ES2769902B2 - Use of secoiridoids for the treatment of optic neuritis. - Google Patents

Description

DESCRIPTION

USE OF SECOIRIDOIDS FOR THE TREATMENT OF OPTIC NEURITIS

The present invention relates to the use of secoiridoids, such as oleacein and oleocanthal, to prevent or treat neuropathies that lead to optic nerve damage, such as optic neuritis (hereinafter, NO). The present invention also relates to pharmaceutical or nutraceutical compositions containing said secoiridoids. Therefore, the present invention belongs to the technical field of medicine as well as to the food industry.

STATE OF THE ART

Optic neuritis (NO) is a demyelinating inflammation that damages the optic nerve, a bundle of nerve fibers that transmits visual information from your eye to your brain. Pathological changes involving various retinal structures can precede this occurrence. It is NOT a common neuro-ophthalmic inflammatory disease that causes persistent vision impairment and generally affects one eye. Symptoms may include: pain, loss of vision in one eye, loss of visual field, loss of color vision, flashing lights (Wu GF et al., Curr Immunol Rev. 2015; 11: 85-92; Beck RW et al., Am J Ophthalmol. 2004; 137: 77-83).

Ocular inflammation usually has an infectious or autoimmune etiology. NO with unilateral involvement is more commonly associated with multiple sclerosis (MS). In fact, at least 50% of patients with MS developed NO, although not all patients with NO developed MS. However, atypical NO may be associated with neuromyelitis optica spectrum disorders (with the presence of anti-aquaporin-4 or anti-myelin oligodendrocyte glycoprotein antibodies, MOG), acute disseminated encephalomyopathy, and other autoimmune conditions, such as sarcoidosis, lupus systemic erythematous, Sjogren's syndrome or Behpet's disease. MOG has been shown to be present in abundance within the optic nerve, and inflammatory cells probably react to the MOG antigen of the optic nerve to cause tissue damage. Thus, the MOG antigen has a high chance of causing optic neuritis.

Pulse therapy with methylprednisolone has been the mainstay of treatment for the acute phase of NO. Corticosteroids speed up the recovery of vision; however, they do not improve visual prognosis. Numerous undesirable side effects are associated with corticosteroids. One study even indicates that methylprednisolone accelerates apoptosis of neurons in the central nervous system. With the serious side effects of corticosteroids and their lack of neuroprotective effect, NO patients urgently need a new drug with anti-inflammatory, immune-regulating, and neuroprotective effects.

The present invention refers to the search for new treatments for disorders related to MS, in particular, preferably optic neuritis, and describes a new pharmacological application of oleacein and oleocanthal as agents that notably reduce the clinical and immunological / oxidative features of the experimental optic neuritis (NOE). Electroretinogram and visual evoked potential studies have shown that visual function is altered in patients with MS and in animals with experimental autoimmune encephalomyelitis (EAE), the animal model that mimics many of the features of MS. Furthermore, since the pathophysiological changes that occur in the spinal cord of EAE also occur in the optic nerve of mice with EAE; the EAE animal model is considered an ideal model for studying NO (Shields et al. Brain Res. 1998; 784: 299-304). As in MS, mice with EAE often develop NO. The well-established mouse model produced by immunizing C57BL / 6 female mice with a MOG peptide fragment, MOG35-55, is used. All C57BL / 6 mice with clinical signs of EAE also have histological evidence of NOE. (Kuerten S. et al., Ann Anat 2008, 190: 1-15; Chaudhary P. et al., J Neuroimmunol. 2011; 233: 90-96).

DESCRIPTION OF THE INVENTION

In the present invention, the inventors have found that secoiridoids, such as oleacein and oleocanthal, possess protective effects on the integrity and function of the blood-brain barrier (BBB), as well as on oxidative and immunoinflammatory events related to optic neuritis.

Secoiridoids are monoterpenoids, derived from iridoids in plants, based on the 7,8-seco-cyclopenta [c] -pyranoid skeleton. Most of the secoiridoids and iridoids have been isolated from plants and approximately 600 different structures are known. Most of the secoiridoids are glycosides. This group of phytochemicals is very widespread in nature, and exhibits a wide range of biological and pharmacological activities, including antibacterial, anti-cancer, anticoagulant, antifungal, antioxidant, anti-protozoal, and hepatoprotective activities (Dinda B. et al. al., Chem Pharm Bull (Tokyo). 2009 Aug; 57 (8): 765-96).

These products are natural substances that can be isolated from olives, these compounds could be used as a dietary supplement or in nutraceutical preparations.

Therefore, a first aspect of the present invention refers to a compound of formula (I):

its pharmaceutically acceptable salts, tautomers and / or solvates thereof

where R1 is -OH or H,

for the treatment and / or prevention of optic neuritis.

The term "pharmaceutically acceptable salts or solvates thereof" refers to salts or solvates which, when administered to the recipient, are capable of providing a compound such as that described herein. The preparation of salts and derivatives can be carried out by methods known in the state of the art. Preferably, "pharmaceutically acceptable" refers to molecular entities and compositions that are physiologically tolerable and do not normally produce an allergic reaction or a similar unfavorable reaction, such as gastric upset, dizziness, and similar side effects, when administered to a human. Preferably, the term "pharmaceutically acceptable" means approved by a regulatory agency of a federal or state government or compiled in the US Pharmacopoeia or other generally recognized pharmacopoeia for use in animals and, more particularly, in humans.

The compounds used in the invention may be present in crystalline form, either as free compounds or as solvates (eg, hydrates), and both forms are understood to be within the scope of the present invention. Solvation methods are generally known in the state of the art. Suitable solvates are pharmaceutically acceptable solvates. In a particular embodiment, the solvate is a hydrate.

"Tautomers" are understood to be the two isomers that differ only in the position of a functional group because between the two forms there is a chemical equilibrium in which a migration of a group or atom occurs.

Unless otherwise indicated, the compounds used in the invention are intended to include compounds that differ only in the presence of one or more atoms. enriched with isotopes. For example, compounds having the present structures, except the substitution of a hydrogen atom with a deuterium atom or a tritium atom, or the substitution of a carbon atom with a carbon atom enriched in 13C or 14C or a 15N-enriched nitrogen atom, fall within the scope of this invention.

In a particular embodiment, the compound of formula (I) is oleacein:

In another particular embodiment, the compound of formula (I) is oleocanthal:

In a particular embodiment, the administered dose of the compound of formula (I), its pharmaceutically acceptable salts, tautomers and / or solvates thereof (hereinafter, the compounds of the present invention) varies between 5 mg / day and 20 mg / day , more in particular 10 mg per kg per day.

The compounds of the present invention can be administered by any suitable route of administration, for example: oral, parenteral (subcutaneous, intraperitoneal, intravenous, intramuscular, etc.), inhaled intranasal, etc.

The invention also relates to a composition comprising the compounds of the present invention for use in the treatment and / or prevention of optic neuritis.

The composition could be a pharmaceutical composition or a nutraceutical composition.

The term "nutraceutical composition" as used herein includes a food product, a food, a dietary supplement, a nutritional supplement, or a supplement composition for a food product or a food.

In a particular embodiment, the composition is a pharmaceutical composition comprising pharmaceutically acceptable excipients, adjuvants and / or vehicles.

The term "excipients, adjuvants and / or vehicles" refers to molecular entities or substances through which the active ingredient is administered. Said excipients, adjuvants or pharmaceutical vehicles can be sterile liquids, such as water and oils, including those of petroleum, animal, animal, vegetable or synthetic origin, such as peanut oil, soybean oil, mineral oil, sesame oil and oils. the like, excipients, disintegrating agents, humectants or dilutions. Suitable pharmaceutical excipients and carriers are described in "Remington's Pharmaceutical Sciences" by E.W. Martin.

The pharmaceutical compositions can be administered by any suitable route of administration, for example: oral, parenteral (subcutaneous, intraperitoneal, intravenous, intramuscular, etc.), etc.

In a particular embodiment, said pharmaceutical compositions can be presented in a pharmaceutical form for oral administration, either solid or liquid. Illustrative examples of dosage forms for oral administration include tablets, capsules, granules, solutions, suspensions, etc., and may contain conventional excipients such as binders, dilutions, disintegrating agents, lubricants, humectants, etc., and may be prepared by methods. conventional. The pharmaceutical compositions can also be adapted for parenteral administration, in the form of, for example, sterile solutions, suspensions or lyophilized products in suitable dosage form; in this case, said pharmaceutical compositions will include suitable excipients, such as buffers, surfactants, etc. In any case, the excipients are chosen according to the selected pharmaceutical form of administration. A review of the different pharmaceutical forms of drug administration and their preparation can be found in the book "Treatise on Galenic Pharmacy" by C. Faulí i Trillo, 10th edition, 1993, Luzán 5, SA de Ediciones, or any book of characteristics similar in each country.

The compounds of the present invention can be used in conjunction with additional drugs to provide combination therapy. Said additional drugs can be part of the same pharmaceutical composition or, alternatively, they can be provided as a separate composition for simultaneous administration or not with the pharmaceutical composition comprising the compounds of the present invention.

For therapeutic use, the compounds of the present invention are in a pharmaceutically acceptable form or are substantially pure, that is, they have a pharmaceutically acceptable level of purity that excludes normal pharmaceutical additives, such as diluents and carriers, and is free of any material considered toxic at normal dosage levels. The purity levels of the active substance are in particular greater than 50%, more particularly greater than 70% and even more particularly greater than 90%. In a particular embodiment, the levels of the compound with formula (I), or its salts or solvates, are above 95%.

As said above, the invention relates to a compound of formula (I), its pharmaceutically acceptable salts, tautomers and / or solvates thereof or compositions for the treatment and / or prevention of optic neuritis. Alternatively, the invention relates to a method of treating and / or preventing optic neuritis which comprises administering a compound or composition of the invention to a subject in need thereof. Alternatively, the invention relates to the use of a compound or a composition of the invention for the preparation of a medicament for the prevention and / or treatment of optic neuritis.

The term "treatment" is broadly understood, referring to reducing the potential of a certain disease, reducing the occurrence of a certain disease and / or reducing the severity of a certain disease in particular, to the extent that the subject no longer suffers discomfort and / or altered function due to it. "Treatment" refers to providing a therapeutic benefit or a desired clinical outcome, which is not necessarily a cure for a particular disease or disorder, but encompasses an outcome that typically includes alleviation of the disease, elimination of the disease, reduction or alleviation of a symptom associated with the disease, prevention of a secondary disease as a result of the occurrence of a primary disease, a decrease in the extent of the disease, the stabilized (i.e. no worsening) state of the disease , the delay or slowing down of the progression of the disease, the improvement or palliation of the disease state and the remission (either partial or total), whether detectable or not detectable of the disease.

"Prevention" is intended to prevent the occurrence of such disease. Prevention can be complete (eg, the total absence of a disease). Prevention can also be partial, such that, for example, the occurrence of a disease in a subject is less than that which would have occurred without the administration of the compounds of the present invention. Prevention also refers to reduced susceptibility to a clinical condition.

Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Methods and materials similar or equivalent to those described herein can be used in the practice of the present invention. Throughout the description and claims, the word "comprise" and its variations are not intended to exclude other technical characteristics, additives, components or steps. Additional objects, advantages, and features of the invention will become apparent to those skilled in the art upon examination of the description or may be learned by 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.

BRIEF DESCRIPTION OF THE DRAWINGS

Fig. 1. Oleacein treatment (OLE) decreases the specific production of autoantibodies in mice with EAE. MOG-specific IgG 1 titers in serum samples at a dilution of 1/60. C, healthy mice. C + OLE, healthy mice treated with OLE. Experimental optic neuritis (hereinafter NOE), induced mice. NOE + OLE, induced mice treated with OLE. Bar graphs represent the mean ± SD of 5 8 animals. tp <0.001 vs control and * p <0.001 vs EAE.

Fig. 2. Oleacein treatment (OLE) reduces inflammation in the optic nerve during NOE. To examine whether OLE treatment prevents inflammatory cell infiltration, optic nerves were isolated from mice at 21-23 days after immunization and stained with H and E (hematoxylin and eosin). Histological analysis of the optic nerve from control mice, C; from control mice treated with OLE, C + OLE; from induced mice, NOE; and from induced mice treated with OLE, NOE + OLE, showed inflammatory infiltration. Oleacein treatment improved all these parameters.

Fig. 3. Oleacein (OLE) treatment reduces inflammation in NOE. The serum expression of parameters related to inflammation was used as a measure protective responses: levels of tumor necrosis factor-a, TNFa (A), granulocyte-macrophage colony-stimulating factor, GM-CSF (B), galectin-3, Gal 3 (C) and interleukin-1p, IL -1 p (D) in serum were used as measures of protective responses. C, healthy mice. C + OLE, healthy mice treated with OLE. NOE, induced mice. NOE + OLE, induced mice treated with OLE. Bar graphs represent the mean ± SD of 5-8 animals. tp <0.001 and t tp <0.01 vs control and ** p <0.01 and *** p <0.05 vs EAE.

Fig. 4. Oleacein treatment (OLE) reduces demyelination in the optic nerve during NOE. To examine whether OLE treatment prevents demyelination, optic nerves were isolated from mice at 21-23 days after immunization and stained with LFB. Histological analysis of the optic nerve from control mice, C; from control mice treated with OLE, C + OLE; from induced mice, NOE; and from induced mice treated with OLE, NOE + OLE, showed demyelination. Oleacein treatment improved all these parameters.

Fig. 5. Oleacein treatment (OLE) reduces oxidative stress on the optic nerve during NOE. To examine whether OLE treatment prevents the generation of superoxide ion, optic nerves were isolated from mice at 21-23 days after immunization and stained with the oxidative fluorescent probe, dihydroethidium, DHE. (A) Representative images of DHE staining, and (B) bar graph showing quantification of red fluorescence from optic nerve sections from control mice, C; from control mice treated with OLE, C + OLE; from induced mice, NOE; and from induced mice treated with OLE, NOE + OLE, demonstrated an accumulation of superoxide anion. Oleacein treatment improved all these parameters.

Fig. 6. Oleacein treatment (OLE) increases neuroprotection in NOE. The expression levels of malondialdehyde, MDA (A), of advanced protein oxidation products, AOPP (B), the antioxidant / iron reducing power, FRAP (C) and the ROS scavenger, sestrin-3 (D), in serum were used as measures of protective responses. C, healthy mice. C + OLE, healthy mice treated with OLE. NOE, induced mice. NOE + OLE, induced mice treated with OLE.

Fig. 7. Oleacein (OLE) treatment protects against blood-brain barrier disruption and decreases molecular extravasation in mice with NOE. Endogenous IgG extravasation was used as a measure of blood-brain barrier disruption and plasma protein extravasation in the brain. A) Representative photomicrographs of immunofluorescence. B) Quantification of fluorescence intensity. C, healthy mice. C + OLE, healthy mice treated with OLE. NOE, induced mice. NOE + OLE, induced mice treated with OLE. The bar graphs represent the mean ± SD of 5 animals. tp <0.001 vs control and *** p <0.05 vs EAE.

Fig. 8. Oleacein (OLE) inhibits the responses of activated microglia , including the production of reactive oxygen species, ROS (A); induction of inflammatory regulators (B); and synthesis and release of inflammatory cytokines (C).

BV-2 microglia cells were pretreated for 30 minutes with the indicated doses of OLE: (A) After 24 hours of stimulation with 0.1 pg / ml of LPS, the production of intracellular ROS was evaluated by cytometric analysis flow. The panel shows the quantification expressed in arbitrary units (UA), (tp <0.001 vs control, ** p <0.01 and * p <0.001 vs stimuli without OLE; n = 3). (B) After 4 or 24 h of stimulation with 0.1 pg / ml of LPS, the expression of COX-2, iNOS, p-p65-NF ^K B and NLRP3 in cell lysates was identified by Western blot; (C) The presence of TNFa and IL-1 p in the 24 h cell culture medium was quantified by commercial ELISA (tp <0.001 vs control, * p <0.001 vs stimuli without OLE; n = 3).

Fig. 9. Oleocanthal inhibits the responses of activated microglia , including the induction of inflammatory regulators (A); and synthesis and release of inflammatory cytokines (B). BV-2 microglia cells were pretreated for 30 minutes with the indicated doses of oleocanthal: (A) After 4 or 24 hours of stimulation with 0.1 pg / ml of LPS, the expression of COX-2, iNOS was identified , p-p65-NF ^K B and NLRP3 in cell lysates by Western blot; and (B) the presence of TNFa and IL-1 p in the 24 h cell culture medium was quantified by commercial ELISA (tp <0.001 vs control, * p <0.001 vs stimuli without OLE; n = 3).

EXAMPLES

The invention will be exemplified, but not necessarily limited by the following experiments.

In the experiments, the inventors used fifteen animals per group. Experimental optic neuritis (NOE) was induced in C57BL mice as described (Quinn TA et al. Front Neurol. 2011; 2:50; Chaudhary P. et al., J Neuroimmunol. 2011; 233: 90-96).

Immunization was carried out with 100 pg of an oligodendrocyte myelin glycoprotein partial peptide (MOG33-55) in Freund's complete adjuvant containing 4 mg of Mycobacterium tuberculosis H37Ra in 1 ml. Mice were immunized by subcutaneous injection of this emulsion on day 0. In addition, on days 0 and 2, 300 ng / 200 µl of Bordetella pertussis toxin was administered intraperitoneally . The administration of 10 mg / kg of oleacein acid (OLE) was performed intraperitoneally once a day, starting on the day of immunization. Oleacein was isolated from an olive oil extract prepared using the extraction method described by Karkoula E. (Karkoula E. et al., J Agric Food Chem. 2012; 60: 11696-11703). Briefly, pure oleacein, as well as oleocanthal, were isolated from an olive oil extract prepared using the extraction method similar to that described below for sample preparation. The olive oil (5.0 g) was mixed with cyclohexane (20 ml) and acetonitrile (25 ml). The mixture was homogenized using a vortex mixer for 30 seconds and centrifuged at 4,000 rpm for 5 minutes. A part of the acetonitrile phase (25 ml) was collected, mixed with 1.0 ml of a solution of syringaldehyde (0.5 mg / ml) in acetonitrile and evaporated under reduced pressure using a rotary evaporator (Buchi, Flawil , Switzerland).

Mice were sacrificed 21-23 days after MOG immunization. The induction of autoantibodies MOG-IgG1 was quantified in the serum of mice from the four experimental groups. As expected, the levels of MOG-IgG1 autoantibodies were significantly increased in the serum of mice with NOE compared to the healthy control group. In mice with NOE treated with OLE, autoantibody levels were significantly lower (Fig. 1).

To determine whether OLE could modulate the inflammatory response in NOE, the inventors first evaluated the presence of inflammatory cell infiltration in optic neuritis (NO) tissues collected on day 21-23 post immunization (Fig. 2 ). Examination of the hematoxylin and eosin stained optic nerve sections (H and E) showed pronounced cellular infiltrates in the tissues of NOE mice compared to healthy control mice. In contrast, in the tissues of mice with NOE treated with OLE the infiltration of cells was markedly reduced, being comparable to that observed in the tissues of untreated control mice. OLE treatment in healthy control mice did not have a significant effect.

The inventors also determine the circulating levels of the inflammatory proteins, tumor necrosis factor-a (TNFa), galectin-3 (Gal-3) and granulocyte-macrophage colony-stimulating factor (GM-CSF) in serum samples. As expected, the expression of inflammatory mediators was significantly increased in the serum of mice with NOE compared to the healthy control group. However, its overexpression was prevented in the serum of mice with EAE treated with OLE: its levels they did not increase significantly compared to those found in the serum of control mice with or without treatment. Furthermore, high levels of active IL-1 p, a mediator connecting innate and adaptive immunity, were detected in the serum of mice with NOE and this increase was reduced by up to 80% in the animals treated with OLE.

Next, it was evaluated whether OLE protects the optic nerve from demyelination. Demyelination of the optic nerve begins after the onset of inflammation and can be detected by fast luxol blue (LFB) staining of myelin. Untreated EAE mice showed a marked increase in demyelination compared to control mice. Unstained regions indicating destruction of the myelin sheath were observed in the optic nerve of mice with NOE. (Fig. 4). The optic nerves of the OLE-treated NOE mice showed greater LFB staining compared to the optic nerves of the untreated EAE mice, signifying a suppression in the demyelination of the optic nerves.

It is well known that inflammation increases the levels of reactive oxygen species (ROS), leading to oxidative stress that mediates tissue damage. To investigate whether prophylactic administration of OLE to mice with NOE also resulted in a reduction in ROS accumulation, the inventors used as an initial indicator of ROS activity the redox-sensitive fluorescent probe DHE that detects the superoxide anion (O2- ). The cryogenic sections of the optic nerve of mice from the different experimental groups were incubated with the DHE probe and evaluated by fluorescence microscopy. The intensity of the fluorescence signals was quantified using Image J software (NIH, Bethesda, MD, USA) (Fig. 5). In healthy control mice, DHE staining was basically undetectable in the optic nerve. In contrast, the optic nerve of mice with NOE showed an increased intensity of red fluorescence throughout the tissue. However, OLE treatment inhibited NOE-induced ROS production: the red fluorescence of ethidium was greatly attenuated, being similar to that observed in tissues of healthy control mice.

Since superoxide anion has also been implicated in lipid peroxidation and protein oxidation, the inventors evaluated serum levels of advanced protein oxidation products (AOPP) and malondialdehyde (MDA), as end products of lipid peroxidation ( Fig. 6A and B). The NOE group showed a significant increase in serum MDA and AOPP levels compared to the control group. Meanwhile, OLE treatment effectively prevented these increases. Similarly, the antioxidant capacity of serum, evaluated by means of an antioxidant / iron reducing power (FRAP) test, as a marker of non-antioxidant status. enzymes, and the levels of sestrin-3, as a ROS disruptor, were significantly reduced in mice with NOE compared to those in the control group (Fig. 6C and D). OLE treatment showed a significant attenuation of this decrease compared to untreated EAE mice.

The optic nerve is considered part of the CNS. During acute optic neuritis, the rupture of the blood-brain barrier (BBB), as well as the rupture of the blood-retinal barrier (BHR) and the activation of the resident microglia in the retina and optic nerve are pathological indicators of the disease. Consequently, the inventors characterized NOE-induced BHR damage in OLE-treated and untreated mice by analyzing extravasation of endogenous IgG from serum to brain using immunohistochemistry. The intensity of the fluorescence signals was quantified using Image J software (NIH, Bethesda, MD, USA). As shown in Fig. 7, the brain of untreated NOE mice showed a marked increase in IgG extravasation (c, g) compared to healthy control animals (a, b). However, this increased permeability was prevented by the administration of OLE. Treatment with OLE did not modify the integrity of BHR in control animals.

Next, the inventors evaluate whether the protective effects found in vivo in mice with NOE treated with OLE involve direct actions on relevant inflammatory cells of the CNS, the inventors studied the effects of OLE on activated BV-2 microglia cells to mimic responses seen in neuroinflammatory disorders (Fig. 8).

Since ROS production can configure specific inflammatory programs, the inventors investigated intracellular ROS accumulation. Flow cytometric analysis showed that the accumulation of intracellular ROS, which was significantly increased in BV-2 cells treated with LPS, was drastically suppressed in cells pre-treated with different doses of OLE (Fig. 8A).

To determine whether OLE was able to directly modulate inflammatory activity in microglia, BV-2 cells were pre-treated with 1, 10, and 20 pM OLE for 30 minutes and then stimulated with LPS (0.1 pg / ml ) for 4 and 24 hours. As shown in Fig. 8B, Western blot analysis of BV2 cells activated by LPS, showed increased expression of iNOS and COX-2 after 4 and 24 h compared to unstimulated control cells. However, cellular pre-treatment with OLE led to a dose-dependent inhibition of LPS-induced iNOs and COX-2 production, while OLE per se did not affect their baseline expression. The The presence of OLE also reduced the ability of LPS to induce TNFα and IL-ip secretion in a dose-dependent manner (Fig. 8C). Likewise, the activation of the mediators / signaling mechanisms that influence these inflammatory responses, such as phosphorylation of p65-NF ^K B and overexpression of NLRP3, was also reduced (Fig. 8B).

This invention also relates to methods of using natural compounds, oleacein and oleocanthal, to inhibit the responses of activated microglia.

Finally, the inventors also evaluate the protective effects of another natural sercoiridoid: oleocanthal on relevant inflammatory cells of the CNS. The inventors studied the effects of oleocanthal on activated BV-2 microglia cells to mimic the responses seen in neuroinflammatory disorders (Fig. 9). Oleocanthal was isolated from an olive oil extract prepared using the extraction method described by Karkoula E. (Karkoula E. et al., J Agric Food Chem. 2012; 60: 11696-11703) as described above.

To determine whether oleocanthal was able to directly modulate inflammatory activity in microglia, BV-2 cells were pre-treated with 1, 10 and 20 pM oleocanthal for 30 minutes and then stimulated with LPS (0.1 pg / ml) for 4 and 24 hours. As shown in Fig. 9A, Western blot analysis of BV2 cells activated with LPS, showed an increase in iNOS and COX-2 expression after 4 and 24 h compared to unstimulated control cells. However, cellular pre-treatment with oleocanthal led to a dose-dependent inhibition of LPS-induced iNOS and COX-2 production, while oleocanthal per se did not affect its baseline expression. The presence of oleocanthal also reduced the ability of LPS to induce TNFα and IL-1 p secretion in a dose-dependent manner (Fig. 9B). Likewise, the activation of the mediators / signaling mechanisms that influence these inflammatory responses, such as phosphorylation of p65-NF ^K B and overexpression of NLRP3, was also reduced (Fig. 9A).

Claims (8)

1. Compound of formula (I):

their pharmaceutically acceptable salts, tautomers and / or solvates, where R1 is -OH or H for the treatment and / or prevention of optic neuritis. 2. Compound for use according to claim 1, wherein the compound of formula (I) is oleacein:

3. Compound for use according to claim 1, wherein the compound of formula (I) is oleocanthal:

4. Compound for use according to any of claims 1-3, wherein the compound is administered orally, parenterally, or intranasally. 5. Compound for use according to any one of claims 1-4, wherein the compound is administered at a dose between 5 mg / day and 20 mg / day. 6. A composition comprising the compound defined in any of claims 1-5, for use in the treatment and / or prevention of optic neuritis. 7. The composition for use according to claim 6, wherein the composition is a pharmaceutical composition or a nutraceutical composition. The composition for use according to claim 6 or 7, further comprising other additional drugs to provide a combination therapy.