KR101512759B1 - Polyethyleneglycol/polyester block copolymers with ionic functional group in side chain or chain-end, and method for preparing the same - Google Patents

Abstract

The present invention relates to a thermosensitive polyethylene glycol / polyester block copolymer having an ionic functional group introduced into a side chain or a terminal thereof, and more particularly to a thermosensitive polyethylene glycol / polyester block copolymer having a caprolactone segment or a caprolactone segment and a lactide segment The present invention relates to a block copolymer in which a cationic, anionic or amphoteric ion is introduced into a polyethylene glycol / polyester block copolymer having a hydrophobic moiety, and a method for producing the same. The thermosensitive polyethylene glycol / polyester block copolymer having an ionic functional group introduced into the side chain or terminal thereof according to the present invention may be used in various fields such as a drug carrier, a gene carrier, and a support for tissue engineering by having various ionic functional groups.

Description

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a thermosensitive polyethylene glycol / polyester block copolymer having an ionic functional group introduced into a side chain or a terminal thereof, and a method for preparing the same. 2. Description of the Related Art Polyethylene glycol / polyester block copolymers,

The present invention relates to a thermosensitive polyethylene glycol / polyester block copolymer having an ionic functional group introduced into a side chain or a terminal thereof, and more particularly to a thermosensitive polyethylene glycol / polyester block copolymer having a cation, an anion or Zwitter ion The present invention relates to thermosensitive polyethyleneglycol / polyester block copolymers and methods for their preparation.

In the case of thermosensitive hydrogel, there are many studies for application to drug delivery system and tissue engineering using the physicochemical properties of polymer responding to external stimulus. In order to apply the thermosensitive hydrogel to injectable drug delivery system and tissue engineering, it should have low viscosity, fast gel formation and biodegradability, and low molecular weight to be easily discharged to the outside of the body. It must also be biocompatible for use in biomaterials and should be free of damage to cells or human metabolic organs during degradation or excretion.

Hydrophilic and hydrophobic parts of a polymer are predominantly hydrogen bonds between hydrophilic groups and water molecules at low temperatures and dissolve in water to form a sol state. However, when the temperature is increased, the bonding power of the hydrophobic part of the polymer becomes higher than that of hydrogen bonding The hydrophobic part of the polymer coagulates and phase transition occurs in gel state. Therefore, the increase of the hydrophobic moiety in the polymer lowers the LCST (lower critical solution temperature), and the LCST is changed by controlling the hydrophilic and hydrophobic molecular chains. When such a polymer solution flows as a fluid in a sol state at a normal temperature, it is possible to simply mix the drug and the polymer hydrogel becomes a gel when heat above the body temperature is applied. In this case, the drug exhibits a sustained release behavior from the gel do. When the polymer is crosslinked, it shows swelling and shrinkage behavior, but when it is not crosslinked, it shows sol - gel phase transition behavior. Poly (N-isopropylacrylamide) is the most widely used because it has LCST near body temperature. Copolymers of butyl methacrylate, polyethylene glycol, polypropylene glycol, etc., . Copolymers of polyethylene oxide-polypropylene oxide (PEO-PPO) are also polymers that show a change in sol-gel properties. Many copolymers are available under the trade names Pluronic, Poloxamers, Tetronic, [U.S. Patent No. 4,188,373].

On the other hand, the sol-gel polymer is disadvantageous in that it is released into the body by metabolism of the human body after it is used in the human body, so that a polylactide-polyglycolide copolymer (PLGA) is introduced as a biodegradable polymer chain A number of sol-gel polymers have also been reported [U.S. Patent Nos. 4,882,168, 4,716,203, 4,942,035, 5,476,909, 5,548,035].

In order to solve this problem, a biocompatible thermosensitive polyethylene glycol is polymerized with caprolactone, lactide, paradoxanone, and trimethylene carbonate, which are biodegradable esters, at a specific ratio, Studies on block copolymers having gel phase transition behavior have been carried out [M. S. Kim, H. Hyun, GS. Khang et al., Macromolecules, 39, 3099-3102 (2006)].

The above-mentioned studies are based on a biodegradable ester-based polymer having high solubility in water and an organic solvent, non-toxic, non-reactive to immune action, hydrophobic, and chemically bonded to form a water- It uses polyethylene glycol which can control the decomposition period by increasing the inflow. It is decomposed into biological metabolites through dissolution and chemical hydrolysis in the human body, and discharged to the outside of the body. Thus, the biodegradable block copolymer is released .

However, since the conventional thermosensitive gel does not have an ionic functional group, its application range is limited when it is used for medical use. Particularly, the biodegradable polyesters so far are limited in their ability to achieve stable drug loading and sustained drug release due to the absence of ionic properties when used as medical materials. As a result of their hydrophobicity, adsorption of proteins in the body to polymers, Drug degeneration in the medium, and side effects due to local accumulation of hydrolysis products in the body.

Accordingly, the present inventors have made efforts to overcome the problems of the prior art as described above, and as a result, they have found that by introducing an ionic functional group into a polyethylene glycol / polyester block copolymer, the thermosensitive behavior of the hydrogel can be controlled, And thus a stable loading of the drug can be realized. As a result, the present invention has been completed.

Accordingly, the present invention is to provide a thermosensitive polyethylene glycol / polyester block copolymer having an ionic functional group introduced into a side chain or a terminal thereof, and a process for producing the same.

The present invention also provides a drug carrier, gene carrier, or tissue engineering support comprising the polyethylene glycol / polyester block copolymer.

The present invention provides a thermosensitive polyethylene glycol / polyester block copolymer having ionic functional groups introduced into the side chain or terminal and a process for preparing the same.

The present invention also provides a drug carrier, gene carrier, or tissue engineering support comprising the polyethylene glycol / polyester block copolymer.

The thermosensitive polyethylene glycol / polyester block copolymer according to the present invention has a variety of ionic functional groups at the side chain or at the terminal thereof, so that the thermosensitive behavior can be easily controlled. When the thermosensitive polyethylene glycol / polyester block copolymer is used as a thermosensitive drug carrier, Or a biologically active ingredient, and thus can be used as an injectable drug delivery system, and can be implemented as a sustained release preparation by inducing interaction between an ionic functional group and a drug. In addition, since it has biodegradability and biocompatibility, it acts as a matrix for controlling drug diffusion and has an advantage in that it can be decomposed by hydrolysis in the human body to regulate the release behavior and rate of drug, and the block copolymer has a positive charge Can be used as a gene carrier by forming a complex with a gene using the characteristic of the gene. In addition, in application to tissue engineering, it can be used as a support for providing a place where cells can be adhered and grown using various types of substrates for cell and tissue culture in the body or in vitro by introducing peptides into the side chain.

Accordingly, the temperature responsive adjustable polyethylene glycol / polyester block copolymer having various biocompatible and biodegradable ionic functional groups according to the present invention is useful as a drug delivery vehicle containing a drug, a gene carrier containing the gene, It can be applied in various ways as a support.

BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a schematic diagram showing a polymer in which cation, anion or zwitter ion is introduced into a side chain or a terminal.
2 is a graph showing the ¹ H-NMR spectrum of the methoxypolyethylene glycol-polycaprolactone copolymer produced in Production Example 1-1.
3 is a graph showing the ¹ H-NMR spectrum of the methoxypolyethylene glycol- (polycaprolactone-co-polylactide) block copolymer produced by Production Example 1-2.
4 is a graph showing a ¹ H-NMR spectrum of a methoxypolyethylene glycol-polycaprolactone copolymer having a terminal functional group and having a carboxyl group prepared in Example 1-1.
FIG. 5 is a graph showing the ¹ H-NMR spectrum of the methoxy polyethylene glycol-polycaprolactone copolymer having an amine functional group prepared according to Example 1-3. FIG.
FIG. 6 is a ¹ H-NMR spectrum of a methoxypolyethylene glycol-polycaprolactone copolymer having amphoteric ion as an end functional group, prepared by Example 1-5. FIG.
7 is a graph showing a ¹ H-NMR spectrum of a methoxypolyethylene glycol- (polycaprolactone-co-polylactide) block copolymer having an amphoteric ion with an end functional group prepared by Example 1-6 .
8 is a graph showing sol-gel phase transition behavior of the block copolymer of the present invention.
9 is a graph showing a change in viscosity value of a block copolymer according to the present invention as a function of temperature.
10 is a view for confirming the formation of a complex with a gene according to a change in the N / P ratio of the block copolymer of the present invention.

The present invention relates to a hydrophilic part composed of polyethylene glycol; And a polyester-based hydrophobic moiety containing a caprolactone segment having an ionic functional group introduced into the side chain or the terminal thereof, and a thermosensitive polyethyleneglycol / polyester block copolymer.

The present invention also relates to a composition comprising a hydrophilic moiety composed of polyethylene glycol; And a polyester-based hydrophobic portion containing a caprolactone segment and a lactide segment to which an ionic functional group is introduced at a side chain or at a terminal thereof. The present invention also provides a thermosensitive polyethylene glycol / polyester block copolymer.

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

The polyethylene glycol (PEG) used as a polymerization initiator of the polyethylene glycol / polyester block copolymer according to the present invention has many merits in the field of drug delivery and tissue engineering, so that it can easily contain and release drugs as a drug delivery vehicle. It has high solubility in solvents, is non-toxic, has no rejection reaction to immunity, exhibits excellent biocompatibility and has been approved for use in the human body by the US Food and Drug Administration. In addition, since PEG has the greatest inhibitory effect on protein adsorption among hydrophilic polymers and improves the biocompatibility of blood contact materials, many applications as biomaterials have been made.

In addition, ester-based biodegradable polymers have the advantage of controlling the decomposition period by controlling molecular weight and chemical composition. Polyethylene glycol (PEG) and polycaprolactone (PCL) -co-polylactide (PLLA) block copolymers, which are basic models in the present invention, are thermosensitive copolymers having sol- It is applied as a biomaterial in the field. The copolymer contained lactide to lower the crystallinity and control the biodegradation period. The present invention is characterized in that biodegradation period and temperature responsive behavior can be controlled by introducing various ionic functional groups to the side chain or terminal.

When the hydrophobic portion of the block copolymer according to the present invention simultaneously contains a caprolactone segment and a lactide segment, it can be represented by the following Formula 1, and the molar ratio of the caprolactone segment and the lactide segment constituting the same is 100: 0.01 To 95: 5.

[Chemical Formula 1]

In Formula 1, m and n are each independently an integer of 1 to 32.

The thermosensitive polyethylene glycol / polyester block copolymer according to the present invention preferably has a molecular weight of 2,400 to 5,000 g / mole. The proportion of hydrophobic groups can be controlled by adjusting the molecular weight of the polyester. When the molecular weight of the polyester is less than 2,000 g / mole, there is a problem that the sol-gel phase transition behavior does not occur in the human body temperature range for the purpose of use of the present invention and the polyester is kept in a sol state. , There is a problem that the molecular weight is large and biodegradation takes a long time, so it is preferable to maintain the above range.

In addition, the thermosensitive polyethylene glycol / polyester block copolymer according to the present invention is a gel phase having a flow characteristic at room temperature or a gel phase at 30 to 60 ° C. When the temperature is above the critical temperature, Shows a new sol - gel phase transition behavior.

The thermosensitive polyethylene glycol / polyester block copolymer according to the present invention can control the temperature responsive behavior by introducing various ionic functional groups to the side chain or the terminal. When used as a drug delivery vehicle, the temperature sensitive polyethylene glycol / And can be applied as an injectable drug delivery system. The ionic functional group may be a cation, an anion, zwitter ion, and the cationic functional group is preferably an amine group, but not limited thereto. The anionic functional group is preferably a carboxyl group but is not limited thereto.

In addition,

(a) drying polyethyleneglycol through azeotropic distillation;

(b) adding caprolactone or caprolactone and lactide monomer to the polyethylene glycol and conducting polymerization to prepare a polyethylene glycol / polyester block copolymer;

(c) reacting the block copolymer with a compound having an ionic functional group to prepare a polyethylene glycol / polyester block copolymer having an ionic functional group introduced into the side chain or terminal of the block copolymer. A method for producing a glycol / polyester block copolymer is provided.

The process for producing the block copolymer of the present invention will be described in detail as follows.

The step (a) is a step of obtaining dried polyethylene glycol. The polyethyleneglycol as an initiator and an organic solvent are placed in a well-dried flask and subjected to azeotropic distillation using a Deanstock trap, followed by distillation to remove the solvent.

The step (b) is a step of preparing a polyethylene glycol / polyester block copolymer, wherein the purified caprolactone or caprolactone and lactide are added to the polyethylene glycol obtained in the step (a), an organic solvent is added, HCl is added as a catalyst and stirred. A typical production process of a polyethylene glycol / polyester block copolymer can be represented by the following reaction formula 1 or 2.

[Reaction Scheme 1]

In the above Reaction Scheme 1, x and m are each independently an integer of 1 to 32.

[Reaction Scheme 2]

In the above Reaction Scheme 2, x, m and n are each independently an integer of 1 to 32.

Reaction 1 is a method for preparing a block copolymer comprising a hydrophilic moiety composed of polyethylene glycol and a polyester hydrophobic moiety composed of caprolactone segment, and a reaction formula 2 is a method for producing a block copolymer comprising a hydrophilic moiety composed of polyethylene glycol and a hydrophilic moiety composed of caprolactone segment and lactide segment. Ester-based hydrophobic moiety of the block copolymer.

The polyethylene glycol / polyester block copolymer thus prepared is capable of controlling the viscosity and biodegradation period as well as controlling the temperature responsive behavior according to the hydrophobicity of the functional group introduced into the side chain.

The step (c) is a step of preparing a polyethylene glycol / polyester block copolymer having an ionic functional group introduced therein. The polyethylene glycol / polyester block copolymer obtained in the step (b) and the organic solvent are dissolved in a well- And azeotropic distillation is carried out using Dean Stark trap. After distillation, the solvent is removed and the block copolymer is reacted with a compound having an ionic functional group to introduce an ionic functional group to the side chain or terminal of the block copolymer.

When the ionic functional group is an anionic functional group, a typical production process of a polyethylene glycol / polyester block copolymer into which an anionic functional group is introduced is represented by the following Reaction Schemes 3-1 or 3-2. Examples of the anionic functional group include a carboxyl group, a sulfate group, a phosphoric acid group, and the like. Examples of the compound having an anionic functional group include glutaric anhydride, acetic acid, 2,7-oxepanedione, dihydro-furan-2,5-dione ( Dihydro-furan-2,5-dione, oxepane-2,7-dione, succinic anhydride, maleic anhydride, citraconic anhydride citraconic anhydride, diethylenetriamine pentaacetic anhydride, 1,4,7-triazacyclononane-N, N ', N' '- triacetic acid, 1,4,7,10-tetraazacyclododecane-N, N ', N' ', N' '' - tetraacacyclododecane- Tetraacetic acid), 1,4,8,11-tetraazacyclotetradodecane-N, N ', N' ', N' '' - tetraacetic acid (1,4,8 , 11-tetraazacyclotetradecane-N, N ', N' ', N' '' - tetraacetic acid, N- (p-isothiocyanatobenzyl) - (p-isothiocyanatobe nzyl) -diethylene-triamine-N, N ', N ", N"' - tetraacetic acid, iodoacetate, chloroacetic acid and the like.

[Reaction Scheme 3-1]

In the above Reaction Scheme 3-1, x and m are each independently an integer of 1 to 32.

[Reaction Scheme 3-2]

In the above Reaction Scheme 3-2, x, m and n are each independently an integer of 1 to 32.

When the ionic functional group is a cationic functional group, a typical production process of a polyethylene glycol / polyester block copolymer into which a cationic functional group is introduced is represented by the following reaction formula 4-1 or 4-2. Specifically, the method further comprises the step of introducing a cationic functional group into the block copolymer into which the anionic functional group is introduced. That is, the obtained polyethylene glycol / polyester block copolymer having anionic functional groups introduced therein and an organic solvent are placed in a well-dried flask and subjected to azeotropic distillation using a Deanstock trap. After the distillation, the solvent is removed, and then the block copolymer into which the anionic functional group is introduced is reacted with the compound having the cationic functional group to introduce the cationic functional group into the side chain or terminal of the block copolymer. The cationic functional group includes, but is not limited to, an amine group and the like. Examples of the compound having a cationic functional group include dimethylamino propylamine, 2-dimethylaminoethylamine, (4-dimethylaminomethyl-benzyl) -methyl-amine ((4-Dimethylaminomethyl-benzyl ) -methyl amine), and the like.

[Reaction Scheme 4-1]

In the above Reaction Scheme 4-1, x and m are each independently an integer of 1 to 32.

[Reaction Scheme 4-2]

In the above Reaction Scheme 4-2, x, m and n are each independently an integer of 1 to 32.

When the ionic functional group is an amphoteric ionic functional group, a typical production process of a poly (ethylene glycol) / polyester block copolymer into which an amphoteric ion functional group is introduced is represented by the following reaction formula 5-1 or 5-2. Specifically, the method further comprises the step of introducing an amphoteric ionic functional group into the copolymer to which the cationic functional group has been introduced. The resulting polyethylene glycol / polyester block copolymer with the cationic functional group introduced therein and an organic solvent are placed in a well-dried flask and azeotropic distillation is carried out using a Deanstock trap. After the distillation, the solvent is removed and then the cationic functional group-introduced block copolymer is reacted with the anionic functional group-containing compound to introduce the amphoteric functional group into the side chain or terminal of the block copolymer. Examples of the anionic functional group include a carboxyl group, a sulfate group, a phosphoric acid group, and the like. Examples of the compound having an anionic functional group include 1,3-propane sultone, 1,4-butane sultone, 1-propene-1,3-sultone 1-Propene-1,3-sultone, 1,8-naphthalene sultone, and the like.

[Reaction Scheme 5-1]

In the above Reaction Scheme 5-1, x and m are each independently an integer of 1 to 32.

[Reaction Scheme 5-2]

In the above Reaction Scheme 5-2, x, m and n are each independently an integer of 1 to 32.

The organic solvent used in the above Reaction Schemes 1 to 5 can be selected from the group consisting of methylene chloride, toluene, tetrahydrofuran (THF), dioxane, acetone, methanol, dimethylacetamide (DMAc), ethyl acetate, dimethylformamide (DMF) (DMSO), chloroform, and the like.

The present invention also provides a drug carrier or a tissue engineering support comprising a polyethylene glycol / polyester block copolymer having an ionic functional group introduced into the side chain or the terminal.

In order to apply the block copolymer of the present invention to an injectable drug delivery system or a tissue engineering support, it should have a low viscosity, a fast gel-forming property, and a low molecular weight for easy excretion to the outside of the body.

The block copolymer according to the present invention can control the temperature responsive behavior and can obtain a molecular weight close to the expected value, so that it can maintain low viscosity and low molecular weight, which are one of the requirements of biocompatible and thermosensitive hydrogels, And hydrogels, the interaction between the ionic functional group and the drug can be induced to realize stable loading and sustained release of the drug. In addition, when the peptide is used as a support for tissue engineering, Affinity can also be increased.

Hereinafter, preferred embodiments of the present invention will be described in order to facilitate understanding of the present invention. However, the following examples are only for the purpose of easier understanding of the present invention, and the present invention is not limited by the examples.

Preparation Example 1: Preparation of polyethylene glycol / polyester block copolymer

1-1: Preparation of Methoxypolyethylene Glycol (MPEG) -Polycaprolactone (PCL) Block Copolymer

To prepare a methoxypolyethylene glycol (MPEG) -polycaprolactone (PCL) block copolymer having a molecular weight of 3,550 g / mole, an initiator, methoxypolyethylene glycol (1 g, 1.3 mmol, Mn = 750 g / mole) and 80 ml of toluene Was placed in a well dried 100 ml round bottom flask and azeotropic distillation was carried out at 140 < 0 > C for 5 hours using a Dean Stark trap. After distillation, 80 ml of toluene was removed, methoxypolyethylene glycol (MPEG) was cooled to 25 ° C, and then preliminarily purified caprolactone (CL) (3.73 g, 32.68 mmol) was added and 28 ml of preliminarily purified methylene chloride , 2.67 ml of HCl as a polymerization catalyst was added, and the mixture was stirred at 25 캜 for 24 hours. All procedures were carried out under high purity nitrogen. After the reaction, the reaction was precipitated by slowly dropping the reaction into 800 ml of hexane to remove unreacted monomers and initiator. The precipitate was dissolved in methylene chloride (MC), filtered with filter paper, the solvent was removed through a rotary evaporator, and dried under reduced pressure to obtain the title compound. A schematic diagram of the title compound is shown in FIG. 1, and its ¹ H-NMR is shown in FIG.

1-2: Preparation of methoxypolyethylene glycol- (polycaprolactone-co-polylactide) block copolymer [MPEG- (PCL-co-PLLA)] (caprolactone: lactide = 95: 5)

To prepare a methoxypolyethylene glycol (MPEG) -polycaprolactone (PCL) -co-polylactide (PLLA) block copolymer having a molecular weight of 3,750 g / mole, methoxypolyethylene glycol (1 g, 1.3 mmol, Mn = 750 g / mole) and 80 ml of toluene were placed in a well-dried 100 ml round-bottomed flask and subjected to azeotropic distillation at 130 ° C for 3 hours using a Dean Stark trap. After distillation, 40 ml of toluene was removed, methoxypolyethylene glycol (MPEG) was cooled to 25 ° C, and then preliminarily purified caprolactone (CL) (3.75 g, 32.85 mmol) and lactide (LA) (0.25 g, 1.73 mmol) 40 ml of preliminarily purified toluene was added as a reaction solvent, 1.6 ml of Sn (Oct) 2 was added as a polymerization catalyst, and the mixture was stirred at 130 ° C for 24 hours. All procedures were carried out under high purity nitrogen. After the reaction, the reaction product was slowly dropped into 800 ml of hexane and 200 ml of ether to remove unreacted monomers and initiator. The precipitate was dissolved in methylene chloride (MC), filtered with filter paper, the solvent was removed through a rotary evaporator, and dried under reduced pressure to obtain the title compound. A schematic diagram of the title compound is shown in Fig. 1, and its ¹ H-NMR is shown in Fig.

Example 1 Preparation of Polyethylene Glycol / Polyester Block Copolymer Having Ionic Functional Group

1-1: Preparation of Methoxy Polyethylene Glycol (MPEG) -Polycaprolactone (PCL) Block Copolymer Having Carboxyl Group as End Functional Group

(MPEG) -polycaprolactone (PCL) block copolymer prepared in Preparation Example 1-1 to replace the terminal functional group of the methoxypolyethylene glycol (MPEG) -polycaprolactone (PCL) block copolymer with the methoxypolyethylene glycol (2 g, 0.56 mmol, Mn = 3,550 g / mole) and 75 ml of toluene were placed in a well-dried 100 ml round flask and azeotropic distillation was carried out at 140 ° C. for 5 hours using a Dean Stark trap. After distillation, 60 ml of toluene was removed, and anhydrous glutaric anhydride (0.19 g, 1.67 mmol, Mn = 114.1 g / mole) and acetic acid (1.5 ml) were added and reacted at 120 ° C for 24 hours. All procedures were carried out under high purity nitrogen. After the reaction, the reaction product was slowly dropped into 800 ml of hexane and 200 ml of ether to remove unreacted monomers and initiator. The precipitate was dissolved in methylene chloride (MC), filtered with filter paper, the solvent was removed through a rotary evaporator, and dried under reduced pressure to obtain the title compound. A schematic diagram of the title compound is shown in Fig. 1, and its ¹ H-NMR is shown in Fig.

1-2: Preparation of methoxypolyethylene glycol (MPEG) - (polycaprolactone-co-polylactide) (PCLA) block copolymer having a carboxyl group as a terminal functional group

(MPEG) -Polyethylene glycol (PC) block copolymer prepared in Preparation Example 1-2 was used in order to replace the terminal functional group of methoxypolyethylene glycol (MPEG) - (polycaprolactone-co-polylactide) (PCLA) Caprolactone (PCL) -co-polylactide (PLLA) block copolymer (2 g) and 75 ml toluene were placed in a well-dried 100 ml round bottom flask and azeotropic distillation was carried out at 140 ° C for 5 hours using a Deanstock trap . After distillation, 60 ml of toluene was removed, and anhydrous glutaric anhydride (0.19 g, 1.67 mmol, Mn = 114.1 g / mole) and acetic acid (1.5 ml) were added and reacted at 120 ° C for 24 hours. All procedures were carried out under high purity nitrogen. After the reaction, the reaction product was gradually dropped into 800 ml of hexane and 200 ml of ether to remove unreacted monomers and initiator. The precipitate was dissolved in methylene chloride (MC), filtered with filter paper, the solvent was removed through a rotary evaporator, and dried under reduced pressure to obtain the title compound. A schematic diagram of the title compound is shown in Fig.

1-3: Preparation of methoxypolyethylene glycol (MPEG) -polycaprolactone (PCL) block copolymer having an amine group as a terminal functional group

Methoxypolyethylene glycol (MPEG) -polycaprolactone (PCL) block copolymer (2 g, Mn = 3679.1 g / mole) having the carboxyl group as a terminal functional group prepared in Example 1-1 and 20 ml of methylene chloride In a dried 100 ml round bottom flask and stirred. After dissolving the copolymer, N, N'-Dicyclohexylcarbodiimide (0.17 g, 0.82 mmol) and N-Hydroxysuccinimide (0.094 g, 0.82 mmol) mmol) were added and reacted at room temperature for 24 hours. After the reaction was completed, the precipitate was removed using a filter. The filtered solution was removed through a rotary evaporator and dried under reduced pressure. The block copolymer obtained after drying and 75 ml of toluene were placed in a well dried 100 ml round-bottomed flask and subjected to azeotropic distillation at 140 ° C for 5 hours using a Dean Stark trap. After distillation, 60 ml of toluene was removed, and dimethylaminopropylamine (0.10 ml, 0.82 mmol) was added thereto, followed by reaction at 110 ° C for 24 hours. After the reaction, the reaction product was slowly dropped into 800 ml of hexane and 200 ml of ether to remove unreacted monomers and initiator. The precipitate was dissolved in methylene chloride (MC), filtered with filter paper, the solvent was removed through a rotary evaporator, and dried under reduced pressure to obtain the title compound. A schematic diagram of the title compound is shown in Fig. 1, and its ¹ H-NMR is shown in Fig.

1-4: Preparation of methoxypolyethylene glycol (MPEG) - (polycaprolactone-co-polylactide) (PCLA) block copolymer having an amine group as a terminal functional group

(MPEG) - (polycaprolactone-co-polylactide) (PCLA) block copolymer (2 g) having a carboxyl group as a terminal functional group prepared in Example 1-2 and 20 ml of methylene chloride (MC) In a well-dried 100 ml round bottom flask and stirred. After dissolving the copolymer, N, N'-Dicyclohexylcarbodiimide (0.17 g, 0.82 mmol) and N-Hydroxysuccinimide (0.094 g, 0.82 mmol) mmol) were added and reacted at room temperature for 24 hours. After the reaction was completed, the precipitate was removed using a filter. The filtered solution was removed through a rotary evaporator and dried under reduced pressure. The block copolymer obtained after drying and 75 ml of toluene were placed in a well dried 100 ml round-bottomed flask and subjected to azeotropic distillation at 140 ° C for 5 hours using a Dean Stark trap. After distillation, 60 ml of toluene was removed, and dimethylaminopropylamine (0.10 ml, 0.82 mmol) was added thereto, followed by reaction at 110 ° C for 24 hours. After the reaction, the reaction product was slowly dropped into 800 ml of hexane and 200 ml of ether to remove unreacted monomers and initiator. The precipitate was dissolved in methylene chloride (MC), filtered through filter paper, the solvent was removed through a rotary evaporator, and dried under reduced pressure to obtain the title compound. A schematic diagram of the title compound is shown in Fig.

1-5: Preparation of methoxypolyethylene glycol (MPEG) -polycaprolactone (PCL) block copolymer having an amphoteric ion as a terminal functional group

Methoxypolyethylene glycol (MPEG) -polycaprolactone (PCL) block copolymer (2g, Mn = 3679.1 g / mole) having an amine group as an end group functional group prepared in Example 1-3 and 75 ml of toluene were added to a well- The mixture was placed in a flask and subjected to azeotropic distillation at 140 ° C for 5 hours using a Dean Stark trap. After distillation, 60 ml of toluene was removed, and 1,3-propane sultone (0.072 g, 0.82 mmol) was added thereto, followed by reaction at 90 ° C for 24 hours. After the reaction, the reaction product was slowly dropped into 800 ml of hexane and 200 ml of ether and 20 ml of methanol to remove unreacted monomers and initiator. The precipitate was dissolved in methylene chloride (MC), filtered with filter paper, the solvent was removed through a rotary evaporator, and dried under reduced pressure to obtain the title compound. A schematic diagram of the title compound is shown in Fig. 1, and its ¹ H-NMR is shown in Fig.

1-6: Preparation of methoxypolyethylene glycol (MPEG) - (polycaprolactone-co-polylactide) (PCLA) block copolymer having an amphoteric ion as a terminal functional group

(MPEG) - (polycaprolactone-co-polylactide) (PCLA) block copolymer (2 g, Mn = 3679.1 g / mole) having an amine group as an end functional group prepared in Example 1-4 and a methoxypolyethylene glycol 75 ml of toluene was placed in a well dried 100 ml round bottom flask and azeotropic distillation was carried out at 140 ° C for 5 hours using a Dean Stark trap. After distillation, 60 ml of toluene was removed, and 1,3-propane sultone (0.072 g, 0.82 mmol) was added thereto, followed by reaction at 90 ° C for 24 hours. After the reaction, the reaction product was slowly dropped into 800 ml of hexane and 200 ml of ether and 20 ml of methanol to remove unreacted monomers and initiator. The precipitate was dissolved in methylene chloride (MC), filtered with filter paper, the solvent was removed through a rotary evaporator, and dried under reduced pressure to obtain the title compound. A schematic diagram of the title compound is shown in Fig. 1, and its ¹ H-NMR is shown in Fig.

Experimental Example 1: Confirmation of introduction of ionic functional groups of MPEG-PCL block copolymer and MPEG-PCLA block copolymer

¹ H-NMR spectroscopy, gel permeation chromatography and DSC analysis of the polyethylene glycol / polyester block copolymer prepared in Preparation Example 1 and the polyethylene glycol / polyester block copolymer to which the ionic functional group introduced in Example 1 was introduced 2 to 7 and Table 1 show the results of the measurement using the above-

M _n , NMR
MPEG-polyester M _n , NMR
MPEG-polyester Yield (%) M _w / M _n T _m H (J / g) MPEG-PCL 2400 750-2400 750-2420 96 1.21 52.76 80.44 MPEG-PCL 2800 750-2800 750-2815 92 1.22 52.51 83.02 MPEG-PCL 3000 750-3000 750-3018 93 1.22 52.47 81.98 MPEG-PCLA 3000 750-2850-150 750-2862-161 95 1.31 52.11 84.03 MPEG-PCL-COOH 3000 750-2850-114 750-2815-114 95 1.28 51.37 83.07 MPEG-PCL-COOH 3000 750-3000-114 750-3018-114 98 1.25 51.88 79.68 MPEG-PCLA-COOH 3000 750-2850-150-114 750-2862-161-114 97 1.34 50.94 78.68 MPEG-PCL-Zwitter 2800 750-2 / 800-114-122 750-2815-114-122 74 1.44 48.07 66.77 MPEG-PCL-Zwitter 3000 750-3000-114-122 750-3018-114-122 62 1.47 50.04 45.74 MPEG-PCLA-Zwitter 3000 750-2850-150-114-122 750-2862-161-114-122 78 1.41 51.02 46.88

In Table 1, M _n is the result of measurement by ¹ H-NMR spectrum, M _w / M _n is the result of gel permeation chromatography based on standard polystyrene, and T _m is the result of DSC measurement.

Experimental Example 2: Sol-Gel Phase Transition Vial Tilt Method

The block copolymer prepared in Example 1 was dissolved in distilled water at a concentration of 20, 25, and 30 wt%, and the sol-gel phase transition behavior was confirmed by vial tilt method. The results are shown in FIG.

As shown in Fig. 8, the sol state was confirmed at 25 占 폚, and the gel state was confirmed at 37 占 폚.

Experimental Example 3: Preparation of methoxypolyethylene glycol (MPEG) -polycaprolactone (PCL) block copolymer having an amphoteric ion as a terminal functional group and methoxypolyethylene glycol- (polycaprolactone-co-polylactide) block copolymer Phase transition control

In order to observe the phase transition behavior of the methoxypolyethylene glycol-polycaprolactone and the methoxypolyethylene glycol- (polycaprolactone-co-polylactide) block copolymer having a bipolar ion as the terminal functional group, Was dissolved in distilled water at a concentration of 25 and 30% by weight, and then stored at 4 ° C for one day in order to maintain the equilibrium of the uniformly dispersed polymer. The prepared polymer solution was visually measured at 10 ° C Gel phase transition behavior was measured at each temperature while the spin rate was fixed at 0.2 rpm while the temperature was increased from 1 to 60 ° C by 1 ° C per 2 minutes. This is shown in Fig.

As shown in FIG. 9, it was confirmed that the sol-gel phase transition can be controlled through the molecular weight control (2800, 3000 g / mole) of the hydrophobic group.

Experimental Example 4: Measurement of charge of the block copolymer of the present invention

In order to compare the charge according to the type of the functional group of the copolymer prepared in Production Example 1 and Example 1, charge measurement was carried out. 1 mg of each copolymer was added to 5 ml vials, 1 ml of distilled water was added thereto, uniformly dispersed, and the charge was measured at 37 ° C.

The measurement results are shown in Table 2 below.

Mobility (cm ² / Vs) Zeta Potential
(mV) Electric Field
(V / cm) MPEG-PCL -2.535e-004 -26.72 -16.56 MPEG-PCL-COOH -2.595e-004 -27.26 -16.49 MPEG-Zwitterionic 3.732e-004 39.38 16.61

As shown in Table 2, it was confirmed that various values of mobility, zeta potential and electric field can be obtained depending on the type of ionic functional group.

Experimental Example 5: Confirmation of formation of a complex between the copolymer of the present invention and a gene

In order to confirm the complex formation between the copolymer prepared in Preparation Example 1 and Example 1 and the plasmid DNA, N / P ratio was prepared from 1 to 64 by the ratio. 181.6 mg of the prepared copolymer was dissolved in 1 ml of distilled water and 64 of N / P ratio was prepared on the basis of 1 μg of plasmid DNA to form a complex. A loading die (1 μl × 10, BlueJuice) was mixed with the sample. After performing experiments on 1.2% agarose gel containing 50 μg of EtBr, the image of the gel was observed using a UV imaging system. This is shown in Fig.

As shown in FIG. 10, it was confirmed that the complex was completely formed at the N / P ratio of 64, and that the complex was not formed in the PCL having negative charge. Based on these results, it was confirmed that the copolymer has a positive charge and can be used as a gene carrier by forming a complex with a gene having a negative charge.

Claims (19)

A hydrophilic moiety composed of polyethylene glycol; And
And a polyester-based hydrophobic portion containing a caprolactone segment having an anionic functional group or a zwitter ion functional group introduced into the side chain or the terminal thereof. The thermosensitive polyethyleneglycol / polyester block copolymer according to claim 1, A hydrophilic moiety composed of polyethylene glycol; And
A thermosensitive polyethylene glycol / polyester block copolymer, comprising a caprolactone segment and a polyester-based hydrophobic portion containing a lactide segment having an anionic functional group or a zwitter ion functional group introduced into the side chain or the terminal. The process of claim 2, wherein the hydrophobic moiety is represented by the following formula (1), and the molar ratio of the caprolactone segment to the lactide segment ranges from 100: 0.01 to 95: 5. coalescence.
[Chemical Formula 1]

In Formula (1), x and m each independently represent an integer of 1 to 32. The thermosensitive polyethylene glycol / polyester block copolymer according to claim 1 or 2, wherein the polyethylene glycol / polyester block copolymer has a molecular weight of 2,400 to 5,000 g / mole. The thermosensitive polyethylene glycol / polyester block copolymer according to claim 1 or 2, wherein the thermosensitive polyethylene glycol / polyester block copolymer is a sol phase at room temperature or a gel phase at 30 to 60 ° C. delete delete The thermosensitive polyethylene glycol / polyester block copolymer of claim 1, wherein the anionic functional group is a carboxyl group. (a) drying polyethyleneglycol through azeotropic distillation;
(b) adding caprolactone or caprolactone and lactide monomer to the polyethylene glycol and conducting polymerization to prepare a polyethylene glycol / polyester block copolymer; And
(c) a thermosensitive polyethylene glycol / polyester block copolymer having an anionic functional group or an amphoteric functional group introduced into the side chain or terminal of the block copolymer by reacting the block copolymer with an anionic functional group or a compound having an amphoteric functional group ≪ / RTI > wherein the thermally responsive polyethylene glycol / polyester block copolymer is prepared from a thermoplastic polyol. delete The method of claim 9, wherein, in step (c), when the ionic functional group is an anionic functional group, the block copolymer produced in step (b) is reacted with a compound having an anionic functional group to form an anionic functional group Wherein the thermosensitive poly (ethylene glycol) / polyester block copolymer is introduced. 12. The method of claim 11, wherein the compound having an anionic functional group is selected from the group consisting of glutaric anhydride, acetic acid, 2,7-oxepanedione, dihydro-furan- Dihydro-furan-2,5-dione, oxepane-2,7-dione, succinic anhydride, maleic anhydride, Citraconic anhydride, diethylenetriamine pentaacetic anhydride, 1,4,7-triazacyclononane-N, N ', N "-triacetic acid (7-triazacyclononane-N , N ', N' '- tetraacetic acid (1,4,7,10-tetraacacyclododecane-N, N' tetraacacyclododecane-N, N ', N' ', N' '' - tetraacetic acid, 1,4,8,11-tetraazacyclododecane- Tetraacetic acid (1,4,8,11-tetraazacyclotetradecane-N, N ', N ", N" "-tetraacetic acid), N- (p-isothiocyanatobenzyl) , N ", N '" -tetraacetic acid Characterized in that it is an acid (N- (p-isothiocyanatobenzyl) -diethylene-triamine-N, N ', N' ', N' '' - tetraacetic acid, iodoacetate or chloroacetic acid , A process for producing a thermosensitive polyethylene glycol / polyester block copolymer. delete delete The method according to claim 9, wherein, in the step (c), when the ionic functional group is an amphoteric ionic functional group, the block copolymer having the cationic functional group introduced therein is reacted with a compound having an anionic functional group, Lt; RTI ID = 0.0 > polyoxyalkylene < / RTI > glycol / polyester block copolymer. 16. The method of claim 15, wherein the compound having an anionic functional group is selected from the group consisting of 1,3-propane sultone, 1,4-butane sultone, , 1-Propene-1,3-sultone, or 1,8-naphthalene sultone. The method of claim 1, wherein the thermosensitive polyethylene glycol / polyester block copolymer is selected from the group consisting of 1-propene-1,3-sultone and 1,8-naphthalene sultone. A drug carrier comprising a polyethylene glycol / polyester block copolymer having an ionic functional group introduced therein according to claim 1 or 2. A gene carrier comprising a polyethylene glycol / polyester block copolymer having an ionic functional group introduced therein according to claim 1 or 2. A tissue engineering support comprising a polyethylene glycol / polyester block copolymer having the ionic functional groups of claim 1 or 2 incorporated therein.

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