• 专利附录
  • 专利说明
  • 权利要求
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    • 专利摘要:
    Use of secoiridoids for the treatment of optic neuritis. The present invention relates to the use of oleacein and oleocantal secoiridoids to prevent or treat neuropathies that lead to injury to the optic nerve such as optic neuritis. Likewise, the present invention relates to a pharmaceutical composition or a nutraceutical composition comprising said secoiridoids. (Machine-translation by Google Translate, not legally binding)
    公开号:ES2769902A1
    申请号:ES201831296
    申请日:2018-12-28
    公开日:2020-06-29
    发明作者:Miranda Beatriz Gutierrez；CALLEJO Mª LUISA NIETO；Prokopios Magiatis
    申请人:Consejo Superior de Investigaciones Cientificas CSIC；National and Kapodistrian University of Athens；
    IPC主号:
    专利说明:

    [0004] The present invention relates to the use of secoiridoids, such as oleacein and oleocantal, to prevent or treat neuropathies that lead to injury to the optic nerve, 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.
    [0006] STATE OF THE ART
    [0008] Optic neuritis (NO) is a demyelinating inflammation that damages the optic nerve, a collection of nerve fibers that transmits visual information from your eye to your brain. Pathological changes involving various retinal structures may precede this occurrence. It is NOT a common neuro-ophthalmologic 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).
    [0010] Ocular inflammation usually has an infectious or autoimmune etiology. NO with unilateral involvement is most commonly associated with multiple sclerosis (MS). In fact, at least 50% of MS patients developed NO, although not all patients with NO developed MS. However, atypical NO may be associated with disorders of the neuromyelitis optica spectrum (with the presence of anti-aquaporin-4 antibodies or oligodendrocyte myelin glycoprotein, MOG), disseminated acute encephalomyopathy, and other autoimmune conditions, such as sarcoidosis, lupus systemic erythematosus, Sjogren's syndrome or Behpet's disease. MOG has been shown to be abundantly present within the optic nerve, and inflammatory cells are likely to react to the optic nerve MOG antigen to cause tissue damage. Thus, the MOG antigen has a high chance of causing optic neuritis.
    [0012] Pulse therapy with methylprednisolone has been the mainstay of treatment for the acute phase of NO. Corticosteroids speed 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, patients with NO urgently need a new drug with anti-inflammatory properties, immune regulation and neuroprotective effects.
    [0014] The present invention relates 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 oleocantal as agents that markedly reduce the clinical and immunological / oxidative traits of experimental optic neuritis (NOE). Electroretinogram and visual evoked potential studies have shown that visual function is impaired in MS patients and in animals with experimental autoimmune encephalomyelitis (EAE), the animal model that mimics many of the characteristics 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). Similar to MS, mice with EAE often develop NO. The well established mouse model produced by immunizing C57BL / 6 female mice with a peptide fragment from MOG, 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).
    [0016] DESCRIPTION OF THE INVENTION
    [0018] In the present invention, the inventors have found that secoiridoids, such as oleacein and oleocantal, have protective effects on the integrity and function of the blood-brain barrier (EHB), as well as on the oxidative and immunoinflammatory events related to optic neuritis.
    [0020] Secoiridoids are monoterpenoids, derived from iridoids in plants, based on the 7,8-dry-cyclopenta [c] -pyranoid skeleton. Most of the secoiridoids and iridoids have been isolated from plants and approximately 600 different structures are known. Almost all 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-carcinogenic, anticoagulant, anti-fungal, antioxidant, anti-protozoal and hepatoprotective activities (Dinda B. et al., Chem Pharm Bull (Tokyo) .August 2009; 57 (8): 765-96).
    [0021] These products are natural substances that can be isolated from olives, these compounds could be used as a dietary supplement or in nutraceutical preparations.
    [0023] Therefore, a first aspect of the present invention relates to a compound of formula (I):
    [0027] their pharmaceutically acceptable salts, tautomers and / or solvates thereof
    [0028] where R1 is -OH or H,
    [0029] for the treatment and / or prevention of optic neuritis.
    [0031] The term "pharmaceutically acceptable salts or solvates thereof" refers to salts or solvates that, 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 cause an allergic reaction or similar unfavorable reaction, such as gastric distress, dizziness, and the like, when administered to a human. Preferably, the term "pharmaceutically acceptable" means approved by a federal or state government regulatory agency or compiled in the US Pharmacopoeia or other generally recognized pharmacopoeia for use in animals and, more particularly, in humans.
    [0033] The compounds used in the invention can be presented 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.
    [0035] "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 migration of a group or atom occurs.
    [0037] 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 replacement of a hydrogen atom with a deuterium atom or a tritium atom, or the replacement of a carbon atom with a carbon atom enriched in 13C or 14C or a 15N-enriched nitrogen atom falls within the scope of this invention.
    [0039] In a particular embodiment, the compound of formula (I) is oleacein:
    [0043] In another particular embodiment, the compound of formula (I) is oleochantal:
    [0048] 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 particularly 10 mg per kg per day.
    [0050] 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.
    [0052] 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.
    [0053] The composition could be a pharmaceutical composition or a nutraceutical composition.
    [0055] 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.
    [0057] In a particular embodiment, the composition is a pharmaceutical composition comprising pharmaceutically acceptable excipients, adjuvants, and / or carriers.
    [0059] 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. similar, excipients, disintegrating agents, humectants or diluted. Suitable pharmaceutical carriers and excipients are described in E.W. "Remington's Pharmaceutical Sciences". Martin.
    [0061] The pharmaceutical compositions can be administered by any suitable route of administration, for example: oral, parenteral (subcutaneous, intraperitoneal, intravenous, intramuscular, etc.), etc.
    [0063] In a particular embodiment, said pharmaceutical compositions can be presented in an oral administration pharmaceutical form, either solid or liquid. Illustrative examples of pharmaceutical forms of oral administration include tablets, capsules, granules, solutions, suspensions, etc., and may contain conventional excipients such as binders, diluents, disintegrating agents, lubricants, humectants, etc., and can 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 the suitable dosage form; In this case, said pharmaceutical compositions will include suitable excipients, such as buffers, surfactants, etc. In either 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 characteristic book similar in each country.
    [0064] The compounds of the present invention can be used in conjunction with other additional drugs to provide combination therapy. Such additional drugs may form part of the same pharmaceutical composition or, alternatively, may be provided as a separate composition for simultaneous or non-simultaneous administration with the pharmaceutical composition comprising the compounds of the present invention.
    [0066] 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 vehicles, and is free of any Material considered toxic at normal dosage levels. The purity levels of the active substance are particularly above 50%, more particularly above 70% and even more particularly above 90%. In a particular embodiment, the levels of the compound with formula (I), or its salts or solvates, are above 95%.
    [0068] 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 comprising 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 composition of the invention for the preparation of a medicament for the prevention and / or treatment of optic neuritis.
    [0070] 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 particular disease, to the extent that the subject no longer suffers discomfort and / or impaired 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 normally includes relief of disease, elimination of disease, reduction or relief of a symptom associated with the disease, prevention of a secondary disease as a result of the occurrence of a primary disease, decrease in the extent of the disease, stabilized (i.e., no worsening) state of the disease , the delay or slowdown of disease progression, improvement or alleviation of disease status, and remission (either partial or total), either detectable or undetectable of the disease.
    [0071] "Prevention" is intended to prevent the occurrence of this disease. Prevention can be complete (eg, the total absence of disease). Prevention can also be partial, so that, for example, the occurrence of a disease in a subject is less than what would have occurred without the administration of the compounds of the present invention. Prevention also refers to reduced susceptibility to a clinical condition.
    [0073] Unless otherwise defined, all technical and scientific terms used herein have the same meaning commonly understood by one skilled in the art to which this invention pertains. Methods and materials similar or equivalent to those described herein can be used in the practice of the present invention. Throughout the description and the claims, the word "comprises" 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 disclosure or can be learned by practicing the invention. The following examples and drawings are provided by way of illustration and are not intended to be limiting of the present invention.
    [0075] BRIEF DESCRIPTION OF THE DRAWINGS
    [0077] Fig. 1. Treatment with oleacein (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.
    [0079] Fig. 2. Treatment with oleacein (OLE) reduces inflammation in the optic nerve during NOE. To examine whether OLE treatment prevents inflammatory cell infiltration, optic nerves were isolated from mice 21-23 days after immunization and stained with H and E (hematoxylin and eosin). Histological analysis of the optic nerve of 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.
    [0081] Fig. 3. Treatment with oleacein (OLE) reduces inflammation in NOE. Serum expression of parameters related to inflammation was used as a measure of 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.
    [0083] Fig. 4. Treatment with oleacein (OLE) reduces demyelination in the optic nerve during NOE. To examine whether OLE treatment prevents demyelination, optic nerves were isolated from mice 21-23 days after immunization and stained with LFB. Histological analysis of the optic nerve of control mice, C; from control mice treated with OLE, C + OLE; from induced mice, NOE; and from induced OLE-treated mice, NOE + OLE, showed demyelination. Oleacein treatment improved all these parameters.
    [0085] Fig. 5. Treatment with oleacein (OLE) reduces oxidative stress on the optic nerve during NOE. To examine whether OLE treatment prevents superoxide ion generation, optic nerves were isolated from mice 21-23 days after immunization and stained with the oxidative fluorescent probe, dihydroetidium, DHE. (A) Representative images of DHE staining, and (B) bar graph showing quantification of red fluorescence of optic nerve sections from control mice, C; from control mice treated with OLE, C + OLE; from induced mice, NOE; and from OLE-treated induced mice, NOE + OLE, demonstrated an accumulation of superoxide anion. Oleacein treatment improved all these parameters.
    [0087] Fig. 6. Treatment with oleacein (OLE) increases neuroprotection in NOE. The expression levels of malondialdehyde, MDA (A), from 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.
    [0089] Fig. 7. Treatment with oleacein (OLE) protects against alteration of the blood-brain barrier and decreases molecular extravasation in mice with NOE. Endogenous IgG extravasation was used as a measure of impaired blood brain barrier and extravasation of plasma proteins in the brain. A) Representative microphotographs of immunofluorescence. B) Quantification of the fluorescence intensity. 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 animals. tp <0.001 vs control and *** p <0.05 vs EAE.
    [0091] 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).
    [0093] 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 LPS, intracellular ROS production was assessed by cytometric analysis flow. The panel shows the quantification expressed in arbitrary units (AU), (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 LPS, the expression of COX-2, iNOS, p-p65-NF K B and NLRP3 was identified in cell lysates 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).
    [0095] Fig. 9. Oleocantal 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 oleocantal: (A) After 4 or 24 hours of stimulation with 0.1 pg / ml LPS, 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).
    [0097] EXAMPLES
    [0099] The invention will be exemplified, but not necessarily limited by the following experiments.
    [0101] 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).
    [0103] Immunization was performed with 100 pg of a partial oligodendrocyte myelin glycoprotein peptide (MOG33-55) in complete Freund's adjuvant containing 4 mg of Mycobacteríum 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 ^ l of Bordetella pertussis toxin was administered intraperitoneally . The administration of 10 mg / kg of oleacein acid (OLE) was carried out intraperitoneally once a day, starting from 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 oleocantal, were isolated from an olive oil extract prepared using the extraction method similar to that described below for sample preparation. 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 spun at 4,000 rpm for 5 minutes. A part of the acetonitrile phase (25 ml) was collected, mixed with 1.0 ml of a syringaldehyde solution (0.5 mg / ml) in acetonitrile and evaporated under reduced pressure using a rotary evaporator (Buchi, Flawil , Switzerland).
    [0105] Mice were sacrificed 21-23 days after immunization with MOG. Induction was self-antibody MOG-IgG1 was quantified in serum of mice from the four experimental groups. As expected, the levels of MOG-IgG1 auto-antibodies increased significantly 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).
    [0107] 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 after immunization (Fig. 2 ). Examination of the hematoxylin and eosin stained sections of the optic nerve (H and E) showed pronounced cell infiltrates in the tissues of mice with NOE compared to that of healthy control mice. In contrast, in the tissues of mice with NOE treated with OLE, the infiltration of cells was remarkably reduced, being comparable to that observed in the tissues of untreated control mice. OLE treatment in healthy control mice had no significant effect.
    [0109] The inventors also determine circulating levels of 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 the inflammatory mediators increased significantly 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: their 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-1p, a mediator that connects innate and adaptive immunity, were detected in the serum of mice with NOE and this increase was reduced by up to 80% in animals treated with OLE.
    [0111] Next, we 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 rapid 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 mice with NOE treated with OLE showed a higher staining of LFB compared to the optic nerves of the mice with untreated EAE, which means a suppression in the demyelination of the optic nerves.
    [0113] It is well known that inflammation increases reactive oxygen species (ROS) levels, leading to oxidative stress that mediates tissue damage. To investigate whether prophylactic administration of OLE to mice with NOE also resulted in reduced ROS accumulation, the inventors used the redox-sensitive fluorescent probe DHE that detects superoxide anion (O2-) as an initial indicator of ROS activity. ). Cryogenic sections of the optic nerve from mice from the different experimental groups were incubated with the DHE probe and evaluated by fluorescence microscopy. Fluorescence signal intensity 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 the NOE mice showed an increase in red fluorescence intensity throughout the tissue. However, OLE treatment inhibited NOE-induced ROS production: ethidium red fluorescence was largely attenuated, being similar to that observed in tissues from healthy control mice.
    [0115] Since the superoxide anion has also been implicated in lipid peroxidation and protein oxidation, the inventors assessed 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, assessed by an antioxidant / iron-reducing power test (FRAP), as a marker of non-antioxidant status Enzymes, and sestrin-3 levels, 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 significant attenuation of this decrease compared to untreated EAE mice.
    [0117] The optic nerve is considered part of the CNS. During acute optic neuritis, the rupture of the blood-brain barrier (EBR), 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. Accordingly, the inventors characterized NOE-induced BHR damage in OLE-treated and untreated mice by analyzing the extravasation of endogenous serum IgG into the brain using immunohistochemistry. Fluorescence signal intensity was quantified using Image J software (NIH, Bethesda, MD, USA). As shown in Fig. 7, the brain of mice with untreated NOE showed a marked increase in IgG extravasation (c, g) compared to healthy control animals (a, b). However, this increase in permeability was prevented by administration of OLE. OLE treatment did not modify the integrity of BHR in control animals.
    [0119] Next, the inventors assess whether the protective effects found in vivo in mice with NOE treated with OLE involve direct actions on relevant inflammatory CNS cells, the inventors studied the effects of OLE on activated BV-2 microglial cells to mimic responses observed in neuroinflammatory disorders (Fig. 8).
    [0121] Since ROS production can configure specific inflammatory programs, the inventors investigated intracellular ROS accumulation. Flow cytometric analysis showed that intracellular ROS accumulation, which was significantly increased in LPS-treated BV-2 cells, was dramatically suppressed in cells pre-treated with different doses of OLE (Fig. 8A).
    [0123] To determine whether OLE was able to directly modulate inflammatory activity in microglia, BV-2 cells were pretreated 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 LPS-activated BV2 cells showed an increase in iNOS and COX-2 expression after 4 and 24 h compared to unstimulated control cells. However, cellular pretreatment with OLE led to a dose-dependent inhibition of LPS-induced iNOs and COX-2 production, while OLE per se did not affect their basal expression. The OLE presence also reduced the ability of LPS to induce TNFa and IL-ip secretion in a dose-dependent manner (Fig. 8C). Similarly, activation of signaling mediators / mechanisms that influence these inflammatory responses, such as phosphorylation of p65-NF K B and overexpression of NLRP3, was also reduced (Fig. 8B).
    [0125] This invention also relates to methods of using natural compounds, oleacein and oleocantal, to inhibit the responses of activated microglia.
    [0127] Finally, the inventors are also evaluating the protective effects of another natural serchoiridoid: oleochantal on relevant inflammatory cells in the CNS. The inventors studied the effects of oleochantal on activated BV-2 microglial cells to mimic the responses observed in neuroinflammatory disorders (Fig. 9). Oleocantal 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.
    [0129] To determine whether oleocantal was able to directly modulate inflammatory activity in microglia, BV-2 cells were pretreated with 1, 10, and 20 pM oleocantal 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 LPS-activated BV2 cells showed an increase in iNOS and COX-2 expression after 4 and 24 h compared to unstimulated control cells. However, cellular pretreatment with oleocantal led to a dose dependent inhibition of LPS-induced iNOS and COX-2 production, while oleocantal per se did not affect its basal expression. The presence of oleochantal also reduced the ability of LPS to induce TNFa and IL-1p secretion in a dose-dependent manner (Fig. 9B). Similarly, activation of signaling mediators / mechanisms that influence these inflammatory responses, such as phosphorylation of p65-NF K B and overexpression of NLRP3, was also reduced (Fig. 9A).
    权利要求:
    Claims (8)
    [1]
    1. Compound of formula (I):

    [2]
    2. Compound for use according to claim 1, wherein the compound of formula (I) is oleacein:

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

    [4]
    4. Compound for use according to any of claims 1-3, wherein the Compound is administered orally, parenterally, or intranasally.
    [5]
    5. Compound for use according to any of claims 1-4, where the compound is administered at a dose between 5 mg / day and 20 mg / day.
    [6]
    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]
    7. The composition for use according to claim 6, wherein the composition is a pharmaceutical composition or a nutraceutical composition.
    [8]
    8. The composition for use according to claim 6 or 7, further comprising other additional drugs to provide a combination therapy.
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    Jin et al.2016|Dose-dependent neuroprotection and neurotoxicity of simvastatin through reduction of Farnesyl Pyrophosphate in mice treated with intracerebroventricular injection of Aβ 1-42
    同族专利:
    公开号 | 公开日
    WO2020136221A1|2020-07-02|
    ES2769902B2|2020-12-04|
    引用文献:
    公开号 | 申请日 | 公开日 | 申请人 | 专利标题
    US20180162800A1|2005-05-09|2018-06-14|Monell Chemical Senses Center|Use of the Irritating Principal Oleocanthal in Olive Oil, as Well as Structurally and Functionally Similar Compounds|
    US20130115202A1|2005-08-31|2013-05-09|Theta Biomedical Consulting & Development Co., Inc|Anti-inflammatory compositions for treating neuro-inflammation|
    WO2007133908A2|2006-05-09|2007-11-22|The Trustees Of The University Of Pennsylvania|Use of the irritating principal oleocanthal in olive oil, as well as structurally and functionally similar compounds|
    US20100173856A1|2006-09-28|2010-07-08|Olive Denk, Inc.|Methods of producing and using nutritional and pharmaceutical compositions that include one or more active substances|
    CA2192209C|1994-06-23|2007-03-13|Evan Benjamin Dreyer|Treatment of optic neuritis|
    EP1575576A2|2002-09-24|2005-09-21|Novartis AG|Organic compounds|
    US7879344B2|2006-06-29|2011-02-01|Kimberly-Clark Worldwide, Inc.|Transdermal delivery of oleocanthal for relief of inflammation|
    US20090082738A1|2007-09-24|2009-03-26|Vad Vijay B|Natural Anti-Inflammatory Agents for Reducing Pain|
    CA3045733A1|2016-12-22|2018-06-28|Novaliq Gmbh|Compositions comprising tacrolimus for the treatment of intraocular inflammatory eye diseases|
    ES2690412B1|2017-03-10|2019-11-13|Univ Sevilla|Procedure for obtaining dialdehyde secairidoids|
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    ES201831296A|ES2769902B2|2018-12-28|2018-12-28|Use of secoiridoids for the treatment of optic neuritis.|ES201831296A| ES2769902B2|2018-12-28|2018-12-28|Use of secoiridoids for the treatment of optic neuritis.|
    PCT/EP2019/087044| WO2020136221A1|2018-12-28|2019-12-26|Use of secoiridoids for the treatment of optic neuritis|
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