ES2704694T3 - Compositions comprising supramolecular complexes of polyanionic polymers and spermidine for use in the treatment of periodontium and damaged oral tissue - Google Patents
Compositions comprising supramolecular complexes of polyanionic polymers and spermidine for use in the treatment of periodontium and damaged oral tissue Download PDFInfo
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- ES2704694T3 ES2704694T3 ES15153105T ES15153105T ES2704694T3 ES 2704694 T3 ES2704694 T3 ES 2704694T3 ES 15153105 T ES15153105 T ES 15153105T ES 15153105 T ES15153105 T ES 15153105T ES 2704694 T3 ES2704694 T3 ES 2704694T3
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- spermidine
- gum
- sulfate
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- ATHGHQPFGPMSJY-UHFFFAOYSA-N spermidine Chemical compound NCCCCNCCCN ATHGHQPFGPMSJY-UHFFFAOYSA-N 0.000 title claims abstract description 77
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- 238000011282 treatment Methods 0.000 title description 6
- 125000000129 anionic group Chemical group 0.000 claims abstract description 13
- 125000002091 cationic group Chemical group 0.000 claims abstract description 6
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- QAOWNCQODCNURD-UHFFFAOYSA-L Sulfate Chemical compound [O-]S([O-])(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-L 0.000 claims description 13
- WCDDVEOXEIYWFB-VXORFPGASA-N (2s,3s,4r,5r,6r)-3-[(2s,3r,5s,6r)-3-acetamido-5-hydroxy-6-(hydroxymethyl)oxan-2-yl]oxy-4,5,6-trihydroxyoxane-2-carboxylic acid Chemical compound CC(=O)N[C@@H]1C[C@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](C(O)=O)O[C@@H](O)[C@H](O)[C@H]1O WCDDVEOXEIYWFB-VXORFPGASA-N 0.000 claims description 6
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Abstract
Composición que comprende un complejo supramolecular formado por lo menos por un polímero polianiónico y espermidina con una proporción de equivalentes aniónicos a equivalentes catiónicos de 10:1 a 107:1 eq/eq, en la que los componentes de dicho complejo supramolecular están mezclados íntimamente, sin ningún enlace covalente entre ellos, siendo dicha composición para la utilización en estomatología para tratar la estomatitis, la úlcera aftosa y la sequedad de boca, y en periodontología para tratar la gingivitis y la periodontitis.Composition comprising a supramolecular complex formed by at least one polyanionic polymer and spermidine with a ratio of anionic equivalents to cationic equivalents of 10: 1 to 107: 1 eq / eq, in which the components of said supramolecular complex are intimately mixed, without any covalent bond between them, said composition being used in stomatology to treat stomatitis, aphthous ulcer and dry mouth, and in periodontology to treat gingivitis and periodontitis.
Description
DESCRIPCIÓNDESCRIPTION
Composiciones que comprenden complejos supramoleculares de polímeros polianiónicos y espermidina para su utilización en el tratamiento de periodonto y tejido oral dañado.Compositions comprising supramolecular complexes of polyanionic polymers and spermidine for use in the treatment of periodontium and damaged oral tissue.
Campo de la invenciónField of the invention
La invención se refiere a composiciones que comprenden complejos supramoleculares formados por polímeros polianiónicos y espermidina para la utilización en el tratamiento, el mantenimiento o la reparación del periodonto y los tejidos orales.The invention relates to compositions comprising supramolecular complexes formed by polyanionic polymers and spermidine for use in the treatment, maintenance or repair of the periodontium and oral tissues.
AntecedentesBackground
La espermidina pertenece al grupo de las poliaminas (PA), policationes metabólicos que unen ADN, ARN, proteínas, fosfolípidos y nucleósidos trifosfato cargados negativamente.Spermidine belongs to the group of polyamines (PA), metabolic polycations that bind negatively charged DNA, RNA, proteins, phospholipids and nucleoside triphosphates.
Esta capacidad podría explicar una contribución a la proliferación y diferenciación celular; ver Heby O. Differentiation 1981;19:1-20 y Cohen SS. A Guide to the Polyamines. New York: Oxford University Press; 1998;185-230.This ability could explain a contribution to cell proliferation and differentiation; see Heby O. Differentiation 1981; 19: 1-20 and Cohen SS. A Guide to the Polyamines. New York: Oxford University Press; 1998; 185-230.
Sin embargo, su papel biológico resulta difícil de definir claramente. Las PA de hecho resultan necesarias para la transcripción de los protooncogenes c-myc y c-fos. En particular, la espermidina preferentemente estimula la transcripción y expresión de c-myc, mientras que la putrescina, de c-fos (Tabib y Bachrach, Int. J. Biochem. Cell Biol. 1999;31:1289-95). Anteriormente se había encontrado un papel en la transducción de señales entre la membrana celular y el núcleo (Tabib & Bachrach. Biochem Biophys Res Commun 1994;202:720-7). La espermidina también participa en la transducción de señales de TGF-p (Blachowski S et al., Int. J. Biochem.However, its biological role is difficult to define clearly. PAs are in fact necessary for the transcription of the proto-oncogenes c-myc and c-fos. In particular, spermidine preferably stimulates transcription and expression of c-myc, while putrescine, from c-fos (Tabib and Bachrach, Int.J. Biochem.Cell Biol. 1999; 31: 1289-95). Previously, a role in signal transduction between the cell membrane and the nucleus had been found (Tabib & Bachrach, Biochem Biophys Res Commun 1994; 202: 720-7). Spermidine also participates in the transduction of TGF-p signals (Blachowski S et al., Int. J. Biochem.
1994;26:891-897) y aparentemente resulta necesaria para la expresión normal del gen de TGF-p durante la migración celular.1994; 26: 891-897) and apparently is necessary for the normal expression of the TGF-p gene during cell migration.
Varias patentes reivindican la espermidina y otras PA en diversos contextos terapéuticos: los documentos WO97/014415 y WO99/051213 en la analgesia local y el eccema; el documento US6555140 en el incremento de la fertilidad y libido masculinas; el documento US06252838 en el antienvejecimiento de la piel; el documento US04242701 en el alcoholismo; el documento WO9852552 como anticancerígeno, y el documento US5432202 como antagonistas de Ca antihipertensivos.Several patents claim spermidine and other PAs in various therapeutic contexts: WO97 / 014415 and WO99 / 051213 in local analgesia and eczema; US6555140 in the increase of male fertility and libido; US06252838 in anti-aging of the skin; document US04242701 on alcoholism; WO9852552 as anticancer, and US5432202 as antihypertensive Ca antagonists.
Un comportamiento típico de la espermidina es la interacción con macromoléculas aniónicas para producir complejos supramoleculares, que, por ejemplo, modulan las interacciones de enzima/ADN (Isobe H. et al., Chem Commun (Camb). 28(12):1549-51,2005.A typical behavior of spermidine is the interaction with anionic macromolecules to produce supramolecular complexes, which, for example, modulate enzyme / DNA interactions (Isobe H. et al., Chem Commun (Camb) .28 (12): 1549- 51,2005.
Los complejos supramoleculares formados por espermidina y grupos fosfato modifican el estado de condensación del ADN y lo protegen de las actividades de las nucleasas (D'Agostino L. et al., IUBMB Life.The supramolecular complexes formed by spermidine and phosphate groups modify the state of DNA condensation and protect it from the activities of the nucleases (D'Agostino L. et al., IUBMB Life.
58(2):75-82, 2006). Se encontró una propiedad similar en los agregados autoensamblados supramoleculares de espermidina de la poliamina, los cuales se informa que se comportan como factores protectores (D'Agostino L. et al., FEBS J. 2009;276(8): 2324-35).58 (2): 75-82, 2006). A similar property was found in the self-assembled supramolecular spermidine aggregates of polyamine, which are reported to behave as protective factors (D'Agostino L. et al., FEBS J. 2009; 276 (8): 2324-35). .
El documento WO2010/049562 da a conocer complejos supramoleculares de polímero polianiónico con espermidina o espermina en proporciones comprendidas entre 1:0,1 y 1:0,5 p/p que claramente se aproximan a la equimolaridad. Su supuesto uso es el transporte de otros agentes bioactivos.WO2010 / 049562 discloses supramolecular polyanionic polymer complexes with spermidine or spermine in proportions comprised between 1: 0.1 and 1: 0.5 w / w that clearly approximate equimolarity. Its supposed use is the transport of other bioactive agents.
La única solicitud conocida son los productos antialopecia comercializados como Bioscalin® por Giuliani SpA (Milan, Italia), con HCl de espermidina comercializado como Biogenina y Cronobiogenina. Las formulaciones y métodos antialopecia se basan en HCl de espermidina han sido patentadas por Giuliani en la patente n° EP1469842, y documentos n° WO03/063851 y n° WO2010/060729.The only known application is antialopecia products marketed as Bioscalin® by Giuliani SpA (Milan, Italy), with spermidine HCl marketed as Biogenin and Cronobiogenin. The formulations and antialopecia methods are based on spermidine HCl and have been patented by Giuliani in the patent n ° EP1469842, and documents n ° WO03 / 063851 and n ° WO2010 / 060729.
Aunque existe una clara necesidad de terapéuticos que puedan estimular la regeneración del periodonto y el tejido oral mediante la proliferación celular, hasta el momento no se ha intentado el diseño complejos de espermidina con una eficacia elevada en este tejido específico.Although there is a clear need for therapies that can stimulate the regeneration of the periodontium and oral tissue through cell proliferation, up to now no design of spermidine complexes with high efficiency in this specific tissue has been attempted.
SumarioSummary
Inesperadamente se ha descubierto que determinados complejos supramoleculares formados por polímeros polianiónicos y espermidina presentan una eficiencia elevada en la regeneración del periodonto y el tejido conectivo oral.Unexpectedly, it has been discovered that certain supramolecular complexes formed by polyanionic polymers and spermidine have a high efficiency in the regeneration of the periodontium and the oral connective tissue.
En un aspecto, la invención se refiere a composiciones que comprenden complejos supramoleculares caracterizados por proporciones de equivalentes aniónicos (del polímero polianiónico) y equivalentes catiónicos (de la espermidina) de entre 10:1 y 107:1 eq/q, caracterizados además por una eficacia elevada en los tratamientos regenerativos.In one aspect, the invention relates to compositions comprising supramolecular complexes characterized by proportions of anionic equivalents (of the polyanionic polymer) and cationic equivalents. (from spermidine) of between 10: 1 and 107: 1 eq / q, also characterized by high efficiency in regenerative treatments.
En otro aspecto, los complejos supramoleculares inventivos presentan unas proporciones preferidas de entre 102:1 y 104:1 eq/eq.In another aspect, the inventive supramolecular complexes have preferred ratios of between 102: 1 and 104: 1 eq / eq.
En todavía otro aspecto, la invención se refiere a composiciones que comprenden los complejos supramoleculares anteriormente indicados para el mantenimiento y reparación de periodonto y tejidos orales senescentes.In yet another aspect, the invention relates to compositions comprising the supramolecular complexes indicated above for the maintenance and repair of periodontium and senescent oral tissues.
Estas y otras características de la invención se describen con mayor detalle a continuación en la presente memoria.These and other features of the invention are described in greater detail hereinafter.
Descripción detalladaDetailed description
La presente invención se define en las reivindicaciones adjuntas.The present invention is defined in the appended claims.
Es una materia objeto de la exposición una composición que comprende un complejo supramolecular formado por como mínimo un polímero polianiónico y espermidina con una proporción de equivalentes aniónicos y equivalentes catiónicos (de la espermidina) de entre 10:1 y 107:1 eq/eq. donde los componentes de dicho complejo supramolecular se mezclan íntimamente, sin ningún enlace covalente entre ellos, siendo dicha composición para la utilización en el mantenimiento y reparación de un tejido conectivo dañado o senescente seleccionado de mucosa oral, encías y periodonto.A subject matter of the disclosure is a composition comprising a supramolecular complex consisting of at least one polyanionic polymer and spermidine with a ratio of anionic equivalents and cationic equivalents (of spermidine) of between 10: 1 and 107: 1 eq / eq. wherein the components of said supramolecular complex are intimately mixed, without any covalent bond between them, said composition being for use in the maintenance and repair of a damaged or senescent connective tissue selected from oral mucosa, gums and periodontium.
La expresión "complejo supramolecular" tal como se utiliza en la presente memoria describe un complejo poliácido/polibásico formado por uno o más polímeros polianiónicos y espermidina caracterizado por una proporción de eq. aniónicos (de polímeros polianiónicos) a eq. catiónicos (de espermidina) de entre aproximadamente 10:1 y aproximadamente 107:1 eq/eq, más preferentemente de entre aproximadamente 102:1 y aproximadamente 104:1 eq/eq.The term "supramolecular complex" as used herein describes a polyacid / polybasic complex formed by one or more polyanionic polymers and spermidine characterized by a ratio of eq. anionic (from polyanionic polymers) to eq. cationic (of spermidine) of between about 10: 1 and about 107: 1 eq / eq, more preferably between about 102: 1 and about 104: 1 eq / eq.
Estos complejos supramoleculares para la utilización según la invención poseen una marcada actividad proliferativa/regenerativa, que es significativamente superior a la espermidina, el polímero polianiónico y la suma de los mismos.These supramolecular complexes for use according to the invention possess a marked proliferative / regenerative activity, which is significantly superior to spermidine, the polyanionic polymer and the sum thereof.
Dichos complejos supramoleculares se caracterizan por una proporción > 10:1 eq/eq, difiriendo de complejos de polímero polianiónico y espermidina que se aproximan a la equimolaridad tanto en sus características estructurales como en una actividad proliferativa significativamente más alta sobre los fibroblastos.Said supramolecular complexes are characterized by a ratio> 10: 1 eq / eq, differing from polyanionic polymer and spermidine complexes that approximate equimolarity both in their structural characteristics and in a significantly higher proliferative activity on the fibroblasts.
La expresión "polímero polianiónico y espermidina que se aproximan a la equimolaridad" tal como se utiliza en la presente memoria se refiere a complejos que presentan proporciones de entre aproximadamente 1:3 y 10:1 eq/eq de equivalentes aniónicos de polímero polianiónico a equivalentes catiónicos de la espermidina.The term "polyanionic polymer and spermidine approaching equimolarity" as used herein refers to complexes having proportions of between about 1: 3 and 10: 1 eq / eq of anionic equivalents of polyanionic polymer to equivalents cationics of spermidine.
La expresión "polímero polianiónico" se refiere, en el sentido más amplio entendido en la técnica, a un material polimérico o polímero que comprende una pluralidad de varias fracciones aniónicas en cada molécula. Incluye polímeros naturales/semisintéticos y polímeros totalmente sintéticos que contienen una pluralidad de fracciones aniónicas, tales como carboxílico (-COO-), sulfato (-OSO3"), sulfonato (-SO3"), fosfato (-OPO32"), fosfonatos (-PO32") y una combinación de los mismos.The term "polyanionic polymer" refers, in the broadest sense understood in the art, to a polymeric material or polymer comprising a plurality of various anionic moieties in each molecule. Includes natural / semi-synthetic polymers and fully synthetic polymers containing a plurality of anionic moieties, such as carboxylic (-COO-), sulfate (-OSO 3 "), sulfonate (-SO 3 "), phosphate (-OPO 3 2 ") , phosphonates (-PO 3 2 ") and a combination thereof.
La expresión "polímero polianiónico" incluye "polímero derivado polianiónicamente", referido a polímeros anteriormente no polianiónicos que se convierten en polímero polianiónico con reactivos de derivación adecuados. Son ejemplos de derivaciones, la carboximetilación, la succinilación o la maleilación para los grupos carboxi; la sulfatación/sulfonación/sulfinilación para los grupos sulfato/sulfonato; la fosfatación/fosforilación para los grupos fosfato/fosfonato.The term "polyanionic polymer" includes "polyanionically derived polymer", referred to previously non-polyanionic polymers that are converted to polyanionic polymer with suitable derivatization reagents. Examples of derivations are carboxymethylation, succinylation or maleylation for carboxy groups; sulfation / sulphonation / sulfinylation for the sulfate / sulfonate groups; phosphating / phosphorylation for the phosphate / phosphonate groups.
Entre los polímeros polianiónicos se incluyen fitopolisacáridos aniónicos, ficopolisacáridos y endopolisacáridos; polímeros polianiónicos semisintéticos y totalmente sintéticos.Polyanionic polymers include anionic phycolysaccharides, polycopolysaccharides and endopolysaccharides; semi-synthetic and fully synthetic polyanionic polymers.
Los polímeros polianiónicos naturales pueden ser fitopolisacáridos y ficopolisacáridos, tales como alginatos, agar, goma gelano, goma ghatti, goma karaya, goma tragacanto, goma wellano, goma xantano, carragenano k, i y A, sulfato de xilomanano, fucoidán y fucogalactano.The natural polyanionic polymers can be phytopylissaccharides and phycopolysaccharides, such as alginates, agar, gellan gum, ghatti gum, karaya gum, tragacanth gum, wellano gum, xanthan gum, k, i and A carrageenan, xylomannan sulfate, fucoidan and fucogalactan.
Otros polímeros polianiónicos naturales pueden ser endopolisacáridos, tales como hialuronato, hialuronato reticulado y otros glucosoaminoglicanos como heparina, heparinas supersulfatadas y modificadas, por ejemplo heparina supersulfatada, fraxiparina, fondaparina, idraparina, condroitín sulfato A, B y C, y los derivados de K5. El hialuronato adecuado, también conocido como hialuranano (HA) puede ser de origen animal o microbiano, con un peso molecular (PM) de entre 5.000 kDa y 10 MDa.Other natural polyanionic polymers can be endopolysaccharides, such as hyaluronate, cross-linked hyaluronate and other glycosaminoglycans such as heparin, supersulfated and modified heparins, for example supersulfated heparin, fraxiparin, fondaparin, idraparin, chondroitin sulfate A, B and C, and the K5 derivatives. The appropriate hyaluronate, also known as hyalurnanan (HA) can be of animal or microbial origin, with a molecular weight (MW) of between 5,000 kDa and 10 MDa.
Los polímeros polianiónicos semisintéticos pueden ser polisacáridos carboximetilados, tales como carboximetilcelulosa, carboximetil-almidón, carboximetil-dextrano, carboximetil-quitosano, carboximetil-quitinas; polisacáridos sulfatados, tales como sulfato de ramnano, sulfato de dextrano, sulfato de celulosa, sulfoquitosanos, sulfato de curdlán, sulfato de gliloide (GP4324), sulfato de goma de algarrobo (GP4327), polisulfato de pentosano (PPS) y polisacáridos fosfatados, tales como fosfocelulosa y fosfoquitosano.The semi-synthetic polyanionic polymers can be carboxymethylated polysaccharides, such as carboxymethylcellulose, carboxymethyl-starch, carboxymethyl-dextran, carboxymethyl-chitosan, carboxymethyl-chitins; sulphated polysaccharides, such as rhamnose sulfate, dextran sulfate, cellulose sulfate, sulfochitosans, curdlan sulfate, gliloid sulfate (GP4324), locust bean gum sulfate (GP4327), pentosan polysulfate (PPS) and phosphate polysaccharides, such as phosphocellulose and phosphochitosan.
Los polímeros polianiónicos semisintéticos también pueden ser derivados de HA, por ejemplo derivados de HA de Fidia, o polisacáridos tiolados, tales como celulosa tiolada, alginatos tiolados, quitosano tiolado, hialuronato tiolado de ThioMatrix-Green River Polymers GmbH (Insbruck, Austria) y similares.Semi-synthetic polyanionic polymers can also be HA derivatives, for example Fidia HA derivatives, or thiolated polysaccharides, such as thiolated cellulose, thiolated alginates, thiolated chitosan, thiolated hyaluronate from ThioMatrix-Green River Polymers GmbH (Innsbruck, Austria) and the like .
El polímero polianiónico sintético puede ser poliacrilato y polimetacrilato, homopolímero lineal y reticulado y copolímeros de los mismos, tales como acrilatos/acrilamidas, acrilatos, acrilatos de alquilo C10-C30, acrilatos/octilacrilamidas, Carbopol™, Carbomer™ y Pemulen™ de Noveon-Lubrizol, ácido butil-poliacrílico, poli(sulfonato de acrilato-co-acrilamidometilpropano sulfonato), copolímeros de poli(acrilato-co-vinilsulfonato) y poli(acrilato-co-vinilbencenosulfonato); metilvinileter/maleico y otros copolímeros de maleato, también conocidos como "polimaleatos", por ejemplo de tipo Gantrez™ (ISP Corp.) y además según el documento n° US2010172861 o US2003021793, así como poli(4-estirenosulfonato sódico), Y-ART-4, suramina, carbómeros tiolados, ácido poli(meta)acrílico tiolado y similares.The synthetic polyanionic polymer can be polyacrylate and polymethacrylate, linear and crosslinked homopolymer and copolymers thereof, such as acrylates / acrylamides, acrylates, C10-C30 alkyl acrylates, acrylates / octylacrylamides, Carbopol ™, Carbomer ™ and Pemulen ™ from Noveon- Lubrizol, butyl-polyacrylic acid, poly (acrylate-co-acrylamidomethylpropane sulphonate sulfonate), copolymers of poly (acrylate-co-vinylsulfonate) and poly (acrylate-co-vinylbenzenesulfonate); methyl vinyl ether / maleic acid and other maleate copolymers, also known as "polymaleate", for example of the Gantrez ™ type (ISP Corp.) and furthermore according to document No. US2010172861 or US2003021793, as well as sodium poly (4-styrenesulfonate), Y- ART-4, suramin, thiolated carbomers, thiolated poly (meta) acrylic acid and the like.
La expresión "polímero polianiónico" comprende además polímeros inorgánicos aniónicos, tales como polifosfatos o polímeros recombinantes, tal como se da a conocer en el documento WO2002/077036.The term "polyanionic polymer" further comprises anionic inorganic polymers, such as polyphosphates or recombinant polymers, as disclosed in WO2002 / 077036.
Los polímeros polianiónicos preferidos en el contexto de la presente invención son hialuronatos lineales y reticulados, alginatos, poliacrilatos lineales y reticulados, y polimaleatos.Preferred polyanionic polymers in the context of the present invention are linear and crosslinked hyaluronates, alginates, linear and crosslinked polyacrylates, and polymaleates.
El polímero polianiónico que debe utilizarse según la presente invención presenta un peso molecular > 5.000 Da y/o una densidad aniónica de entre 0,1 y 18 meq/g, preferentemente de entre 1 y 14 meq/g.The polyanionic polymer to be used according to the present invention has a molecular weight> 5,000 Da and / or an anionic density of between 0.1 and 18 meq / g, preferably between 1 and 14 meq / g.
En el contexto de la presente invención, puede utilizarse preferentemente la espermidina, N-(3-aminopropil)-1,4-butanodiamina o "Spd" a continuación en la presente memoria, es la sustancia de fórmula NH2 (CH2 )aNH(CH2 )4 NH2 como tal o como sal de la misma, de origen sintético y, también preferentemente, de pureza > 99% en peso seco.In the context of the present invention, spermidine, N- (3-aminopropyl) -1,4-butanediamine or "Spd" hereinafter may be used, is the substance of formula NH 2 (CH 2 ) aNH (CH 2 ) 4 NH 2 as such or as a salt thereof, of synthetic origin and, also preferably, of purity> 99% by dry weight.
Los complejos supramoleculares para la utilización según la invención se producen mediante intercambio ácidobase, por ejemplo mediante combinación de una sal de polímero polianiónico (típicamente Na) con sal de espermidina (por ejemplo, 3HCl) o mediante la combinación de polímeros polianiónicos en la forma de ácido con espermidina como base libre.The supramolecular complexes for use according to the invention are produced by acid-base exchange, for example by combining a polyanionic polymer salt (typically Na) with a spermidine salt (for example, 3HCl) or by combining polyanionic polymers in the form of acid with spermidine as a free base.
Los equivalentes aniónicos restantes en el polímero polianiónico dentro del complejo pueden encontrarse en forma de ácido o parcialmente neutralizados, por ejemplo con iones alcalinos o alcalino-térreos, tales como Na, K, Li, Ca y Mg, o aminas, tales como NH4+, monoetanolamina, dietanolamina y trietanolamina, trometamina, isopropilamina, lisina, aminas eterocíclicas, tales como piperazina y similares.The remaining anionic equivalents in the polyanionic polymer within the complex can be in the form of an acid or partially neutralized, for example with alkaline or alkaline earth ions, such as Na, K, Li, Ca and Mg, or amines, such as NH 4 +, monoethanolamine, diethanolamine and triethanolamine, tromethamine, isopropylamine, lysine, ethercyclic amines, such as piperazine and the like.
La combinación de polímero polianiónico y espermidina puede llevarse a cabo en estado solvatado, preferentemente en agua y/o solventes solubles en agua, tales como alcoholes inferiores, etc. el complejo supramolecular formado de esta manera puede secarse, por ejemplo Mediante liofilización, secado por pulverización, secado al vacío, y similar, o utilizarse sin modificación en forma de solución, con la condición de que los solventes sean fisiológicamente aceptables y compatibles con el uso final deseado.The combination of polyanionic polymer and spermidine can be carried out in solvated state, preferably in water and / or water-soluble solvents, such as lower alcohols, etc. the supramolecular complex formed in this way can be dried, for example by lyophilization, spray drying, vacuum drying, and the like, or used without modification in the form of a solution, with the proviso that the solvents are physiologically acceptable and compatible with the use desired end
En una forma de realización, el complejo supramolecular para la utilización según la invención se forma mediante la combinación de uno o más polímeros polianiónicos con espermidina directamente dentro del tanquemezclador en la preparación galénica. Puede llevarse a cabo, por ejemplo, mediante mezcla de la solución acuosa de los reactivos, opcionalmente junto con cosolventes, excipientes, diluyentes, portadores y adyuvantes seleccionados.In one embodiment, the supramolecular complex for use according to the invention is formed by the combination of one or more polyanionic polymers with spermidine directly within the mixer tank in the galenic preparation. It can be carried out, for example, by mixing the aqueous solution of the reagents, optionally together with cosolvents, excipients, diluents, carriers and selected adjuvants.
Alternativamente, el complejo supramolecular para la utilización según la invención se prepara directamente en estado sólido, por ejemplo mediante pulverización de una solución de espermidina sobre el polímero o, alternativamente, mediante la mezcla o trituración de los componentes en formas secas o parcialmente humectadas, y procedimientos similares.Alternatively, the supramolecular complex for use according to the invention is prepared directly in the solid state, for example by spraying a solution of spermidine on the polymer or, alternatively, by mixing or grinding the components in dry or partially wetted forms, and similar procedures.
A pesar del procedimiento aplicado, los complejos supramoleculares obtenidos para la utilización según la invención pueden mezclarse con varios excipientes, diluyentes, portadores y adyuvantes adicionales para producir una composición adecuada para la reparación y mantenimiento del periodonto y tejidos orales. Despite the procedure applied, the supramolecular complexes obtained for use according to the invention can be mixed with various excipients, diluents, carriers and additional adjuvants to produce a composition suitable for repair and maintenance of the periodontium and oral tissues.
Según un aspecto de la presente invención, se proporcionan composiciones y métodos para regenerar tejidos periodontales y orales conectivos mediante la inducción/estimulación de la proliferación celular.According to one aspect of the present invention, compositions and methods are provided for regenerating connective periodontal and oral tissues by induction / stimulation of cell proliferation.
Las composiciones para la utilización según la invención se administran para mejorar el periodonto y tejidos orales senescentes o para reparar el periodonto y tejidos orales dañados, incluyendo:The compositions for use according to the invention are administered to improve the periodontium and senescent oral tissues or to repair damaged periodontium and oral tissues, including:
• la mucosa oral, es decir, en estomatología, para tratar la estomatitis, las úlceras aftosas y la xerostomía, y el síndrome de la boca seca y/o de Sjogren;• the oral mucosa, that is, in stomatology, to treat stomatitis, aphthous ulcers and xerostomia, and dry mouth and / or Sjogren's syndrome;
• encías y periodonto, es decir, en periodontología, para tratar la gingivitisperiodontitis.• gums and periodontium, that is, in periodontology, to treat gingivitisperiodontitis.
Las composiciones de la invención comprenden complejos supramoleculares de proporción entre 103 y 107 eq/eq en una cantidad de entre aproximadamente 0,01% y aproximadamente 10% p/p, y complejos supramoleculares de proporción entre 10 y 102 eq/eq y cantidad entre aproximadamente 0,0001% y aproximadamente 10% p/p de la composición.The compositions of the invention comprise supramolecular complexes of between 103 and 107 eq / eq in an amount between about 0.01% and about 10% w / w, and supramolecular complexes of between 10 and 102 eq / eq and amount between about 0.0001% and about 10% w / w of the composition.
Preferentemente se diseña una composición para proporcionar una cantidad de espermidina de entre aproximadamente 10 jmoles y 0,1 nmoles por dosis unitaria, más preferentemente de entre aproximadamente 1 |jmol y 1 nmol por dosis unitaria, todavía más preferentemente de entre aproximadamente 100 nmoles y 10 nmoles por dosis unitaria.Preferably a composition is designed to provide an amount of spermidine of between about 10 jmoles and 0.1 nmoles per unit dose, more preferably between about 1 | jmole and 1 nmole per unit dose, still more preferably between about 100 nmoles and 10. nmol per unit dose.
Se recomiendan unos niveles de dosis bajos en composiciones absorbentes, en las que resulta suficiente un periodo de tiempo largo para liberar una cantidad terapéutica de espermidina sobre el periodonto y tejido oral diana. Los niveles de dosis superiores por el contrario se recomiendan en composiciones enjuagables u otras con un contacto breve a fin de proporcionar un nivel suficiente de espermidina para inducir la regeneración del periodonto y tejidos orales. La cantidad y duración del tratamiento se determinarán según la diana y condición del paciente, típicamente durante un periodo de 30 a 60 días o más, hasta conseguir el alivio, seguido posiblemente de la parada, reducción gradual o reducción durante un periodo indefinido.Low dose levels are recommended in absorbent compositions, where a long period of time is sufficient to release a therapeutic amount of spermidine over the target oral tissue and periodontium. Higher dose levels, on the other hand, are recommended in rinse-off or other compositions with brief contact in order to provide a sufficient level of spermidine to induce regeneration of the periodontium and oral tissues. The amount and duration of treatment will be determined according to the target and condition of the patient, typically for a period of 30 to 60 days or more, until relief is achieved, possibly followed by stopping, gradual reduction or reduction for an indefinite period.
Cabe destacar que la composición para la utilización según la invención difiere de composiciones, en caso de existir, que comprenden ocasionalmente uno o más polímeros polianiónicos y espermidina mezclados separadamente y que, de esta manera, pueden no producir, o sólo producir parcialmente, el complejo supramolecular correspondiente.It should be noted that the composition for use according to the invention differs from compositions, if any, which occasionally comprise one or more polyanionic polymers and spermidine mixed separately and which, in this way, may not produce, or only partially produce, the complex corresponding supramolecular
La composición para la utilización según la invención puede producirse según técnicas conocidas con ingredientes y portadores fisiológicamente aceptables con el fin de proporcionar el mejor perfil de beneficio/riesgo, por ejemplo los indicados en INCI-CTFA apéndice 93/35/ECC y/o en las farmacopeas.The composition for use according to the invention can be produced according to known techniques with physiologically acceptable ingredients and carriers in order to provide the best benefit / risk profile, for example those indicated in INCI-CTFA appendix 93/35 / ECC and / or in the pharmacopoeias.
Una composición líquida puede encontrarse en diferentes presentaciones, incluyendo gel, lipogel, aerosol, espray, loción, leche, espuma, crema, emulsión agua-en-aceite, emulsión aceite-en-agua o emulsiones multifase, parches mucoadhesivos y similares, junto con excipientes, portadores o propelentes adecuados.A liquid composition can be found in different presentations, including gel, lipogel, aerosol, spray, lotion, milk, foam, cream, water-in-oil emulsion, oil-in-water emulsion or multiphase emulsions, mucoadhesive patches and the like, together with suitable excipients, carriers or propellants.
Las composiciones para las mucosas orales pueden formularse de muchas maneras, por ejemplo según ADA/PDR: Guide to Dental Therapeutics, 4a ed, como colutorio solución oral, espray, gel sobre película, dentífricos, polvo dentífrico, comprimidos dentales, cremas y geles, goma de mascar, comprimidos y pastillas masticables, gel, película, gel sobre película, gránulos, pasta, polvos esparcibles, etc., para la aplicación sobre la cavidad oral directamente o mediante un dispositivo externo.Oral mucosal compositions can be formulated in many ways, for example according to ADA / PDR: Guide to Dental Therapeutics, 4th ed., As an oral solution mouthwash, spray, gel on film, toothpastes, toothpaste, dental tablets, creams and gels, chewing gum, chewable tablets and tablets, gel, film, gel on film, granules, paste, spreadable powders, etc., for application on the oral cavity directly or by an external device.
EjemplosExamples
Ejemplo 1. Patrón metabólico de HA-Spd 50:1 eq/eq en fibroblastos gingivales nativosExample 1. Metabolic pattern of HA-Spd 50: 1 eq / eq in native gingival fibroblasts
Se obtuvieron especímenes de tejido de tejido periodontal gingival no inflamado de la zona premolar durante cirugía oral (seis mujeres, 20 a 30 años de edad). Se lavó cada biopsia con D-PBS 0,1 M e inmediatamente se trituró con tijeras estériles. Los fragmentos de tejido se transfirieron a matraces Nunc de 25 cm2 y, tras la adherencia, se complementaron con 5 ml de DMEM que contenía FBS al 10%, 100 Ul/ml de penicilina, estreptomicina 10 ng/ml y 25 jg/ml de anfotericina-B. Los cultivos se mantuvieron en una atmósfera de humedad saturada (5% de CO2 , 37°C) y se subcultivaron rutinariamente después de la utilización con tripsina al 0,1%, EDTA al 0,02% para desprender las células. En tiempos dados se recogieron sobrenadantes de cultivo celular y se lavaron los fibroblastos en PBS, se tripsinizaron y se recolectaron mediante centrifugación (100xg, 5 min). Tratamiento: 4 dosis, 1 punto temporal (24 o 48 h) con la muestra tratada con HA-Spd 104:1, mientras que el espécimen no tratado sirvió de control.Tissue specimens from non-inflamed gingival periodontal tissue were obtained from the premolar area during oral surgery (six women, 20 to 30 years of age). Each biopsy was washed with 0.1 M D-PBS and immediately ground with sterile scissors. Tissue fragments were transferred to 25 cm2 Nunc flasks and, after adherence, were supplemented with 5 ml DMEM containing 10% FBS, 100 IU / ml penicillin, 10 ng / ml streptomycin and 25 jig / ml amphotericin-B. The cultures were kept in an atmosphere of saturated humidity (5% CO 2 , 37 ° C) and routinely subcultured after use with 0.1% trypsin, 0.02% EDTA to detach the cells. In given times cell culture supernatants were collected and the fibroblasts were washed in PBS, trypsinized and harvested by centrifugation (100xg, 5 min). Treatment: 4 doses, 1 temporary point (24 or 48 h) with the sample treated with HA-Spd 104: 1, while the untreated specimen served as control.
Niveles de ARNm para TGF-p1, LH2B, TIMP-1, TIMP-2 y GAPDH mediante RT-PCR en tiempo real. Se aisló el ARN mediante una modificación del método de tiocianato de guanidinio ácido-fenolcloroformo (reactivo TRI, Sigma). Se transcribió inversamente 1 |jg de ARN total en 20 |jl de volumen final de mezcla de reacción (Biorad). Las secuencias de los cebadores para los genes diana eran las del software Beacon Designer 6.0 (BioRad). Se utilizó GAPDH para normalizar para las diferencias en cantidad de ARN total en cada muestra. Se llevó a cabo la amplificación en un volumen final de 20 j l en cada pocillo con 10 j l de 1x SYBR-verde Supermix (BioRad), 2 j l de molde, 300 pmoles de cada cebador, y se analizó cada muestra por triplicado en amplificaciones por duplicado. El ciclo umbral y los niveles de expresión génica respecto a GApDH se calcularon mediante el método de 2"ññCt.Levels of mRNA for TGF-p1, LH2B, TIMP-1, TIMP-2 and GAPDH by RT-PCR in real time. The RNA was isolated by a modification of the guanidinium thiocyanate acid-phenolchloroform method (TRI reagent, Sigma). 1 [mu] g of total RNA was reverse transcribed in 20 [mu] l of final volume of reaction mixture (Biorad). The sequences of the primers for the target genes were those of the Beacon Designer 6.0 software (BioRad). GAPDH was used to normalize for the differences in total RNA amount in each sample. Amplification was carried out in a final volume of 20 μl in each well with 10 μl of 1x SYBR-green Supermix (BioRad), 2 μl of template, 300 μmol of each primer, and each sample was analyzed in triplicate in amplifications by duplicate. The threshold cycle and the levels of gene expression with respect to GApDH were calculated by the method of 2 "ññCt.
Niveles de las proteínas COL-I y COL-III mediante transferencia en ranura. Para evaluar COL-1 y COL-III secretados por fibroblastos de paladar y tuberosidad, se concentraron los medios de cultivo celular 20 veces con columnas Centricon 10 (Amicon Y10, Millipore). Se determinó el contenido de proteínas mediante un ensayo colorimétrico (ensayo DC Protein, BioRad); se aplicaron manchas de 100 jg de proteínas totales de cada muestra en un vol. final de 200 j l de solución salina-tampón Tris (TBS) sobre una membrana de nitrocelulosa en un aparato Bio-Dot SF (Bio-Rad). Las membranas se bloquearon durante 1 h con leche desnatada al 5% en TBST (TBS que contenía Tween-20 al 0,05%), pH 8, y se incubaron durante 1 h a temperatura ambiente en anticuerpo monoclonal de COL-1 (1:1.000 en TBST) (Sigma) o de COL-III (1:2.000 en t BsT) (Sigma). Tras el lavado, las membranas se incubaron en suero de conejo antirratón conjugado con HRP (1:80.000 en TBST) (Sigma) durante 1 h. Las bandas inmunorreactivas se encontraban sobre sustrato Amplified Opti-4CN (BioRad) y se escanearon (UVBand, Eppendorf).Levels of COL-I and COL-III proteins by slot transfer. To evaluate COL-1 and COL-III secreted by palatal fibroblasts and tuberosity, the cell culture media was concentrated 20 times with Centricon 10 columns (Amicon Y10, Millipore). The protein content was determined by a colorimetric assay (DC Protein assay, BioRad); stains of 100 jg of total proteins of each sample were applied in one vol. final of 200 μl of saline-Tris buffer (TBS) on a nitrocellulose membrane in a Bio-Dot SF apparatus (Bio-Rad). The membranes were blocked for 1 h with 5% skimmed milk in TBST (TBS containing 0.05% Tween-20), pH 8, and incubated for 1 h at room temperature on COL-1 monoclonal antibody (1: 1,000 in TBST) (Sigma) or COL-III (1: 2,000 in t BsT) (Sigma). After washing, the membranes were incubated in rabbit anti-mouse serum conjugated with HRP (1: 80,000 in TBST) (Sigma) for 1 h. The immunoreactive bands were on Amplified Opti-4CN substrate (BioRad) and were scanned (UVBand, Eppendorf).
Niveles y actividad de la proteína MMP-1 medidos mediante transferencia western. Se diluyeron unos medios de cultivo concentrados (5 jg de proteínas totales) en SDS-tampón para muestras, se cargaron en gel de SDS-poliacrilamida al 10%, se separaron bajo condiciones reductoras y desnaturalizantes a 80 V y se transfirieron a 90 V a una membrana de nitrocelulosa en Tris 0,025 M, glicina 192 mM y metanol al 20%, pH 8,3. Tras la electrotransferencia, las membranas se secaron al aire y se bloquearon durante 1 h. Tras el lavado, las membranas se incubaron durante 1 h a temperatura ambiente con anticuerpo monoclonal de MMP-1 (1 jg/ml en TBST, Oncogene Research) y, tras el lavado, en suero de conejo antirratón conjugado con HRP (dilución 1:40.000, Sigma-Aldrich). Las bandas inmunorreactivas se revelaron con sustrato Amplified Opti-4CN (BioRad) y se escanearon (UVBand, Eppendorf).Levels and activity of the MMP-1 protein measured by western blot. Concentrated culture media (5 jg of total proteins) were diluted in SDS-buffer for samples, loaded on 10% SDS-polyacrylamide gel, separated under reducing and denaturing conditions at 80 V and transferred to 90 V a a nitrocellulose membrane in 0.025 M Tris, 192 mM glycine and 20% methanol, pH 8.3. After electroblotting, the membranes were air dried and blocked for 1 h. After washing, the membranes were incubated for 1 h at room temperature with MMP-1 monoclonal antibody (1 jg / ml in TBST, Oncogene Research) and, after washing, in rabbit serum anti-mouse conjugated with HRP (1: 40,000 dilution). , Sigma-Aldrich). The immunoreactive bands were developed with Amplified Opti-4CN substrate (BioRad) and scanned (UVBand, Eppendorf).
Gelatinasas (actividad de MMP-2 y MMP-9) mediante zimografía en SDS. Los medios de cultivo concentrados se mezclaron 3:1 con tampón para muestras (que contenía SDS al 10%). Las muestras (5 jg de proteínas totales de cada muestra) se hicieron migrar bajo condiciones no reductoras sin desnaturalización por calor en SDS-PAGE al 10% copolimerizado con 1 mg/ml de gelatina de tipo I. Los geles migraron a 4°C. Tras el SDS-PAGE, los geles se lavaron dos veces en Triton X-100 al 2,5% durante 30' cada uno y se incubaron durante la noche a 37°C. La actividad gelatinolítica de MMP se detectó tras la tinción de los geles con azul brillante de Coomassie R250 como bandas claras sobre un fondo azul.Gelatinases (activity of MMP-2 and MMP-9) by zymography in SDS. The concentrated culture media was mixed 3: 1 with sample buffer (containing 10% SDS). Samples (5 jg of total proteins from each sample) were migrated under non-reducing conditions without heat denaturing on 10% SDS-PAGE copolymerized with 1 mg / ml type I gelatin. The gels migrated at 4 ° C. After SDS-PAGE, the gels were washed twice in 2.5% Triton X-100 for 30 'each and incubated overnight at 37 ° C. The gelatinolytic activity of MMP was detected after staining the gels with Coomassie bright blue R250 as light bands on a blue background.
Recuento de fibroblastos. Se evaluó el efecto de proliferación a partir de la capacidad de las células de excluir el azul de tripán, de acuerdo con Patterson MK Jr , en: Jacob & Pastan, eds. Methods in Enzymology, vol. 58. New York: Academic Press; 1977:141.Fibroblasts count. The proliferation effect was evaluated from the ability of the cells to exclude trypan blue, according to Patterson MK Jr, in: Jacob & Pastan, eds. Methods in Enzymology, vol. 58. New York: Academic Press; 1977: 141.
Los resultados revelaron un patrón complejo de actividades reguladoras de los enzimas de degradación, tales como las colagenasas y MMP, y la modulación de TGF-p1, LH2B, TIMP-1/2 y GAPDH, que revertían resultados anteriores de Gagliano N at al. J Periodontol 2005; 76:443-9.The results revealed a complex pattern of regulatory activities of the degradation enzymes, such as collagenases and MMP, and the modulation of TGF-p1, LH2B, TIMP-1/2 and GAPDH, which reversed previous results of Gagliano N at al. J Periodontol 2005; 76: 443-9.
Ejemplo 2. Gel de HA-Spd simpleExample 2. Simple HA-Spd gel
Se disolvieron 10 g de HA sódico (PM: 1,2 MDa) en 800 ml de agua hasta la hidratación total; después se añadieron 10 ml de Spd 1 mM bajo agitación durante 5', proporcionando un complejo HA-Spd 102:1 eq/eq. A continuación, se mezclaron 0,9 g de alcohol bencílico y se agitó, y el volumen final se completó a 1 litro con agua purificada. El gel se cargó en tubos de PE de 60 ml adecuados para la mayoría de aplicaciones, tales como las mucosas urogenital, oral, auditiva, nasal y de la garganta, piel, y similares.10 g of sodium HA (MW: 1.2 MDa) were dissolved in 800 ml of water until total hydration; then 10 ml of 1 mM Spd was added under stirring for 5 ', providing a HA-Spd complex 102: 1 eq / eq. Next, 0.9 g of benzyl alcohol was mixed and stirred, and the final volume was completed to 1 liter with purified water. The gel was loaded into 60 ml PE tubes suitable for most applications, such as urogenital, oral, auditory, nasal and throat mucosa, skin, and the like.
Ejemplo 3. Gel de polimaleato-SpdExample 3. Polymaleate-Spd Gel
Se preparó un gel con los ingredientes indicados en la Tabla I, a continuación.A gel was prepared with the ingredients indicated in Table I, below.
Tabla ITable I
Se dispersó Gantrez S97BF, un copolímero de polimaleato de ISP Corp. (Wayne, NJ, USA) en agua y 8 ml de NaOH 1 N. Se añadió solución de espermidina y se agitó durante 5', proporcionando complejo de polimaleato-Spd 3x104:1 eq/eq. Se mezclaron los demás componentes para acabar con un gel consistente adecuado para la mayoría de aplicaciones indicadas en la presente memoria.Gantrez S97BF, a polymeleate copolymer from ISP Corp. (Wayne, NJ, USA) in water and 8 ml of 1 N NaOH was dispersed. Spermidine solution was added and stirred for 5 ', yielding 3 × 10 4 polymaleate-Spd complex: 1 eq / eq. The other components were mixed to finish with a consistent gel suitable for most of the applications indicated herein.
Ejemplo 4. Gel de polianiones mixtos-SpdExample 4. Gel of mixed polyanions-Spd
Se preparó un gel con los ingredientes indicados en la Tabla II, a continuación.A gel was prepared with the ingredients indicated in Table II, below.
Tabla IITable II
Se disolvió HA sódico, Blanose 7HXF (CMC) y policarbófilo Noveon AA-1 en agua hasta la hidratación oral, proporcionando un gel viscoso. A continuación, se añadió la solución de espermidina y se agitó durante 15'. Se añadieron individualmente los demás ingredientes y se mezclaron, proporcionando un gel de color verde homogéneo útil para la reparación de las mucosas.Noveon AA-1 was dissolved in sodium hydroxide, Blanose 7HXF (CMC) and polycarbophil in water until oral hydration, providing a viscous gel. Next, the spermidine solution was added and stirred for 15 '. The other ingredients were added individually and mixed, providing a homogeneous green gel useful for mucosal repair.
Ejemplos 5-6. Gel de polimaleato-Spd con NAC e inhibidor de la mucoadhesiónExamples 5-6. Polyaleate-Spd gel with NAC and mucoadhesion inhibitor
Se preparó un gel con los ingredientes indicados en la Tabla III, a continuación.A gel was prepared with the ingredients indicated in Table III, below.
Tabla IIITable III
Se suspendió polimaleato (Gantrez S97BF) en agua y se tituló con NaOH 1 N hasta pH 6. A continuación, se mezcló espermidina, proporcionando un complejo de polimaleato-Spd 103:1 eq/eq.Polymaleate (Gantrez S97BF) was suspended in water and titrated with 1 N NaOH until pH 6. Then, spermidine was mixed, yielding a polymeleate-Spd complex 103: 1 eq / eq.
La adición de los siguientes ingredientes finalizó con un gel homogéneo adecuado para la utilización sobre mucosas dañadas, con una actividad auxiliar de disruptor de biopelícula.The addition of the following ingredients ended with a homogeneous gel suitable for use on damaged mucous membranes, with an auxiliary biofilm disruptor activity.
Se concibió una formulación todavía más mejorada con la adición de 5% a 15% p/p de D-manosa, proporcionando la inhibición de la adhesión bacteriana a las mucosas tratadas de esta manera.An even more improved formulation was conceived with the addition of 5% to 15% w / w of D-mannose, providing inhibition of bacterial adhesion to mucous membranes treated in this manner.
Ejemplo 7. Colutorio modificado de Carbopol-SpdExample 7. Modified Carbopol-Spd mouthwash
Se disolvió 1 g de Carbopol Ultrez 20 (Noveon-Lubrisol) en 500 ml del colutorio comercial Iodosan Antiplacca (lodosan SpA, grupo GSK). Se corrigió el pH a 6,5 con NaOH 1 N. A continuación, se añadieron 5 ml de espermidina 3HCl 1 mM y se mezclaron hasta obtener una solución homogénea adecuada para el cuidado oral, problemas estomatológicos y gingivitis.1 g of Carbopol Ultrez 20 (Noveon-Lubrisol) was dissolved in 500 ml of the commercial mouthwash Iodosan Antiplacca (lodosan SpA, group GSK). The pH was corrected to 6.5 with 1 N NaOH. Then, 5 ml of 1 mM spermidine 3 HCl was added and mixed until obtaining a homogeneous solution suitable for oral care, stomatological problems and gingivitis.
Ejemplo 8. Goma de mascar con HA-SpdExample 8. Chewing gum with HA-Spd
Se suministró una mezcla que comprendía xilitol, HA sódico y espermidina en una proporción 100:10:1 p/p/p obtenida mediante mezcla en seco de dichos componentes, a Gum Base Co Srl (Lainate, Italia) con instrucciones de fabricar una goma de mascar con 0,8% de dicha mezcla. El producto resultante se caracterizaba por una agradable palatabilidad que se aplica útilmente para la curación gingival.A mixture comprising xylitol, sodium HA and spermidine in a proportion of 100: 10: 1 w / w / w obtained by dry blending said components was supplied to Gum Base Co. Srl (Lainate, Italy) with instructions to make a rubber chew with 0.8% of said mixture. The resulting product was characterized by a pleasant palatability that is usefully applied for gingival healing.
Ejemplo 9. Evaluación in vivo en la estomatitis (estudio de caso)Example 9. In vivo evaluation in stomatitis (case study)
Se proporcionó el gel de Ejemplo 21 a una mujer de 46 años con úlceras aftosas recurrentes en la boca y se le dio las instrucciones de aplicarlo por lo menos dos veces al día. El sujeto refirió la resolución en aproximadamente una semana, frente a las 3-4 semanas generalmente necesarias. El producto resultó bien tolerado, excepto por el sabor amargo, probablemente debido al alcohol bencílico.The gel of Example 21 was given to a 46-year-old woman with recurrent aphthous ulcers in her mouth and she was instructed to apply it at least twice a day. The subject referred the resolution in about a week, compared to the usual 3-4 weeks. The product was well tolerated, except for the bitter taste, probably due to the benzyl alcohol.
Ejemplo 10. Dentífrico de gel de CMC-SpdExample 10. CMC-Spd Gel Toothpaste
Se preparó una pasta dentífrica con los ingredientes indicados en la Tabla IV, a continuación.A toothpaste was prepared with the ingredients indicated in Table IV, below.
Tabla IVTable IV
El procedimiento siguió el método ordinario para la producción de un dentífrico, excepto en que la CMC en primer lugar se disolvió en agua y se añadió con la solución de espermidina; después se incorporaron los ingredientes restantes, proporcionando un gel traslúcido azul. The procedure followed the ordinary method for the production of a dentifrice, except that the CMC was first dissolved in water and added with the spermidine solution; then the remaining ingredients were incorporated, providing a blue translucent gel.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| ITMI2010A001491A IT1405498B1 (en) | 2010-08-04 | 2010-08-04 | SUPRAMOLECULAR COMPLEXES OF SPERMIDINE FOR REGENERATION AND TISSUE TROPHISM. |
| IT002277A ITMI20102277A1 (en) | 2010-12-14 | 2010-12-14 | GYNECOLOGICAL COMPOSITIONS FOR USE IN ATROPHICAL VAGINITIS AND LUBRICATION DISORDERS |
| ITMI2010A002308A IT1403262B1 (en) | 2010-12-16 | 2010-12-16 | TOPIC COMPOSITIONS FOR THE CURE OF INJURIES OF THE ORAL CABLE |
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