KR20120035775A - Preparing of ph-sensitive polymersome - Google Patents

Abstract

PURPOSE: A manufacturing method of pH-sensitive polymersome is provided to accept hydrophobic and hydrophilic drug at the same time. CONSTITUTION: A manufacturing method of pH-sensitive polymersome comprises: a step of forming a poly(β-aminoester) by reacting a bisacrylate compound, and an amine based compound having hydroxy group; a step of forming hydroxyl group in the side chain of the poly (β - amino ester) compound; a step of polymerizing the (β - amino ester) compound, and the lactide; a step of copolymerizing the polyethylene glycol compound to the polymer.

Description

Manufacturing method of pH sensitive polymersome

The present invention relates to a method for producing PH-sensitive polymersomes, and more particularly to a method for producing a polymersome that can function as a receptor regardless of the nature of the drug in the human body.

In general, micelles (micelles) refers to a thermodynamically stable and uniform spherical structure formed by low molecular weight materials having amphiphilic, for example, hydrophilic and hydrophobic groups. Since the inside is composed of hydrophobic groups, when the hydrophobic drug is dissolved and injected, the drug is present in the inside of the micelle, so that the target-oriented drug release can be performed in response to changes in temperature or pH in the body, whereas in the case of hydrophilic drugs, the polymer micelle is used. I have felt the limitation in delivering using.

On the other hand, micelle aggregates obtained from amphiphilic block copolymers are of continuing interest due to various applications such as drug delivery, nanoreactors, and nanotemplates. Compared with micelles of traditional surfactants, polymeric self-assemblies have been in the spotlight recently for their advantages such as their excellent stability, toughness, and micellization depending on selective solvents.

Moreover, polymer self-assembly has many possibilities because it can control physical chemical and biological properties by conjugation with biomolecules or by changing copolymer chemical structure. The control of the morphology of the assembled structure from the block copolymer is of great practical value. The polymer micelle morphology and size can be designed with a choice of suitable properties and different block lengths.

The research on self-assembly of straight coil-coil and rod-coil block copolymers is progressing considerably. However, the introduction of dendritic structures as building blocks in the design of new block copolymers confers some interesting properties. The dendritic component consists of a well-organized branch structure and a well-organized internal part. Most studies deal with the formation of spherical micelles from straight-dendritic block copolymers. On the other hand, there are not many studies on morphologies such as cylinder micelles and polymersomes.

  Specifically, US Pat. No. 5955509, "pH dependent polymer micelles," shows that a block copolymer of poly (vinyl N-heterocycle) and poly (alkylene oxide) forms micelles at a pH of 6.0 or higher, and at a pH of 2-6. A method for preparing a collapsible pH-sensitive polymer micelle was described. Japanese Unexamined Patent Publication No. 2002-179556, "Block copolymer-anticancer complex pharmaceutical preparation," is a block of a hydrophilic polyethylene glycol compound and a hydrophobic polyamino acid compound. Copolymers are described as forming micelles at certain pH.

As such, there has been a demand for the development of a polymer material as a drug carrier which is not affected by hydrophobicity and hydrophilicity with micelles.

It is an object of the present invention to provide a method for preparing a polymersome which can accommodate both of them in delivering hydrophobic and hydrophilic drugs.

In another aspect, the present invention provides a method for producing a polymersome that can express sustained release in the human body.

In another aspect, the present invention is to provide a diagnostic and therapeutic composition using the polymersome.

The present invention comprises the steps of reacting a bisacrylate compound with an amine compound having a hydroxyl group to form a poly (β-amino ester) compound; Forming a hydroxyl group on the side chain of the poly (β-amino ester) compound; Polymerizing the poly (β-amino ester) and lactide having a hydroxyl group in the side chain; And copolymerizing a polyethylene glycol compound to the polymer.

The present invention also provides a method wherein the poly (ㅯ -amino ester) comprises a tertiary amine group which is ionized at a pH below 7.0.

In addition, the present invention forms a polymersome when the copolymer is in the range of pH 7.0 to 7.4 (preferably 7.2 to 7.4), the polymersome is collapsed when the pH is in the range of 6.0 to less than 7.0, the (unimer) It provides a way to be.

In the present invention, the bisacrylate is ethylene glycol diacrylate, 1,4-butane diol diacrylate, 1,3-butane diol diacrylate, 1,6-hexane diol diacrylate, 1, 6-hexane diol ethoxylate diacrylate, 1,6-hexane diol propoxylate diacrylate, 3-hydroxy-2,2-dimethylpropyl3-hydroxy-2,2-dimethylprop Cypionate diacrylate, 1,7-heptane diol diacrylate, 1,8-octane diol diacrylate, 1,9-nona diol diacrylate, 1,10-tecan diol diacrylate, Provided are a method selected from the group consisting of neopentylglycol diacrylate, pentaerythritol diacrylate, trimetholpropane or mixtures thereof.

In the present invention, the amine-based compound is ethanol amine, 3-amino-1-propanol, 2-amino-1-propanol, 1-amino-2-propanol, 3-amino-1,2-propanediol, 2- Amino-1,3-propanediol, serinol, 2- (2-aminoethoxy) ethanol, 2-amino-2-methyl-1,3-propanediol, L-Treninol , 2-amino-1-butanol, 2-amino-2-methyl-1-propanol, 4-amino-1-propanol, 4-amino-2-butanol, 2-amino-1-pentanol, 2-amino- 3-methyl-1-butanol, 2-amino-1-pentanol, 4-amino-1-butanol, 5-amino-1-pentanol, valinol, 6-amino-1-hexanol, 7 -Amino-1-heptanol, 8-amino-1-octanol, 10-amino-1-decanol, 2-amino-2-ethyl-1,3-propanediol, N, N-bis (2- Provided are a method selected from the group consisting of hydroxyethyl) ethylene diamine.

In another aspect, the present invention provides a method wherein the molar ratio of the bisacrylate compound and the amine-based compound having a hydroxyl group is 1: 0.5-1.6 when preparing poly (β-amino ester).

In another aspect, the present invention provides a method for performing a poly (β-amino ester) and lactide having a hydroxyl group in the side chain by a ring-opening polymerization method.

The present invention also provides a method in which the number average molecular weight of the polylactic acid compound of the poly (β-amino ester) -polylactic acid copolymer is 150 to 1000.

The present invention also provides a method wherein the polyethylene glycol compound has a number average molecular weight (Mn) of 500 to 5000.

The present invention also provides a process wherein the poly (β-amino ester) -polylactic acid-polyethylene glycol copolymer is of the formula:

상기 식에서,

Where

R ₁ , R ₂ is an alkyl group having 1 to 12 carbon atoms, wherein x, y and n, m are natural numbers ranging from 1 to 200.

The present invention also provides a method wherein the copolymer has a number average molecular weight of 10,000 to 20,000.

In the present invention, the content of poly (β-amino ester) is 10 to 70% by weight, the content of polylactic acid (B) is 30 to 80% by weight, the content of polyethylene glycol is less than 1% by weight or 10% by weight To provide.

The present invention also provides a method wherein the molar ratio of the poly (β-amino ester) compound, the polylactic acid compound and the polyethylene glycol compound is 10: 15: 1 to 10: 5: 1.

The present invention also provides a method wherein the polyethylene glycol compound includes a group in which the terminal group is substituted with a functional group selected from the group consisting of a carboxyl group, an amine group and a hydroxyl group.

The present invention also provides a method wherein the diameter of the polymersome ranges from 10 to 100 nm.

The present invention also provides a polymersome drug composition comprising a polymersome prepared by the above method and an optical imaging marker or molecular imaging contrast agent for the diagnosis of a disease that may be chemically bound to the polymersome, or a therapeutic substance for the treatment of the disease. To provide.

In another aspect, the present invention is pyrene, RITC (rhodamine B isothiocyanate), FITC (fluorescein isothiocyanate), phycoerythrin (PE), ICG (indocyanine green), as a marker for molecular imaging for the diagnosis of diseases that can be chemically bound to the polymersome Prostate-specific antibody (PSA), alpha-fetoprotein (AFP), human chorionic gonadotropin (HCG), cancer antigen 125 (CA 125), cancer antigen 15-3 (CA 15-3), and carcinoembryonic antigen (CEA) At least one selected composition is provided.

The present invention also provides a composition selected from the group consisting of paramagnetic materials iron oxide, manganese oxide, zinc oxide, gadolinium oxide as a molecular imaging contrast agent for diagnosing a disease that can be chemically bound to the polymersome.

In another aspect, the present invention is an anticancer agent, antibacterial agent, steroids such as paclitaxel (paclitaxel, PTX), doxorubicin (DOX), docetaxel (docetaxel, DOCE) as a therapeutic agent for the treatment of diseases that can be chemically bound to the polymersome , Anti-inflammatory analgesic, sex hormone, immunosuppressant, antiviral, anesthetic, antiemetic, antihistamine and protein as bovine serum albumin, human serum albumin, human growth hormone Provided at least one composition selected from the group consisting of.

In another aspect, the present invention provides a method for producing a polymersome is a compound represented by the following scheme.

Where

R ₁ and R ₂ are alkyl groups having 1 to 12 carbon atoms, wherein x, y and n, m are natural numbers ranging from 1 to 200.

The polymersome according to the present invention has the advantage of being able to accommodate both hydrophobic and hydrophilic drugs without being restricted in the properties of the drug.

In addition, the present invention is characterized by having a solubility in water according to the pH, but can not form a micelle by the self-assembly as a polymersome to express the sustained release according to the pH conditions in the human body.

In addition, the present invention can induce a hydrophilic drug encapsulation inside the polymersomes through which there is an effect that can be used as a target-oriented drug carrier and diagnostic use in the body pH change.

1 is a schematic diagram of a polymersome consisting of poly (β-amino-ester) (PAE), polylactic acid and methoxy polyethylene glycol (MPEG). Or the release of hydrophilic drugs is shown schematically.
Figure 2 is a graph showing the molecular weight distribution of the copolymer prepared in Examples 1 and 4 and the retention time by GPC measurement of the polyethylene glycol substituted terminal carboxyl group as a comparative sample.
Figure 3 is a ¹ H-NMR graph of the copolymer prepared in Example 4, a Michael reaction of 3-amino 1-propanol and 1,4-butanediol diacrylate as precursors of poly (β-amino ester) The results of the synthesis of poly (β-amino esters) are shown.
Figure 4 is a graph showing the critical concentration measured by the fluorescence spectrometer of the pH sensitive copolymer prepared in Example 4.
Figure 5 is a graph showing the size of the polymersomes with the pH change measured by DLS of the pH sensitive copolymer prepared in Example 4.
6 is a graph showing the number of polymersomes according to the pH change measured by DLS of the pH sensitive copolymer prepared in Example 4. FIG.
7 is a polymersome photograph measured by TEM of a pH-sensitive copolymer prepared in Example 4, characterized in that it has a donut form in the middle of the hollow.
8 is a graph showing permeability according to pH change as a result of TNS of the copolymer prepared in Example 4. FIG.

DETAILED DESCRIPTION Hereinafter, exemplary embodiments of the present invention will be described in detail with reference to the accompanying drawings. First, it should be noted that, in the drawings, the same components or parts have the same reference numerals as much as possible. In describing the present invention, detailed descriptions of related well-known functions or configurations are omitted in order not to obscure the subject matter of the present invention.

The terms "about "," substantially ", etc. used to the extent that they are used herein are intended to be taken to mean an approximation of, or approximation to, the numerical values of manufacturing and material tolerances inherent in the meanings mentioned, Accurate or absolute numbers are used to help prevent unauthorized exploitation by unauthorized intruders of the referenced disclosure.

Polymersomes according to the present invention include (i) a poly (β-amino ester) compound (A); (ii) polylactic acid compound (B); And (iii) a polyethylene glycol-based compound (C).

Specifically, the present invention copolymerizes a poly (β-amino ester) compound which is sensitive to pH, a polylactic acid compound having a hydrophobicity, and a polyethylene glycol-based compound having hydrophilicity, thereby being sensitive to pH changes in the body and polymer polymers in a specific pH range. It is characterized by providing a copolymer capable of chemically binding a molecular imaging marker for diagnosing a disease and a contrast agent and a drug for treating a disease through a hydrosil group formed on the side chain of the poly (β-amino ester).

The present inventors recognize that when poly (β-amino ester) is used alone, it exhibits pH dependence but does not form micelles due to self assembly phenomenon, and is hydrophilic to the poly (β-amino ester). Polyethylene glycol-based compounds were copolymerized to confirm that the hydrophilic polyethylene glycol-based compound was present outside of micelles and hydrophobic poly (β-amino ester) was present inside of micelles to form micelles.

Furthermore, a new copolymer was formed for the purpose of chemically bonding hydrophilic drugs as therapeutic agents, molecular imaging markers and contrast agents for the diagnosis of diseases, and various therapeutic agents to these polymers. In addition, it has been found to form polymersome structures in the form of hollow donuts formed with hydrophilic groups inside, and have been found to be applicable as target-oriented drug release carriers for the diagnosis and treatment of diseases capable of targeted release at specific pH.

The pH-sensitive polymersomes of the present invention form stable polymersomes at certain pH, such as pH 7.0-7.4, preferably 7.2-7.4, which is the pH range of normal cells in the body, and have a pH range of less than 7.0, indicated by abnormal cells such as cancer cells. When the polymersome structure is collapsed, cancer cells can be targeted for diagnosis due to the release of a diagnostic agent chemically bound to the polymersome and thus used as a carrier for targeted drug release of the therapeutic agent for treatment. That is, due to the increased degree of ionization of the tertiary amines present in the poly (β-amino ester) at low pH (below pH 7.0), the entire PAE becomes water soluble and polymersomes cannot be formed. The degree of ionization is lowered to show hydrophobic characteristics, thereby forming polymersomes by self-assembly.

In addition, the copolymer capable of forming the pH-sensitive polymersomes may be applied to applications in which diagnosis and treatment are simultaneously performed by delivering a substance for diagnosis and treatment of a disease as well as gene delivery and drug delivery to abnormal cells. .

In addition, in the present invention, a polymersome is formed in the range of pH 7.0 to 7.4, which is the same as a normal body condition, and the polymersome is designed and applied to a cancer cell target-oriented polymersome that is degraded under the pH 7.0, which is an abnormal condition such as cancer cells. By appropriately modifying the constituents of the copolymer, their molar ratios, molecular weights and / or side chain functionalities, the target-oriented polymersomes can be designed and applied to cancer cells as well as to genetic variations or other applications.

Poly (β-amino ester) compound (A) having both hydrophobicity and pH sensitivity among the components of the copolymer forming the pH sensitive polymersome according to the present invention.

The poly (β-amino ester) compound has ionization characteristics such that its solubility in water varies depending on pH due to the tertiary amine groups present in itself, thereby forming and / or forming a polymersome structure according to the pH change in the body as described above. Can collapse. The poly (β-amino ester) compound may be synthesized by polymerizing an amine-based compound having a bisacrylate compound and a hydroxyl group as a non-limiting example.

Bisacrylate may be represented as follows [Formula 1], non-limiting examples thereof include ethylene glycol diacrylate, 1,4-butane diol diacrylate, 1,3-butane diol diacrylate, 1,6-hexane diol diacrylate, 1,6-hexane diol ethoxylate diacrylate, 1,6-hexane diol propoxylate diacrylate, 3-hydroxy-2,2-dimethyl Propyl3-hydroxy-2,2-dimethylpropionate diacrylate, 1,7-heptane diol diacrylate, 1,8-octane diol diacrylate, 1,9-nona diol diacrylate Latex, 1,10-tecan diol diacrylate, neopentylglycol diacrylate, pentaerythritol diacrylate, trimetholpropane or mixtures thereof and the like.

Where

R is an alkyl group having 1 to 20 carbon atoms.

In addition, the amine-based compound having a hydrosil group may be represented by the following [Formula 2].

Where

R is an alkyl group having 1 to 20 carbon atoms.

Non-limiting examples of the amine-based compound include ethanol amine, 3-amino-1-propanol, 2-amino-1-propanol, 1-amino-2-propanol, 3-amino-1,2-propanediol, 2 -Amino-1,3-propanediol, serinol, 2- (2-aminoethoxy) ethanol, 2-amino-2-methyl-1,3-propanediol, L-treninol (Threninol ), 2-amino-1-butanol, 2-amino-2-methyl-1-propanol, 4-amino-1-propanol, 4-amino-2-butanol, 2-amino-1-pentanol, 2-amino -3-methyl-1-butanol, 2-amino-1-pentanol, 4-amino-1-butanol, 5-amino-1-pentanol, valinol, 6-amino-1-hexanol, 7-amino-1-heptanol, 8-amino-1-octanol, 10-amino-1-decanol, 2-amino-2-ethyl-1,3-propanediol, N, N-bis (2 -Hydroxyethyl) ethylene diamine and the like.

In preparing the poly (β-amino ester), the molar ratio of the bisacrylate compound and the amine compound having a hydroxyl group is preferably in the range of 1: 0.5 to 1.6. When the molar ratio of the amine-based compound is less than 0.5 or more than 1.6, the reaction does not occur properly or forms branched polymers, making it difficult to obtain pH sensitivity under the biological conditions (pH 7.4) of the human body to be obtained in the present invention. do.

The poly (β-amino ester) of the present invention formed through the reaction of the aforementioned bisacrylate compound with an amine compound having a hydroxyl group may be represented by [Formula 3].

 Where

R ₁ , R ₂ , is an alkyl group having 1 to 12 carbon atoms, wherein x is a natural number in the range of 1 to 200.

Lactide may be copolymerized through ring-opening polymerization after molecular design to have hydroxyl groups as side chains in addition to the pH sensitive group originally possessed in the poly (β-amino ester) compound.

The poly (β-amino ester) -polylactic acid pH sensitive copolymer formed by ring-opening polymerization of poly (β-amino ester) having a hydroxyl group on the side chain and lactide may be represented by [Formula 4].

Where

R ₁ , R ₂ is an alkyl group having 1 to 12 carbon atoms, where x and m are natural numbers ranging from 1 to 200.

The molecular weight of the polylactic acid compound of the poly (β-amino ester) -polylactic acid copolymer is in the range of 150 to 1000.

Meanwhile, according to the present invention, for example, a methoxy polyethyleneglycol compound may be used as the biodegradable compound having a hydrophilic block which is one of the components of the copolymer forming the pH-sensitive polymersomes. In particular, it is preferable to have a single functional group (monofunctional), such as a carboxyl group, to react with a poly (β-amino ester) or a polylactic acid, which is a pH sensitive polymer, at the terminal of the methoxy polyethylene glycol compound, for example, [Formula 5] As such, there is a polyethylene glycol compound in which the molecular terminal portion is substituted with a carboxyl group.

Where

      R is a hydrogen atom or an alkyl group having 1 to 6 carbon atoms, where x is 1 to

It is a natural number in the range of 200.

The alkyl group refers to a linear or branched lower saturated aliphatic hydrocarbon, for example, methyl, ethyl, n -propyl, isopropyl, n -butyl, s -butyl, isobutyl, t -butyl and n -pentyl groups Etc.

The number average molecular weight (Mn) of the polyethylene glycol compound is not particularly limited, but is preferably in the range of 500 to 5000. When the number average molecular weight (Mn) of the polyethylene glycol compound is less than 500 and more than 5000, it is difficult to control the molecular weight of the final copolymer, and it is not easy to form polymersomes using the copolymer.

PH-sensitive poly (β-amino ester) -polylac last synthesized through the reaction of a poly (β-amino ester) -polylactic acid copolymer with a carboxyl group at the end of the polylactic acid and a carboxyl group at the end of the polyethylene glycol The ticic acid-polyethylene glycol polymer is represented by [Formula 6].

Where

R ₁ , R ₂ is an alkyl group having 1 to 12 carbon atoms, wherein x, y and n, m are natural numbers ranging from 1 to 200.

The copolymer represented by [Formula 6] is self-assembled according to the pH change due to the amphiphilic properties composed of the hydrophilic block of the polyethylene glycol and the hydrophobic block of polylactic acid and the pH sensitivity of the poly (β-amino ester) ( self-assembly) to form or disintegrate the polymersomes, and preferably form polymersomes when the pH is in the range of 7.0 to 7.4 and more preferably in the range of 7.2 to 7.4 and the pH is in the range of 6.0 to less than 7.0. If so, the polymersome will collapse. In particular, the copolymers of the present invention may exhibit good sensitivity within the pH 0.3 range.

The molecular weight range of the copolymer is not particularly limited, but is preferably in the range of 10,000 to 20,000. If the molecular weight is less than 10,000, the copolymer polymersome is not only difficult to form at a specific pH, but also easily dissolves in water and disintegrates even when formed. In addition, when the molecular weight exceeds 20,000, the balance of hydrophilicity / hydrophobicity may be broken and polymersome may not be formed at a specific pH and may precipitate.

The content of poly (β-amino ester) in the pH-sensitive copolymer according to the present invention is 10 to 70% by weight, the content of polylactic acid (B) is preferably 30 to 80% by weight, preferably 45 to 70% by weight. %to be. When the content of the polylactic acid is less than 30% by weight or more than 80% by weight, the hydrophilic-hydrophobic balance is broken, and thus, polymersomes cannot be formed and precipitation occurs. In addition, the content of polyethylene glycol (C) in the pH sensitive copolymer is suitably 1 to 10% by weight, preferably 3 to 6% by weight. When the content of polyethylene glycol is less than 1% by weight or more than 10% by weight, the hydrophilic-hydrophobic balance may be broken, thereby preventing the formation of polymersomes.

In addition, the copolymer can form various types of copolymers by controlling the reaction molar ratio of poly (β-amino ester), polylactic acid, and polyethylene glycol compounds.

For example, a method of preparing a poly (β-amino ester) -polylactic acid-polyethyleneglycol pH sensitive copolymer according to the present invention may be synthesized by the route of [Scheme 1] or [Scheme 2]. have.

Scheme 1

Where

x is a natural number ranging from 1 to 200.

For example, a bisacrylate having an acrylate end group and 3-amino 1-propanol may be used as an example of the preparation method illustrated in [Scheme 1], which is an addition known as a Michael reaction commonly known in the art. The reaction forms a poly (β-amino ester), and the poly (β-amino ester) formed has a hydroxyl group in a side chain with a tertiary amine which is a pH sensitive group. As shown in [Scheme 2], a poly (β-amino ester) -polylactide copolymer can be prepared by adding lactide and ring-opening polymerization.

Scheme 2

Where

 x and n are natural numbers ranging from 1 to 200.

For example, one embodiment of the manufacturing method illustrated in [Scheme 2], a lactide is added to a poly (β-amino ester) having a hydroxyl group as a side chain and a tertiary amine, which is a pH sensitive group formed in the above [Scheme 1]. Through ring-opening polymerization, a poly (β-amino ester) -polylactic acid copolymer is prepared, wherein in the preparation of the copolymer, dichloromethane, tetrahydrofuran, dimethyl sulfoxide, dimethyl formamide and the like are included in pure water. Organic solvents may be used.

The molecular weight of the polylactic acid compound of the poly (β-amino ester) -polylactic acid copolymer is in the range of 150 to 1000.

In the following [Scheme 3], a poly (β-amino ester) -polylactic acid copolymer prepared in [Scheme 2] was put into a polyethylene glycol substituted with a carboxyl group by a hydrophilic block and polymerized. Amino ester) -polylactic acid-polyethyleneglycol copolymers could be prepared.

Scheme 3

Where

R ₁ , R ₂ is an alkyl group having 1 to 12 carbon atoms, wherein x, y and n, m are natural numbers ranging from 1 to 200.

Meanwhile, the molar ratio of the poly (β-amino ester) compound, polylactic acid compound, and polyethylene glycol compound may be 10: 15: 1 to 10: 5: 1.

Since the polylactic acid terminal of the above formula is a carboxyl group, various diagnostic agents such as optical image markers or molecular imaging contrast agents can be modified, and various therapeutic drugs such as anticancer drugs and protein drugs can be modified.

In the present invention, gel permeation chromatography (GPC) was used to measure the molecular weight of the copolymer synthesized as above, and ¹ H-NMR was used to calculate the molar ratio of the grafted hydrophilic, hydrophobic and cancer cell targets. Fluorescence spectrometer (FL) and Dynamic Light Scattering (DLS) were used to measure the concentration change of micelle and change of micelle size according to pH change. The possibility was confirmed.

In addition, the present invention (a) the copolymer described above to form a polymersome in accordance with the pH change; And (b) a molecular imaging marker or contrast agent for diagnosing a disease capable of inducing chemical binding to the copolymer, or a therapeutic agent material for treating a disease.

The polymersome-type drug composition forms a polymersome when injected into the body, and the polymersome collapses when it reaches a locally low pH, such as cancer cells, thereby providing molecular imaging markers for diagnosing chemically bound diseases. Targeted drug delivery can be achieved through the release of contrast agents and therapeutic agents for the treatment of diseases.

Diagnostic and therapeutic agent materials that can chemically bind to the polymersome form of the present invention can be used without particular limitation, and non-limiting examples thereof include pyrene, RITC, fluorescein isothiocyanate (FITC), phycoerythrin (PE) ), Indocyanine green (ICG), prostate-specific antibody (PSA), alpha-fetoprotein (AFP), human chorionic gonadotropin (HCG), cancer antigen 125 (CA 125), cancer antigen 15-3 (CA 15-3), CEA (carcinoembryonic antigen), contrast agents include paramagnetic substances iron oxide, manganese oxide, zinc oxide, gadolinium oxide, etc., and therapeutic drugs include paclitaxel (PTX), doxorubicin (DOX), docetaxel (docetaxel, DOCE), etc. Anticancer drugs, antibacterial agents, steroids, anti-inflammatory drugs, sex hormones, immunosuppressants, antiviral drugs, anesthetics, antiemetic drugs or antihistamines, proteins as bovine serum albumin human serum albumin (hunman se) rum albumin), human growth hormone, etc. In addition to the aforementioned ingredients, conventional additives known in the art such as excipients, stabilizers, pH adjusters, antioxidants, preservatives, binders or disintegrants, etc. It may include.

The method for preparing the polymer polymersomes according to the present invention may be used alone or in combination with methods such as agitation, heating, ultrasonic scanning, solvent evaporation using an emulsification method, matrix formation, or dialysis using an organic solvent.

The diameter of the prepared polymersome is not particularly limited, but is preferably in the range of 10 to 100 nm. In addition, the polymersome-type drug composition may be used in the form of oral or parenteral preparations, and may be prepared by intravenous, intramuscular or subcutaneous injection.

Hereinafter, the present invention will be described in more detail with reference to Examples and Experimental Examples. However, the following examples are only for illustrating the present invention, and the scope of the present invention is not limited thereto.

[Examples 1 to 10. Synthesis of pH Sensitive Copolymer]

Example 1 Preparation of Poly (β-Amino Ester) Polymer

1 mole of 3-amino-1-propanol and 1 mole of 1,4-butane diol diacrylate were placed in a two-necked round flask and reacted at 100 ° C. for 5 hours to give a number average molecular weight (Mn) having a hydrosil group in the side chain. This 7,200 poly (β-amino ester) was prepared.

Example 2. Preparation of Poly (β-Amino Ester) -Polylactic Acid Polymers

1 mole of lactide was added to 1 mole of poly (β-amino ester) prepared in Example 1, and the ring-opening reaction was carried out at 135 ° C. for 5 hours using Sn (Oct) ₂ , a tin-based catalyst, to give poly (β-amino ester). A poly (β-amino ester) -polylactic acid copolymer was prepared in which a hydrophobic polylactic acid was connected to the side chain of C) and a terminal of the polylactic acid was substituted with a carboxyl group (Mn = 6,000).

Example 3.

A poly (β-amino ester) -polylactic acid copolymer having a number average molecular weight (Mn) of 8,300, except that 2 moles instead of 1 mole of lactide of Example 2 were used. Got.

Example 4 Preparation of Poly (β-Amino Ester) -Polylactic Acid-Polyethylene Glycol Copolymer

To 1 mole of the poly (β-amino ester) -polylactic acid copolymer prepared in Example 2, 1 mole of polyethylene glycol methyl ether (MPEG2000, Mn = 2,000) substituted with a carboxyl group was added to room temperature using pure water as a solvent. After reacting for 24 hours at, the cyclourea formed as a by-product was separated and precipitated in ethyl ether to prepare a poly (β-amino ester) -polylactic acid-polyethylene glycol copolymer (Mn = 10,100).

Example 5.

Except for using 2 moles instead of 1 mole of the poly (β-amino ester) -polylactic acid copolymer of Example 4, a polyamide having a number average molecular weight (Mn) of 20,000 was carried out in the same manner as in Example 4. (β-amino ester) -polylactic acid-polyethylene glycol copolymer was obtained.

Experimental Example 1. Determination of molecular weight of pH sensitive copolymer

In order to measure the molecular weight of the pH sensitive copolymer prepared according to the present invention, the following analysis was carried out.

Poly (β-amino ester) prepared in Examples 1 to 4 alone and a copolymer using the same were used, and Gel Permeation Chromatography (GPC, Waters, Inc.) analysis was performed to investigate their molecular weight control possibilities. As shown in FIG. 2, a GPC diagram was obtained in the order of polyethylene glycol, a poly (β-amino ester) substituted with a carboxyl group, and a copolymer using the same (Example 4), and the molecular weight could be calculated from the retention time. there was.

Experimental Example 2 Measurement of Critical Concentration Concentration

In order to observe the behavior according to the pH change of the pH-sensitive copolymer micelles prepared according to the present invention, the following experiment was carried out.

Since the aggregate aggregation concentration (CGC) of the graft copolymers prepared in Examples 4 and 5 was not directly known through the Fluorescence Spectrometer, the behavior of the polymersomes was not changed. (pyrene) was used.

A buffer solution of pH 7.0 containing ^10-6 M pyrene was prepared, and the samples prepared in Examples 4 and 5 were dissolved in the buffer solution at a concentration of 2 mg / ml, and then diluted 1/5. Fluorescence spectroscopy measured the change in energy emitted due to the change in concentration of the polymersomes.

Experimental Example 3. Measurement of pH sensitivity of the copolymer

The pH sensitivity of the pH sensitive copolymers prepared according to the present invention was measured by dynamic light scattering (DLS). As shown in FIG. 6, the copolymers prepared in Examples 4 and 5 appeared in the low pH region as the molar ratio of the polylactic acid in which the polymersome-uniformer transition was changed to a hydrophobic block by pH change.

It will be apparent to those skilled in the art that various modifications and variations can be made in the present invention without departing from the spirit or scope of the inventions. It will be clear to those who have knowledge of.

Claims (20)

Reacting a bisacrylate compound with an amine compound having a hydroxyl group to form a poly (β-amino ester) compound;
Forming a hydroxyl group on the side chain of the poly (β-amino ester) compound;
Polymerizing the poly (β-amino ester) and lactide having a hydroxyl group in the side chain; And
A method of preparing a PH-sensitive polymersome comprising copolymerizing a polyethylene glycol compound to the polymer. The method of claim 1,
Wherein said poly (ㅯ -amino ester) comprises a tertiary amine group that is ionized at less than pH 7.0. The method of claim 1,
The copolymer forms a polymersome when the pH is in the range of 7.0 to 7.4 (preferably 7.2 to 7.4), and the polymersome collapses and becomes a unimer when the pH is in the range of 6.0 to 7.0. The method of claim 1,
The bisacrylates are ethylene glycol diacrylate, 1,4-butane diol diacrylate, 1,3-butane diol diacrylate, 1,6-hexane diol diacrylate, 1,6-hexane di All ethoxylate diacrylate, 1,6-hexane diol propoxylate diacrylate, 3-hydroxy-2,2-dimethylpropyl3-hydroxy-2,2-dimethylpropionate diacryl Late, 1,7-heptane diol diacrylate, 1,8-octane diol diacrylate, 1,9-nona diol diacrylate, 1,10-tecan diol diacrylate, neopentylglycol di A method selected from the group consisting of acrylate, pentaerythritol diacrylate, trimetholpropane or mixtures thereof. The method of claim 1,
The amine compound is ethanol amine, 3-amino-1-propanol, 2-amino-1-propanol, 1-amino-2-propanol, 3-amino-1,2-propanediol, 2-amino-1, 3-propanediol, serinol, 2- (2-aminoethoxy) ethanol, 2-amino-2-methyl-1,3-propanediol, L-Treninol, 2-amino -1-butanol, 2-amino-2-methyl-1-propanol, 4-amino-1-propanol, 4-amino-2-butanol, 2-amino-1-pentanol, 2-amino-3-methyl- 1-butanol, 2-amino-1-pentanol, 4-amino-1-butanol, 5-amino-1-pentanol, valinol, 6-amino-1-hexanol, 7-amino-1 -Heptanol, 8-amino-1-octanol, 10-amino-1-decanol, 2-amino-2-ethyl-1,3-propanediol, N, N-bis (2-hydroxyethyl) The method selected from the group consisting of ethylene diamine. The method of claim 1,
In preparing a poly (β-amino ester), the molar ratio of the bisacrylate compound and the amine compound having a hydroxyl group is 1: 0.5 to 1.6. The method of claim 1,
A method in which the poly (β-amino ester) and lactide having a hydroxyl group in the side chain are carried out by a ring-opening polymerization method. The method of claim 7, wherein
The number average molecular weight of the polylactic acid compound of the poly (β-amino ester) -polylactic acid copolymer is 150 to 1000. The method of claim 1,
The number average molecular weight (Mn) of the polyethylene glycol compound is 500 to 5000 method. The method of claim 1,
The poly (β-amino ester) -polylactic acid-polyethylene glycol copolymer is of the formula

Where
R ₁ , R ₂ is an alkyl group having 1 to 12 carbon atoms, wherein x, y and n, m are natural numbers ranging from 1 to 200. The method of claim 1,
The number average molecular weight of the copolymer is 10,000 to 20,000. The method of claim 1,
The content of poly (β-amino ester) is 10 to 70% by weight, the content of polylactic acid (B) is 30 to 80% by weight, the content of polyethylene glycol is less than 1% by weight or 10% by weight. The method of claim 1,
The molar ratio of the group poly (β-amino ester) compound, polylactic acid compound and polyethylene glycol compound is 10: 15: 1 to 10: 5: 1. The method of claim 1,
The polyethylene glycol compound comprises a group in which the terminal group is substituted with a functional group selected from the group consisting of a carboxyl group, an amine group and a hydroxyl group. The method of claim 1,
The diameter of the polymersomes ranges from 10 to 100 nm. A polymersome prepared by the method according to any one of claims 1 to 15 and
A polymersome drug composition comprising an optical imaging marker or molecular imaging contrast agent for treating a disease that may be chemically bound to the polymersome or a therapeutic agent for treating a disease. 17. The method of claim 16,
Molecular imaging markers for diagnosing diseases that may be chemically bound to the polymersome include pyrene, RITC (rhodamine B isothiocyanate), FITC (fluorescein isothiocyanate), phycoerythrin (PE), ICG (indocyanine green), PSA (prostate-) 1 or more selected from the group consisting of specific antibody (alpha) -fetoprotein (AFP), human chorionic gonadotropin (HCG), cancer antigen 125 (CA 125), cancer antigen 15-3 (CA 15-3), and carcinoembryonic antigen (CEA) Composition. 17. The method of claim 16,
Molecular imaging contrast agent for diagnosing a disease that can be chemically bound to the polymersome as a composition selected from the group consisting of paramagnetic material iron oxide, manganese oxide, zinc oxide, gadolinium oxide. 17. The method of claim 16,
Therapeutic materials for the treatment of diseases that may be chemically bound to the polymersome body include anti-cancer agents such as paclitaxel (paclitaxel, PTX), doxorubicin (DOX), docetaxel (docetaxel, DOCE), antibacterial agents, steroids, anti-inflammatory drugs, Sex hormones, immunosuppressants, antiviral agents, anesthetics, antiemetics, antihistamines and proteins in the group consisting of bovine serum albumin, human serum albumin, and human growth hormone At least one selected composition. A method for producing a polymersome, which is a compound represented by the following scheme.

Where
R ₁ and R ₂ are alkyl groups having 1 to 12 carbon atoms, wherein x, y and n, m are natural numbers ranging from 1 to 200.

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