WO2003075905A1 - Use of derivatives of 2,4-disubstituted phenols as inhibitors of the expression of l-selectins and of the isoform that is inducible from nitric oxide synthase - Google Patents

Use of derivatives of 2,4-disubstituted phenols as inhibitors of the expression of l-selectins and of the isoform that is inducible from nitric oxide synthase Download PDF

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WO2003075905A1
WO2003075905A1 PCT/ES2002/000119 ES0200119W WO03075905A1 WO 2003075905 A1 WO2003075905 A1 WO 2003075905A1 ES 0200119 W ES0200119 W ES 0200119W WO 03075905 A1 WO03075905 A1 WO 03075905A1
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terbutyl
isopropyl
phenyl
use according
methyl
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PCT/ES2002/000119
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Spanish (es)
French (fr)
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Jesús LOPEZ-BELMONTE PASCUAL
Juan Lopez-Belmonte Pascual
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Bobel 246, S.L.
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Priority to AU2002246135A priority Critical patent/AU2002246135A1/en
Priority to PCT/ES2002/000119 priority patent/WO2003075905A1/en
Publication of WO2003075905A1 publication Critical patent/WO2003075905A1/en

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/045—Hydroxy compounds, e.g. alcohols; Salts thereof, e.g. alcoholates
    • A61K31/05—Phenols
    • A61K31/055—Phenols the aromatic ring being substituted by halogen

Definitions

  • the present invention relates to the use in medicine and veterinary of certain derivatives of 2,4-disubstituted phenols which, due to a potent activity as an inhibitor of the inducible isoform of nitric oxide synthase (iNOS) and as an inhibitor of the expression of The adhesion molecule L-selectin on the surface of the plasma membrane of leukocytes, are very useful in the preparation of drugs for the treatment of therapeutic indications mediated by the action of iNOS, by the action of L expression -selectin or both.
  • iNOS inducible isoform of nitric oxide synthase
  • NO nitric oxide
  • NO is of a gaseous nature and is synthesized from the amino acid L-arginine thanks to the action of a cytosolic emzyme called nitric oxide synthase.
  • This enzyme belongs to the family of flavoproteins, has a certain homology with cytochrome P450 reductases and from it three different isoforms have been described: endothelial nitric oxide synthase (eNOS), neuronal nitric oxide synthase (nNOS) and inducible nitric oxide synthase (iNOS).
  • Both eNOS and nNOS are expressed constitutively, are dependent on calcium and calmodulin and insensitive to the effects of glucocorticoids, release small amounts of NO intermittently for short periods, the constitutive NO acts as a mechanism of signal transduction in multiple physiological processes and is involved in the maintenance of vascular tone.
  • the biochemical characteristics of the constitutive NOS are similar, but they differ in their location and in the function performed by the NO produced by them.
  • the eNOS is preferentially located in renal endothelial cells, platelets and mesangial, osteoblasts and osteoclasts and is involved in the regulation of vascular homeostasis and platelet function.
  • the nNOS is of nerve location, both central and peripheral, and is a producer of a NO that acts as a central and peripheral neurotransmitter.
  • the iNOS is not calcium dependent and inflammatory stimuli of endotoxins (such as lipopolysaccharide) and certain cytokines (interleukin-1, tumor necrosis factor (TNF-alpha) or interferon gamma (INF-gamma)), induce the formation of iNOS in a variety of cells, including epithelial cells, macrophages and neutrophils. Inducible nitric oxide synthase produces much greater amounts of NO for long periods of time compared to the other isoforms.
  • endotoxins such as lipopolysaccharide
  • cytokines interleukin-1, tumor necrosis factor (TNF-alpha) or interferon gamma (INF-gamma)
  • the NO synthesized by the iNOS plays a fundamental role in the defense against external agents such as the cytotoxic molecule and is involved in vasodilation in inflammatory processes. It is known to those skilled in the art that large amounts of NO produced by iNOS have antimicrobial (J. Clin. Invest., 1989, 81: 1129-1136) and antitumor (Science, 1987, 235: 473-476) functions. The NO is involved in the elimination of parasites, bacteria, tumor cells and viruses. The high concentration of NO synthesized by iNOS explains the cytotoxic and cytostatic effects of this enzyme. Therefore, iNOS plays an important role in the immune response.
  • additional states in which there is an advantage of inhibiting the production of NO to Starting from L-arginine, by inhibiting the expression of iNOS include autoimmune and / or inflammatory joint and musculoskeletal states, inflammatory diseases of the gastrointestinal tract, cardiovascular ischemia, diabetes, hyperalgesia, cerebral ischemia, systemic hypotension associated with septic shock and / or toxic induced by a wide variety of agents, cytokine therapy, as an adjuvant for short-term immunosuppression in transplant therapy, as well as NO-mediated CNS disorders.
  • Inflammation is the response of vascularized tissues to aggressions.
  • the main objective of inflammation is to locate and eradicate irritation and repair surrounding tissue.
  • the inflammatory response involves 3 important stages: dilation of the capillaries to increase blood flow, microvascular structural changes and release of plasma proteins from the bloodstream, and thirdly leukocyte transmigration through the endothelium and accumulation in the damaged area.
  • Its regulation involves different humoral factors, such as prostaglandins, vasoactive amines, cytokines, etc., but in the last two decades it has been found that certain cytoplasmic membrane molecules present in both leukocytes and endothelial cells play an important role. in the regulation of leukocyte infiltration of inflamed tissues and, therefore, in the development and maintenance of the inflammatory response.
  • the leukocyte adhesion cascade is a sequence of adhesion and activation events that ends with the leukocyte extravasation. At least 5 steps appear to be necessary for the effective recruitment of leukocytes: capture, rotation, slow rotation, firm adhesion and transmigration. Blocking any of these steps can significantly reduce the accumulation of leukocytes in the tissue.
  • the recruitment of leukocytes to the foci of inflammation is directed by a series of adhesion molecules present both on the surface of the plasma membrane of the leukocytes themselves, as in the endothelial cells of the capillaries and venules of inflamed tissues.
  • the adhesion molecules involved in the inflammatory response belong mainly to 3 large families: selectins, integrins and the immunoglobulin superfamily.
  • Selectins are glycoproteins characterized by containing an amino terminal region with structural homology to calcium dependent lectins (LC), followed by another structure similar to epidermal growth factor (EGF); from 2 to 9 complement regulatory protein (SCR) regions; a transmembrane zone; and a cytoplasmic region.
  • LC calcium dependent lectins
  • EGF epidermal growth factor
  • SCR complement regulatory protein
  • L-selectin is constitutively expressed in virgin neutrophils, monocytes, T and B lymphocytes and in a subpopulation of K cells.
  • This adhesion molecule contributes greatly to the capture of leukocytes during the early stages of the adhesion cascade, demonstrating its involvement in said recruitment of leukocytes to foci of inflammation.
  • L-selectin is removed from the surface of the leukocytes, and released into the medium, in response to different activating stimuli, such as cytokines, calcium ionophore, forbol esters, etc. This mechanism of regulation of its expression is shared with other adhesion molecules or cytokine receptors.
  • the lectin-like region of L-selectin participates in the recognition of ligands; these domains bind L-selectin to ligands through a calcium-dependent interaction of the lectin-like amino-terminal region of L-selectin with complex carbohydrates (sialylated, sulphated and / or fucosylated) present in different glycoproteins of endothelial cells.
  • NK cells are lymphocytes of the innate immune system that recognize and induce lysis of a variety of cells, including virus-infected cells, tumor cells, and allogeneic cells without prior sensitization.
  • NK cells make a variety of cytokines such as interferon-gamma, tumor necrosis factor alpha (TNF-alpha), IL-l ⁇ , IL-10 and also produce chemokines such as RANTES, l ⁇ macrophage inflammatory protein and l ⁇ macrophage inflammatory protein, which are involved in the elimination of intracellular pathogens in vivo, as well as in the generation of antigen-specific immune response.
  • NK cells The differentiation of NK cells is accompanied by the expression on the cell surface of a glycoprotein (PEN5), which is a post-translational modification of the selectin P ligand (PSGL-1).
  • PEN5 glycoprotein
  • PSGL-1 selectin P ligand
  • the PEN5 epitope creates on PSGL-1 a single binding site for L-selectin.
  • the selectivity of PEN5 expression in NK cells, and their absence in antigen-specific T and B lymphocytes suggests that the specific functions of NK cells depend on the acquisition of this ligand for L-selectin. Therefore, the control of interactions of NK cells via PEN5 can have important implications for the induction and maintenance of immunity to tumor and infectious diseases, as well as amplifying the immune response to an inflammatory stimulus.
  • the present invention provides a group of products derived from 2,4-disubstituted phenols, known chemically in the literature. Of these 2,4-disubstituted phenols, their inhibitory activity of the inducible isoform of nitric oxide synthase, nor their ability to inhibit the expression of the adhesion molecule L-selectin on the membrane surface had never been described before. leukocyte plasma
  • Patent ES2087019 mentions the inhibitory activity of 5-lipoxygenase (5-LO) of the included products in the present invention, but at no time the possibility is mentioned that said products may exhibit inhibitory activity of the inducible isoform of nitric oxide synthase, nor of its ability to inhibit the expression of the adhesion molecule L-selectin in the surface of the plasma membrane of leukocytes.
  • 5-LO 5-lipoxygenase
  • the object of the present invention is the use of a 2,4-disubstituted phenol derivative of formula (I),
  • R2 is (C1-C4) -alkyl, (C1-C4) -acrylic, CF 3 , Cl, Br or I;
  • R3 is H, (C 1 -C 4 ) -alkyl, (C 1 -C 4 ) -acrylic, (C 1 -C 4 ) -alkoxy, CF 3 / F, Cl, Br, phenyl, OH, SH, NH 2 or amino mono- or disubstituted by (Ci-C 3 ) -alkyl;
  • R4 is (C 1 -C 4 ) -alkyl, (C1-C4) -acrylic, CF 3 , F, Cl, Br, I, carboxyl, (C 1 -C 4 ) -alkoxycarboxyl, cyano, nitro or phenyl; Y
  • R6 is H, (C 1 -C 4 ) -alkyl, (C 1 -C 4 ) -acrylic, CF 3 , Cl, Br or I;
  • R2 is (C 1 -C 4) -alkyl, CF 3 , Cl, Br or I
  • R3 is H, (C 1 -C 4 ) -alkyl, CF 3 , F, Cl, Br, phenyl, OH, SH, NH 2 or dimethylamino
  • R4 is (C 1 -C 4 ) -alkyl, CF 3 , F, Cl, Br, I, carboxyl, cyano, nitro or phenyl
  • R6 is H, (C1-C4) -alkyl, CF 3 , Cl, Br or I.
  • R2 is I;
  • R3 is H, methyl, ethyl, isopropyl, terbutyl, CF 3 , F, Cl, Br, phenyl or OH;
  • R4 is methyl, ethyl, isopropyl, terbutyl, CF 3 , F, Cl, Br, I, carboxyl or phenyl;
  • R6 is H, isopropyl, terbutyl, CF 3 , Cl, Br or I.
  • R3 is H, methyl, ethyl, isopropyl, terbutyl, CF 3 , Cl, Br or phenyl
  • R4 is isopropyl, terbutyl, CF 3 , Br, I, carboxyl or phenyl
  • R6 is H, isopropyl, terbutyl, CF 3 or I.
  • R3 is H; R4 is terbutyl, CF 3 , I or phenyl; and R6 is terbutyl, CF 3 or I. Still more preferred is that in which R4 and R6 are I.
  • a 2,4-disubstituted phenol derivative of formula (I) wherein: R2 is CF 3 or Br; R3 is H, methyl, ethyl, isopropyl, terbutyl, CF 3 , F, Cl, Br, phenyl or OH; R4 is methyl, ethyl, isopropyl, terbutyl, CF 3 , F, Cl, Br, I, carboxyl or phenyl; and R6 is H, isopropyl, terbutyl, CF 3 , Cl, Br or I.
  • R3 is H, methyl, ethyl, isopropyl, terbutyl, CF 3 , Cl, Br or phenyl
  • R4 is isopropyl, terbutyl, CF 3 , Br, I, carboxyl or phenyl
  • R6 is H, isopropyl, terbutyl, CF 3 or I.
  • R3 is H; R4 is terbutyl, CF 3 , I or phenyl; and R6 is terbutyl, CF 3 or I.
  • a 2,4-disubstituted phenol derivative of formula (I) wherein: R2 is isopropyl or terbutyl; R3 is H, methyl, ethyl, isopropyl, terbutyl, CF 3 , F, Cl, Br, phenyl or OH; R4 is methyl, ethyl, isopropyl, terbutyl, CF 3 , F, Cl, Br, I, carboxyl or phenyl; and R6 is H, isopropyl, terbutyl, CF 3 , Cl, Br or I.
  • R3 is H, methyl, ethyl, isopropyl, terbutyl, CF 3 , Cl, Br or phenyl
  • R4 is isopropyl, terbutyl, CF 3 , Br, I, carboxyl or phenyl
  • R6 is H, isopropyl, terbutyl, CF 3 or I.
  • R3 is H; R4 is terbutyl, CF 3 , I or phenyl; and R6 is terbutyl, CF 3 or I.
  • Some of the products of formula (I) have stereoisomers.
  • the use of these isomers, separately or mixed, as inhibitors of the inducible isoform of nitric oxide synthase and inhibitors of the expression of the L-selectin adhesion molecule on the surface of the plasma membrane of leukocytes, is also the subject of the present invention.
  • the products of formula (I) or their pharmaceutically acceptable salts or solvates are useful in the preparation of medicaments for the treatment of diseases mediated by the action of the inducible isoform of nitric oxide lasintase, by the action of L-selectin expression or both, with the preferences outlined above.
  • Derivatives of 2,4-disubstituted phenols or their pharmaceutically acceptable salts can be administered orally, parenterally or topically, in the form of injectables, tablets, dragees , syrups, lotions, shampoos, ointments, suppositories, eye drops, etc.
  • Example 1 Effect on the activity of nitric oxide synthase (NOS) in a model of TNB-induced ulcerative colitis in rats.
  • NOS nitric oxide synthase
  • TLB trinitrobenzene sulfonic acid
  • the animals were given different anti-inflammatory drugs, 24 hours after the administration of TNB.
  • the anti-inflammatory drugs that were used were: prednisolone (0.5mg kg "1 ), L-NAME (30mg kg " 1 ) and 2,4,6-triodophenol (BOBEL-24, l-25mg kg "1 ), administered orally for 3 consecutive days.
  • nitric oxide synthase activity in colonic tissue was carried out by means of the citrulline test, in which the conversion of monohydrochloric L- [14C] -argenine to [14C] -citrulline is measured, according to the method described by Knowles et al. (Biochem. Biophys. Res. Commun., 1990, v.172, 1042-1048).
  • Colonic tissue samples were obtained 24 hours after TNB administration and 2, 3 and 4 days after TNB administration. These tissues were homogenized in a buffer solution with HEPES, sucrose, DL-dithiothreitol, leupeptin, soy trypsin inhibitor and aprotinin. After centrifuging the homogenates, a sample of the supernatant was added to a tube containing 10 ⁇ l of preheated incubation buffer at 37 ° C which in turn contained potassium phosphate, L-valine, NADPH, MgCl 2 and CaCl 2 , L- argenine and L- [14C] -monohydric chloride and was incubated for 10min at 37 ° C.
  • the reaction was terminated by the addition of 500 ⁇ l of a mixture of H 2 0: Dowex-AG50.
  • the mixture of resin and incubated solution of the sample was dispersed and diluted by the addition of 860 ⁇ l of distilled water. After precipitation of the resin, 975 ⁇ l of supernatant was subtracted to count in a liquid scintillation counter (2ml of EcoScint H liquid; Packard).
  • BOBEL-24 prednisolone and L-NAME once a day for a period of 3 days, on the induced colonic damage after a single injection of trinitrobenzene sulfonic acid (TNB; 80 mg kg "1 administered on day 0), the results show the damage score as a function of the time elapsed after the administration of TNB.
  • TNB trinitrobenzene sulfonic acid
  • L-citrulline that can be inhibited by in vitro incubation with L-NMMA is taken as an index of NOS activity, expressed as nmol min "1 g " 1 of tissue.
  • cNOS constitutive NOS
  • iNOS inducible and independent form of calcium from NOS
  • Figure 2 shows the effect of BOBEL-24 treatment on the inhibition of the increase in total NOS activity. Finding a 68 + 7% inhibition of NOS activity.
  • Figure 3 shows the effect that treatment with BOBEL-24 produced on the activity of calcium-independent NOS (iNOS), which could be observed 2 and 4 days after the administration of TNB.
  • the inhibition of iNOS activity was 82 ⁇ 11% and 45 ⁇ 14% at 2 and 4 days after TNB administration, respectively.
  • Example 2 Study of the effect on the activity of polymorphonuclear leukocytes in vivo in the rat by intravital microscopy.
  • the objective of the study was to determine if the substance
  • mesenteric venules without branches of 25-40 ⁇ m in diameter, and their associated arterioles (15-25 ⁇ m) were selected for study.
  • polymorphonuclear leukocytes The activation of polymorphonuclear leukocytes was performed by administering a dose of 3mg kg "1 iv of endotoxin (LPS E. Coli 0.111: B4) to the animals once they have been anesthetized and prepared for intravital observation. To determine the effect of the test and reference substances on the activation of leukocytes, these were administered orally as a pre-treatment lh before the administration of endotoxin. The administration of endotoxin resulted in an increase in the number of adherent neutrophils determined in a section of lOO ⁇ m of venule over an observation period of 1 hour.
  • pre-treatment with BOBEL-24 also resulted in an inhibition of the reduction of endotoxin-induced neutrophil rolling, with an increase in the speed of rolling that is related to the dose of BOBEL. -24 administered (see Figure 5).

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Abstract

The invention relates to the use of certain derivatives of 2,4-disubstituted phenols in medicine and veterinary medicine. Due to the powerful activity of said derivatives as an inhibitor of the isoform inducible from nitric oxide synthase (iNOS) and as an inhibitor of the expression of the L-selectin adhesion molecule on the surface of the plasma membrane of the leukocytes, they can be used in the preparation of medicaments for the treatment of pathologies induced by the action of the iNOS, by the action of the expression of the L-selectin or by both.

Description

USO DE DERIVADOS DE FENOLES 2 , 4 -DISUBSTITUIDOS COMO INHIBIDORES DE LA EXPRESIÓN DE L-SELECTINAS Y DE LA ISOFORMA INDUCIBLE DE LA SINTASA DEL OXIDO NÍTRICOUSE OF PHENOL DERIVATIVES 2, 4 -DISUBSTITUTES AS INHIBITORS OF THE EXPRESSION OF L-SELECTINES AND THE INDUCIBLE ISOFORM OF THE SYNTHESIS OF NITRIC OXIDE
La presente invención se refiere al uso en medicina y veterinaria de ciertos derivados de fenoles 2,4- disubstituidos que, debido a una potente actividad como inhibidor de la isoforma inducible de la sintasa del óxido nítrico (iNOS) y como inhibidor de la expresión de la molécula de adhesión L-selectina en la superficie de la membrana plasmática de los leucocitos, resultan de gran utilidad en la preparación de medicamentos para el tratamiento de indicaciones terapéuticas mediadas por la acción de la iNOS, por la acción de la expresión de la L-selectina o por las dos.The present invention relates to the use in medicine and veterinary of certain derivatives of 2,4-disubstituted phenols which, due to a potent activity as an inhibitor of the inducible isoform of nitric oxide synthase (iNOS) and as an inhibitor of the expression of The adhesion molecule L-selectin on the surface of the plasma membrane of leukocytes, are very useful in the preparation of drugs for the treatment of therapeutic indications mediated by the action of iNOS, by the action of L expression -selectin or both.
ESTADO DE LA TÉCNICA ANTERIORSTATE OF THE PREVIOUS TECHNIQUE
La excesiva producción de óxido nítrico (NO) ha sido implicado en la respuesta inmune e inflamatoria. El papel del NO, como mediador fisiológico o como radical citotóxico, viene determinado por las condiciones y cantidad de su síntesis.Excessive production of nitric oxide (NO) has been implicated in the immune and inflammatory response. The role of NO, as a physiological mediator or as a cytotoxic radical, is determined by the conditions and amount of its synthesis.
El NO es de naturaleza gaseosa y es sintetizado a partir del aminoácido L-arginina gracias a la actuación de una emzima citosólica denominada sintasa del óxido nítrico. Esta enzima pertenece a la familia de las flavoproteínas, tiene cierta homología con las reductasas del citocromo P450 y de ella se han descrito tres diferentes isoformas: la sintasa del óxido nítrico endotelial (eNOS) , la sintasa del óxido nítrico neuronal (nNOS) y la sintasa del óxido nítrico inducible (iNOS) . Tanto eNOS como nNOS son expresadas de forma constitutiva, son dependientes de calcio y calmodulina e insensibles a los efectos de los glucocorticoides, liberan pequeñas cantidades de NO de forma intermitente por periodos cortos, el NO constitutivo actúa como un mecanismo de transducción de señales en múltiples procesos fisiológicos y está involucrado en el mantenimiento del tono vascular.NO is of a gaseous nature and is synthesized from the amino acid L-arginine thanks to the action of a cytosolic emzyme called nitric oxide synthase. This enzyme belongs to the family of flavoproteins, has a certain homology with cytochrome P450 reductases and from it three different isoforms have been described: endothelial nitric oxide synthase (eNOS), neuronal nitric oxide synthase (nNOS) and inducible nitric oxide synthase (iNOS). Both eNOS and nNOS are expressed constitutively, are dependent on calcium and calmodulin and insensitive to the effects of glucocorticoids, release small amounts of NO intermittently for short periods, the constitutive NO acts as a mechanism of signal transduction in multiple physiological processes and is involved in the maintenance of vascular tone.
Las características bioquímicas de las NOS constitutivas son similares, pero se diferencian en su localización y en la función realizada por el NO producido por ellas. La eNOS está localizada preferentemente en las células endoteliales, plaquetas y mesangiales renales, osteoblastos y osteoclastos y está involucrada en la regulación de la homeostasia vascular y la función plaquetaria. La nNOS es de localización nerviosa, tanto central como periférica, y es productora de un NO que actúa como neurotransmisor central y periférico.The biochemical characteristics of the constitutive NOS are similar, but they differ in their location and in the function performed by the NO produced by them. The eNOS is preferentially located in renal endothelial cells, platelets and mesangial, osteoblasts and osteoclasts and is involved in the regulation of vascular homeostasis and platelet function. The nNOS is of nerve location, both central and peripheral, and is a producer of a NO that acts as a central and peripheral neurotransmitter.
Por el contrario, la iNOS no es expresada bajo condiciones fisiológicas sino que requiere ser inducida. La iNOS no es calcio dependiente y los estímulos inflamatorios de endotoxinas (como el lipopolisacárido) y ciertas citoquinas (interleucina-1 , factor de necrosis tumoral (TNF-alpha) o interferon gamma (INF-gamma) ) , inducen la formación de iNOS en una variedad de células, incluyendo células epiteliales, macrófagos y neutrófilos. La sintasa del óxido nítrico inducible produce cantidades mucho mayores de NO durante largos periodos de tiempo en comparación con las otras isoformas. El NO sintetizado por la iNOS desarrolla un papel fundamental en la defensa contra agentes externos como la molécula citotóxica y está involucrado en la vasodilatación en los procesos inflamatorios . Es conocido por los expertos en la materia que grandes cantidades de NO producido por iNOS presenta funciones antimicrobianas (J. Clin. Invest . , 1989, 81: 1129-1136) y antitumorales (Science, 1987, 235: 473-476). El NO está involucrado en la eliminación de parásitos, bacterias, células tumorales y virus. La alta concentración de NO sintetizado por iNOS explica los efectos citotóxicos y citostáticos de este enzima. Por lo tanto, el iNOS desempeña un importante papel en la respuesta inmune.On the contrary, the iNOS is not expressed under physiological conditions but needs to be induced. The iNOS is not calcium dependent and inflammatory stimuli of endotoxins (such as lipopolysaccharide) and certain cytokines (interleukin-1, tumor necrosis factor (TNF-alpha) or interferon gamma (INF-gamma)), induce the formation of iNOS in a variety of cells, including epithelial cells, macrophages and neutrophils. Inducible nitric oxide synthase produces much greater amounts of NO for long periods of time compared to the other isoforms. The NO synthesized by the iNOS plays a fundamental role in the defense against external agents such as the cytotoxic molecule and is involved in vasodilation in inflammatory processes. It is known to those skilled in the art that large amounts of NO produced by iNOS have antimicrobial (J. Clin. Invest., 1989, 81: 1129-1136) and antitumor (Science, 1987, 235: 473-476) functions. The NO is involved in the elimination of parasites, bacteria, tumor cells and viruses. The high concentration of NO synthesized by iNOS explains the cytotoxic and cytostatic effects of this enzyme. Therefore, iNOS plays an important role in the immune response.
Altas concentraciones de NO debido a una crónica, inapropiada o excesiva expresión de iNOS ha sido implicado en el daño tisular e inflamación, especialmente en ciertas enfermedades autoinmunes. El iNOS encontrado en macrófagos, monocitos, etc., produce cantidades micromolares de NO el cual contribuye al daño tisular de forma local y a la hipertensión sistémica la cual acompaña al shoc séptico y otros procesos inflamatorios. Del mismo modo, la interacción del NO con el metabolismo del hierro, tiene como efecto adverso el desarrollo de una enfermedad crónica, debido a la inhibición de la ALA sintasa y al secuestro de hierro que inducen a una inhibición tanto de la síntesis de hem como de la eritropoyesis . También puede ser responsable directo de alguno de los cuadros patológicos de la malaria grave, ya que la hiperproducción de NO que se da en el curso de la malaria grave, alteraría la función sináptica e induciría áreas locales de hipoglucemia y acidosis. Del mismo modo, puede estar linvolucrado en la inmunodeficiencia asociada a la malaria.High concentrations of NO due to chronic, inappropriate or excessive expression of iNOS has been implicated in tissue damage and inflammation, especially in certain autoimmune diseases. The iNOS found in macrophages, monocytes, etc., produces micromolar amounts of NO which contributes to tissue damage locally and to systemic hypertension which accompanies septic shoc and other inflammatory processes. Similarly, the interaction of NO with iron metabolism, has the adverse effect of the development of a chronic disease, due to the inhibition of ALA synthase and iron sequestration that induce an inhibition of both heme synthesis and of erythropoiesis. It may also be directly responsible for any of the pathological conditions of severe malaria, since the hyperproduction of NO that occurs in the course of severe malaria would alter synaptic function and induce local areas of hypoglycemia and acidosis. Similarly, it may be involved in the immunodeficiency associated with malaria.
De acuerdo con esto, estados adicionales en los que existe una ventaja de inhibir la producción de NO a partir de L-arginina, mediante la inhibición de la expresión de la iNOS, incluyen estados autoinmunes y/o inflamatorios articulares y musculoesqueléticos, enfermedades inflamatorias del tracto gastrointestinal, isquemia cardiovascular, diabetes, hiperalgesia, isquemia cerebral, hipotensión sistémica asociada con el choque séptico y/o tóxico inducido por una amplia variedad de agentes, terapia de citoquinas, como adyuvante para la inmunosupresión a corto plazo en terapia de transplantes, así como trastornos del SNC mediados por NO.Accordingly, additional states in which there is an advantage of inhibiting the production of NO to Starting from L-arginine, by inhibiting the expression of iNOS, include autoimmune and / or inflammatory joint and musculoskeletal states, inflammatory diseases of the gastrointestinal tract, cardiovascular ischemia, diabetes, hyperalgesia, cerebral ischemia, systemic hypotension associated with septic shock and / or toxic induced by a wide variety of agents, cytokine therapy, as an adjuvant for short-term immunosuppression in transplant therapy, as well as NO-mediated CNS disorders.
La inflamación es la respuesta de los tejidos vascularizados frente a las agresiones. El principal objetivo de la inflamación es localizar y erradicar la irritación y reparar el tejido circundante. La respuesta inflamatoria implica 3 importantes etapas: dilatación de los capilares para incrementar el flujo sanguíneo, cambios estructurales microvasculares y liberación de proteínas plasmáticas del torrente sanguíneo, y en tercer lugar la transmigración leucocitaria a través del endotelio y acumulación en la zona dañada. En su regulación se encuentran implicados diferentes factores humorales, como las prostaglandinas, aminas vasoactivas, citoquinas, etc., pero en las dos últimas décadas se ha podido constatar que ciertas moléculas de membrana citoplasmática presentes tanto en leucocitos como en células endoteliales desempeñas un papel importante en la regulación de la infiltración leucocitaria de los tejidos inflamados y, por lo tanto, en el desarrollo y mantenimiento de la respuesta inflamatoria.Inflammation is the response of vascularized tissues to aggressions. The main objective of inflammation is to locate and eradicate irritation and repair surrounding tissue. The inflammatory response involves 3 important stages: dilation of the capillaries to increase blood flow, microvascular structural changes and release of plasma proteins from the bloodstream, and thirdly leukocyte transmigration through the endothelium and accumulation in the damaged area. Its regulation involves different humoral factors, such as prostaglandins, vasoactive amines, cytokines, etc., but in the last two decades it has been found that certain cytoplasmic membrane molecules present in both leukocytes and endothelial cells play an important role. in the regulation of leukocyte infiltration of inflamed tissues and, therefore, in the development and maintenance of the inflammatory response.
La cascada de adhesión de leucocitos es una secuencia de eventos de adhesión y activación que finaliza con la extravasación del leucocito. Al menos 5 pasos parecen ser necesarios para el reclutamiento efectivo de leucocitos: captura, rotación, rotación lenta, adhesión firme y transmigración. El bloqueo de cualquiera de estos pasos puede reducir de forma significativa la acumulación de leucocitos en el tejido.The leukocyte adhesion cascade is a sequence of adhesion and activation events that ends with the leukocyte extravasation. At least 5 steps appear to be necessary for the effective recruitment of leukocytes: capture, rotation, slow rotation, firm adhesion and transmigration. Blocking any of these steps can significantly reduce the accumulation of leukocytes in the tissue.
El reclutamiento de leucocitos a los focos de inflamación está dirigido por una serie de moléculas de adhesión presentes tanto en la superficie de la membrena plasmática de los propios leucocitos, como en las células endoteliales de los capilares y vénulas de tejidos inflamados. Las moléculas de adhesión implicadas en la respuesta inflamatoria pertenecen principalmente a 3 grandes familias: selectinas, integrinas y la superfamilia de inmunoglobulinas .The recruitment of leukocytes to the foci of inflammation is directed by a series of adhesion molecules present both on the surface of the plasma membrane of the leukocytes themselves, as in the endothelial cells of the capillaries and venules of inflamed tissues. The adhesion molecules involved in the inflammatory response belong mainly to 3 large families: selectins, integrins and the immunoglobulin superfamily.
Señales específicas producidas en respuesta a la herida e infección, controlan la expresión y activación de ciertas moléculas de adhesión. Las interacciones y respuestas entonces iniciadas por la unión de estas moléculas de adhesión a sus receptores/ligandos juegan un papel importante en la mediación de las reacciones inflamatoria e inmune que constituyen una línea de la defensa del cuerpo contra estos ataques .Specific signals produced in response to injury and infection control the expression and activation of certain adhesion molecules. The interactions and responses then initiated by the binding of these adhesion molecules to their receptors / ligands play an important role in mediating the inflammatory and immune reactions that constitute a line of the body's defense against these attacks.
Los miembros de la familia de selectinas están involucradas en la interacción inicial entre los leucocitos que se encuentran en el torrente sanguíneo, y las células endoteliales activadas en los focos de inflamación. Las selectinas son glicoproteínas caracterizadas por contener una región amino terminal con homología estructural a lectinas calcio dependientes (LC) , seguida de otra estructura similar al factor de crecimiento epidérmico (EGF) ; de 2 a 9 regiones tipo proteína reguladora del complemento (SCR) ; una zona transmembrana; y una región citoplasmática .Members of the selectin family are involved in the initial interaction between leukocytes found in the bloodstream, and activated endothelial cells in the foci of inflammation. Selectins are glycoproteins characterized by containing an amino terminal region with structural homology to calcium dependent lectins (LC), followed by another structure similar to epidermal growth factor (EGF); from 2 to 9 complement regulatory protein (SCR) regions; a transmembrane zone; and a cytoplasmic region.
La L-selectina se expresa de forma constitutiva en neutrófilos, monocitos, linfocitos T y B vírgenes y en una subpoblación de células K. Esta molécula de adhesión contribuye en gran medida a la captura de leucocitos durante las primeras etapas de la cascada de adhesión, demostrándose su implicación en dicho reclutamiento de leucocitos a focos de inflamación. Después de la captura, la L-selectina es eliminada de la superficie de los leucocitos, y liberada al medio, en respuesta a diferentes estímulos activadores, como citoquinas, ionóforo de calcio, esteres de forbol , etc. Este mecanismo de regulación de su expresión es compartido con otras moléculas de adhesión o receptores de citoquinas.L-selectin is constitutively expressed in virgin neutrophils, monocytes, T and B lymphocytes and in a subpopulation of K cells. This adhesion molecule contributes greatly to the capture of leukocytes during the early stages of the adhesion cascade, demonstrating its involvement in said recruitment of leukocytes to foci of inflammation. After capture, L-selectin is removed from the surface of the leukocytes, and released into the medium, in response to different activating stimuli, such as cytokines, calcium ionophore, forbol esters, etc. This mechanism of regulation of its expression is shared with other adhesion molecules or cytokine receptors.
La región tipo lectina de la L-selectina participa en el reconocimiento de los ligandos; estos dominios unen la L-selectina a los ligandos mediante una interacción dependiente de calcio de la región amino terminal tipo lectina de la L-selectina con carbohidratos complejos (sialilados, sulfatados y/o fucosilados) presentes en diferentes glicoproteinas de las células endoteliales.The lectin-like region of L-selectin participates in the recognition of ligands; these domains bind L-selectin to ligands through a calcium-dependent interaction of the lectin-like amino-terminal region of L-selectin with complex carbohydrates (sialylated, sulphated and / or fucosylated) present in different glycoproteins of endothelial cells.
Las células NK son linfocitos del sistema inmune innato que reconocen e inducen la lisis de una variedad de células, incluyendo células infectadas por virus, células tumorales, y células alogénicas sin sensibilización previa. Además, las células NK elaboran una variedad de citoquinas como interferon-gamma, factor de necrosis tumoral alfa (TNF-alpha) , IL-lβ, IL- 10 y también producen quimioquinas como RANTES, proteína inflamatoria del macrófago lα y proteína inflamatoria del macrófago lβ, las cuales están involucradas en la eliminación de patógenos intracelulares in vivo, así como en la generación de respuesta inmune antígeno-específica.NK cells are lymphocytes of the innate immune system that recognize and induce lysis of a variety of cells, including virus-infected cells, tumor cells, and allogeneic cells without prior sensitization. In addition, NK cells make a variety of cytokines such as interferon-gamma, tumor necrosis factor alpha (TNF-alpha), IL-lβ, IL-10 and also produce chemokines such as RANTES, lα macrophage inflammatory protein and lβ macrophage inflammatory protein, which are involved in the elimination of intracellular pathogens in vivo, as well as in the generation of antigen-specific immune response.
La diferenciación de las células NK está acompañada por la expresión en la superficie celular de una glicoproteína (PEN5) , la cual es una modificación post-translacional del ligando de la selectina P (PSGL- 1) . El epítopo PEN5 crea sobre PSGL-1 un único sitio de unión para la L-selectina. La selectividad de la expresión de PEN5 en las células NK, y su ausencia en linfocitos T y B antígeno-específicos, sugiere que las específicas funciones de las células NK dependen de la adquisición de este ligando para L-selectina. Por lo tanto, el control de interacciones de las células NK vía PEN5 puede tener importantes implicaciones para la inducción y mantenimiento de la inmunidad a enfermedades tumorales e infecciosas, así como amplificando la respuesta inmune a un estímulo inflamatorio .The differentiation of NK cells is accompanied by the expression on the cell surface of a glycoprotein (PEN5), which is a post-translational modification of the selectin P ligand (PSGL-1). The PEN5 epitope creates on PSGL-1 a single binding site for L-selectin. The selectivity of PEN5 expression in NK cells, and their absence in antigen-specific T and B lymphocytes, suggests that the specific functions of NK cells depend on the acquisition of this ligand for L-selectin. Therefore, the control of interactions of NK cells via PEN5 can have important implications for the induction and maintenance of immunity to tumor and infectious diseases, as well as amplifying the immune response to an inflammatory stimulus.
La presente invención proporciona un grupo de productos derivados de fenoles 2 , 4 -disubstituidos, conocidos químicamente en la literatura. De estos fenoles 2,4- disubstituidos nunca se había descrito antes su actividad inhibitoria de la isoforma inducible de la sintasa del óxido nítrico, ni de su capacidad de inhibición de la expresión de la molécula de adhesión L-selectina en la superficie de la membrana plasmática de los leucocitos.The present invention provides a group of products derived from 2,4-disubstituted phenols, known chemically in the literature. Of these 2,4-disubstituted phenols, their inhibitory activity of the inducible isoform of nitric oxide synthase, nor their ability to inhibit the expression of the adhesion molecule L-selectin on the membrane surface had never been described before. leukocyte plasma
La patente ES2087019 menciona la actividad inhibitoria de la 5-lipoxigenasa (5-LO) de los productos incluidos en la presente invención, pero en ningún momento se menciona la posibilidad de que dichos productos puedan presentar actividad inhibitoria de la isoforma inducible de la sintasa del óxido nítrico, ni de su capacidad de inhibición de la expresión de la molécula de adhesión L-selectina en la superficie de la membrana plasmática de los leucocitos.Patent ES2087019 mentions the inhibitory activity of 5-lipoxygenase (5-LO) of the included products in the present invention, but at no time the possibility is mentioned that said products may exhibit inhibitory activity of the inducible isoform of nitric oxide synthase, nor of its ability to inhibit the expression of the adhesion molecule L-selectin in the surface of the plasma membrane of leukocytes.
En la solicitud de patente EP147892 se menciona cierta actividad analgésica y antiinflamatoria de un producto incluido en la presente invención, pero dicha actividad analgésica y antiinflamatoria podría deberse a alguna actividad inhibitoria de la cicloxigenasa, puesto que los ensayos comparativos descritos se realizaron frente a típicos inhibidores de dicho enzima.In patent application EP147892 some analgesic and anti-inflammatory activity of a product included in the present invention is mentioned, but said analgesic and anti-inflammatory activity could be due to some inhibitory activity of cycloxygenase, since the comparative tests described were performed against typical inhibitors. of said enzyme.
Por lo tanto, nada en el conocimiento técnico general, enseña o hace obvio que los productos incluidos entre los derivados de la presente invención, puedan presentar actividad inhibitoria de la isoforma inducible de la sintasa del óxido nítrico, ni de su capacidad de inhibición de la expresión de la molécula de adhesión L-selectina en la superficie de la membrana plasmática de los leucocitos, enseñanza ésta que es parte integrante de la presente invención.Therefore, nothing in the general technical knowledge, teaches or makes it obvious that the products included among the derivatives of the present invention, can exhibit inhibitory activity of the inducible isoform of nitric oxide synthase, nor of its ability to inhibit expression of the L-selectin adhesion molecule on the surface of the leukocyte plasma membrane, this teaching being an integral part of the present invention.
DESCRIPCIÓN DETALLADA DE LA INVENCIÓNDETAILED DESCRIPTION OF THE INVENTION
Es objeto de la presente invención el uso de un derivado de fenol 2 , 4-disubstituido de fórmula (I), The object of the present invention is the use of a 2,4-disubstituted phenol derivative of formula (I),
Figure imgf000010_0001
Figure imgf000010_0001
donde :where :
R2 es (C1-C4) -alquilo, (C1-C4) -acilo, CF3, Cl , Br o I;R2 is (C1-C4) -alkyl, (C1-C4) -acrylic, CF 3 , Cl, Br or I;
R3 es H, (C1-C4) -alquilo, (C1-C4) -acilo, (C1-C4) - alcoxilo, CF3/ F, Cl , Br, fenilo, OH, SH, NH2 o amino mono- o disubstituido por (Ci-C3) -alquilo;R3 is H, (C 1 -C 4 ) -alkyl, (C 1 -C 4 ) -acrylic, (C 1 -C 4 ) -alkoxy, CF 3 / F, Cl, Br, phenyl, OH, SH, NH 2 or amino mono- or disubstituted by (Ci-C 3 ) -alkyl;
R4 es (C1-C4) -alquilo, (C1-C4) -acilo, CF3, F, Cl , Br, I, carboxilo, (C1-C4) -alcoxicarboxilo, ciano, nitro o fenilo; yR4 is (C 1 -C 4 ) -alkyl, (C1-C4) -acrylic, CF 3 , F, Cl, Br, I, carboxyl, (C 1 -C 4 ) -alkoxycarboxyl, cyano, nitro or phenyl; Y
R6 es H, (C1-C4) -alquilo, (C1-C4) -acilo, CF3, Cl , Br o I;R6 is H, (C 1 -C 4 ) -alkyl, (C 1 -C 4 ) -acrylic, CF 3 , Cl, Br or I;
o de las sales o solvatos farmacéuticamente aceptables de dicho derivado, en la preparación de un medicamento inhibidor de la isoforma inducible de la sintasa del óxido nítrico e inhibidor de la expresión de la molécula de adhesión L-selectina en la superficie de la membrana plasmática de los leucocitos.or of the pharmaceutically acceptable salts or solvates of said derivative, in the preparation of an inducible drug for the inducible isoform of nitric oxide synthase and inhibitor of the expression of the adhesion molecule L-selectin on the surface of the plasma membrane of leukocytes
Son preferidos aquéllos en los que: R2 es (C1-C4) - alquilo, CF3, Cl , Br o I; R3 es H, (C1-C4) -alquilo, CF3, F, Cl, Br, fenilo, OH, SH, NH2 o dimetilamino; R4 es (C1-C4) -alquilo, CF3, F, Cl , Br, I, carboxilo, ciano, nitro o fenilo; y R6 es H, (C1-C4) -alquilo, CF3, Cl , Br o I. En una realización particular de la presente invención, R2 es I; R3 es H, metilo, etilo, isopropilo, terbutilo, CF3, F, Cl, Br, fenilo o OH; R4 es metilo, etilo, isopropilo, terbutilo, CF3, F, Cl, Br, I, carboxilo o fenilo; y R6 es H, isopropilo, terbutilo, CF3, Cl , Br o I. Siendo preferidos aquéllos en los que: R3 es H, metilo, etilo, isopropilo, terbutilo, CF3, Cl , Br o fenilo; R4 es isopropilo, terbutilo, CF3, Br, I, carboxilo o fenilo; y R6 es H, isopropilo, terbutilo, CF3 o I. Aún se prefiere más aquellos en los que: R3 es H; R4 es terbutilo, CF3, I o fenilo; y R6 es terbutilo, CF3 o I . Todavía se prefiere más aquél en el que R4 y R6 son I .Preferred are those in which: R2 is (C 1 -C 4) -alkyl, CF 3 , Cl, Br or I; R3 is H, (C 1 -C 4 ) -alkyl, CF 3 , F, Cl, Br, phenyl, OH, SH, NH 2 or dimethylamino; R4 is (C 1 -C 4 ) -alkyl, CF 3 , F, Cl, Br, I, carboxyl, cyano, nitro or phenyl; and R6 is H, (C1-C4) -alkyl, CF 3 , Cl, Br or I. In a particular embodiment of the present invention, R2 is I; R3 is H, methyl, ethyl, isopropyl, terbutyl, CF 3 , F, Cl, Br, phenyl or OH; R4 is methyl, ethyl, isopropyl, terbutyl, CF 3 , F, Cl, Br, I, carboxyl or phenyl; and R6 is H, isopropyl, terbutyl, CF 3 , Cl, Br or I. Those preferred in which: R3 is H, methyl, ethyl, isopropyl, terbutyl, CF 3 , Cl, Br or phenyl; R4 is isopropyl, terbutyl, CF 3 , Br, I, carboxyl or phenyl; and R6 is H, isopropyl, terbutyl, CF 3 or I. Even more preferred are those in which: R3 is H; R4 is terbutyl, CF 3 , I or phenyl; and R6 is terbutyl, CF 3 or I. Still more preferred is that in which R4 and R6 are I.
En otra realización particular de la presente invención, se usa un derivado de fenol 2,4- disubstituido de fórmula (I) donde: R2 es CF3 o Br; R3 es H, metilo, etilo, isopropilo, terbutilo, CF3, F, Cl , Br, fenilo o OH; R4 es metilo, etilo, isopropilo, terbutilo, CF3, F, Cl, Br, I, carboxilo o fenilo; y R6 es H, isopropilo, terbutilo, CF3, Cl, Br o I. De entre estos, se prefiere usar aquéllos en los que: R3 es H, metilo, etilo, isopropilo, terbutilo, CF3, Cl , Br o fenilo; R4 es isopropilo, terbutilo, CF3, Br, I, carboxilo o fenilo; y R6 es H, isopropilo, terbutilo, CF3 o I. Y todavía se prefiere más aquellos en los que: R3 es H; R4 es terbutilo, CF3, I o fenilo; y R6 es terbutilo, CF3 o I.In another particular embodiment of the present invention, a 2,4-disubstituted phenol derivative of formula (I) is used wherein: R2 is CF 3 or Br; R3 is H, methyl, ethyl, isopropyl, terbutyl, CF 3 , F, Cl, Br, phenyl or OH; R4 is methyl, ethyl, isopropyl, terbutyl, CF 3 , F, Cl, Br, I, carboxyl or phenyl; and R6 is H, isopropyl, terbutyl, CF 3 , Cl, Br or I. Among these, it is preferred to use those in which: R3 is H, methyl, ethyl, isopropyl, terbutyl, CF 3 , Cl, Br or phenyl ; R4 is isopropyl, terbutyl, CF 3 , Br, I, carboxyl or phenyl; and R6 is H, isopropyl, terbutyl, CF 3 or I. And still more preferred are those in which: R3 is H; R4 is terbutyl, CF 3 , I or phenyl; and R6 is terbutyl, CF 3 or I.
En otra realización particular de la presente invención, se usa un derivado de fenol 2,4- disubstituido de fórmula (I) donde: R2 es isopropilo o terbutilo; R3 es H, metilo, etilo, isopropilo, terbutilo, CF3, F, Cl , Br, fenilo o OH; R4 es metilo, etilo, isopropilo, terbutilo, CF3, F, Cl, Br, I, carboxilo o fenilo; y R6 es H, isopropilo, terbutilo, CF3, Cl, Br o I. De entre los anteriores se prefiere usar aquéllos en los que: R3 es H, metilo, etilo, isopropilo, terbutilo, CF3, Cl , Br o fenilo; R4 es isopropilo, terbutilo, CF3, Br, I, carboxilo o fenilo; y R6 es H, isopropilo, terbutilo, CF3 o I. Y todavía se prefiere usar más aquél los en los que: R3 es H; R4 es terbutilo, CF3, I o fenilo; y R6 es terbutilo, CF3 o I.In another particular embodiment of the present invention, a 2,4-disubstituted phenol derivative of formula (I) is used wherein: R2 is isopropyl or terbutyl; R3 is H, methyl, ethyl, isopropyl, terbutyl, CF 3 , F, Cl, Br, phenyl or OH; R4 is methyl, ethyl, isopropyl, terbutyl, CF 3 , F, Cl, Br, I, carboxyl or phenyl; and R6 is H, isopropyl, terbutyl, CF 3 , Cl, Br or I. Among the above it is preferred to use those in which: R3 is H, methyl, ethyl, isopropyl, terbutyl, CF 3 , Cl, Br or phenyl ; R4 is isopropyl, terbutyl, CF 3 , Br, I, carboxyl or phenyl; and R6 is H, isopropyl, terbutyl, CF 3 or I. And it is still more preferred to use those in which: R3 is H; R4 is terbutyl, CF 3 , I or phenyl; and R6 is terbutyl, CF 3 or I.
En particular, son especialmente útiles los productos de la Tabla 1 adjunta, donde se indican los códigos internos (códigos de la compañía) , los nombres químicos y los número de registro (RN = registry number) de Chemical Abstracts Service (CAS) o mediante la mención del volumen y número de abstracts donde se han publicado en la revista Chemical Abstracts (CA) o el número de patente donde se describen por primera vez, de algunos de los derivados de fenoles 2,4- disubstituidos de fórmula (I) que han sido preparados a modo de ejemplo. In particular, the products in the attached Table 1 are particularly useful, where internal codes (company codes), chemical names and registration numbers (RN = registry number) of Chemical Abstracts Service (CAS) are indicated or by the mention of the volume and number of abstracts where they have been published in the journal Chemical Abstracts (CA) or the patent number where they are described for the first time, of some of the 2,4-disubstituted phenol derivatives of formula (I) that They have been prepared by way of example.
Tabla 1Table 1
Derivados de fenoles 2 , 4 -disubstituidosPhenol derivatives 2, 4 -disubstituted
Cód. Int. Nombre químico RN/Ref .CA/Pat .Code Int. Chemical name RN / Ref .CA / Pat.
Bobel - -1 4- -fluoro-2 -yodofenol [2713-29-3]Bobel - -1 4- -fluoro-2-iodophenol [2713-29-3]
Bobel- -2 4- -terbutil-2-yodofenol [38941-98-9]Bobel- -2 4- -butyl-2-iodophenol [38941-98-9]
Bobel- -3 2- -terbutil-4 -yodofenol [60803-25-0]Bobel- -3 2- -butyl-4-iodophenol [60803-25-0]
Bobel- -4 2 6-diisopropil-4-yodofenol ES2087019Bobel- -4 2 6-diisopropyl-4-iodophenol ES2087019
Bobel- -5 2 4 -diyodofenol CA 24:5289Bobel- -5 2 4 -diiodophenol CA 24: 5289
Bobel- -6 2- -metil-4 , 6 -diyodofenol [4186-52-1]Bobel- -6 2- -methyl-4,6-diiodophenol [4186-52-1]
Bobel - -7 2- -etil-4 , 6-diyodofenol ES2087019Bobel - -7 2- -ethyl-4, 6-diiodophenol ES2087019
Bobel- -8 2- -isopropil-4 , 6-diyodofenol [127502-66-3]Bobel- -8 2- -isopropyl-4,6-diiodophenol [127502-66-3]
Bobel- -9 2- -terbutil-4 , 6-diyodofenol [60803-26-1]Bobel- -9 2- -butyl-4, 6-diiodophenol [60803-26-1]
Bobel- -10 2- -cloro-4 , 6 -diyodofenol [15459-49-1]Bobel- -10 2- -chloro-4, 6-diiodophenol [15459-49-1]
Bobel- -11 2- -bromo-4 , 6 -diyodofenol [89466-01-3]Bobel- -11 2- -bromo-4,6-diiodophenol [89466-01-3]
Bobel - -12 2- -trifluorometil-4 , 6-diyodofenol [61494-84-6]Bobel - -12 2- -trifluoromethyl-4, 6-diiodophenol [61494-84-6]
Bobel - -13 4- -metil-2, 6 -diyodofenol [2432-18-0]Bobel - -13 4- -methyl-2, 6-diiodophenol [2432-18-0]
Bobel- -14 4- -isopropil-2 , 6-diyodofenol [2432-19-1]Bobel- -14 4- -isopropyl-2, 6-diiodophenol [2432-19-1]
Bobel - -15 4- -terbutil-2 , 6-diyodofenol [75908-75-7]Bobel - -15 4- -butyl-2, 6-diiodophenol [75908-75-7]
Bobel - -16 4- -fenil-2 , 6 -diyodofenol CA 55:5845dBobel - -16 4- -phenyl-2, 6-diiodophenol CA 55: 5845d
Bobel - -17 4- -fluoro-2 , 6-diyodofenol [392-72-3]Bobel - -17 4- -fluoro-2, 6-diiodophenol [392-72-3]
Bobel - -18 4- -cloro-2 , 6 -diyodofenol [15459-50-4]Bobel - -18 4- -chloro-2, 6-diiodophenol [15459-50-4]
Bobel - -19 4- -bromo-2 , 6 -diyodofenol [15459-51-5]Bobel - -19 4- -bromo-2, 6-diiodophenol [15459-51-5]
Bobel - -20 4- -trifluorometil-2 , 6 -diyodofenol ES2087019Bobel - -20 4- -trifluoromethyl-2, 6-diiodophenol ES2087019
Bobel - -21 3- -trifluorometil-2 , 6-diyodofenol ES2087019Bobel - -21 3- -trifluoromethyl-2, 6-diiodophenol ES2087019
Bobel - -22 4- -nitro-2, 6-diyodofenol [305-85-1]Bobel - -22 4- -nitro-2, 6-diiodophenol [305-85-1]
Bobel - -23 3- -terbutil-2 , 4-diyodofenol ES2087019Bobel - -23 3- -butyl-2, 4-diiodophenol ES2087019
Bobel - -24 2 4 , 6-triyodofenol [609-23-4]Bobel - -24 2 4, 6-triiodophenol [609-23-4]
Bobel - -25 3- -metil-2, 4-6-triyodofenol [2109-12-8] Tabla 1 (Continuación)Bobel - -25 3- -methyl-2, 4-6-triiodophenol [2109-12-8] Table 1 (Continued)
Cód. Int. Nombre químico RN/Ref .CA/PatCode Int. Chemical name RN / Ref .CA / Pat
Bobel-26 3-etil-2 , 4 , 6 -triyodofenol [124311-20-2]Bobel-26 3-ethyl-2, 4, 6 -triodophenol [124311-20-2]
Bobel-27 3-isopropil-2 , 4-6-triyodofenol ES2087019Bobel-27 3-isopropyl-2, 4-6-triiodophenol ES2087019
Bobel-28 3-fenil-2 , 4 , 6 -triyodofenol [91353-81-0]Bobel-28 3-phenyl-2, 4, 6 -triodophenol [91353-81-0]
Bobel-29 3-fluoro-2 , 4 , 6-triyodofenol [444-07-5]Bobel-29 3-fluoro-2, 4, 6-triiodophenol [444-07-5]
Bobel-30 3-cloro-2 , 4 , 6 -triyodofenol [89465-75-8]Bobel-30 3-Chloro-2, 4, 6 -triodophenol [89465-75-8]
Bobel-31 3 -bromo-2 , 4 , 6 -triyodofenol [124311-19-9]Bobel-31 3 -bromo-2, 4, 6 -triodophenol [124311-19-9]
Bobel-32 3 -trifluorometil-2 ,4,6- triyodofenol ES2087019Bobel-32 3 -trifluoromethyl-2, 4,6- triiodophenol ES2087019
Bobel-33 3-hidroxi-2 , 4 , 6 -triyodofenol [19403-92-0]Bobel-33 3-hydroxy-2, 4, 6 -triodophenol [19403-92-0]
Bobel-34 4-hidroxi-3 , 5-diyodobenzonitrilo [1689-83-4]Bobel-34 4-hydroxy-3, 5-diiodobenzonitrile [1689-83-4]
Bobel-35 2,4, 6-triterbutilfenol [732-26-3]Bobel-35 2,4,6-triterbutylphenol [732-26-3]
Bobel-36 ácido 4 -hidroxi-3 , 5- diyodobenzoico [618-76-8]Bobel-36 4-hydroxy-3, 5-diiodobenzoic acid [618-76-8]
Algunos de los productos de fórmula (I) presentan estereoisómeros . El uso de éstos isómeros, por separado o mezclados, como inhibidores de la isoforma inducible de la sintasa del óxido nítrico e inhibidores de la expresión de la molécula de adhesión L-selectina en la superficie de la membrana plasmática de los leucocitos, también es objeto de la presente invención.Some of the products of formula (I) have stereoisomers. The use of these isomers, separately or mixed, as inhibitors of the inducible isoform of nitric oxide synthase and inhibitors of the expression of the L-selectin adhesion molecule on the surface of the plasma membrane of leukocytes, is also the subject of the present invention.
Como inhibidores de la isoforma inducible de la sintasa del óxido nítrico e inhibidores de la expresión de la molécula de adhesión L-selectina en la superficie de la membrana plasmática de los leucocitos, los productos de fórmula (I) o sus sales o solvatos farmacéuticamente aceptables, son útiles en la preparación de medicamentos para el tratamiento de enfermedades mediadas por la acción de la isoforma inducible de lasintasa del óxido nítrico, por la acción de la expresión de la L-selectina o por las dos, con las preferencias señaladas anteriormente.As inhibitors of the inducible isoform of nitric oxide synthase and inhibitors of the expression of the L-selectin adhesion molecule on the surface of the leukocyte plasma membrane, the products of formula (I) or their pharmaceutically acceptable salts or solvates , are useful in the preparation of medicaments for the treatment of diseases mediated by the action of the inducible isoform of nitric oxide lasintase, by the action of L-selectin expression or both, with the preferences outlined above.
Los derivados de fenoles 2 , 4 -disubstituidos o sus sales farmacéuticamente aceptables, cuyo uso es objeto de la presente invención, convenientemente asociados a aditivos o excipientes conocidos, pueden administrarse por vía oral, parenteral o tópica, en forma de inyectables, comprimidos, grageas, jarabes, lociones, champúes, pomadas, supositorios, colirios, etc.Derivatives of 2,4-disubstituted phenols or their pharmaceutically acceptable salts, the use of which is the subject of the present invention, conveniently associated with known additives or excipients, can be administered orally, parenterally or topically, in the form of injectables, tablets, dragees , syrups, lotions, shampoos, ointments, suppositories, eye drops, etc.
EJEMPLOSEXAMPLES
La presente invención se ilustra mediante los siguientes ejemplos no limitativos.The present invention is illustrated by the following non-limiting examples.
Ejemplo 1. Efecto sobre la actividad de la sintasa del óxido nítrico (NOS) en un modelo de colitis ulcerosa inducida por TNB en rata .Example 1. Effect on the activity of nitric oxide synthase (NOS) in a model of TNB-induced ulcerative colitis in rats.
Mediante el presente ejemplo se pretendió demostrar la posible actividad inhibitoria del BOBEL-24 sobre la sintasa del óxido nítrico en un modelo de colitis ulcerosa inducida por ácido sulfónico de trinitrobenzeno (TNB) en la rata.By means of the present example, it was intended to demonstrate the possible inhibitory activity of BOBEL-24 on nitric oxide synthase in a model of ulcerative colitis induced by trinitrobenzene sulfonic acid (TNB) in the rat.
Se emplearon ratas istar macho, las cuales fueron anestesiadas y se les indujo una inflamación crónica del colon mediante la aplicación de una única dosis intracolónica de ácido sulfónico de trinitrobenzeno (TNB, 80mg kg"1, disuelto en etanol 30% y administrado en un volumen de lml kg"1) .Male istar rats were used, which were anesthetized and chronic inflammation of the colon was induced by applying a single intracolonic dose of trinitrobenzene sulfonic acid (TNB, 80mg kg "1 , dissolved in 30% ethanol and administered in one volume of lml kg "1 ).
A los animales se les administraron diferentes drogas antiiflamatorias, 24 horas tras la administración de TNB. Las drogas antiinflamatorias que se emplearon fueron: prednisolona (0.5mg kg"1) , L-NAME (30mg kg"1) y 2,4, 6 -triyodofenol (BOBEL-24, l-25mg kg"1) , administrándose oralmente durante 3 días consecutivos.The animals were given different anti-inflammatory drugs, 24 hours after the administration of TNB. The anti-inflammatory drugs that were used were: prednisolone (0.5mg kg "1 ), L-NAME (30mg kg " 1 ) and 2,4,6-triodophenol (BOBEL-24, l-25mg kg "1 ), administered orally for 3 consecutive days.
La medición de la actividad de la sintasa del óxido nítrico en el tejido colónico se llevó a cabo mediante el ensayo de la citrulina, en le que se mide la conversión de L- [14C] -argenina monohidroclorhídrica a [14C] -citrulina, según el método descrito por Knowles et al. (Biochem. Biophys. Res. Commun. , 1990, v.172, 1042-1048) .The measurement of nitric oxide synthase activity in colonic tissue was carried out by means of the citrulline test, in which the conversion of monohydrochloric L- [14C] -argenine to [14C] -citrulline is measured, according to the method described by Knowles et al. (Biochem. Biophys. Res. Commun., 1990, v.172, 1042-1048).
Se obtuvieron muestras de tejido colónico a las 24 horas tras la administración de TNB y a los 2, 3 y 4 días tras la administración de TNB. Estos tejidos se homogeneizaron en una solución tampón con HEPES, sucrosa, DL-dithiothreitol , leupeptina, inhibidor de tripsina de soja y aprotinina. Tras centrifugar los homogenizados, se añadió una muestra del sobrenadante a un tubo que contenía lOOμl de solución tampón de incubación precalentado a 37 °C que a su vez contenía fosfato de potasio, L-valina, NADPH, MgCl2 y CaCl2, L- argenina y L- [14C] -monohidriclorídrica y fue incubado durante lOmin a 37 °C. La reacción se terminó mediante la adición de 500μl de una mezcla de H20 : Dowex-AG50 . La mezcla de resina y solución incubada de la muestra se dispersó y se diluyó mediante la adición de 860μl de agua destilada. Tras la precipitación de la resina se sustrayeron 975μl de sobrenadante para contar en un contador líquido de centelleo (2ml de líquido EcoScint H; Packard) .Colonic tissue samples were obtained 24 hours after TNB administration and 2, 3 and 4 days after TNB administration. These tissues were homogenized in a buffer solution with HEPES, sucrose, DL-dithiothreitol, leupeptin, soy trypsin inhibitor and aprotinin. After centrifuging the homogenates, a sample of the supernatant was added to a tube containing 10 µl of preheated incubation buffer at 37 ° C which in turn contained potassium phosphate, L-valine, NADPH, MgCl 2 and CaCl 2 , L- argenine and L- [14C] -monohydric chloride and was incubated for 10min at 37 ° C. The reaction was terminated by the addition of 500μl of a mixture of H 2 0: Dowex-AG50. The mixture of resin and incubated solution of the sample was dispersed and diluted by the addition of 860μl of distilled water. After precipitation of the resin, 975μl of supernatant was subtracted to count in a liquid scintillation counter (2ml of EcoScint H liquid; Packard).
En la Figura 1 se muestra el efecto del tratamiento conFigure 1 shows the effect of treatment with
BOBEL-24, prednisolona y L-NAME una vez al día durante un periodo de 3 días, sobre el daño colónico inducido tras una sola inyección de ácido sulfónico de trinitrobenzeno (TNB; 80 mg kg"1 administrado en el día 0) , los resultados muestran la puntuación del daño en función del tiempo transcurrido tras la administración de TNB. Encontramos que BOBEL-24 inhibió el daño inducido por TNB en un 46±14% a los tres días tras la administración de TNB en dosis de lOmg kg"1, mientras que prednisolona y L-NAME no tuvieron ningún efecto significativo sobre el daño inducido por TNB en ninguno de los tiempos estudiados.BOBEL-24, prednisolone and L-NAME once a day for a period of 3 days, on the induced colonic damage after a single injection of trinitrobenzene sulfonic acid (TNB; 80 mg kg "1 administered on day 0), the results show the damage score as a function of the time elapsed after the administration of TNB. We found that BOBEL-24 inhibited the damage induced by TNB in 46 ± 14% at three days after administration of TNB in doses of 10 mg kg "1 , while prednisolone and L-NAME had no significant effect on TNB-induced damage at any of the times studied .
La formación de L-citrulina que se puede inhibir mediante la incubación in vitro con L-NMMA se toma como índice de actividad de NOS, expresada como nmol min"1g"1 de tejido. Mediante incubación con EGTA, se determinó la dependencia de la actividad enzimática de la presencia de calcio, siendo la actividad que depende de la presencia del calcio bajo condiciones de control la considerada como NOS constitutiva (cNOS) , mientras que la que no es inhibida por EGTA se toma como la forma inducible e independiente del calcio de NOS (iNOS) .The formation of L-citrulline that can be inhibited by in vitro incubation with L-NMMA is taken as an index of NOS activity, expressed as nmol min "1 g " 1 of tissue. By incubation with EGTA, the dependence of the enzymatic activity on the presence of calcium was determined, being the activity that depends on the presence of calcium under control conditions considered as constitutive NOS (cNOS), while the one that is not inhibited by EGTA is taken as the inducible and independent form of calcium from NOS (iNOS).
En la figura 2 se puede observar el efecto del tratamiento con BOBEL-24 sobre la inhibición en el incremento en la actividad total de NOS. Encontrándose una inhibición del 68+7% de la actividad de NOS.Figure 2 shows the effect of BOBEL-24 treatment on the inhibition of the increase in total NOS activity. Finding a 68 + 7% inhibition of NOS activity.
En la figura 3 se puede observar el efecto que el tratamiento con BOBEL-24 produjo sobre la actividad de NOS independiente del calcio (iNOS) , la cual se pudo observar a los 2 y 4 días tras la administración de TNB. La inhibición de la actividad de iNOS fue de un 82±11% y de un 45±14% a los 2 y 4 días tras la administración de TNB respectivamente. Ejemplo 2. Estudio del efecto sobre la actividad de los leucocitos polimorfonucleares in vivo en la rata mediante microscopía intravital .Figure 3 shows the effect that treatment with BOBEL-24 produced on the activity of calcium-independent NOS (iNOS), which could be observed 2 and 4 days after the administration of TNB. The inhibition of iNOS activity was 82 ± 11% and 45 ± 14% at 2 and 4 days after TNB administration, respectively. Example 2. Study of the effect on the activity of polymorphonuclear leukocytes in vivo in the rat by intravital microscopy.
El objetivo del estudio era determinar si la sustanciaThe objective of the study was to determine if the substance
BOBEL-24, posee algún efecto sobre la actividadBOBEL-24, has some effect on the activity
(velocidad de "rolling", adherencia) de los leucocitos polimorfonucleares activados in vivo en la rata, utilizando la circulación mesentérica como modelo y la técnica de microscopía intravital .(speed of "rolling", adhesion) of polymorphonuclear leukocytes activated in vivo in the rat, using mesenteric circulation as a model and the technique of intravital microscopy.
Tras la inducción de anestesia general conpentotal sódico (60mg kg"1, i.p.), se procedió a la exteriorización de un segmento de la porción ileo-cecal del mesenterio para su observación en el microscopio intravital. El resto del intestino que queda expuesto tras esta operación es cubierto con una gasa mojada en solución fisológica para prevenir la evaporación.Following the induction of general conpentotal sodium anesthesia (60mg kg "1 , ip), a segment of the ileo-cecal portion of the mesentery was performed for observation in the intravital microscope. The rest of the intestine that is exposed after this operation is covered with gauze dipped in physiological solution to prevent evaporation.
Una vez transcurrido un periodo de 30 minutos de estabilización tras la preparación del mesenterio, se seleccionaron vénulas mesentéricas sin ramificaciones de 25-40μm de diámetro, y sus arteriolas asociadas (15- 25μm) para su estudio.After a period of 30 minutes of stabilization after the preparation of the mesentery, mesenteric venules without branches of 25-40μm in diameter, and their associated arterioles (15-25μm) were selected for study.
La activación de los leucocitos polimorfonucleares se realizó mediante la administración de una dosis de 3mg kg"1 i.v. de endotoxina (LPS E. Coli 0.111:B4) a los animales una vez han sido anestesiados y preparados para la observación intravital. Para determinar el efecto de las sustancias de ensayo y referencia sobre la activación de los leucocitos, éstas se administraron oralmente como un pre-tratamiento lh antes de la administración de la endotoxina. La administración de la endotoxina resultó en un incremento en el número de neutrófilos adherentes determinado en una sección de lOOμm de vénula a lo largo de un periodo de observación de 1 hora.The activation of polymorphonuclear leukocytes was performed by administering a dose of 3mg kg "1 iv of endotoxin (LPS E. Coli 0.111: B4) to the animals once they have been anesthetized and prepared for intravital observation. To determine the effect of the test and reference substances on the activation of leukocytes, these were administered orally as a pre-treatment lh before the administration of endotoxin. The administration of endotoxin resulted in an increase in the number of adherent neutrophils determined in a section of lOOμm of venule over an observation period of 1 hour.
La administración de una dosis intravenosa de endotoxina resultó en una reducción de la velocidad del "rolling" de los neutrófilos en las vénulas post- capilares del mesenterio de la rata.The administration of an intravenous dose of endotoxin resulted in a reduction in the speed of "rolling" of the neutrophils in the post-capillary venules of the rat mesentery.
La administración oral de 3 dosis crecientes de 2,4,6- triyodofenol (BOBEL-24) , 1 hora antes de la administración de endotoxina resultó en una reducción, relacionada con la dosis administrada, en el aumento de la adherencia de neutrófilos inducido por la endotoxina en las vénulas post-capilares del mesenterio de la rata (ver Figura 4) .The oral administration of 3 increasing doses of 2,4,6-triiodophenol (BOBEL-24), 1 hour before the administration of endotoxin resulted in a reduction, related to the administered dose, in the increase in neutrophil adhesion induced by the endotoxin in the post-capillary venules of the rat mesentery (see Figure 4).
Del mismo modo, el pre-tratamiento con BOBEL-24 también resultó en una inhibición de la reducción del "rolling" de los neutrófilos inducida por la endotoxina, observándose un incremento en la velocidad de "rolling" que está relacionado con la dosis de BOBEL-24 administrada (ver Figura 5) . Similarly, pre-treatment with BOBEL-24 also resulted in an inhibition of the reduction of endotoxin-induced neutrophil rolling, with an increase in the speed of rolling that is related to the dose of BOBEL. -24 administered (see Figure 5).

Claims

REIVINDICACIONES
1. Uso de un derivado de fenol 2 , 4 -disubstituido de fórmula (I) ,1. Use of a 2,4-substituted phenol derivative of formula (I),
Figure imgf000020_0001
Figure imgf000020_0001
caracterizado porque:characterized in that:
R2 es (Cι-C4) -alquilo, (C1-C4) -acilo, CF3, Cl , Br o I;R2 is (Cι-C 4 ) -alkyl, (C 1 -C 4 ) -acrylic, CF 3 , Cl, Br or I;
R3 es H, (C1-C4) -alquilo, (Cι-C4) -acilo, (C1-C4) - alcoxilo, CF3, F, Cl , Br, fenilo, OH, SH, NH2 o amino mono- o disubstituido por (Cι-C3) -alquilo;R3 is H, (C 1 -C 4 ) -alkyl, (Cι-C 4 ) -acrylic, (C 1 -C 4 ) -alkoxy, CF 3 , F, Cl, Br, phenyl, OH, SH, NH 2 or amino mono- or disubstituted by (Cι-C 3 ) -alkyl;
R4 es (C1-C4) -alquilo, (C1-C4) -acilo, CF3, F, Cl , Br, I, carboxilo, (Cα-C4) -alcoxicarboxilo, ciano, nitro o fenilo; yR4 is (C 1 -C 4 ) -alkyl, (C 1 -C4) -acrylic, CF 3 , F, Cl, Br, I, carboxyl, (Cα-C 4 ) -alkoxycarboxyl, cyano, nitro or phenyl; Y
R6 es H, (Cι-C4) -alquilo, (Cι-C4) -acilo, CF3, Cl , Br o I;R6 is H, (Cι-C 4 ) -alkyl, (Cι-C 4 ) -acrylic, CF 3 , Cl, Br or I;
o de una sal o solvato farmacéuticamente aceptable de dicho derivado, en la preparación de un medicamento inhibidor de la isoforma inducible de lasintasa del óxido nítrico e inhibidor de la expresión de la molécula de adhesión L-selectina en la superficie de la membrana plasmática de los leucocitos . or of a pharmaceutically acceptable salt or solvate of said derivative, in the preparation of a drug inducible of nitric oxide inducible isoform and inhibitor of the expression of the adhesion molecule L-selectin on the surface of the plasma membrane of the leukocytes
2. Uso según la reivindicación 1, caracterizado porque en el derivado de fenol 2 , 4 -disubstituido : R2 es (Ci-C4) -alquilo, CF3, Cl , Br o I; R3 es H,2. Use according to claim 1, wherein the phenol derivative 2, 4 -disubstituido: R2 is (Ci-C 4) -alkyl, CF 3, Cl, Br or I; R3 is H,
(Cx-C4) -alquilo, CF3, F, Cl , Br, fenilo, OH, SH, NH2 o dimetilamino; R4 es (Cι-C4) -alquilo, CF3, F, Cl , Br, I, carboxilo, ciano, nitro o fenilo; y R6 es H, (d-C4) -alquilo, CF3/ Cl , Br o I .(C x -C 4 ) -alkyl, CF 3 , F, Cl, Br, phenyl, OH, SH, NH 2 or dimethylamino; R4 is (Cι-C 4 ) -alkyl, CF 3 , F, Cl, Br, I, carboxyl, cyano, nitro or phenyl; and R6 is H, (dC 4 ) -alkyl, CF 3 / Cl, Br or I.
3. Uso según la reivindicación 2, caracterizado porque en el derivado de fenol 2 , 4 -disubstituido :3. Use according to claim 2, characterized in that in the phenol derivative 2,4 -disubstituted:
R2 es I; R3 es H, metilo, etilo, isopropilo, terbutilo, CF3, F, Cl , Br, fenilo o OH; R4 es metilo, etilo, isopropilo, terbutilo, CF3, F, Cl , Br, I, carboxilo o fenilo; y R6 es H, isopropilo, terbutilo, CF3, Cl , Br o I .R2 is I; R3 is H, methyl, ethyl, isopropyl, terbutyl, CF 3 , F, Cl, Br, phenyl or OH; R4 is methyl, ethyl, isopropyl, terbutyl, CF 3 , F, Cl, Br, I, carboxyl or phenyl; and R6 is H, isopropyl, terbutyl, CF 3 , Cl, Br or I.
4. Uso según la reivindicación 3, caracterizado porque: R3 es H, metilo, etilo, isopropilo, terbutilo, CF3, Cl , Br o fenilo; R4 es isopropilo, terbutilo, CF3, Br, I, carboxilo o fenilo; y R6 es H, isopropilo, terbutilo, CF3 o I.4. Use according to claim 3, characterized in that: R3 is H, methyl, ethyl, isopropyl, terbutyl, CF 3 , Cl, Br or phenyl; R4 is isopropyl, terbutyl, CF 3 , Br, I, carboxyl or phenyl; and R6 is H, isopropyl, terbutyl, CF 3 or I.
5. Uso según la reivindicación 4, caracterizado porque: R3 es H; R4 es terbutilo, CF3, I o fenilo; y R6 es terbutilo, CF3 o I.5. Use according to claim 4, characterized in that: R3 is H; R4 is terbutyl, CF 3 , I or phenyl; and R6 is terbutyl, CF 3 or I.
6. Uso según la reivindicación 5, caracterizado porque : R4 y R6 son I .6. Use according to claim 5, characterized in that: R4 and R6 are I.
7. Uso según la reivindicación 2, caracterizado porque en el derivado de fenol 2 , 4 -disubstituido : R2 es CF3 o Br; R3 es H, metilo, etilo, isopropilo, terbutilo, CF3, F, Cl , Br, fenilo o OH; R4 es metilo, etilo, isopropilo, terbutilo, CF3, F, Cl, Br, I, carboxilo o fenilo; y R6 es H, isopropilo, terbutilo, CF3, Cl, Br o I .7. Use according to claim 2, wherein the phenol derivative 2, 4 I -disubstituido: R2 is CF 3 or Br; R3 is H, methyl, ethyl, isopropyl, terbutyl, CF 3 , F, Cl, Br, phenyl or OH; R4 is methyl, ethyl, isopropyl, terbutyl, CF 3 , F, Cl, Br, I, carboxyl or phenyl; and R6 is H, Isopropyl, terbutyl, CF 3 , Cl, Br or I.
8. Uso según la reivindicación 7, caracterizado porque: R3 es H, metilo, etilo, isopropilo, terbutilo, CF3, Cl , Br o fenilo; R4 es isopropilo, terbutilo, CF3, Br, I, carboxilo o fenilo; y R6 es H, isopropilo, terbutilo, CF3 o I.8. Use according to claim 7, characterized in that: R3 is H, methyl, ethyl, isopropyl, terbutyl, CF 3 , Cl, Br or phenyl; R4 is isopropyl, terbutyl, CF 3 , Br, I, carboxyl or phenyl; and R6 is H, isopropyl, terbutyl, CF 3 or I.
9. Uso según la reivindicación 8, caracterizado porque: R3 es H; R4 es terbutilo, CF3 , I o fenilo; y R6 es terbutilo, CF3 o I.9. Use according to claim 8, characterized in that: R3 is H; R4 is terbutyl, CF 3 , I or phenyl; and R6 is terbutyl, CF 3 or I.
10. Uso según la reivindicación 2, caracterizado porque en el derivado de fenol 2 , 4 -disubstituido : R2 es isopropilo o terbutilo; R3 es H, metilo, etilo, isopropilo, terbutilo, CF3, F, Cl , Br, fenilo o OH; R4 es metilo, etilo, isopropilo, terbutilo, CF3, F, Cl , Br, I, carboxilo o fenilo; y R6 es H, isopropilo, terbutilo, CF3, Cl , Br o I .10. Use according to claim 2, characterized in that in the 2,4-disubstituted phenol derivative: R2 is isopropyl or terbutyl; R3 is H, methyl, ethyl, isopropyl, terbutyl, CF 3 , F, Cl, Br, phenyl or OH; R4 is methyl, ethyl, isopropyl, terbutyl, CF 3 , F, Cl, Br, I, carboxyl or phenyl; and R6 is H, isopropyl, terbutyl, CF 3 , Cl, Br or I.
11. Uso según la reivindicación 10, caracterizado porque: R3 es H, metilo, etilo, isopropilo, terbutilo, CF3, Cl , Br o fenilo; R4 es isopropilo, terbutilo, CF3, Br, I, carboxilo o fenilo; y R6 es H, isopropilo, terbutilo, CF3 o I.11. Use according to claim 10, characterized in that: R3 is H, methyl, ethyl, isopropyl, terbutyl, CF 3 , Cl, Br or phenyl; R4 is isopropyl, terbutyl, CF 3 , Br, I, carboxyl or phenyl; and R6 is H, isopropyl, terbutyl, CF 3 or I.
12. Uso según la reivindicación 11, caracterizado porque: R3 es H; R4 es terbutilo, CF3 , I o fenilo; y R6 es terbutilo, CF3 o I.12. Use according to claim 11, characterized in that: R3 is H; R4 is terbutyl, CF 3 , I or phenyl; and R6 is terbutyl, CF 3 or I.
13. Uso de un derivado de fenol 2 , 4 -disubstituido según cualquiera de las reivindicaciones 1 a 12, o de una sal o solvato farmacéuticamente aceptable, en la preparación de un medicamento para el tratamiento de una enfermedad mediada por la acción de la isoforma inducible de lasintasa del óxido nítrico.13. Use of a 2, 4-substituted phenol derivative according to any one of claims 1 to 12, or a pharmaceutically acceptable salt or solvate, in the preparation of a medicament for the treatment of a disease mediated by the action of the inducible isoform of nitric oxide lasintase.
14. Uso de un derivado de fenol 2 , 4 -disubstituido según cualquiera de las reivindicaciones 1 a 12 , o de una sal o solvato farmacéuticamente aceptable, en la preparación de un medicamento para el tratamiento de una enfermedad mediada por la acción de la expresión de la L-selectina.14. Use of a derivative of phenol 2,4 -disubstituted according to any of claims 1 to 12, or a pharmaceutically acceptable salt or solvate, in the preparation of a medicament for the treatment of a disease mediated by the action of expression of L-selectin.
15. Uso según cualquiera de las reivindicaciones anteriores 13 a 14, caracterizado porque la enfermedad pertenece al grupo de estados autoinmunes y/o inflamatorios articulares y musculoesqueléticos, enfermedades inflamatorias del tracto gastrointestinal, isquemia cardiovascular, diabetes, hiperalgesia, isquemia cerebral, hipotensión sistémica asociada con el choque séptico y/o tóxico inducido por una amplia variedad de agentes, inmunosupresión a corto plazo en terapia de transplantes, trastornos del SNC, enfermedades tumorales e infecciosas. 15. Use according to any of the preceding claims 13 to 14, characterized in that the disease belongs to the group of autoimmune and / or inflammatory joint and musculoskeletal states, inflammatory diseases of the gastrointestinal tract, cardiovascular ischemia, diabetes, hyperalgesia, cerebral ischemia, associated systemic hypotension with septic and / or toxic shock induced by a wide variety of agents, short-term immunosuppression in transplant therapy, CNS disorders, tumor and infectious diseases.
PCT/ES2002/000119 2002-03-14 2002-03-14 Use of derivatives of 2,4-disubstituted phenols as inhibitors of the expression of l-selectins and of the isoform that is inducible from nitric oxide synthase WO2003075905A1 (en)

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Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP1577289A1 (en) * 2004-03-18 2005-09-21 Revotar Biopharmaceuticals AG Non-glycosylated/-glycosidic/-peptidic small molecule selectin inhibitors for the treament of inflammatory disorders
ES2258415A1 (en) * 2006-04-12 2006-08-16 Fundacion Universitaria San Pablo Ceu Human and animal cryptosporidiosis medicine ingredient comprises an anti-parasite agent inhibiting L selection
EP1764093A1 (en) * 2005-09-20 2007-03-21 Revotar Biopharmaceuticals AG Novel aromatic compounds and their use in medical applications
US7851501B2 (en) 2005-09-20 2010-12-14 Revotar Biopharmaceuticals Ag Aromatic nitrocatechol compounds and their use for modulating processes mediated by cell adhesion molecules
US7919532B2 (en) 2005-09-20 2011-04-05 Revotar Biopharmaceuticals Ag Hydroxylated aromatic compounds

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0147892A2 (en) * 1983-12-16 1985-07-10 Bislak, S.A. Process for preparing and therapeutical applications of the 2,4,6-triiodophenol
WO1994025047A1 (en) * 1993-05-03 1994-11-10 Genentech, Inc. Inhibition of leukocyte adhesion
ES2087019A1 (en) * 1994-02-08 1996-07-01 Bobel246 S L Use of derivatives of 2,4-bisubstituted phenols as 5-lipoxigenase inhibitors
WO1996030012A1 (en) * 1995-03-24 1996-10-03 Defeudis Francis V Methods for treating conditions associated with excess nitric oxide

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0147892A2 (en) * 1983-12-16 1985-07-10 Bislak, S.A. Process for preparing and therapeutical applications of the 2,4,6-triiodophenol
WO1994025047A1 (en) * 1993-05-03 1994-11-10 Genentech, Inc. Inhibition of leukocyte adhesion
ES2087019A1 (en) * 1994-02-08 1996-07-01 Bobel246 S L Use of derivatives of 2,4-bisubstituted phenols as 5-lipoxigenase inhibitors
WO1996030012A1 (en) * 1995-03-24 1996-10-03 Defeudis Francis V Methods for treating conditions associated with excess nitric oxide

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
LEE C.S. ET AL.: "Inhibition of expression of P-selectine by antioxidant in cholesterol-fed rats", J. KOREAN MED. SCI., vol. 14, no. 1, 1999, pages 8 - 14 *
THERIAULT A. ET AL.: "Tocotrienol is the most effective vitamin E for reducing endothelial expression of adhesion molecules and adhesion to monocytes", ATHEROSCLEROSIS, vol. 160, January 2002 (2002-01-01), pages 21 - 30 *
XIA L. ET AL.: "A proteosome inhibitor, an antioxidant, or a salicylate but not a glucocorticoid blocks constitutive and cytokine-inducible expression of P-selectine in human endothelial cells", BLOOD, vol. 91, no. 5, 1998, pages 1625 - 1632 *

Cited By (11)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP1577289A1 (en) * 2004-03-18 2005-09-21 Revotar Biopharmaceuticals AG Non-glycosylated/-glycosidic/-peptidic small molecule selectin inhibitors for the treament of inflammatory disorders
WO2005090284A1 (en) * 2004-03-18 2005-09-29 Revotar Biopharmaceuticals Ag Non-glycosylated/non-glycosidic/non-peptidic small molecule psgl-1 mimetics for the treatment of inflammatory disorders
US8367677B2 (en) 2004-03-18 2013-02-05 Revotar Biopharmaceuticals Ag Non-glycosylated/non-glycosidic/non-peptidic small molecule PSGL-1 mimetics for the treatment of inflammatory disorders
EP1764093A1 (en) * 2005-09-20 2007-03-21 Revotar Biopharmaceuticals AG Novel aromatic compounds and their use in medical applications
WO2007039113A1 (en) * 2005-09-20 2007-04-12 Revotar Biopharmaceuticals Ag Novel aromatic compounds and their use in medical applications
US7851501B2 (en) 2005-09-20 2010-12-14 Revotar Biopharmaceuticals Ag Aromatic nitrocatechol compounds and their use for modulating processes mediated by cell adhesion molecules
US7919532B2 (en) 2005-09-20 2011-04-05 Revotar Biopharmaceuticals Ag Hydroxylated aromatic compounds
US7923473B2 (en) 2005-09-20 2011-04-12 Revotar Biopharmaceuticals Ag Aromatic compounds and their use in medical applications
US8394835B2 (en) 2005-09-20 2013-03-12 Revotar Biopharmaceuticals Ag Aromatic compounds and their use in medical applications
US8461207B2 (en) 2005-09-20 2013-06-11 Revotar Biopharmaceuticals Ag Phloroglucinol derivatives having selectin ligand activity
ES2258415A1 (en) * 2006-04-12 2006-08-16 Fundacion Universitaria San Pablo Ceu Human and animal cryptosporidiosis medicine ingredient comprises an anti-parasite agent inhibiting L selection

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