KR20100122881A

KR20100122881A - A gamma-cyclodextrin derivative and polyelectrolyte multilayer comprising gamma-cyclodextrin derivative layer

Info

Publication number: KR20100122881A
Application number: KR1020100045166A
Authority: KR
Inventors: 양성윤; 진혜승
Original assignee: 충남대학교산학협력단
Priority date: 2009-05-13
Filing date: 2010-05-13
Publication date: 2010-11-23

Abstract

PURPOSE: A gamma-cyclodextrin derivative and the polymer electrolyte multi-layered membrane containing thereof are provided to secure the properties of gamma-cyclodextrin, and to form the multi-layered membrane with the excellent biocompatibility. CONSTITUTION: A gamma-cyclodextrin derivative is obtained by reacting gamma-cyclodextrin, epichlorohydrin, and choline chloride. The gamma-cyclodextrin derivative has eight glucopyranose units combining into an alpha-(1,4) linkage. The glucopyranose units have a substituent marked with chemical formula 2.

Description

A polyelectrolyte multilayer film comprising a gamma-cyclodextrin derivative and the gamma-cyclodextrin derivative layer {A GAMMA-CYCLODEXTRIN DERIVATIVE AND POLYELECTROLYTE MULTILAYER COMPRISING GAMMA-CYCLODEXTRIN DERIVATIVE LAYER}

The present invention relates to a polymer electrolyte multilayer film comprising a gamma-cyclodextrin derivative and the gamma-cyclodextrin derivative layer.

Recently, bioengineering has emerged. Bioengineering refers to the characteristics of cells through interaction between the support and the cells where cells are attached and grow, unlike conventional methods of controlling hormones, growth factors, and serum contained in cell culture in studying or inducing cell functions. It refers to the regulation of phosphorus attachment, proliferation, differentiation and extracellular matrix secretion. This requires the development of materials with biocompatibility and the development of chemical surface modification.

Therefore, a variety of multilayer membranes including a multilayer membrane having not only biocompatibility but also improving cell adhesion, and a multilayer membrane having both capturing ability and cell adhesion for a specific material, and technologies for biosensors including the same Development is required.

In order to meet the above requirements, the present invention provides a novel gamma-cyclodextrin derivative for laminating a gamma-cyclodextrin having a high biocompatibility and a selective capturing ability to a material into a layer constituting a multilayer. An object of the present invention is to provide a polymer electrolyte multilayer film in which a polymer and a gamma-cyclodextrin derivative are laminated.

In order to achieve the above object, the present invention provides a novel gamma-cyclodextrin derivative for the production of a stackable polymer electrolyte.

The present invention also provides polymers of the novel gamma-cyclodextrin derivatives.

In addition, the present invention provides a multilayer membrane having an alternating laminated structure of an anionic polymer electrolyte layer and a cationic polymer electrolyte layer, wherein at least one layer of the cationic polymer electrolyte layer is a polymer layer of the novel gamma-cyclodextrin derivative. .

In addition, the present invention provides a biosensing device comprising the multi-layer film in order to achieve the above object.

In addition, the present invention provides a chemical sensitive device including the multilayer film in order to achieve the above object.

The inventors of the present invention prepare a multilayer membrane using a layer-by-layer (LBL) deposition technique using a polymer electrolyte that is safest in terms of immunity and toxicity, and secures antimicrobial and antibacterial properties, and has a cell adsorption capacity and Studies have been conducted to add gamma-cyclodextrins that can provide the ability to capture certain substances depending on the application conditions. In the above research process, by simply adsorbing gamma-cyclodextrin to the film to prepare a multi-layered film containing gamma-cyclodextrin, it was confirmed that it is not easy to control the amount of adsorption of gamma-cyclodextrin and the like. In the course of research, the gamma-cyclodextrin can be solved in the case of stacking the problem, a novel gamma-cyclodextrin derivative and a polymer of the derivative is prepared as a means for laminating the gamma-cyclodextrin In addition, when the polymer of the derivative is used, it was confirmed that gamma-cyclodextrin and / or derivative thereof can be laminated like the polymer electrolyte, thereby completing the present invention. In addition, when the polymer of the derivative of the gamma-cyclodextrin is laminated in place of the cationic polymer in the multilayer film, the multilayer structure in which the polymer of the derivative is laminated, that is, the trapping ability to capture a specific material is improved, and The present invention was completed by further confirming that not only the optical activity of the material can be induced, but also the degree of lamination can be used to control the change of the multilayer film properties according to gamma-cyclodextrin.

Hereinafter, the present invention will be described in more detail.

In the present specification, cyclodextrin refers to a cyclic oligosaccharide in which glucopyranose units have α- (1,4) bonds. The cyclodextrin is obtained by degrading starch by an enzyme called cyclodextrin glucanotransferase (CGTase), followed by an intermolecular transglycosylation.

In addition, in the present specification, gamma-cyclodextrin (γ-cyclodextrin) means a cyclic oligosaccharide in which eight glucopyranose units are bonded.

As shown in FIG. 1, a hydroxyl group bonded to carbon 2 (C 2) and carbon 3 (C 3) of the cyclodextrin is expanded to the outside of the ring, and is bonded to carbon 6 (C 6). The bound hydroxyl groups also extend outwards in the opposite direction, making the outer ring hydrophilic. In addition, the oxygen of hydrogen and ether bonded to carbon 3 (C3) and carbon 5 (C5) are located in the inner direction of the ring, and the cavity of the ring is hydrophobic. . Cyclodextrins have hydrophilic properties with high solubility in polar solvents such as water, and the hydrophobic properties of the ring form molecular size pores within the molecules, and thus have different properties within one molecule. You have a micro heterogeneous environment.

In the present specification, a polymer electrolyte (Polyelectrolyte) refers to a polymer that can be dissolved in a solvent and dissociate, specifically, a polymer that can be dissolved in water and can be positively charged or negatively charged. The polymer electrolyte includes a cationic polymer electrolyte (polyanion). And anionic polymer electrolytes (polycation) may be included.

The cationic polymer electrolyte may mean a polymer electrolyte in which the chemical functional group of the polymer chain loses an anion or obtains hydrogen ions in an aqueous solution, and thus may be positively charged. The degree of ionization may be affected by the acidity or basicity of the surrounding environment. In addition to various weakly electrolytic cationic polymers, neutrally water-soluble polymers include neutral water-soluble polymers such as polyacrylamide, which are not ionized well in the pH range of general water and have a positive charge when the acidity of the surroundings is very high.

The anionic polymer electrolyte may mean a polymer electrolyte in which a polymer functional group of the polymer chain loses a cation or loses hydrogen ions, and thus may be negatively charged. The degree of ionization may vary depending on the acidity or basicity of the surrounding environment. Weak electrolyte anionic polymers are included.

In the present specification, 1 bilayer means a multilayer film in which one layer of each of two polymer electrolyte layers having different kinds is stacked. For example, the three bilayers are repeatedly immersed three times in a polymer electrolyte solution containing an anionic polymer electrolyte and a polymer electrolyte solution containing a cationic polymer electrolyte, thereby anionic polymer electrolyte layer and cationic polymer electrolyte layer. This means a multilayer film in which three double films stacked one by one are laminated.

In one aspect of the invention, the invention relates to novel gamma-cyclodextrin derivatives. The present invention also relates to polymers of the novel gamma-cyclodextrin derivatives.

The novel gamma-cyclodextrin derivatives of the present invention are prepared by reacting gamma-cyclodextrin (γ-CycloDextrin, γ-CD), epichlorohydrin (EP) and choline chloride (CC). Cyclodextrin derivatives. The gamma-cyclodextrin derivative may be prepared by first reacting gamma-cyclodextrin with choline chloride and then adding epiclohydrin to the reaction product.

The content of the gamma-cyclodextrin, epichlorohydrin and choline chloride is 1: 12 to 17: 0.5 to 2, preferably 1: 14 to 16: 0.8 to 1.2, more preferably 1: 1: based on molar ratio. 14.5 to 15.5: to 0.9 to 1.1, gamma-cyclodextrin derivatives.

More specifically, the gamma-cyclodextrin derivative is a gamma-cyclodextrin derivative having 8 glucopyranose units having a structural formula of Formula 1 to which α- (1,4) is bonded. At least one or more of the glucopyranose units having the structure of Formula 1 includes a substituent of Formula 2 below, wherein X in the substituent of Formula 2 in the gamma-cyclodextrin derivative is at least one gamma- Cyclodextrin derivatives.

[Formula 1]

R 1, R 2, and R 3 are each independently any one selected from the group consisting of hydrogen and a substituent of Formula 2 below.

[Formula 2]

X is hydrogen or a substituent of formula (3).

(3)

In addition, in R1, R2, and R3 of Chemical Formula 1, a position which is preferentially substituted with a substituent of Chemical Formula 2 may be R1. In this aspect, the gamma-cyclodextrin derivative may include one or more glucopyranose units having the structural formula of Formula 4, and preferably 8 glucopyranose units having the structural formula of Formula 4 are α-. (1,4) is a gamma-cyclodextrin derivative having a bond, and at least one or more of the glucopyranose units having the structural formula of Formula 4 may include a substituent of Formula 5 below.

[Formula 4]

R2 and R3 are each independently any one selected from the group consisting of hydrogen and a substituent of the formula (5)

[Chemical Formula 5]

The present invention also relates to polymers of gamma-cyclodextrin derivatives.

The gamma-cyclodextrin derivative may include the novel gamma-cyclodextrin derivative as a monomer.

Specifically, the polymer may include a gamma-cyclodextrin derivative having eight glucopyranose units having a structural formula of Formula 1 having α- (1,4) linkage as a monomer.

In the gamma-cyclodextrin derivative having 8 glucopyranose units having the structural formula of Formula 1, having α- (1,4) linkage, at least one or more of the glucopyranose units having the structural formula of Formula 1 may be represented by Formula 2 below: X of the substituent of Formula 2 in the gamma-cyclodextrin derivative, including at least one substituent may be a gamma-cyclodextrin derivative having at least one formula (3).

[Formula 1]

[Formula 2]

X is hydrogen or a substituent of formula (3).

(3)

The polymer is preferably 10% to 30%, preferably 12% to 25%, more preferably 15% to 20% of hydrogen, 70% of R1, R2 and R3 included in the structural formula of Formula 1 To 90%, preferably 75% to 88%, more preferably 80% to 85% may be a polymer of the formula (2).

In addition, the polymer is 20% to 40%, preferably 25% to 37%, more preferably 30% to 35% of R1, R2 and R3 included in the structural formula of Formula 1 includes a substituent of Formula 3 It may be a polymer.

In addition, the polymer may be one comprising the novel gamma-cyclodextrin derivative and gamma-cyclodextrin as monomers.

In addition, in one aspect of the present invention, the present invention relates to a method for producing the polymer.

The method for preparing the polymer may include preparing an aqueous NaOH solution to which gamma-cyclodextrin is added; Preparing a mixed solution by adding choline chloride to an aqueous NaOH solution to which gamma-cyclodextrin is added; It may be a method comprising the step of adding the epichlorohydrin to the mixed solution and the reaction by adding heat to the mixed solution to which the epichlorohydrin is added.

The preparing of the NaOH aqueous solution to which the gamma-cyclodextrin is added may be performed by adding and stirring gamma-cyclodextrin to the NaOH aqueous solution, and the NaOH aqueous solution is preferably 20 ° C. to 30 ° C., more preferably. It may be 23 ℃ to 27 ℃, the stirring time may be 12 hours to 36 hours or 18 hours to 30 hours or 20 hours to 28 hours.

In the step of preparing the mixed solution, the content of choline chloride with the added gamma-cyclodextrin is 1: 0.5 to 1: 2 (gamma-cyclodextrin: choline chloride) or 1: 0.8 to 1: 1.2 based on the molar ratio. (Gamma-cyclodextrin: choline chloride) or 1: 0.9 to 1: 1.1.

Adding epichlorohydrin to the mixed solution may be performed by adding epichlorohydrin at a rate of 0.05 ml / min to 0.15 ml / min or 0.075 ml / min to 0.125 ml / min. Epichlorohydrin has a content ratio of 1: 12 to 1: 17 (gamma-cyclodextrin: epichlorohydrin) or 1: 14 to 1: 16 (gamma-cyclo) based on the molar ratio with the added gamma-cyclodextrin. Dextrin: epichlorohydrin) or 1: 14.5 to 1: 15.5.

The step of advancing the reaction by adding heat to the epichlorohydrin-added mixture may be carried out at 40 ℃ to 80 ℃ or 50 ℃ to 70 ℃ or 55 ℃ to 65 ℃ , The mixture may be stirred by stirring at a speed of 300 rpm to 1,000 rpm or 400 rpm to 800 rpm or 500 rpm to 700 rpm, and the reaction time is 30 minutes to 4 hours or 1 hour to 3 hours or 90 minutes to 150 It may be minutes.

In addition, the preparation method may further proceed to terminate the reaction by adding an aqueous hydrochloric acid after the step of proceeding the reaction. The aqueous hydrochloric acid solution may be a 2N hydrochloric acid aqueous solution to 4N hydrochloric acid aqueous solution or 2.5N hydrochloric acid aqueous solution to 3.5N hydrochloric acid aqueous solution.

In addition, the production method may further perform a process of concentrating by filtering or dialysis the reaction solution after the reaction is completed.

In another aspect of the present invention, the present invention relates to a multilayer film.

The multilayer film may be a polymer electrolyte multilayer film, and specifically, the multilayer film may be a multilayer film including an alternating layered structure of an anionic polymer electrolyte layer and a cationic polymer electrolyte layer, preferably the anionic polymer electrolyte layer and a cationic It includes an alternating layered structure of a polymer electrolytically etched layer, wherein the cationic polymer electrolyte layer is a poly allylamine hydrochloride, poly acrylamide, a polymer comprising the novel gamma-cyclodextrin derivative and the gamma-cyclodextrin derivative as a monomer It may be a multilayer film consisting of one or more selected from the group consisting of.

The multilayer film is preferably a multilayer film made of one or more selected from the group consisting of at least one layer of the cationic polymer electrolyte layer and a polymer comprising the novel gamma-cyclodextrin derivative and the gamma-cyclodextrin derivative as monomers. Can be.

In addition, the multilayer film may preferably be a multilayer film made of a polymer in which at least one layer of the cationic polymer electrolyte layer includes the gamma-cyclodextrin derivative as a monomer. Preferably, the multilayer membrane is formed of at least one selected from the group consisting of polyallylamine hydrochloride, polyacrylamide, and a polymer comprising the gamma-cyclodextrin derivative as a monomer in the cationic polymer electrolyte layer. Any one or both layers of the outermost layer of the structure may be a multilayer film made of a polymer containing the gamma-cyclodextrin derivative as a monomer.

In addition, the multilayer film is a polymer in which one or both of the outermost layers of the alternating layered structure of the anionic polymer electrolyte layer and the cationic polymer electrolyte layer contains the novel gamma-cyclodextrin derivative and the gamma-cyclodextrin derivative as monomers. It may be made of one or more selected from the group consisting of.

The multilayer film may preferably be a multilayer film in which one or both layers of the outermost layer is made of a polymer including the gamma-cyclodextrin derivative as a monomer.

The anionic polymer electrolyte layer is preferably poly methacrylic acid (PMA), poly acrylacid (PAA), polystyrene sulfonate (PSS) and hyaluronic acid (HA) It may be made of any one selected from the group consisting of, more preferably may be made of polymethacrylic acid or polyacrylic acid.

The cationic polymer electrolyte layer is poly (allylamine hydrochloride (PAH), poly acrylamide (PAAm), poly 4-vinylbenzlytri-methyl ammonium chloride , PVTAC), the novel gamma-cyclodextrin derivative and the polymer comprising the gamma-cyclodextrin derivative as a monomer may be one or more selected from the group consisting of, polyallylamine hydrochloride, polyacryl As the amide, the novel gamma-cyclodextrin derivative and the gamma-cyclodextrin derivative may be one or more selected from the group consisting of polymers containing monomers.

The anionic polyelectrolyte and polyallylamine hydrochloride, poly acrylamide, poly 4-vinylbenzyltrimethylammonium chloride, any one selected from the group consisting of polymethacrylic acid, polyacrylic acid, polystyrene sulfonate and hyaluronic acid The cationic polymer electrolyte, which is any one selected from the group consisting of gamma-cyclodextrin derivatives and the gamma-cyclodextrin derivatives, is a weak polyelectrolyte in which the charge density is sensitively changed depending on pH, and the pH of the polymer electrolyte The change directly affects the chain structure of the polymer electrolyte. The change in the chain structure affects various properties such as the structure or morphology of the polymer membrane or the wettability. Therefore, the anionic polymer electrolyte layer and the cationic polymer electrolyte layer corresponding to the polymer weak electrolyte can easily modify the structure of the electrolyte layer by adjusting the pH to form a multilayer film having a desired structure, or a desired material on the multilayer film. Has the advantage of being easy to adsorb.

The polymethacrylic acid may include not only polymethacrylic acid itself but also salts thereof. Specifically, at least one selected from the group consisting of the polymethacrylic acid and polymethacrylic acid salts. The molecular weight of the polymethacrylic acid is not particularly limited, but may be, for example, 10,000 to 200,000 or 50,000 to 150,000.

The polyacrylic acid may include not only polyacrylic acid itself but also a salt thereof. Specifically, it may be at least one selected from the group consisting of the polyacrylic acid and polyacrylic acid salt. The molecular weight of the polyacrylic acid is not particularly limited, but may be, for example, 10,000 to 100,000 or 30,000 to 90,000.

The hyaluronic acid is a kind of glucosamide glycan consisting of a disaccharide unit in which D-glucuronic acid and N-acetylglucosamine are connected by β (1 → 3) glycosidic bonds, and the hyaluronic acid has a chemical and physical structure. There are no species differences, humans have a metabolic system and are the safest biomaterials in terms of immunity and toxicity.

The hyaluronic acid may include not only hyaluronic acid itself but also all salts thereof. Specifically, the hyaluronic acid may be one or more selected from the group consisting of hyaluronic acid and hyaluronic acid salts. The hyaluronic acid salt includes both inorganic salts such as sodium hyaluronate, magnesium hyaluronate, zinc hyaluronate, cobalt hyaluronic acid and the like and organic salts such as tetrabutylammonium hyaluronic acid. The molecular weight of the hyaluronic acid is not particularly limited, but may be, for example, 100,000 to 10,000,000.

The polyallylamine hydrochloride may be chemically bonded with hyaluronic acid or polyacrylic acid, and the molecular weight of the polyallylamine hydrochloride is not particularly limited, but may be, for example, 10,000 to 100,000 or 30,000 to 80,000.

The poly acrylamide may be chemically bonded with hyaluronic acid or polyacrylic acid, and the molecular weight of the poly acrylamide is not particularly limited, but may be, for example, 100,000 to 10,000,000.

When the cationic polyelectrolyte is polyacrylamide, the multi-layered film may have cross-linking formed by performing a crosslinking reaction between the polymer electrolytes in order to impart stability to the biological environment. The crosslinking reaction may be performed by, for example, a chemical reaction or heat treatment through EDAC.

The multilayer membrane may be a multilayer membrane having at least one cationic polymer electrolyte layer and one anionic polymer electrolyte layer. For example, the multilayer film may be 1 bilayer or more, and may be two or more layers, respectively.

In the present invention, the layer-by-layer (LBL) refers to a method of controlling the thickness of the polymer electrolyte multilayer film or the film by repeating as many times as desired so that the polymer electrolyte having a relative charge is adsorbed on the substrate surface. .

In addition, when the cationic polymer electrolyte layer or the anionic polymer electrolyte layer is two or more, the electrolyte layer may be laminated with the same polymer electrolyte layer or different polymer electrolyte layers, respectively. Specifically, when the anionic polymer electrolyte layer is two or more layers, specific anionic polymer electrolyte layers may be alternately stacked with the cationic polymer electrolyte layer, or different types of anionic polymer electrolyte layers may be alternately stacked with the cationic polymer electrolyte layer. Can be. The cationic polymer electrolyte layer may be the same as above.

The gamma-cyclodextrin is a cyclic oligosaccharide in which eight glucopyranose units have α- (1,4) bonds. In the multilayer film including the gamma-cyclodextrin, a compound having a predetermined size that can be trapped in the gamma-cyclodextrin, in particular an optically inert molecule, is collected in the pore portion of the gamma-cyclodextrin, Specifically, when coordination is formed with gamma-cyclodextrin, the coordinated conjugated compound and gamma-cyclodextrin interact with each other to induce optical activity. The induced optical activity imparts a beneficial effect to the multilayer film, in particular chiral selectivity in the analysis and separation of a specific material, so that the multilayer film has a remarkable effect of performing analysis and separation of a specific material according to optical properties. .

In addition, since the diameter and volume of the pores formed in the gamma-cyclodextrin may form an inclusion complex through host-guest interaction with a large molecule such as a compound having a steroid or a macrocycle, the gamma- The multilayer membrane of the present invention in which cyclodextrin is laminated is characterized by having a remarkable effect in the analysis and separation of macromolecules such as steroids.

In addition, the novel gamma-cyclodextrin derivatives may be polymerized as described above and stacked on a multilayer membrane with a cationic polymer electrolyte layer such as polyallylaminehydrochloride, poly acrylamide and poly 4-vinylbenzyltrimethylammonium chloride. Therefore, while maintaining the advantages of the gamma-cyclodextrin, it is not adsorbed or contained in a difficult-to-control form in the multilayer film, it can be laminated in a form that can control the function of the gamma-cyclodextrin, if necessary, The properties and functions of the gamma-cyclodextrin may be controlled to impart various effects to the multilayer film.

Moreover, in one aspect of this invention, this invention relates to the manufacturing method of the said multilayer film.

The method of manufacturing the multilayer film may be a method including forming an alternate stacked structure by alternately stacking an anionic polymer electrolyte layer and a cationic polymer electrolyte layer.

Forming the alternate stacking structure may be performed by alternately stacking the anionic polymer electrolyte layer and the cationic polymer electrolyte layer.

The method of alternately stacking the anionic polymer electrolyte layer and the cationic polymer electrolyte layer may be performed by, for example, a layer-by-layer (LBL) deposition technique. The layer-by-layer (LBL) deposition technique may be performed by the method as shown in FIG. 2. In the layer-by-layer (LBL) deposition technique, the adsorption step is repeated as desired while alternately adsorbing a polymer electrolyte having a relative charge, thereby controlling the thickness of the polymer electrolyte layer, that is, the thickness of a multilayer film or a film. desirable.

As an example, the step of forming the alternating stacked structure may include a polymer electrolyte having a relative charge between a colloid having a charge (for example, a colloid having a negative charge) or a substrate (for example, a substrate having a negative charge) as in the method of FIG. 2. A process of forming a polymer electrolyte layer by immersing in a solution (positive charge polymer electrolyte solution), adsorbing a polymer electrolyte to a substrate, and a polymer electrolyte solution (negative charge) having a substrate on which the polymer electrolyte layer is formed has a relative charge with the polymer electrolyte of the polymer electrolyte solution. Or immersing the polymer electrolyte in a colloid or a substrate on which the polymer electrolyte layer is formed to form another polymer electrolyte layer on the polymer electrolyte layer, or the polymer electrolyte layer Process and another polymer electrolyte It can be carried out by repeating the process of forming the layer.

The process of forming the polymer electrolyte layer and the process of forming another polymer electrolyte layer may further include a process of washing the substrate on which the electrolyte layer is formed after each process.

The step of forming the alternate stacking structure may be performed by alternately stacking the anionic polymer electrolyte layer and the cationic polymer electrolyte layer on the colloid or the substrate.

The colloid may mean colloidal particles, and the colloidal particles may be particles having carboxylic acid functional groups on the surface thereof, for example, polystyrene particles or magnetic particles having carboxylic acid functional groups, for example Fe ₃ O ₄ .

In the process of forming the multilayer film on the colloidal particles, as shown in FIG. 3, the polymer electrolyte layer having the opposite inversion to the charged colloidal particles is laminated, and then the polymer electrolyte layer is laminated with the polymer electrolyte layer having the relative charge. The method may be performed by repeatedly stacking the multilayer film of the 1 bilayer.

The substrate is not particularly limited, but may be, for example, a glass slide made of glass, a metal substrate made of silicon or gold, or a substrate made of polystyrene. Specifically, the substrate may be a substrate that tends to be positively charged, such as mica, and may be a substrate that tends to be negatively charged, such as metal substrates such as glass, silicon, gold, and steel. It may be an organic substrate having no charge such as plastics (polystyrene, acrylic, Teflon, etc.).

In the case of the charged substrate, the polymer electrolyte layer may be formed using the polymer electrolyte of the opposite charge, and in the case of the organic substrate having the non-charge, the polymer electrolyte layer may be formed by using hydrophobic adsorption between the polymer materials. Can be formed.

The step of alternately stacking the anionic polymer electrolyte layer and the cationic polymer electrolyte layer may be performed by alternately immersing the polymer electrolyte solution containing the anionic polymer electrolyte and the polymer electrolyte solution containing the cationic polymer electrolyte. have.

The anionic polymer electrolyte solution used in the step of forming the electrolyte layer is pH 1.0 to pH 11.0, preferably pH 2.0 to pH 10.0, more preferably pH 3.0 to pH 5.0, even more preferably pH 3.5 to pH 4.5 PH may be adjusted to.

In addition, in the case of the cationic polymer electrolyte solution used in forming the electrolyte layer, when the cationic polymer is polyallylamine hydrochloride or poly acrylamide, the pH of the electrolyte solution is preferably pH 1.0 to pH 11.0. , Preferably pH 2.0 to pH 10.0, more preferably pH 7.5 to pH 9.5, even more preferably pH adjusted to pH 8.0 to pH 9.0, the cationic polymer is the gamma-cyclodextrin derivative In the case of the polymer including the monomer, the pH of the electrolyte solution may be pH 1.0 to pH 11.0 or pH 2.0 to pH 10.0 or pH 3.0 to pH 5.0 or pH 3.5 to pH 4.5.

When laminating using the anionic polymer electrolyte solution of pH and the cationic polymer electrolyte solution of pH, the gamma-cyclodextrin or its derivatives can be effectively adsorbed or included in the multilayer membrane, The pH is preferably in the above pH range.

When the cationic polyelectrolyte is polyacrylamide, polyallylamine hydrochloride is immersed in a polymer electrolyte solution containing polyallylamine hydrochloride, which is a positively charged polyelectrolyte, in the case of a weak silicon substrate having a surface adhesion. After the layer is formed, an alternating layered structure can be formed using the anionic polymer electrolyte solution and polyacrylamide solution.

The step of forming the alternating stacked structure is, when the cationic polyelectrolyte is poly acrylamide, in order to give stability to the biological environment, to perform cross-linking reaction between the polymer electrolyte to form cross-linking (cross-linking) It may further comprise a step. The forming of the crosslinking may be performed by a method of forming a crosslinking reaction through chemical reaction or heat treatment through EDAC. The heat treatment may be performed by heating at 70 ° C. to 90 ° C. or 80 ° C. for 8 to 12 hours or 10 hours, and the reaction time or temperature may be adjusted by a tendency to reduce when using a vacuum oven.

In addition, any one or both of the outermost layers of the alternating layered structure of the multilayer film is a multilayer film made of one or more selected from the group consisting of a polymer comprising the novel gamma-cyclodextrin derivative and the gamma-cyclodextrin derivative as monomers. May be prepared by using a polymer electrolyte solution containing a polymer including a gamma-cyclodextrin derivative as a monomer as a polymer electrolyte solution used first or for the first and last time in the manufacture of the laminated structure.

In addition, the multilayer membrane may be used as a tissue engineering material, for example, a support material for growing cells, and a device or apparatus requiring various polymer multilayer membranes including a chemical sensitive device such as a biosensing device or a chemical sensor. It can be applied to.

In one embodiment, the present invention provides a biosensing device including the multilayer film.

In the case of the multilayer membrane, the polymer electrolyte may be patterned for purposes such as selective adsorption of cells or for application of a biosensing device.

A biomaterial fixing material may be combined with the multilayer pattern layer formed through the patterning. For example, the biomaterial may be any one selected from the group consisting of cells, monosaccharides, disaccharides, oligosaccharides, fatty acids, polypeptides or proteins and polynucleotides, and preferably cells. The cell may be any cell including prokaryotic and eukaryotic cells, for example, fibroblasts, hepatocytes, neurons, cancer cells (eg HeLa cells), B cells, white blood cells (white blood cell, eg, Raw 264.7), and the like, and immune cells and embryonic cells.

The biomaterial-fixing material may be a biomaterial-fixing biomaterial. The biomaterial for fixing the biomaterial may be any oligopeptide including an oligopeptide sequence having an amino acid sequence of RGD and a protein including the oligopeptide and other receptor peptides, and the protein including the oligopeptide may be fibronectin ( fibronectin or fibrin, and the like, and the protein having the receptor peptide may be laminin, collagen, or the like.

In addition, the multilayer film may further include a thin film on the pattern layer, the thin film may include the biomaterial, preferably cells, the biomaterial is a biomaterial for fixing the biomaterial, preferably Preferably, the polypeptide may include an oligopeptide sequence having an amino acid sequence of RGD. The thin film may be attached to a polymer electrolyte pattern layer including a polymer of a polymer electrolyte or gamma-cyclodextrin or a derivative thereof. Preferably, the cell included in the thin film is a polymer electrolyte or gamma combined with a cell biomaterial. It may be attached to a pattern layer including a cyclodextrin or a derivative thereof.

When the multilayer film is used for a DNA chip, a protein chip, or a cell-based biosensor, the outermost layer not including the pattern layer may be a substrate adhesive layer bonded to a substrate. And when the multilayer film is used as a surface facing the damaged tissue for tissue regeneration inducing function, such as used for induced tissue regeneration, or as a dressing material for skin or mucosal tissue, the outermost layer not including the pattern layer Each layer may be a tissue adhesion layer that bonds to biological tissue.

In another aspect of the invention, the invention relates to a bioreactor.

The bioreactor may be a biosensing device including the multilayer film.

Specifically, the bioreactor is

It may be a biosensing device including a substrate and the multilayer film.

The biosensing device is combined with a positively charged polymer electrolyte containing an amide group in a cell-fixing biomaterial, and thus can be applied to a cell-based biosensing device because it has excellent chemical stability and can provide conditions suitable for cell culture. . The biosensing device may be, for example, a biosensor.

The substrate is a substrate that can be applied to the biosensing device, the material is not particularly limited, for example, made of glass (glass slide) or metal substrate such as silicon (silicon) or gold (gold) It may have been. Specifically, the substrate may be a substrate that tends to be positively charged, such as mica, and may be a substrate that tends to be negatively charged, such as metal substrates such as glass, silicon, gold, and steel. It may be an organic substrate having no charge such as plastics (polystyrene, acrylic, Teflon, etc.).

In the case of the charged substrate, the multilayer film may be coated using a polymer electrolyte of opposite charge, and in the case of the organic substrate in the absence of the charge, the multilayer film may be coated using hydrophobic adsorption between polymer materials. .

The multilayer membrane may be one in which an anionic polymer electrolyte layer and a cationic polymer electrolyte layer are manufactured by using a layer-by-layer (LBL) deposition technique, and the cationic polymer electrolyte layer may be polyallylamine hydrochloride or polyacrylamide. Thus, the novel gamma-cyclodextrin derivatives and the gamma-cyclodextrin derivatives are composed of at least one selected from the group consisting of polymers comprising monomers, and at least one layer of the cationic polymer electrolyte layer is the new gamma It may be composed of one or more selected from the group consisting of a cyclodextrin derivative and a polymer comprising the gamma-cyclodextrin derivative as a monomer.

In addition, the multilayer film may be applied to a device or a device requiring various polymer multilayer films including a chemical sensitive device.

In another aspect of the invention, the invention relates to a chemical sensitive device. For example, the present invention may be a chemical sensitive device including the multilayer film.

Specifically, the chemical sensitizer may be a chemical sensitizer including a substrate and a multilayer membrane including gamma-cyclodextrin or a derivative thereof in an alternating stacked structure of the anionic polymer electrolyte layer and the cationic polymer electrolyte layer.

The novel gamma-cyclodextrin derivatives of the present invention not only retain the characteristics of gamma-cyclodextrins but can also form polymers, which can be laminated in a form that can control the function of gamma-cyclodextrins, thereby gamma It is advantageous in that the properties and functions of cyclodextrin can be controlled to impart various effects to the multilayered film.

Accordingly, the multilayer membrane and the biosensing device of the present invention, in which the polymer of the gamma-cyclodextrin derivative is stacked, can easily control the adsorption of specific cells as compared to the laminated structure in which the existing cationic polymer layer and the anionic polymer layer are alternately stacked. And, it can adsorb a specific material inside the multilayer film, and can improve the optical activity of the specific material, it can be used for various purposes including a biosensor, as well as for experiment or diagnosis in combination with biological tissue, It can also be used for medical treatment or disease treatment, so the industrial effect will be very large.

1 is a schematic diagram showing the structure of a cyclodextrin analyzed by x-ray structure analysis according to an embodiment of the present invention.
2 is a process diagram illustrating a method of alternately stacking an anionic polymer electrolyte layer and a cationic polymer electrolyte layer by a layer-by-layer (LBL) deposition technique according to an embodiment of the present invention.
3 is a flowchart illustrating a method of alternately stacking a cationic polymer electrolyte layer and an anionic polymer electrolyte layer on colloidal particles according to an embodiment of the present invention.
Figure 4 is a graph showing the results of performing NMR to analyze the polymer of the gamma-cycle polymer derivative prepared by the present invention, according to an embodiment of the present invention.

Hereinafter, preferred embodiments of the present invention are described. However, the following examples are only examples for illustrating the present invention, and thus the protection scope of the present invention is not limited to the following examples.

Example 1: Gamma Cyclodextrin Containing Multilayer Produce

Example 1-1. Materials and reagents

Poly (methacrylic acid) (PMA, Mw = 100,000) and Poly (allylamine hydrochloride) (PAH, Mw = 60,000) were purchased from Polysciences, Inc. (UK) and gamma-cyclodextrin (γ-cyclodextrin) was SUPELCO. The carboxylated γ-cyclodextrin purchased from Inc., USA was used. Methylene blue Hydrate (MeB) was used as a product of Fluka (Germany).

The polymer electrolyte (PMA, PAH and gamma-cyclodextrin) and MeB were used without further purification. Deionized water used in the preparation and washing process of the aqueous solution used in the experiment was performed using deionized water (> 18 MΩcm) produced by Millipore's Milli-Q water purifier (Millipore Co., USA).

In addition, Dulbecco's Phosphate Buffered Saline (DPBS), 0.25% Tripsin-EDTA, and Dulbecco's modified eagle medium (DMEM) used Gibco (USA) products, and also used fetal bovine serum (FBS) and Antibiotic-Antimycotic used in the preparation of DMEM. Gibco (USA) product was used.

Example 1-2. gamma- Cyclodextrin Preparation of Derivatives and Polymers

The amount of gamma-cyclodextrin, EP and CC used in the preparation of the cationic gamma-cyclodextrin was 1: 15: 1 based on the molar ratio. First, 1 g of NaOH was dissolved in 20 ml of deionized water to prepare an aqueous NaOH solution, and then 1.135 g of gamma-cyclodextrin was added to the aqueous NaOH solution.

The NaOH aqueous solution to which gamma-cyclodextrin was added was stirred at 25 ° C. for 24 hours, and then 0.140 g of CC was added to the aqueous solution, and 1.388 g of EP was added at a rate of about 0.1 ml / min. After the EP was added, heat was applied at 60 ° C., and the conditions of 60 ° C. and 600 rpm were maintained for 2 hours while the polymerization was in progress. After 2 hours had elapsed, the polymerization was terminated by adding 3N aqueous hydrochloric acid solution. The final reactant was dialyzed for 24 hours using the molecular weight cut-off 1000 membrane, and lyophilization was performed on the dialyzed product to obtain a pure powder.

In order to identify the synthesized gamma-cyclodextrin derivative, GPC and NMP were performed on the final product.

The GPC sample for measuring the molecular weight of the synthesized gamma-cyclodextrin derivative (cationic γ-CyD polymer) was prepared in a concentration of 3 mg / ml of the powder, was carried out using PEG as the GPC standard, The result Mn was 2211.5 g / mol, Mw was 2306.4 g / mol, and D: 1.0429 was obtained.

In addition, the NMR performed to analyze the structure of the synthesized gamma-cyclodextrin derivative (cationic γ-CyD polymer) was performed at 400 MHz resonance frequency using JNM-AL400, to perform NMR. For the preparation of the solution, Aldrich's distilled water (dH ₂ O) was used. The NMR results are shown in FIG. 4.

As shown in FIG. 4, the peaks of EP and CC were confirmed, and the results of the 2-OH, 3-OH, 6-H, and the like were confirmed. As a result, Nos. 2, 3, and 6 of glucopyranose were identified. It was found that OH bonded to carbon at position was substituted (modified).

Example 1-3. Multilayer Produce

The polymer multilayer thin film was carried out by the LBL method using the solution prepared in Example 1-1 in a concentration of 0.01M in the deionized water. More specifically, the manufacturing process of the polymer multilayer film is as follows.

The PAH aqueous solution, the PMA aqueous solution, and the polymer of the gamma-cyclodextrin derivative prepared in Example 1-2, the polymer of the PAH, PMA and the gamma-cyclodextrin derivative prepared in Example 1-2 were added to the deionized water. It was used to make a concentration of M, the pH of the aqueous solution was adjusted using 0.1M HCl aqueous solution or 0.1M NaOH aqueous solution. The aqueous PAH solution, which is the cationic polymer electrolyte solution, and the aqueous polymer solution of the gamma-cyclodextrin derivatives prepared in Examples 1-2 were selected and used according to the production of a multilayer film of a desired form.

The substrate used in the LBL method was a glass or silicon substrate (slide glass or silicon wafer). The substrate was put together with a detergent diluted in a washing bottle, washed by ultrasonication (Ultrasonication) for 15 minutes, and then rinsed three times with the deionized water. The substrate on which the rinsing process was performed was dried using nitrogen gas.

The dried substrate was immersed in a pH-controlled cationic electrolyte (PAH) aqueous solution for 20 minutes and then taken out, followed by rinsing twice for 2 minutes using the deionized water. After the rinsing process, the solution was immersed in an anionic electrolyte (PMA) solution for 15 minutes and then taken out, and the rinsing process was performed using the deionized water in the same manner as described above. The thin film produced by the above process was called 1 bilayer, and the above process was repeated in order to increase the number of bilayers by a desired thickness.

Through the above process, a multilayer thin film of a desired thickness was prepared, and then dried in the same manner as above using nitrogen gas.

Claims

Gamma-cyclodextrin derivatives prepared by reacting gamma-cyclodextrin, epichlorohydrin and choline chloride.

The method of claim 1,
Gamma-cyclodextrin, epichlorohydrin and choline chloride content of the gamma-cyclodextrin derivative is 1: 12 to 17: 0.5 to 2 based on the molar ratio.

The method of claim 1,
The gamma-cyclodextrin derivative is a gamma-cyclodextrin derivative having 8 glucopyranose units having a structural formula of Formula 1 to which α- (1,4) is bound, and the gamma-cyclodextrin derivative is a structural formula of Formula 1 Wherein at least one of the glucopyranose units having a substituent includes a substituent of Formula 2, wherein X of the substituent of Formula 2 in the gamma-cyclodextrin derivative is at least one gamma-cyclodextrin derivative of Formula 3 .
[Formula 1]

R 1, R 2, and R 3 are each independently any one selected from the group consisting of hydrogen and a substituent of Formula 2 below.
[Formula 2]

X is hydrogen or a substituent of formula (3).
(3)

The method of claim 3,
The gamma-cyclodextrin derivative is a gamma-cyclodextrin derivative in which eight glucopyranose units having a structural formula of Formula 4 are α- (1,4) bonds, and a glucopyranose unit having a structural formula of Formula 4 At least one of the gamma-cyclodextrin derivative comprising a substituent of the formula (5).
[Chemical Formula 4]

R2 and R3 are each independently any one selected from the group consisting of hydrogen and a substituent of the formula (5)
[Chemical Formula 5]

A polymer comprising as a monomer a gamma-cyclodextrin derivative according to any one of claims 1 to 4.

The method of claim 5,
R1, R2 and R3 included in the gamma-cyclodextrin derivative of claim 3 are 10% to 30% of the total R1, R2 and R3 are hydrogen, and 70% to 90% of the total R1, R2 and R3 are the substituents of the formula (2). Phosphorus polymer.

The method of claim 5,
R 1, R 2 and R 3 included in the gamma-cyclodextrin derivative of claim 3 wherein 20% to 40% of the total R 1, R 2 and R 3 comprises a substituent of Formula 3.

The method of claim 5,
The polymer is a polymer in which the gamma-cyclodextrin derivative of claim 3 and the gamma-cyclodextrin are polymerized with monomers.

The gamma-cyclodextrin derivative according to any one of claims 1 to 4 and the gamma-cyclodextrin according to any one of claims 1 to 4, wherein any one or both layers of the outermost layer of the alternating laminated structure of the anionic polymer electrolyte layer and the cationic polymer electrolyte layer A multilayer film comprising one or more selected from the group consisting of polymers comprising derivatives as monomers.

10. The method of claim 9,
Any one or both layers of the outermost layer is a multilayer film made of a polymer comprising the gamma-cyclodextrin derivative of claim 5.

An anionic polymer electrolyte layer and a cationic polymer electrolyte layer have an alternating stacked structure, wherein the cationic polymer electrolyte layer is polyallylamine hydrochloride, poly acrylamide, and gamma according to any one of claims 1 to 4 The cyclodextrin derivatives and the gamma-cyclodextrin derivatives of claim 5 consisting of at least one member selected from the group consisting of a polymer comprising a monomer, wherein at least one of the cationic polymer electrolyte layer of claim 1 to claim 4 A multilayer film comprising at least one selected from the group consisting of a gamma-cyclodextrin derivative according to any one of claims and a polymer comprising the gamma-cyclodextrin derivative according to claim 5 as a monomer.

The method of claim 11,
The cationic polymer electrolyte layer is at least one selected from the group consisting of polyallylamine hydrochloride, poly acrylamide and a polymer comprising the gamma-cyclodextrin derivative of claim 5 as a monomer, and the outermost layer of the alternating layered structure A multilayer film, wherein any one or both layers are made of a polymer comprising the gamma-cyclodextrin derivative of claim 5 as a monomer.