RES), 5 - [(E) -2- ( 4-hydroxyphenyl) ethenyl] benzen-1,3-diol. Both compounds can be obtained synthetically but are found naturally in more than 70 species of plants, fruits and vegetables.

As for its clinical application, both polyphenols in general and those derived from stilbene or stilbeneides in particular are molecules used for different treatments. In this regard, some studies indicate that RES has several health benefits, such as the prevention of cardiovascular, cancer and inflammatory problems (Das S., Das DK, 2007, Inflamm Allergy Drug Targets, 6 (3): 168-173 ). For these reasons, some ophthalmic compositions comprising polyphenolic compounds for the treatment of ocular inflammation have been described, such as that described in US20060127505.

However, because a large number of eye diseases and lesions frequently occur with inflammatory processes, oxidative stress and / or ocular neovascularization, the identification of ophthalmic compounds and compositions that allow treating and / or ophthalmic compositions continues to exist as an objective in the field of ophthalmology. or efficiently prevent such pathological processes.

DESCRIPTION OF THE INVENTION

The present invention proposes the use of quercetin (QCT), or any of its salts or derivatives, for the treatment and / or prevention of inflammation, oxidative stress and / or ocular neovascularization, preferably of the anterior segment, more preferably of the ocular surface, and even more preferably of the corneal epithelium. Furthermore, the present invention provides a composition, hereinafter "composition of the invention", comprising QCT, or any of its salts or derivatives, and at least one other polyphenolic compound, or any of its salts or derivatives, preferably resveratrol ( RES), which is also useful in such clinical application.

The QCT, or any of its salts or derivatives, as well as the composition of the invention, are therefore useful for the

5 treatment and / or prevention of inflammation, oxidative stress and ocular neovascularization, preferably of the anterior segment, more preferably of the ocular surface, and even more preferably of the corneal epithelium. As the examples of the present invention demonstrate, both QCT alone and said composition are capable of reducing the levels of, for example, but not limited to, cytokines, chemokines, cyclooxygenase 2 (COX-2) and oxidative species in, for example, but not limited to, human corneal epithelium (HCE) cells stimulated with TNF-a or

10 irradiated with ultraviolet light. They are also able to significantly decrease the amount of CD4 + lymphocytes in conjunctiva in animal models of dry eye syndrome (SOS).

Therefore, a first aspect of the invention relates to the use of QCT, or any of its salts or derivatives, for the preparation of a medicament for the treatment and / or prevention of ocular inflammation, preferably of the

15 ocular surface, more preferably of the corneal epithelium. Alternatively, this aspect of the invention relates to QCT, or any of its salts or derivatives, for use as a medicine in the treatment and / or prevention of ocular inflammation, preferably of the ocular surface, more preferably of the corneal epithelium.

"Quercetin" or "QCT" or "3,3 ', 4', 5,7-pentahydroxy-2-phenylchromen-4-one" is a flavonoid polyphenolic compound and

20 natural, which is found in high concentrations in a wide variety of plant species and, therefore, in foods such as, for example, but not limited to, fruit, vegetables, cereals, tea or red wine; although it can also be chemically synthesized. Its molecular formula is C15H10O7 and its chemical structure (I) is as follows:

(I)

Thus, the medicine comprising QCT, or any of its salts or derivatives, is useful for the treatment and / or prevention of diseases or injuries that occur with ocular inflammation.

Another aspect of the invention relates to the use of QCT, or any of its salts or derivatives, for the preparation of a medicament for the treatment and / or prevention of ocular oxidative stress, preferably of the ocular surface, more preferably of the corneal epithelium Alternatively, this aspect of the invention relates to QCT, or any of its salts or derivatives, for use as a medicine in the treatment and / or prevention of stress.

Ocular oxidative, preferably of the ocular surface, more preferably of the corneal epithelium.

Thus, the medicine comprising QCT, or any of its salts or derivatives, is useful for the treatment and / or prevention of diseases or lesions that occur with ocular oxidative stress.

Another aspect of the invention relates to the use of QCT, or any of its salts or derivatives, for the preparation of a medicament for the treatment and / or prevention of ocular neovascularization, preferably of the anterior segment, more preferably of the ocular surface, even more preferably of the cornea. Alternatively, this aspect of the invention relates to QCT, or any of its salts or derivatives, for use as a medicine in the treatment and / or prevention of ocular neovascularization, preferably of the anterior segment, more

45 preferably of the ocular surface, even more preferably of the cornea.

Thus, the medicine comprising QCT, or any of its salts or derivatives, is useful for the treatment and / or prevention of diseases or lesions that occur with ocular neovascularization.

In another preferred embodiment, the medicament comprising QCT, or any of its salts or derivatives, is for the treatment and / or prevention of dry eye syndrome (SOS), blepharitis, dysfunction of the Meibomian glands, meibomitis, allergic processes ocular, corneal dystrophies, conjunctivitis, alteration of the ocular surface by the use of contact lenses, autoimmune diseases that affect the anterior segment of the eye, post-surgery of the anterior segment, burns (both chemical or “caustic” or thermal) or injuries or pathologies eyepieces produced by ultraviolet radiation. More preferably, the medicament comprising QCT, or any of its salts or derivatives, is for the treatment and / or prevention of SOS.

Another aspect of the invention relates to the use of QCT, or any of its salts or derivatives, as an antimicrobial, antioxidant and anti-inflammatory agent in contact lens cleaning and maintenance solutions. Another aspect of the invention relates to a solution for cleaning and maintaining contact lenses comprising QCT or any of its salts or derivatives.

A second aspect of the invention relates to a composition, "composition of the invention", comprising QCT,

or any of its salts or derivatives, and at least one other polyphenolic compound, or any of its salts or derivatives.

The term "other polyphenolic compound" used in the present invention refers to a secondary metabolite.

15 biosynthesized by, but not limited to, organisms from the plant kingdom that can be found in, for example, foods derived from plant sources, but which can also be obtained synthetically, and which has more than one phenol group. Polyphenols are a group of molecules whose main characteristic is the presence of benzyl and hydroxyl groups in their structure. These polyphenols can be broadly classified as: phenolic acids, flavonoids, stilbenes and lignans. In turn, flavonoid compounds can be subclassified as chalconas, flavonols,

20 flavones, flavanones, anthocyanidins and isoflavonoids. The other polyphenolic compound of the composition of the invention may be, but not limited to, any polyphenolic compound within the four families. Therefore, in a preferred embodiment, the other polyphenolic compound comprised in the composition of the invention is a stilbenoid, more preferably the RES, or any of its salts or derivatives.

A "stilbenoid compound" is any compound whose principal structure is the stilbene (II) molecule, such

(II)

"Resveratrol", "RES" or "5 - [(E) -2- (4-hydroxyphenyl) ethenyl] benzene-1,3-diol" is found in a wide variety of plants, although it can also be chemically synthesized. Its CAS number is 501-36-0, its molecular formula is C14H12O3 and

35 (III)

As used herein, the term "derivative" includes pharmaceutically acceptable compounds, that is, derivatives of the QCT or the other polyphenolic compound comprised in the composition of the invention, preferably RES, which can be used in making the composition. of the invention, already pharmaceutically acceptable derivatives, since these may be useful in the preparation of pharmaceutically acceptable derivatives.

In a more preferred embodiment, the composition of the invention further comprises a pharmaceutically acceptable carrier. In addition, said composition may comprise one or more excipients.

The term "excipient" refers to a substance that helps the absorption of the elements of the composition of the invention, stabilizes said elements, activates or aids the preparation of the composition in the sense of giving it consistency or providing flavors that make it nicer. Thus, the excipients could have the function of keeping the ingredients together, such as, for example, starches, sugars or cellulose, the sweetening function, the coloring function, the protective function of the composition, for example, to isolate it from air and / or moisture, the filling function of a pill, capsule or any other form of presentation, the disintegrating function to facilitate the dissolution of the components and their absorption in the intestine, without excluding other types of excipients. mentioned in this paragraph.

The "pharmaceutically acceptable carrier", like the excipient, is a substance that is used in the composition to dilute any of the components included therein to a certain volume or weight. The pharmaceutically acceptable carrier is an inert substance or action analogous to any of the elements included in the composition of the present invention. The function of the vehicle is to facilitate the incorporation of other elements, allow a better dosage and administration or give consistency and form to the composition.

Preferably, the composition of the invention comprises QCT, or any of its salts or derivatives, and at least one other polyphenolic compound, or any of its salts or derivatives, preferably RES, in a therapeutically effective amount, meaning "therapeutically effective amount" the level, quantity or concentration of QCT, or of any of its salts or derivatives, and of at least one other polyphenolic compound, or of any of its salts or derivatives, preferably of RES, that produces the desired effect by treating and / or preventing inflammation , oxidative stress and / or ocular neovascularization, without causing adverse effects. The dosage to obtain a therapeutically effective amount depends on a variety of factors, such as, for example, the age, weight, sex or tolerance of the individual to whom the composition of the invention is to be administered.

The composition of the present invention can be formulated for administration in a variety of ways known in the state of the art. Examples of preparations include any solid composition (tablets, pills, capsules, granules, etc.) or liquid (solutions, suspensions or emulsions) for oral, topical or parenteral administration. The composition of the present invention may also be in the form of sustained release formulations of drugs or any other conventional release system, so it may be contained, but not limited to, in nanoparticles, liposomes or nanospheres, in a polymeric material, in a polymeric material. Biodegradable or non-biodegradable implant or in biodegradable microparticles, such as biodegradable microspheres.

Such a composition and / or its formulations may be administered to an animal, including a mammal and, therefore, to man, in a variety of ways, including, but not limited to, intraperitoneal, intravenous, intradermal, intraspinal, intrastromal, intraarticular, intrasynovial, intrathecal, intralesional, intraarterial, intramuscular, intranasal, intracranial, subcutaneous, intraorbital, intracapsular, topical, using transdermal patches, percutaneous, nasal spray, surgical implant, internal surgical paint or infusion pump.

In an even more preferred embodiment, the composition of the invention is formulated for ophthalmic administration. The term "formulated for ophthalmic administration" refers to a formulation that allows the composition of the invention to be administered ocularly, for example, but not limited to, topically

or intraocularly, without such administration negatively affecting the properties, for example, optical and / or physiological, of the eye. Examples of the composition of the invention formulated for ophthalmic administration are, but not limited to, said composition associated with water, salts, a polymeric or semi-solid liquid carrier, a phosphate buffer or any other ophthalmically acceptable liquid carrier of known in the state of the art.

As explained above, the composition of the invention is capable of reducing the levels of, for example, but not limited to, cytokines, chemokines and COX-2, factors involved in the inflammatory process, in ocular cells that have an inflamed phenotype, as well as oxidative species in eye cells that have been induced to suffer oxidative stress. Therefore, the composition of the invention is useful for the treatment and / or prevention of inflammation, oxidative stress and ocular neovascularization, preferably of the anterior segment, more preferably of the ocular surface, and even more preferably of the corneal epithelium.

Thus, another aspect of the invention relates to the use of the composition of the invention for the preparation of a medicament or, alternatively, to the composition of the invention for use as a medicament, hereinafter, "medicament of the invention" .

The "medicaments" referred to in the present invention may be for human or veterinary use. The "medicine for human use" is any substance or combination of substances that is presented as having properties for the treatment or prevention of diseases in humans or that can be used in humans or administered to humans in order to restore, correct or modify physiological functions by exerting a pharmacological, immunological or metabolic action, or establishing a medical diagnosis. The "veterinary medicinal product" means any substance or combination of substances that is presented as having curative or preventive properties with respect to animal diseases or that can be administered to the animal in order to restore, correct or modify its physiological functions by exercising a pharmacological, immunological or metabolic action, or to establish a veterinary diagnosis.

In a preferred embodiment of this aspect of the invention, the medicament is for the treatment and / or prevention of ocular inflammation.

By "ocular inflammation" is meant the inflammation produced in any ocular structure, including all structures of the anterior segment, the posterior segment of the eye and the eye appendages, for example, but not limited to, in the optic nerve, eyelids, main tear gland , hialoid duct, retina, choroid, sclera, ocular musculature or in any structure that is part of the anterior segment, including the ocular surface. “Anterior segment” means any structure selected from the list that includes: conjunctiva, cornea, sclerocorneal limbus, iris, pupil, lens, Zinn zonula, ciliary body, anterior chamber, aqueous humor, posterior chamber, Meibomian glands, glands mucous or lacrimal apparatus (lacrimal glands, except for the main lacrimal gland, nasolacrimal duct and lacrimal sac). The ocular inflammation presents with symptoms such as, for example, but not limited to, pain, redness and swelling of the affected tissue, as well as with the overexpression of factors involved in the inflammatory process such as, for example, but not limited to, IL-6, IL -8, IP-10, VEGF, TNF-a, COX-1 or COX-2.

In a more preferred embodiment, the medicament of the invention is for the treatment and / or prevention of inflammation of the anterior segment and / or of the main lacrimal gland and / or eyelids. In an even more preferred embodiment, the medicament of the invention is for the treatment and / or prevention of inflammation of the ocular surface. By "ocular surface" is meant any structure selected from the list comprising: epithelia of the cornea, sclero-corneal or conjunctive limbus, overlying tear film or stroma underlying these structures. In an even more preferred embodiment, the medicament of the invention is for the treatment and / or prevention of inflammation of the corneal epithelium.

Eye inflammation is associated with a variety of eye diseases and injuries, such as, but not limited to, Limbic Insufficiency Syndrome (SIL), SOS, blepharitis, dysfunction of the Meibomian glands, meibomitis, allergic eye processes, conjunctivitis, alteration of the ocular surface, preferably of the corneal epithelium, caused by the use of contact lenses and their cleaning and maintenance systems, autoimmune diseases, preferably affecting the anterior segment of the eye, such as, but not limited to, Sjögren's syndrome , post-surgery, preferably of the anterior segment of the eye, burns (both chemical or caustic, and thermal) or ocular lesions or pathologies caused by ultraviolet radiation. Therefore, the medicament of the invention is useful for the treatment and / or prevention of diseases or lesions that occur with ocular inflammation, preferably, of those indicated in this paragraph.

In another preferred embodiment, the medicament of the invention is for the treatment and / or prevention of ocular oxidative stress.

By "ocular oxidative stress" is understood the cytotoxicity condition that is the result of an imbalance between the production of free radicals and the ability of the cell to defend against them, which is why it is caused by an increase in the formation of said free radicals. or by a decrease in agents that act as antioxidants, or for both reasons together. Ocular oxidative stress can occur in any ocular structure, including all structures of the anterior segment, the posterior segment of the eye and the eye appendages, for example, but not limited to, in the optic nerve, main lacrimal gland, hialoid duct, retina, choroid, sclera, ocular musculature or any structure that is part of the anterior segment, including the ocular surface. In a more preferred embodiment, the medicament of the invention is for the treatment and / or prevention of oxidative stress of the anterior segment. In an even more preferred embodiment, the medicament of the invention is for the treatment and / or prevention of oxidative stress of the ocular surface. In an even more preferred embodiment, the medicament of the invention is for the treatment and / or prevention of oxidative stress of the corneal epithelium.

Ocular oxidative stress is associated with a variety of eye injuries and diseases, such as, but not limited to, pterygium, cataracts, corneal dystrophies, glaucoma, diabetic retinopathy, macular degeneration, SOS, SIL, blepharitis, gland dysfunction Meibomial, meibomitis, ocular allergic processes, conjunctivitis, alteration of the ocular surface, preferably of the corneal epithelium, caused by the use of contact lenses and their cleaning and maintenance systems, autoimmune diseases, preferably affecting the anterior segment of the eye, such as, but not limited to, Sjögren's syndrome, post-surgery, preferably of the anterior segment of the eye, burns (both chemical or heat, or thermal) or ocular lesions or pathologies caused by ultraviolet radiation or any other endogenous factor, such as although without limiting ourselves, inflammatory reactions, or exogenous, as po r example but without limiting ourselves, tobacco smoke or environmental pollutants, capable of inducing formation of radical species. Therefore, the medicament of the invention is useful for the treatment and / or prevention of diseases or injuries that occur with ocular oxidative stress, preferably, of those indicated in this paragraph.

In another preferred embodiment, the medicament of the invention is for the treatment and / or prevention of ocular neovascularization. By "ocular neovascularization" is meant the formation of new blood vessels in the ocular structures, for example, but not limited to, in the retina, optic papilla, choroid or in any structure that is part of the anterior segment, including the ocular surface, preferably in cornea, conjunctiva, sclerocorneal limbus or iris. In a more preferred embodiment, the medicament of the invention is for the treatment and / or prevention of neovascularization of the anterior segment. In an even more preferred embodiment, the medicament of the invention is for the treatment and / or prevention of neovascularization of the ocular surface. In an even more preferred embodiment, the medicament of the invention is for the treatment and / or prevention of corneal neovascularization.

Ocular neovascularization is associated with a variety of ocular lesions and diseases, such as, for example, but not limited to conjunctivitis, alteration of the ocular surface, preferably of the corneal epithelium, caused by the use of contact lenses and their cleaning systems. and maintenance, autoimmune diseases (such as Stevens-Johnson syndrome, Lyell, mucous membrane pemphigoid, etc.), preferably affecting the anterior segment, post-surgery, preferably the anterior segment, burns (both chemical or heat, or thermal) or eye lesions or pathologies caused by ultraviolet radiation. Therefore, the medicament of the invention is useful for the treatment and / or prevention of diseases or lesions that occur with ocular neovascularization, preferably, of those indicated in this paragraph.

In an even more preferred embodiment, the medicament of the invention is for the treatment and / or prevention of SOS, blepharitis, dysfunction of the Meibomian glands, meibomitis, ocular allergic processes, corneal dystrophies, conjunctivitis, alteration of the ocular surface by the use of contact lenses, autoimmune diseases that affect the anterior segment of the eye, post-surgery of the anterior segment, burns (both chemical or "caustic" and thermal) or ocular lesions or pathologies caused by ultraviolet radiation. Even more preferably, the medicament of the invention is for the treatment and / or prevention of SOS (dry keratoconjunctivitis, dry keratitis or xerophthalmia), which has been defined as a multifactorial disease of the ocular surface, including the tear film, which causes symptoms of discomfort, visual disturbance and tear instability, with potential damage to the ocular surface, accompanied by an increase in the osmolarity of the tear film and inflammation of the ocular surface. Such a syndrome can be diagnosed, for example, but not limited to, by a tear film slit lamp examination, during which a dye can be placed in the eye, such as fluorescein, to make said film more visible and thus be able to assess its stability, or by the Schirmer test, which measures the rate of tear production using a strip of filter paper that is placed at the end of the eyelid and measures the amount of tear produced by the eye.

On the other hand, the cleaning and maintenance of contact lenses is essential to avoid possible ocular pathologies derived from their use. The cleaning and maintenance systems must meet a series of requirements such as not altering or irritating the eye tissues, not altering or interfering with the normal physiology of the eye, not altering or damaging contact lenses, avoiding contamination by lens microorganisms Contact and keep these as clean as possible. Both the QCT alone and the composition of the invention meet all these requirements, and also have the characteristics that such a disinfectant agent must meet: pH and tonicity similar to tear, bacteriostatic and / or bactericidal, soluble in water and stable in aqueous solution and in plastic bottles.

Therefore, another aspect of the invention relates to the use of the composition of the invention as an antimicrobial, antioxidant and anti-inflammatory agent in contact lens cleaning and maintenance solutions. Another aspect of the invention relates to a solution for cleaning and maintaining contact lenses comprising the composition of the invention.

An "antimicrobial agent" is that chemical compound, or mixture of compounds, that inhibits growth or kills microorganisms. The antimicrobial agent, as understood in the present invention, may be, but not limited to, antibacterial (directed against bacteria), antifungal (directed against fungi) or antiviral (directed against viruses) and may be static, which inhibits growth of the microorganism without causing its death, for example, bacteriostatic or fungistatic, or it can destroy microorganisms, for example, bactericidal or fungicidal.

The “cleaning solutions and maintenance of contact lenses” include, but are not limited to, cleaning solutions, conservative or moisturizing solutions, whose function is to store and store contact lenses when they are not in use, conditioning solutions , peroxide systems and single solution systems, which perform a mechanical cleaning and disinfectant and in turn act as preservatives and lens moisturizers.

As used in the present invention, the term "contact lenses" refers to both hard contact lenses, including rigid and semi-rigid and permeable gas, and soft or hydrophilic ones.

Throughout the description and the claims the word "comprises" and its variants are not intended to exclude other technical characteristics, additives or components. For those skilled in the art, other objects, advantages and features of the invention will be derived partly from the description and partly from the practice of the invention. The following examples and drawings are provided by way of illustration, and are not intended to be limiting of the present invention.

DESCRIPTION OF THE FIGURES

Fig. 1. Shows the amount of IL-6 produced in human corneal epithelium (HCE) cells stimulated with 25 ng / mL of TNF-  and treated with vehicle (0.5% ethanol (EtOH)), 50 IM of resveratrol (RES), 25 IM of quercetin (QCT), 25 IM of RES + 25 IM of QCT or 50 IM of RES + 25 IM of QCT, for 24 hours. NOTE: the cells were treated before stimulation for 2 hours with the corresponding treatment. Control: HCE cells not stimulated with TNF-a and treated with vehicle. *: p <0.05.

Fig. 2. Shows the amount of IL-8 produced in human corneal epithelial cells (HCE) stimulated with 25 ng / mL of TNF-a and treated with vehicle (0.5% ethanol (EtOH)), 50 IM of resveratrol (RES), 25 IM of quercetin (QCT), 25 IM of RES + 25 IM of QCT or 50 IM of RES + 25 IM of QCT, for 24 hours. NOTE: the cells were treated before stimulation for 2 hours with the corresponding treatment. Control: HCE cells not stimulated with TNF-a and treated with vehicle. *: p <0.05.

Fig. 3. Shows the amount of IP-10 produced in human corneal epithelial cells (HCE) stimulated with 25 ng / mL of TNF-a and treated with vehicle (0.5% ethanol (EtOH)), 50 IM of resveratrol (RES), 25 IM of quercetin (QCT), 25 IM of RES + 25 IM of QCT or 50 IM of RES + 25 IM of QCT, for 24 hours. NOTE: the cells were treated before stimulation for 2 hours with the corresponding treatment. Control: HCE cells not stimulated with TNF-a and treated with vehicle. *: p <0.05.

Fig. 4. Shows the amount of VEGF produced in human corneal epithelial cells (HCE) stimulated with 25 ng / mL of TNF-a and treated with vehicle (0.5% ethanol (EtOH)), 50 IM of resveratrol ( RES), 25 IM of quercetin (QCT), 25 IM of RES + 25 IM of QCT or 50 IM of RES + 25 IM of QCT, for 24 hours. NOTE: the cells were treated before stimulation for 2 hours with the corresponding treatment. Control: HCE cells not stimulated with TNF-a and treated with vehicle. *: p <0.05.

Fig. 5. Shows the amount of cyclooxygenase 2 (COX-2), in standardized units, produced in human corneal epithelial cells (HCE) stimulated with 25 ng / mL of TNF-a and treated with vehicle (0.5% of ethanol (EtOH)), 50 IM of resveratrol (RES), 25 IM of quercetin (QCT), 25 IM of RES + 25 IM of QCT or 50 IM of RES + 25 IM of QCT, for 24 hours. NOTE: the cells were treated before stimulation for 2 hours with the corresponding treatment. Control: HCE cells not stimulated with TNF-a and treated with vehicle. *: p <0.05.

Fig. 6. Shows the amount of fluorescence in units normalized to the total amount of oxidized H2DCF-DA probe protein, as a measure of the intracellular generation of relative oxygen species, in human corneal epithelium (HCE) cells exposed to radiation UVB and vehicle treated (0.5% ethanol (EtOH)), 50 IM of resveratrol (RES), 25 IM of quercetin (QCT), 25 IM of RES + 25 IM of QCT or 50 IM of RES + 25 IM of QCT, for one hour. NOTE: the cells were treated before stimulation for 1 hour with the corresponding treatment. Control: HCE cells not irradiated with UVB and treated with vehicle. *: p <0.05.

Fig. 7. Shows the average of CD4 + T cells in the conjunctiva of C57BL / 6 mice in a mouse SOS model (Dursun et al., 2002, Invest Ophthalmol Vis Sci .; 43 (3): 632-638) and treated with vehicle, 0.01% resveratrol (RES), 0.01% quercetin (QCT) and 0.01% QCT + 0.01% RES. Control: mice not exposed to induced dry eye (OSI) and not treated topically. Without treatment: mice exposed to OSI and not treated topically. *: p <0.05.

EXAMPLES OF REALIZATION

The invention will now be illustrated by tests carried out by the inventors, which show the effectiveness of the composition of the invention, as well as quercetin (QCT) and resveratrol (RES) individually, in prevention and / or treatment of inflammation, oxidative stress and ocular neovascularization. These specific examples provided serve to illustrate the nature of the present invention and are included for illustrative purposes only, and therefore should not be construed as limitations on the invention claimed herein. Therefore, the examples described below illustrate the invention without limiting its scope of application.

EXAMPLE 1. Anti-inflammatory effect of quercetin (QCT), resveratrol (RES) and its mixture on a human corneal epithelial cell line stimulated with TNF- .

Cell culture

For all experiments a cell line (HCE) derived from human corneal epithelium was used. The cells were cultured in DMEM / F12 medium supplemented with 15% fetal bovine serum (FBS), 0.5% DMSO, 0.1 mg / mL choleric toxin, 10 ng / mL EGF, 5 mg / mL insulin obtained from bovine pancreas and antibiotics (100 U / mL of penicillin and 0.1 mg / mL of streptomycin). To carry out the experiments, the culture medium was replaced with DMEM medium free of NaHCO3, phenol red and pyruvate.

RES, QCT and their mixture were dissolved in ethanol (EtOH). Stock solutions were prepared to have a constant concentration of vehicle in all wells.

Cellular stimulation with TNF-a

The cells were planted in 24-well culture plates and allowed to grow to preconfluence. Subsequently, they were pretreated with 50 IM of RES, 25 IM of QCT, 25 IM of RES + 25 IM of QCT or 50 IM of RES + 25 IM of QCT, for 2 hours. After this time, the supernatants were removed and the cells were stimulated for 24 hours with 25 ng / mL of TNF-a in the presence of RES, OCT or OCT + RES (returned to

nadir). As controls, non-stimulated cells with TNF-a and stimulated cells were used and treated both with vehicle alone. After the incubation time had elapsed, the supernatants were collected, centrifuged and frozen at -80 ° C for later analysis. The cells adhered to the culture plates were also frozen at -80 ° C for later analysis.

Determination of cytokine and chemokine production

The analysis of the production of the IL-6 and VEGF cytokines and the IL-8 and IP-10 chemokines was carried out in the supernatants collected after stimulation with TNF-a. This analysis was carried out using X-MAP technology in a Luminex IS-100 with a commercial multi-analyte assay (Milliplex, Millipore), following the manufacturer's instructions. The production values, in pg / mL, of cytokines / chemokines (obtained after interpolation of the fluorescence values in standard curves generated in the assay) were subsequently normalized with respect to the amount of corresponding total protein in each sample, determined by a commercial BCA assay (Pierce, USA) in cells attached to the well.

Determination of the production of cyclooxygenase 2 (COX-2)

For this test a commercial "Cell-based ELISA" (R&D, USA) was used following the manufacturer's instructions. In this test, the amounts of COX-2 and GAPDH of each well are determined simultaneously. The cells were planted on the assay plate and treated and stimulated in the same manner as described above. Once the incubation time was over, the fluorescence values corresponding to the amount of COX-2 and GAPDH were measured in a spectrophotometer (SpectraMAX M5, Molecular Devices, Inc. USA). The COX-2 values in each well were normalized to the corresponding amount of GAPDH.

Results of cytokine and chemokine production

Figure 1 shows the amount of IL-6 stimulated with TNF-a and its variation with respect to treatments with QCT, RES and QCT + RES. It can be seen how the stimulation of HCE cells with TNF-a significantly increased the production of IL-6 (p <0.05). QCT, RES and QCT + RES dramatically decreased the values of IL-6 (p <0.05). The compounds studied and their vehicle, at the concentrations tested, were not cytotoxic. Figure 2 shows the amount of IL-8 stimulated with TNF-a and its variation with respect to treatments with QCT, RES and QCT + RES. Stimulation of HCE cells with TNF-a significantly increased the production of IL-8 (p <0.05). QCT, RES and QCT + RES significantly decreased the values of IL-8 (p <0.05).

Figure 3 shows the amount of IP-10 stimulated with TNF-a and its variation with respect to treatments with QCT, RES and QCT + RES. Stimulation of HCE cells with TNF-a significantly increased the production of IP -10 (p <0.05). QCT, RES and QCT + RES significantly decreased IP-10 values to baseline levels (p <0.05).

Figure 4 shows the amount of VEGF stimulated with TNF-a and its variation with respect to treatments with QCT, RES and QCT + RES. Stimulation of HCE cells with TNF-a significantly increased the production of VEGF (p <0.05). QCT, RES and QCT + RES significantly decreased VEGF values to baseline levels (p <0.05).

Results of the production of COX-2

Figure 5 shows the total amount of COX-2 (normalized with GAPDH) stimulated with TNF-a and its variation with respect to treatments with QCT, RES and QCT + RES. Stimulation of HCE cells with TNF-a significantly increased the production of COX-2 (p <0.05). QCT, RES and QCT + RES significantly decreased the amount of COX-2 stimulated by TNF- a (p <0.05).

EXAMPLE 2. Antioxidant effect of resveratrol (RES), quercetin (QCT) and its mixture on a human corneal epithelial cell line stimulated with ultraviolet radiation.

Cell culture

As in the previous example, the HCE cell line derived from human corneal epithelium was used. The cells were cultured in DMEM / F12 medium supplemented with 15% fetal bovine serum (FBS), 0.5% DMSO, 0.1 mg / mL choleric toxin, 10 ng / mL EGF, 5 mg / mL insulin obtained from bovine pancreas and antibiotics (100 U / mL of penicillin and 0.1 mg / mL of streptomycin). To carry out the experiments, the culture medium was replaced with DMEM medium free of NaHCO3, phenol red and pyruvate.

QCT, RES and its mixture were dissolved in ethanol (EtOH). Stock solutions were prepared to have a constant concentration of vehicle in all wells.

Intracellular oxidative stress caused by stimulation with ultraviolet light

The cells were planted in 24-well culture plates and allowed to grow to preconfluence. Subsequently, they were pretreated with 50 µM RES, 25 µM OCT, 25 µM RES +25 µM OCT or 50 µM RES +25 µM OCT, for 1 hour. Next, the cells were loaded with 10-dichlorodihydrofluorescein

! M of 2 ', 7' diacetate (H2DCF-DA) for 30 minutes. The cells were then treated with QCT, RES or QCT + RES, exposed to UVB light (302 nm; 107 mJ / cm2) and left in incubation at 37 ° C for 1 hour. As controls, non-irradiated cells and irradiated cells were used, and both treated with vehicle only. Finally, fluorescence was read at 522 nm. Fluorescence values were normalized with respect to the amount of total protein in each sample, determined by a commercial BCA assay (Pierce, USA) in the cells attached to the well.

Results of oxidative stress caused by UVB radiation

Figure 6 shows the amount of relative fluorescence produced by exposure to UVB radiation and its variation with respect to treatments with QCT, RES and QCT + RES. Stimulation of HCE cells with UVB radiation produced a significant increase in oxidative species (p <0.05). QCT, RES and QCT + RES acted as antioxidants decreasing the levels of oxidative species generated by exposure to UVB radiation, a decrease that in the case of QCT and QCT + RES was significant (p <0.05).

EXAMPLE 3. Anti-inflammatory effect of quercetin (QCT), resveratrol (RES) and its mixture, administered topically, in an animal model of dry eye.

Dry eye animal model

Induced dry eye (OSI) was caused in C57BL / 6 mice by subcutaneous injection of 200 µL of scopolamine (5mg / mL) three times a day for 10 days, and exposure to a constant air flow for 24 hours. The humidity in the room was maintained between 30% and 35%, at a constant temperature of 25 ° C (Dursun et al. Invest Ophthalmol Vis Sci. 2002; 43 (3): 632-8).

Mice and topical treatments

C57BL / 6 mice were divided into 6 groups according to the treatment followed:

a) Control group: mice not exposed to OSI and not treated topically.

b) OSI Group: mice exposed to OSI and not treated topically.

c) OSI + vehicle group: mice exposed to OSI and treated topically with the vehicle.

d) OSI + RES Group: mice exposed to OSI and treated topically with 0.01% RES.

e) OSI + QCT Group: mice exposed to OSI and treated topically with 0.01% QCT.

f) OSI + QCT + RES Group: mice exposed to OSI and treated topically with 0.01% QCT + 0.01% RES.

Topical treatments began 1 day before inducing dry eye, instilling in both eyes 5 μL of the corresponding treatment for each group three times a day.

Determination of CD4 + T lymphocytes in conjunctiva

The eyes embedded in OCT were sectioned in a cryostat. Samples were fixed with cold acetone at -20 ° C. Subsequently, the endogenous peroxidase was blocked with hydrogen peroxide. Afterwards, the joints were not blocked

5 specific with serum and the samples were incubated with the primary antibody at room temperature. Consequently, a biotinylated secondary antibody followed by ABC reagent was used. Finally, the NovaRed vector chromogen was used to color the IMQ labeling, followed by nuclear contratinction with hematoxylin. Three sections of each mouse were examined under an optical microscope and the results were expressed as the average of CD4 + T cells in conjunctiva.

10 Results of the anti-inflammatory properties of QCT, RES and their mixture in a dry eye animal model

Figure 7 shows the average of CD4 + T lymphocytes in conjunctiva with respect to topical treatments with vehicle, QCT, RES and QCT + RES. The dry eye model produced an increase in the amount of CD4 + cells in conjunctiva

15 with respect to the control group (p <0.05). RES showed a non-significant decrease (p> 0.05) of CD4 + lymphocytes, compared with the OSI + vehicle group. QCT and QCT + RES significantly decreased the amount of CD4 + cells in conjunctiva produced by OSI in mice, compared to the OSI + vehicle group (p <0.05).

Claims (16)

one.

Composition comprising quercetin, or any of its salts or derivatives, and at least one other stilbenoid compound, or any of its salts or derivatives.

2.

Composition according to claim 1 wherein the other stilbenoid compound is resveratrol, or any of its salts or derivatives.

3.

Composition according to any one of claims 1 or 2, further comprising a pharmaceutically acceptable carrier.

Four.

Use of the composition according to any of claims 1 to 3 for the preparation of a medicament.

5.

Use of the composition according to claim 4 wherein the medicament is formulated for ophthalmic administration.

6.

Use of the composition according to any of claims 4 or 5 wherein the medicament is for the treatment and / or prevention of ocular inflammation.

7.

Use of the composition according to claim 6 wherein the medicament is for the treatment and / or prevention of inflammation of the anterior segment and / or of the main lacrimal gland.

8.

Use of the composition according to claim 7 wherein the medicament is for the treatment and / or prevention of inflammation of the ocular surface.

9.

Use of the composition according to any of claims 4 or 5 wherein the medicament is for the treatment and / or prevention of ocular oxidative stress.

10.

Use of the composition according to claim 9 wherein the medicament is for the treatment and / or prevention of oxidative stress of the anterior segment.

eleven.

Use of the composition according to claim 10 wherein the medicament is for the treatment and / or prevention of oxidative stress of the ocular surface.

12.

Use of the composition according to any of claims 4 or 5 wherein the medicament is for the treatment and / or prevention of ocular neovascularization.

13.

Use of the composition according to claim 12 wherein the medicament is for the treatment and / or prevention of neovascularization of the anterior segment.

14.

Use of the composition according to claim 13 wherein the medicament is for the treatment and / or prevention of corneal neovascularization.

fifteen.

Use of the composition according to any one of claims 6 to 14 wherein the medicament is for the treatment and / or prevention of dry eye syndrome, blepharitis, dysfunction of the Meibomian glands, meibomitis, allergic ocular processes, corneal dystrophies, conjunctivitis, alteration of the ocular surface by the use of contact lenses, autoimmune diseases that affect the anterior segment of the eye, post-surgery of the anterior segment, burns or injuries or ocular pathologies caused by ultraviolet radiation.

FIG. one FIG. 2 FIG. 3 FIG. 4 FIG. 5 FIG. 6 FIG. 7 SPANISH OFFICE OF THE PATENTS AND BRAND Application no .: 201230664 SPAIN Date of submission of the application: 04.05.2012 Priority Date: REPORT ON THE STATE OF THE TECHNIQUE 51 Int. Cl.: See Additional Sheet RELEVANT DOCUMENTS

Category

56 Documents cited Claims Affected

X

WO 2007103960 A1 (ALLERGAN INC) 13.09.2007, the whole document. 1-15

X

WO 2005099721 A2 (THE REGENTS OF THE UNIVERSITY OF CALIFORNIA) 27.10.2005 1-4

Y

5-15

Y

LIU Q. et al. Oxidative Stress Is an Early Event in Hydrostatic Pressure – Induced Retinal Ganglion Cell Damage. Investigative Ophthalmology & Visual Science, 10.2007, Vol. 48, No. 10, pages 4580-4589, the entire document. 5-15

TO

TSUBOTA K et al. The Era of Antiaging Ophthalmology Comes of Age: Antiaging Approach for Dry Eye Treatment. Ophthalmic Res. 09.09.2010, Vol. 44, pages 146-154, the whole document. 1-15

TO

CHUNG S et al. Regulation of SIRT1 in cellular functions: Role of polyphenols. Archives of Biochemistry and Biophysics. 01.09.2010, Vol. 501 (1), pages 79-90, the whole document. 1-15

TO

MIYAMOTO N et al. Quercetin Induces the Expression of Peroxiredoxins 3 and 5 via the Nrf2 / NRF1 Transcription Pathway. Investigative Ophthalmology & Visual Science. 02.2011, Vol. 52, No. 2, pages 1055-1063, the whole document. 1-15

Category of the documents cited X: of particular relevance Y: of particular relevance combined with other / s of the same category A: reflects the state of the art O: refers to unwritten disclosure P: published between the priority date and the date of priority submission of the application E: previous document, but published after the date of submission of the application

This report has been prepared • for all claims • for claims no:

Date of realization of the report 29.04.2013

Examiner M. Cumbreño Galindo Page 1/5

REPORT OF THE STATE OF THE TECHNIQUE Application number: 201230664 CLASSIFICATION OBJECT OF THE APPLICATION A61K31 / 05 (2006.01) A61K31 / 353 (2006.01) A61P27 / 02 (2006.01) Minimum documentation sought (classification system followed by classification symbols) A61K, A61P Electronic databases consulted during the search (name of the database and, if possible, terms of search used) INVENES, EPODOC, WPI, MEDLINE, NPL, EMBASE, BIOSIS State of the Art Report Page 2/5  WRITTEN OPINION Application number: 201230664 Date of Completion of Written Opinion: 04.29.2013 Statement

Novelty (Art. 6.1 LP 11/1986)

Claims Claims 7, 8, 10, 11 and 13-15 1-6, 9 and 12 IF NOT

Inventive activity (Art. 8.1 LP11 / 1986)

Claims Claims 1-15 IF NOT

The application is considered to comply with the industrial application requirement. This requirement was evaluated during the formal and technical examination phase of the application (Article 31.2 Law 11/1986).  Opinion Base.- This opinion has been made on the basis of the patent application as published. State of the Art Report Page 3/5  WRITTEN OPINION Application number: 201230664  1. Documents considered.- The documents belonging to the state of the art taken into consideration for the realization of this opinion are listed below.

Document

Publication or Identification Number publication date

D01

WO 2007103960 A1 13.09.2007

D02

WO 2005099721 A2 10/27/2005

D03

LIU Q. et al. Investigative Ophthalmology & Visual Science. Vol. 48, No. 10, pages 4580-4589. 10.2007

D04

TSUBOTA K et al. Ophthalmic Res. Vol. 44, pages 146-154. 09.09.2010

D05

CHUNG S et al. Archives of Biochemistry and Biophysics. Vol. 501 (1), pages 79-90. 01.09.2010

D06

MIYAMOTO N et al. Investigative Ophthalmology & Visual Science. Vol. 52, No. 2, pages 1055-1063. 02.2011

 2. Statement motivated according to articles 29.6 and 29.7 of the Regulation of execution of Law 11/1986, of March 20, on Patents on novelty and inventive activity; quotes and explanations in support of this statement The present invention aims at a composition comprising quercetin and a stilbenoid compound, in particular resveratrol, as well as its use in the prevention and / or treatment of inflammation, oxidative stress and ocular neovascularization (claims 1 to 15). D01 discloses an intraocular implant for the treatment of eye diseases comprising an ophthalmic composition. D02 anticipates compositions comprising polyphenolic compounds and inhibitors of reactive oxygen species for the treatment of inflammatory diseases and cancer. D03 demonstrates the potential use of antioxidants such as resveratrol and quercetin in the inhibition of oxidative damage that occurs in the eye prior to the development of glaucoma. D04 exposes the role of caloric restriction as well as reactive oxygen species in aging and analyzes how they can be applied in the prevention and treatment of eye diseases, especially in the case of dry eye. He mentions that resveratrol stimulates the activity of sirtuins in the eye by suppressing inflammation in the retina and that the use of resveratrol in the treatment of dry eye syndrome can be useful because of this anti-inflammatory capacity and can be combined with another anti-inflammatory compound. D05 reviews the functions of sirtuin 1 (SIRT1) and how it is regulated by polyphenols. The determination of their role and mechanism can help identify pharmacological agents for possible use as nutraceuticals in the treatment of chronic diseases. Thus, polyphenols such as resveratrol or quercetin act by inhibiting inflammation and oxidative stress due to their activity as antioxidants. D06 verifies that quercetin, thanks to its antioxidant capacity, protects against oxidative stress caused by various eye diseases. NOVELTY D01 discloses an intraocular implant for the treatment of eye diseases comprising an ophthalmic composition. Said composition comprises sirtuin activating agents such as resveratrol that reduces neovascularization and inflammation and, in addition, may contain another therapeutic agent for activating sirtuins, quercetin, in combination with resveratrol. Therefore, claims 1-6, 9 and 12 cannot be considered new in view of the state of the art or present inventive activity. D02 anticipates compositions comprising polyphenolic compounds and inhibitors of reactive oxygen species for the treatment of inflammatory diseases and cancer. It demonstrates that the combination of quercetin with resveratrol causes a synergistic effect on the activation of caspase-3 and on the release of cyclochrome c resulting in an antioxidant activity. In the event that ocular administration of the composition of the invention is required, an ophthalmic vehicle will be used that allows contact with the surface of the eye for a sufficient time for the composition to penetrate the cornea, as well as in the anterior chamber or posterior eye. Therefore, claims 1-4 cannot be considered new in view of the state of the art or present inventive activity. State of the Art Report Page 4/5  WRITTEN OPINION Application number: 201230664 INVENTIVE ACTIVITY D01 discloses an intraocular implant for the treatment of eye diseases comprising an ophthalmic composition. Said composition comprises sirtuin activating agents such as resveratrol that reduces neovascularization and inflammation and, in addition, may contain another therapeutic agent for activating sirtuins, quercetin, in combination with resveratrol. The composition serves to treat diseases of the posterior segment of the eye. Thus, this document anticipates compositions comprising quercetin and resveratrol and their use in the preparation of a medicament for treating diseases of the posterior ocular segment. In addition, the activity of both compounds as antioxidants, anti-inflammatories and neovascularization inhibitors and their use in the treatment of various eye diseases such as dry eye syndrome is well known in the prior art. Therefore, claims 7, 8, 10, 11 and 13-15 are new but have no inventive activity. D02 anticipates compositions comprising polyphenolic compounds and inhibitors of reactive oxygen species for the treatment of inflammatory diseases and cancer. It demonstrates that the combination of quercetin with resveratrol produces a synergistic effect on the activation of caspase-3 and on the release of cyclochrome c resulting in an antioxidant activity. In the event that ocular administration of the composition of the invention is required, an ophthalmic vehicle will be used that allows contact with the surface of the eye for a sufficient time so that the composition penetrates the cornea as well as the anterior chamber or posterior eye. The difference between D02 and the present invention is that the use of quercetin and resveratrol in the treatment of eye diseases is not mentioned. D03 demonstrates the potential use of antioxidants such as resveratrol and quercetin in the inhibition of oxidative damage that occurs in the eye prior to the development of glaucoma. For a person skilled in the art, it would be evident to use the combined use of quercetin and resveratrol (D02) for the treatment of ocular diseases (D02), and the activity of both compounds as antioxidants, anti-inflammatories, and e neovascularization inhibitors. Therefore, claims 5-15 have no inventive activity. State of the Art Report Page 5/5