KR100949850B1 - TEMPERATURE AND pH-SENSITIVE BLOCK COPOLYMER HAVING EXCELLENT LONG-TIME DRUG DELIVERY AND USING THE SAME - Google Patents

Abstract

A temperature and pH sensitive block copolymer comprising: (a) a polyethylene glycol series compound (A); And (b) provides a temperature- and pH-sensitive block copolymer having excellent long-term drug release properties, including a coupling agent of a poly (amido amine) oligomer (B) alone, and a method for preparing the same and a hydrogel-type drug carrier using the same.

pH sensitive block copolymers, drug carriers, PEG, PAA

Description

TEMPERATURE AND pH-SENSITIVE BLOCK COPOLYMER HAVING EXCELLENT LONG-TIME DRUG DELIVERY AND USING THE SAME}

The present invention relates to a pH-sensitive block copolymer hydrogel and a drug carrier using the same, and in particular, in order to drastically improve the toxicity of biodegradable degradation products during injection in human body, it has biodegradability, but accumulates in the human body and causes various side effects. Target-oriented drug release is possible according to changes in body temperature and pH without using a substance that can be injected, and the components are completely released into the body by biodegradation at the time of injection in the human body. It relates to a copolymer hydrogel and a drug carrier using the same.

Various research and development using stimulus (temperature, pH, light, electric field, magnetic field, etc.) sensitized block copolymers have been actively conducted to develop drug carriers using hydrogels.

US Pat. No. 4,942,035 uses a copolymer of a hydrophilic polymer, polyalkylene glycol, and a biodegradable polyester polymer, polylactide or polyglycolide, and polycaprolactone, to provide a conventional polyethylene glycol called Pluronic. Excessive polyethylene oxide-polypropylene oxide-polyethylene oxide block copolymer has been improved the problem that does not decompose in the body.

U.S. Patent No. 5,476,909 describes triblock copolymers in the form of biodegradable polyester polymer ABA, wherein the hydrophobic block (A) is limited to polylactide (PLA), polyglycolide (PGA) and their copolymers. The hydrophilic block (B) is also limited to polyethylene glycol (PEG) and derivatives thereof.

In addition, the Republic of Korea Patent Publication No. 2000-0012970 proposed a pH-sensitive polymer containing a sulfonamide group and a method for producing the same, mainly linear polymers formed by random copolymerization of the sulfonamide monomer and dimethyl acrylamide or isopropyl acrylamide The solubility change or the swelling degree of the crosslinked polymer is described.

Meanwhile, US Patent No. 6,476,156 describes the preparation of a biodegradable and temperature sensitive triple block copolymer consisting of PEG-COO-PL (PG) -COO-PEG.

The above-described conventional techniques are to show a sol-gel transition phenomenon according to temperature using a block copolymer of a hydrophobic polymer and a hydrophilic polymer, and the block copolymer is in the form of an aqueous solution in a sol state. In the case of injection into the body, it was used as a sustained-release drug carrier that gradually transferred to a gel (gel) state by body temperature, so that the drug was stably loaded in the body and the drug was gradually released. However, when using a block copolymer that exhibits a temperature-sensitive sol-gel transition property, the temperature of the needle reaches a thermal equilibrium with the temperature in the body during the injection into the body, thereby completely injecting it into the body. Problems such as clogging of the injection needle caused by gelation occurred before. In addition, although hydrophobic moieties consisting of PLA, PGA, PLGA, PCL, or PCLA are reported to exhibit pH sensitivity, a block copolymer prepared using poly (b-amino ester), which is a pH sensitive component, is also used. As a drug delivery carrier which releases a drug constantly for a long time, the block copolymer backbone is ester-bonded, and thus it is not suitable for a long-term drug release requiring more than one month to three months.

In the case of US Patent No. 6,476,156, there is a complicated problem in that the steps for coupling PEG and PLA, PGA, or PCL are complicated, and there is no consideration of pH change in the human body as mentioned above.

In order to solve the above problems, an object of the present invention is to provide a block copolymer temperature and pH sensitive block copolymer sensitive to not only the temperature in the body but also pH and a method for producing the same.

It is also an object of the present invention to provide a novel temperature and pH sensitive block copolymer composed only of components harmless to the human body, which are biodegraded in the human body and completely discharged to the outside of the body, and a method for preparing the same.

It is also an object of the present invention to suppress the initial burst release of the drug so that it can be used as a hydrogel drug delivery system and a new temperature and pH molecular designed to release the drug at a constant concentration for 1 to 3 months The present invention provides a sensitive block copolymer, a method for preparing the same, and a drug carrier using the same.

It is also an object of the present invention to provide a hydrogel-type drug carrier comprising a bioactive material that can be encapsulated in the block copolymer.

It is also an object of the present invention to appropriately modify the components of the block copolymer, their molar ratios, molecular weights and / or functional groups in the block to target-oriented design not only for cancer cells but also for genetic variation or other applications, which are usefully applied in the body. It is to provide a high temperature and pH sensitive block copolymer, a method for preparing the same and a drug carrier using the same.

The present invention for achieving the above object in the temperature and pH sensitive block copolymer, (a) polyethylene glycol-based compound (A); And (b) provides a temperature- and pH-sensitive block copolymer with excellent long-term drug release properties comprising a coupling of a mixture of poly (amido amine) oligomer (B).

In another aspect, the present invention provides a block copolymer wherein the copolymer comprises a triple block of B-A-B.

In another aspect, the present invention provides a block copolymer of the polyethylene glycol compound.

Wherein R is a hydrogen atom or an alkyl group having 1 to 5 carbon atoms, and n is a natural number in the range of 11 to 45.

In another aspect, the present invention provides a block copolymer having a molecular weight (Mn) of the polyethylene glycol-based compound is 500 to 8,000.

In another aspect, the present invention provides a block copolymer having a molecular weight (Mn) of 1,000 to 2,000 when R in the chemical formula of polyethylene glycol (PEG).

In another aspect, the present invention provides a block copolymer having a molecular weight (Mn) of 1,000 to 5,000 when R in the chemical formula of the compound is methoxy polyethylene glycol (MPEG) is a methyl group.

In another aspect, the present invention further comprises one or more substituents selected from the group consisting of a primary amine group, a secondary amine group and an alkylene diacrylamide capable of reacting the poly (amido amine) series precursor with a polyethylene glycol series compound. A block copolymer is provided.

The present invention also provides a block copolymer in which the poly (amido amine) oligomer further comprises a functional group of a hydroxy group (-OH), a carboxy group (-COOH) or an amine group (-NH ₂ ).

The present invention also provides a block copolymer comprising a tertiary amine group in which the poly (amido amine) -based oligomer is mild at pH less than 7.0.

In another aspect, the present invention provides a block copolymer having a molecular weight of 1,000 to 5,000 of the poly (amido amine) -based oligomer.

In another aspect, the present invention provides a block copolymer consisting of the following formula.

Wherein k is a natural number in the range of 4 to 10, n is a natural number in the range of 11 to 45, and m is a natural number in the range of 10 to 100.

In another aspect, the present invention provides a block copolymer having a molecular weight of 2,500 to 21,000.

In another aspect, the present invention provides a block copolymer of the A and B molecular weight ratio of 1: 1 to 4.

The present invention also provides a block copolymer in which the block copolymer forms a hydrogel at pH 7.0 to 7.4, and forms a sol state at pH 3.0 to less than 7.0.

In another aspect, the present invention provides a method for producing a temperature and pH sensitive block copolymer, comprising the steps of: a) forming a polyethylene glycol-based block precursor; b) provides a method for producing a temperature- and pH-sensitive block copolymer having excellent long-term drug release property comprising the step of coupling the resulting polyethylene glycol-based block precursor and poly (amido amine) oligomer.

The present invention also provides a method for producing a block copolymer in which the sock end of polyethylene is imidazole so that the polyethylene glycol-based block precursor can react with the precursors of the poly (amido amine) -based block.

In another aspect, the present invention provides a method for producing a block copolymer of the polyethylene glycol-based compound of formula (1).

Wherein R is a hydrogen atom or an alkyl group having 1 to 5 carbon atoms, and n is a natural number in the range of 11 to 45.

In another aspect, the present invention provides a method for producing a block copolymer having a molecular weight (Mn) of the polyethylene glycol-based compound is 500 to 8,000.

In another aspect, the present invention provides a method for producing a block copolymer having a molecular weight (Mn) of 1,000 to 2,000 when R in the chemical formula of polyethylene glycol (PEG).

In another aspect, the present invention provides a method for producing a block copolymer having a molecular weight (Mn) of 1,000 to 5,000 when R is a methoxy polyethylene glycol (MPEG) in the formula of the compound.

In another aspect, the present invention provides a method for producing a block copolymer consisting of the following reaction formula to form the polyethylene glycol-based precursor reacts the polyethylene glycol-based compound and carbonyldiimidazole (N, N-carbonyldiimidasole).

Where n is a natural number ranging from 11 to 45.

The poly (amido amine) -based precursors in the block air further comprises one or more substituents selected from the group consisting of a primary amine group, a secondary amine group and alkylene diacrylamide can react with the block of the polyethylene glycol-based compound Provided are methods for preparing coalescing.

In another aspect, the present invention provides a method for producing a block copolymer formed by polymerizing the alkylene diacrylamide compound and the amine-based compound of the poly (amido amine) -based oligomer.

In another aspect, the present invention provides a method for producing a block copolymer of the alkylene diacrylamide compound of the formula.

In the above formula, R is an alkyl group having 1 to 20 carbon atoms.

In the present invention, the alkylene diacrylamide compound is N, N-methylene bisacrylamide (MDA), N, N-ethylene bisacrylamide, N-isopropyl acrylamide, 1,4 butylene diacrylamide, Provided is a method for producing at least one block copolymer selected from the group consisting of 1,6 hexylenediacrylamide, 1,8 octylene diacrylamide, 1,10 decanadiacrylamide, or mixtures thereof.

The present invention also provides a method for producing a block copolymer wherein the amine-based compound is a primary amine or a secondary amine-containing diamine compound or a mixture thereof.

In the present invention, the primary amine compound is 4,4'-trimethyldipiperidine, 3-methyl-4- (3-methylphenyl) piperazine, 3 methylpiperazine, 4- (bis- (fluorophenyl) methyl Piperazine, 4- (ethoxycarbonylmethyl) piperazine, 4- (phenylmethyl) piperazine, 4- (1-phenylethyl) piperazine, 4- (1,1-dimethoxycarbonyl) piperazine , 4- (2- (bis- (2-propenyl) amino) ethyl) piperazine, methylamine, ethylamine, butylamine, hexylamine, 2-ethylhexylamine, 2-piperidin-1-yl-ethyl Amine, C-aziridin-1-yl-methylamine, 4-aminomethylpiperidine (AMPD), N-methylethylenediamine (MEDA), N-ethylethylenediamine (EEDA), N-hexylethylenediamine ( HEDA) and 1- (2-aminoethyl) piperazine (AEPZ) or mixtures thereof.

In addition, the present invention, the secondary amine-containing diamine compound is piperazine, 4,4'- trimethylene dipiperidine, N, N'- dimethyl ethylene diamine, N, N'- diethyl ethylene diamine, imida Provided are a process for preparing at least one block copolymer selected from the group consisting of zolidine and diazephan.

The present invention also provides a method for producing a block copolymer in which the molar ratio of the alkylene diacrylamide compound and the amine compound is 1: 0.5 to 4.0.

The present invention also provides a method for producing a block copolymer having a molecular weight of 1,000 to 5,000 of the poly (amido amine) -based oligomer.

The present invention also provides a method for preparing a block copolymer comprising a tertiary amine group in which the poly (amido amine) -based oligomer is ionized at pH less than 7.0.

In another aspect, the present invention provides a method for producing a block copolymer of the block copolymer of the following formula.

Wherein k is a natural number in the range of 4 to 10, n is a natural number in the range of 11 to 45, and m is a natural number in the range of 10 to 100.

In another aspect, the present invention provides a method for producing a block copolymer having a molecular weight of 2,500 to 21,000 of the block copolymer.

The present invention also provides a method for producing a block copolymer having a molecular weight ratio of 1: 1 to 4 of the polyethylene glycol-based compound and the poly (amidoamine) -based oligomer.

The present invention also provides a method for producing a block copolymer in which the block copolymer forms a hydrogel at pH 7.0 to 7.4, and forms a sol at pH 3.0 to less than 7.0.

In addition, the present invention is a mixture of an amine group (-NH) and alkylene diacrylamide by mixing an amine-based compound and an alkylene diacrylamide, a precursor of a poly (amido amine) oligomer as a pH-sensitive compound to the imidazole polyethylene glycol compound Provided is a method for preparing a block copolymer prepared by coupling with a group (CH ₂ = CH-CONH-R-NHCO-CH = CH ₂ ).

In another aspect, the present invention provides a method for producing a block copolymer prepared in the reaction scheme to the imidazole polyethylene glycol compound.

Wherein k is a natural number in the range of 4 to 10, n is a natural number in the range of 11 to 45, and m is a natural number in the range of 10 to 100.

In another aspect, the present invention provides a method for producing a temperature- and pH-sensitive block copolymer, comprising: adding a carbonyldiimidazole to a polyethylene glycol having a molecular weight of 500 to 8,000 and forming a block copolymer having an imidazole bond; After the polyethylene glycol having an imidazole bond prepared above was put in a reactor at room temperature, chloroform was added to dissolve, and then trimethylenedipiperidine and 1,10 dekilene diol acrylamide were formed to form amidoamine blocks. A PAA-PEG-PAA triple block copolymer is prepared by adding at room temperature, wherein the polyethylene glycol block and the poly (amido amine) block have a molecular weight ratio of 1: 1 to 1: 4 and a long-term drug of 2,500 to 21,000. Process for producing temperature- and pH-sensitive block copolymers having excellent release properties.

In another aspect, the present invention provides a polymer hydrogel made of the block copolymer or a block copolymer prepared by the method.

The present invention also provides a drug carrier consisting of the block copolymer or a block copolymer prepared by the method.

In addition, the present invention is a block copolymer or a block copolymer prepared by the method; And it provides a drug carrier comprising a bioactive material that can be encapsulated in the block copolymer.

In addition, the present invention is a bioactive material that can be encapsulated in the block copolymer is insulin (insulin), leukocyte proliferation factor (G-SCF), human growth hormone (hGH), EPO, simlin (Symlin), exendin (Exendin) , Proteins such as Somatokine, Paclitaxal, Chlorambucil Interferon, Monoclonal antibodies, Vaccines, etc. In addition to the aforementioned ingredients, immunosuppressants, antiviral agents, anesthetics, antiemetics or antihistamines, and the like, conventional additives known in the art, such as excipients, stabilizers, pH adjusters, antioxidants, preservatives, binders or disintegrants, etc. In this case, the composition provides a drug carrier that may additionally include other additives, solvents and the like known in the art.

The present invention also provides a drug delivery agent in the form of the carrier is oral or parenteral, preferably in the form of intravenous, intramuscular or subcutaneous injection.

Hereinafter, exemplary embodiments of the present invention will be described in detail with reference to the accompanying drawings. First of all, it should be noted that in the drawings, the same components or parts denote 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.

As used herein, the terms "about", "relatively", and the like, are used at, or in close proximity to, numerical values when manufacturing and material tolerances inherent in the meanings indicated are intended to aid the understanding of the invention. Accurate or absolute figures are used to assist in the prevention of unfair use by unscrupulous infringers.

The present inventors have described the above-described case of using a temperature-sensitive block copolymer hydrogel composed of a polyethylene glycol series compound as a hydrophilic block and a hydrophobic block alone with polylactide, polyglycolide and polycaflolactone, and copolymers thereof. As described above, by-products decomposed in the process of biodegradation at the time of human injection are sufficient to cause various side effects in the process of accumulating in the body or discharged out of the body. Temperature- and pH-sensitive blocks consisting solely of glycol-based compounds and poly (amido amines) (PAAs), which exhibit hydrophilicity and hydrophobicity by the degree of ionization according to pH changes, and are completely released in vitro by biodegradation during human injection And the block polymer by synthesizing a copolymer address the above problems found in the temperature-sensitive hydrogel in the foregoing, thereby completing the present invention. Indeed, the prepared block copolymer hydrogels exhibit sol-gel transition properties at specific pH, thereby gelling around pH 7.0 to 7.4, similar to the body, and sollation below these ranges, resulting in conventional temperature sensitive hydrogels when injected into the body. It was found that the gel formed safely in the body without the occurrence of clogging of the needle, which can be seen in the present invention, and the prepared block copolymer can be applied as a drug release carrier capable of target release at a specific temperature and a specific pH. Found that it can. On the other hand, the pH-sensitive polymer is biodegraded in the human body so that the main chain is biodegraded in the human body by coupling the components consisting of ester bonds with pH sensitivity without directly using hydrophobic polymers so as to be completely discharged in vitro. It was possible to produce a hydrogel that is harmless to the human body, but it is limited to be applied to drug carriers that require long-term sustained release from 1 month to 3 months. Therefore, poly (amidoamine) having an amide bond in the main chain is used. Coupling could solve this problem.

It can be formed of a triblock of the temperature and pH sensitive block copolymer PAA-PEG-PAA according to the invention.

One of the components of the temperature and pH sensitive block copolymers according to the invention is a polyethylene glycol (PEG) family. PEG-based compounds constituting the copolymer (A) can be used without limitation conventional PEG-based compounds known in the art, in particular, PEG-based compounds represented by the formula (1) is preferred.

Wherein R is a hydrogen atom or an alkyl group having 1 to 5 carbon atoms, and n is a natural number in the range of 11 to 45.

The molecular weight (Mn) of the polyethylene glycol-based compound is not particularly limited, but is preferably in the range of 500 to 8,000. In particular, when R is hydrogen in polyethylene glycol (PEG) in Formula 1, the molecular weight range of 1,000 to 2,000 is preferable, and the molecular weight of methoxy polyethylene glycol (MPEG) in which R is a methyl group is preferably in the range of 1,000 to 5,000. When the molecular weight (Mn) of the polyethyleneglycol-based compound is outside the above range, that is, less than 500 and more than 8,000, the gel formation is not well achieved, and even if the gel is formed, the actual drug may be due to weakening of the gel strength or the like. It can be difficult to apply as a carrier for delivery.

The above-mentioned polyethylene glycol-based compound is not particularly limited as long as it contains the above components, but if possible, the sock end may be imidazole to react with the primary amine group, secondary amine group, etc. of the poly (amido amine) series block. Preferably formed.

As one of the components of the temperature and pH sensitive block copolymer according to the present invention, any compound exhibiting various ionization degrees depending on pH may be used without particular limitation, and in particular, a poly (amido amine) compound having both hydrophobicity and pH sensitivity Oligomers formed from are preferred.

On the other hand, the poly (amido amine) oligomer of the precursor further comprises one or more substituents selected from the group consisting of a primary amine group, a secondary amine group and alkylene diacrylamide can react with the block of the polyethylene glycol-based compound It can react with a polyethyleneglycol type compound in the form.

Poly (amido amine) compounds may exhibit sensitivity to pH in the body by including tertiary amine groups that ionize below pH 7.0. The poly (amido amine) -based oligomers more preferably include functional groups such as hydroxy group (-OH), carboxy group (-COOH) or amine group (-NH ₂ ), which is a block according to the present invention by polymerization In order to manufacture a copolymer easily.

The oligomers of the poly (amido amine) (PAA) series have ionization characteristics of varying solubility in water according to pH due to the tertiary amine groups present in them, thereby forming a hydrogel according to the pH change in the body as described above. Or sol. The compounds may be prepared according to conventional methods known in the art, for example, by polymerizing an amine-based compound to an alkylene diacrylamide compound having a double bond through a Michael reaction, Oligomers of the (amido amine) series can be obtained.

The alkylene diacrylamide compound used herein may be represented by the following Chemical Formula 2, and non-limiting examples thereof include N, N-methylene bisacrylamide (MDA), N, N-ethylene bisacrylamide, N-yi At least one in the group consisting of propyl acrylamide, 1,4 butylene diacrylamide, 1,6 hexylene diacrylamide, 1,8 octylene diacrylamide, 1,10 decaned diacrylamide, or mixtures thereof It may be selected.

In the above formula, R is an alkyl group having 1 to 20 carbon atoms.

In addition, the amine-based compound may be used without limitation as long as it has an amine group, and particularly preferred is a primary amine represented by the following formula (3), a secondary amine-containing diamine compound represented by the formula (4), or a mixture thereof.

H ₂ NR ₁ -NH ₂

In the above formula, R ₁ and R ₂ are alkyl groups having 1 to 20 carbon atoms.

Non-limiting examples of the primary amine compound include 4,4-trimethyldipiperidine, 3-methyl-4- (3-methylphenyl) piperazine, 3 methylpiperazine, 4- (bis- (fluorophenyl) methyl Piperazine, 4- (ethoxycarbonylmethyl) piperazine, 4- (phenylmethyl) piperazine, 4- (1-phenylethyl) piperazine, 4- (1,1-dimethoxycarbonyl) piperazine , 4- (2- (bis- (2-propenyl) amino) ethyl) piperazine, methylamine, ethylamine, butylamine, hexylamine, 2-ethylhexylamine, 2-piperidin-1-yl-ethyl It may be one or more selected from the group consisting of an amine, C-aziridin-1-yl-methylamine, and non-limiting examples of the secondary amine-containing diamine compound include piperazine, 4,4-trimethylene dipiperidine, It may be one or more selected from the group consisting of N, N'-dimethyl ethylene diamine, N, N'-diethyl ethylene diamine, imidazolidine and diazepan.

In preparing a poly (amido amine) oligomer exhibiting pH sensitivity, the reaction molar ratio of the alkylene diacrylamide compound and the amine compound is preferably in the range of 1: 0.5 to 4.0. When the molar ratio of the amine-based compound is less than 0.5 or more than 4.0, the molecular weight distribution is widened, the pH sensitivity is lowered, and the block length of the block copolymer is difficult to control.

The molecular weight of the poly (amido amine) oligomer is not particularly limited but is preferably in the range of 1,000 to 5,000. If the molecular weight is less than 1,000, the sol-gel transition behavior does not appear according to the pH change, and if it exceeds 5,000, the pH sensitivity is deteriorated.

The block copolymer of the present invention formed by coupling the aforementioned components, that is, a PEG-based compound (A) and a poly (amido amine) -based oligomer (B), is preferably a triple block copolymer. In particular, it may be represented by the following formula (5).

Wherein k is a natural number in the range of 4 to 10, n is a natural number in the range of 11 to 45, and m is a natural number in the range of 10 to 100.

The block copolymer represented by Chemical Formula 5 may form a hydrogel or maintain a sol state according to pH change due to the above-described amphipathicity and pH sensitivity, and in particular, uses that require sensitivity according to pH change in the body, such as drug release. A satisfactory result can be obtained, for example, for a carrier or diagnostic application.

The molecular weight range of the block copolymer is not particularly limited, but about 2,500 to 21,000 is suitable. In order to prepare a temperature-sensitive block copolymer, polyethylene glycol having a molecular weight of 500 to 8,000 is used. In this case, the molecular weight ratio of the hydrophilic block and the hydrophobic block is preferably 1: 1. If the ratio is less than 1: 1, the gel is not formed. If the ratio is greater than 1: 4, the strength of the hydrophobicity increases, which causes a problem that the block copolymer is not dissolved in water.

 The temperature and pH sensitive block copolymers according to the present invention may include other components or additives commonly used in addition to the aforementioned components.

In order to prepare the temperature and pH sensitive block copolymer according to the present invention using the PEG-based compound and the poly (amido amine) -based oligomer, sugar techniques such as Mitchell polymerization, radical polymerization, cationic polymerization, anionic polymerization, condensation polymerization, etc. Any one of the various polymerization methods known in the art or a combination method can be used.

Hereinafter, when PEG is described as an example in the method for preparing a temperature and pH sensitive block copolymer according to the present invention, a) reacting a PEG-based compound with carbonyldiimidazole (N, N'-carbonyldiimidazole); b) coupling the resulting diimidazole PEG to a poly (amido amine) oligomer.

First, a PEG precursor having an imidazole group introduced therein for coupling a PEG-based compound and a poly (amido amine) oligomer was prepared, and the reaction thereof may be illustrated by Scheme 1 below. In other words, a PEG-based compound is reacted with carbonyldiimidazole to prepare an imidazole PEG precursor having an imidazole bond at the sock end.

Where n is a natural number ranging from 11 to 45.

The amine group is mixed with the diimidazole PEG compound prepared as in Scheme 1 by mixing an amine-based compound (for example, dipiperidine) and alkylene diacrylamide as a precursor of a poly (amido amine) oligomer which is a pH sensitive compound. The temperature and pH sensitive triple block copolymers according to the present invention were prepared through coupling between (-NH) and an alkylenediacrylamide group (CH ₂ = CH-CONH-R-NHCO-CH = CH ₂ ). It may be prepared, the reaction thereof may be exemplified by Scheme 2.

Wherein k is a natural number in the range of 4 to 10, n is a natural number in the range of 11 to 45, and m is a natural number in the range of 10 to 100.

There is no particular limitation on the reaction temperature and time, which are reaction conditions of the reaction.

The primary amine compound and the secondary amine containing diamine compound used for preparing the poly (amido amine) oligomer are the same as described above. In addition, the alkylene diacrylamide-based compound that reacts with this to form an amido amine block includes N, N-methylene bisacrylamide (MDA), N, N-ethylene bisacrylamide, N-isopropylacrylamide, Preference is given to compounds of the same class as 1,4 butylene diacrylamide, 1,6 hexylenediacrylamide, 1,8 octylene diacrylamide, 1,10 decane diacrylamide or derivatives of these compounds.

As described above, the triple block copolymer prepared by the above method is a hydrophilic block, a hydrophobic block, and a poly (amido amine) -based oligomer which shows various ionization degrees according to pH change, and thus is partially bonded. At the same time pH sensitivity can be shown.

Indeed, the poly (amido amine) -polyethyleneglycol-poly (amido amine) triple block copolymers prepared in this way utilize FT-IR and ¹ H-NMR to introduce the respective functional groups and The reaction was confirmed, and it was confirmed that the copolymer of the PEG-based compound and the biodegradable polymer and the beta amino ester-based oligomer were coupled by increasing the molecular weight of the block copolymer using GPC (Gel Permeation Chromatography). In addition, in order to confirm the pH sensitivity, the change in the sol-gel transition properties was measured while changing the pH according to the temperature, it was confirmed that the triblock copolymer of the present invention has a pH sensitive properties. (See Figure 1)

In addition, the present invention provides a composition comprising (a) a temperature and pH sensitive block copolymer; And (b) it may be prepared in the form of a polymer hydrogel (hydrogel) drug composition comprising a bioactive material that can be encapsulated in the block copolymer.

Bioactive materials that can be encapsulated in the polymer hydrogel block copolymer may be used without particular limitation, and non-limiting examples thereof include insulin, leukocyte proliferation factor (G-SCF), human growth hormone (hGH), and EPO. Anti-cancer agents, such as proteins such as Symlin, Exendin, Somatokine, Paclitaxal, Chlorambucil Interferon, Monoclonal antibodies, Vaccines, etc. , Steroids, anti-inflammatory drugs, sex hormones, immunosuppressants, antiviral agents, anesthetics, antiemetic or antihistamines. In addition to the aforementioned components, it may also include conventional additives known in the art such as excipients, stabilizers, pH adjusters, antioxidants, preservatives, binders or disintegrants and the like. In this case, the composition may additionally include other additives, solvents, and the like known in the art.

In addition, the polymer hydrogel drug composition may be used in the form of oral or parenteral preparations, and may be prepared by intravenous, intramuscular or subcutaneous injection.

In addition, the present invention provides a method of using the temperature and pH sensitive block copolymer as a carrier for drug delivery or disease diagnosis. At this time, the material contained in the block copolymer is not particularly limited as long as it is a material for the treatment, prevention or diagnosis of the disease.

As described above, the injection-type pH-sensitive block copolymer hydrogel having excellent human safety according to the present invention and a method for preparing the same have biodegradability in order to drastically improve the toxicity of the biodegradable degradation products upon injection in human body, Compounds composed of poly (amido amine) (PAA) which show hydrophilicity and hydrophobicity by the degree of ionization according to the pH change to the hydrophilic polymer without using any polyester-based hydrophobic polymer which accumulates enough to cause various side effects. By the coupling reaction, in addition to the temperature sensitivity, the pH sensitivity is also given at the same time to solve the above-mentioned problems of the temperature sensitive hydrogel, as well as to form a more stable hydrogel.

In addition, the hydrogel according to the present invention and a method for preparing the same form a stable hydrogel at a specific pH, for example, pH 7.0 to 7.4, which is a pH range of normal cells in the body, and a pH range of 3.0 to 7.0 indicated by abnormal cells such as cancer cells. If less, it can be used as a target-oriented drug release carrier for cancer cells by keeping it in a sol state. In other words, due to the increased degree of ionization of the tertiary amine present in the poly (amido amine) (PAA) at low pH (less than pH 7.0), the entire PAA becomes water-soluble and remains in a sol state. The degree of ionization is lowered to show hydrophobic properties, forming a gel state. Due to the above properties, the multiblock copolymers of the present invention can exhibit sol-gel transition properties that are sensitive to temperature as well as pH.

In addition, the block copolymer according to the present invention and a method for preparing the same may be formed of a poly (amido amine) -polyethylene glycol-poly (amido amine) triple block copolymer, thereby controlling the biodegradation rate by controlling the molecular weight and the like of each compound. The poly (amido amine) block having an amide bond in the main chain can be used as a drug carrier that shows long-term orthotropic drug release behavior of 1 month to 3 months.

In addition, the block copolymer and the method for preparing the same according to the present invention can be applied as a sustained release drug carrier for medical, gene delivery, drug delivery, in particular, drug loading and release because the temperature and pH sensitive block copolymer is safe in the body. In addition, by delivering a substance for diagnosis of a disease to abnormal cells, it can be applied to diagnostic applications such as diagnostic imaging (diagnostic imaging).

In addition, the block copolymer according to the present invention and a method for preparing the same form a hydrogel in the pH range of 7.0 to 7.4, which is the same as a normal body condition, and maintain a sol state at pH 7.0, which is an abnormal condition such as cancer cells, to achieve cancer cell target-oriented design. Although applied, by appropriately changing the components of the block copolymer, their molar ratios, molecular weight and / or functional groups in the block can be usefully applied to target-oriented design not only cancer cells but also genetic variation or other applications.

Hereinafter, the present invention will be described as examples.

[ Example  1 to 8. Temperature and pH  Sensitive Triblock Copolymer Synthesis]

Example  One. Poly (Amido amine) Polyethylene glycol - Poly (Amido amine) block copolymer ( PAA -PEG-PAA) Manufacturing

10 g of polyethylene glycol (PEG 2,000, Mn = 2,000) and 0.2 g of stannous octoate as a catalyst were placed in a reactor and vacuum dried at 110 ° C. for 4 hours to remove moisture. 0.2g of carbonyldiimidazole was added to 0.1g of vacuum-dried polyethyleneglycol, and polyethyleneglycol having imidazole bond was obtained at the sock end. The reaction was carried out in an ice-bath in a nitrogen environment and the reaction time was 24 hours. After completion of the reaction, precipitated in ethyl ether to remove the unreacted product, filtered and dried in vacuo at room temperature to obtain polyethylene glycol having imidazole bond at the end group, yield was more than 70%.

Meanwhile, polyethylene glycol having an imidazole bond prepared above was put in a reactor at room temperature, and then dissolved by addition of chloroform, followed by trimethylenedipiperidine and 1,10 to form an amido amine (Mn = 2,000) block. Tequylenediacrylamide was added and dissolved at room temperature, and then reacted at 60 ° C. for 72 hours. After completion of the reaction, precipitated in excess ethyl ether to remove unreacted material and then filtered to block poly (amidoamine). The molecular weight of 4,000 was obtained, and the molecular weight of the polyethylene glycol block was 2,000 to obtain a triple block copolymer having a number average molecular weight of 6,000 of the overall PAA-PEG-PAA block copolymer, and the yield was more than 70%.

Example  2

A PAA-PEG-PAA block copolymer having a molecular weight of 5,000 was obtained by performing the same method as Example 1, except that the molecular weight of the PEG block was 1,000, and the yield was 70% or more.

Example  3

A PAA-PEG-PAA block copolymer having a molecular weight of 7,000 was obtained in the same manner as in Example 1, except that the molecular weight of the PEG block was 3,000, and the yield was 70% or more.

Example  4

A PAA-PEG-PAA block copolymer having a molecular weight of 8,000 was obtained by performing the same method as Example 1 except that the molecular weight of the PEG block was 4,000, and the yield was 65% or more.

Example  5

A PAA-PEG-PAA block copolymer having a molecular weight of 9,000 was obtained by performing the same method as Example 1 except that the molecular weight of the PEG block was 5,000, and the yield was 65% or more.

Example  6

A PAA-PEG-PAA block copolymer having a molecular weight of 5,000 was obtained by following the same method as Example 1, except that the poly (amido amine) block had a molecular weight of 3,000, and the yield was 70% or more.

Example  7

A PAA-PEG-PAA block copolymer having a molecular weight of 7,000 was obtained in the same manner as in Example 1, except that the poly (amido amine) block had a molecular weight of 5,000, and the yield was 65% or more.

Example  8

Except for having a molecular weight of 7,000 poly (amido amine) block, the same method as in Example 1 was carried out to obtain a PAA-PEG-PAA block copolymer having a molecular weight of 9,000, the yield was 65% or more.

Comparative example  One

A PAA-PEG-PAA block copolymer having a molecular weight of 4400 was obtained by following the same method as Example 1 except that the PEG molecular weight was 400, and the yield was 45% or more.

Comparative example  2

Except for having a PEG molecular weight of 9,000, PAA-PEG-PAA block copolymer having a molecular weight of 13,000 was obtained in the same manner as in Example 1, and the yield was 45% or more.

Comparative example  3

A PAA-PEG-PAA block copolymer having a molecular weight of 3,000 was obtained in the same manner as in Example 1, except that the poly (amido amine) block had a molecular weight of 1,000, and the yield was 50% or more.

Comparative example  4

A PAA-PEG-PAA block copolymer having a molecular weight of 7,000 was obtained in the same manner as in Example 1 except that a poly (amido amine) molecular weight of 3,000 was used, and the yield was 50% or more.

Comparative example  5

The same method as in Example 1 was performed except that polyethyleneimine having a molecular weight of 2,000 was used to compare the cytotoxicity of poly (amido amine), a pH sensitive component.

Comparative Example 6

In order to verify the effect of the initial over-release of the drug by the ionic complex of the drug, the drug was released in a PLGA-PEG-PLGA triple block copolymer containing no ionic groups to form the ionic complex with the drug. Was performed

Experimental Example  One. pH  Sol-gel according to the change Transition behavior  evaluation

Evaluation of the sol-gel transition behavior according to the temperature and pH change of the block copolymer prepared according to the present invention was performed.

10 wt% of the triple block copolymer (PAA-PEG-PAA) prepared in Examples 1 to 8 was dissolved in a buffer solution, and then titrated with NaOH solution at 50 ° C. to pH 5.5, 6.0, 6.5, 7.0, and 7.5, respectively. Adjusted to. Tri-block copolymer solutions having respective pHs were allowed to equilibrate under constant temperature for 10 minutes while raising 2 ° C., and then sol-gel transition behavior was measured by tilting each solution. The sol-gel transition behavior of the block copolymers with temperature and pH changes is shown in FIG. 1.

As shown in FIG. 1, the block copolymer of the present invention has a sol-gel transition behavior depending on pH as well as temperature due to a change in pH and hydrophobicity of the poly (amidoamine) -based oligomer in the copolymer. It was confirmed that this is done reversibly.

Experiment 2. Example  And Comparative Example  Cytotoxicity Assessment

 Cell line NIH 3T3 (fibroblast) method was used, and medium was used as DMEM (90% Dulbecco's modified Eagle's medium, 10% fetal calf serum, penicillin 100 units / mL, streptomycin 100 ㎍ / mL). XTT analysis revealed 2,3-bis (2-methoxy-4-nitro-5-suspophenyl) -2H-tetrazolib-5-5-carboxanilide (2,3-bis (2-methoxy-4-nitro). -5-susfophenyl) -2H-tetrazolium-5-carbox anilide) and analyzed by a microplate reader using 96-well plates, incubator.

As a result of the evaluation, as shown in FIG. 2, cell survival rate was improved by 10 to 50% compared to the comparative examples.

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.

1 is a sol- according to the temperature and pH change of the triple block copolymer composed of a polyethylene glycol-based compound having a temperature sensitivity and a biodegradable poly (amido amine) compound having a pH sensitivity according to an embodiment of the present invention Graph showing gel transition behavior.

Figure 2 is a graph showing the cytotoxicity analysis results of Examples 1, 3 and 6 and Comparative Example 5.

3 is a graph showing the results of decreasing the concentration of the drug with time indicating the drug release rate of Examples 1, 3, 6 and Comparative Example 6

Claims (43)

For temperature and pH sensitive block copolymers, (a) polyethylene glycol series compounds (A); And (b) A temperature and pH sensitive block copolymer having excellent long term drug release properties comprising a coupling body of a poly (amido amine) based oligomer (B) mixture. The method of claim 1, The copolymer is a block copolymer comprising a triblock of B-A-B. The method of claim 1, The polyethylene glycol-based compound is a block copolymer of the following formula.

Wherein R is a hydrogen atom or an alkyl group having 1 to 5 carbon atoms, and n is a natural number in the range of 11 to 45. The method of claim 1, The molecular weight (Mn) of the polyethylene glycol compound is a block copolymer of 500 to 8,000. The method of claim 3, Block copolymer having a molecular weight (Mn) of 1,000 to 2,000 when R in the formula of the compound is polyethylene glycol (PEG) is hydrogen. The method of claim 3, Block copolymer having a molecular weight (Mn) of 1,000 to 5,000 when R in the formula of the compound is methoxy polyethylene glycol (MPEG) is a methyl group. The method of claim 1, The poly (amido amine) -based precursor is a block copolymer further comprises one or more substituents selected from the group consisting of a primary amine group, a secondary amine group and alkylene diacrylamide can react with the polyethylene glycol-based compound. The method of claim 1, The poly (amido amine) -based oligomer further comprises a functional group of a hydroxyl group (-OH), carboxy group (-COOH) or amine group (-NH ₂ ). The method of claim 1, The poly (amido amine) -based oligomer is a block copolymer comprising a tertiary amine group that is ionized at less than pH 7.0. The method of claim 1, Block copolymer of the poly (amido amine) series oligomer has a molecular weight of 1,000 to 5,000. The method of claim 1, The block copolymer is a block copolymer consisting of the following formula.

Wherein k is a natural number in the range of 4 to 10, n is a natural number in the range of 11 to 45, and m is a natural number in the range of 10 to 100. The method of claim 1, The block copolymer has a molecular weight of 2,500 to 21,000. The method of claim 1, The molecular weight ratio of the A and B is 1: 1 to 4 block copolymer. The method of claim 1, The block copolymer forms a hydrogel at pH 7.0 to 7.4, and forms a sol state at a pH of 3.0 to less than 7.0. In the method of producing a temperature and pH sensitive block copolymer, a) forming a polyethylene glycol based block precursor; b) a process for producing a temperature- and pH-sensitive block copolymer having excellent long-term drug release property comprising the step of coupling the produced polyethylene glycol-based block precursor and poly (amido amine) -based oligomer. The method of claim 15, The polyethylene glycol-based block precursor is a method for producing a block copolymer in which the end of the polyethylene imidazole so as to react with the precursors of the poly (amido amine) block. The method of claim 15, The polyethylene glycol-based compound is a method for producing a block copolymer of the formula.

Wherein R is a hydrogen atom or an alkyl group having 1 to 5 carbon atoms, and n is a natural number in the range of 11 to 45. The method of claim 15, The molecular weight (Mn) of the polyethylene glycol-based compound is a method of producing a block copolymer of 500 to 8,000. The method of claim 17, Method of producing a block copolymer having a molecular weight (Mn) of 1,000 to 2,000 when R in the formula of the compound is polyethylene glycol (PEG) is hydrogen. The method of claim 17, Method of producing a block copolymer having a molecular weight (Mn) of 1,000 to 5,000 when R in the chemical formula of the compound is methoxy polyethylene glycol (MPEG) is a methyl group. The method of claim 15, Forming the polyethylene glycol-based precursor is a method for producing a block copolymer consisting of the following reaction formula to react the polyethylene glycol-based compound and carbonyldiimidazole (N, N-carbonyldiimidasole).

Where n is a natural number ranging from 11 to 45. The method of claim 15, The poly (amido amine) -based precursors in the block air further comprises one or more substituents selected from the group consisting of a primary amine group, a secondary amine group and alkylene diacrylamide can react with the block of the polyethylene glycol-based compound Process for the preparation of coalescing. The method of claim 15 The poly (amido amine) -based oligomer is a method for producing a block copolymer formed by polymerizing an alkylene diacrylamide compound and an amine-based compound. 24. The method of claim 23, The alkylene diacrylamide compound is a method of producing a block copolymer made of a chemical formula.

In the above formula, R is an alkyl group having 1 to 20 carbon atoms. The method of claim 24, The alkylene diacrylamide compound is N, N-methylene bisacrylamide (MDA), N, N-ethylene bisacrylamide, N-isopropylacrylamide, 1,4 butylene diacrylamide, 1,6 hex A method for producing a block copolymer selected from the group consisting of silenediacrylamide, 1,8 octylene diacrylamide, 1,10decanediacrylamide, or mixtures thereof. 24. The method of claim 23, The amine-based compound is a method for producing a block copolymer is a primary amine or a secondary amine-containing diamine compound or a mixture thereof. The method of claim 26, The primary amine compound may be 4,4'-trimethyldipiperidine, 3-methyl-4- (3-methylphenyl) piperazine, 3 methylpiperazine, 4- (bis- (fluorophenyl) methyl) piperazine, 4- (ethoxycarbonylmethyl) piperazine, 4- (phenylmethyl) piperazine, 4- (1-phenylethyl) piperazine, 4- (1,1-dimethoxycarbonyl) piperazine, 4- (2- (bis- (2-propenyl) amino) ethyl) piperazine, methylamine, ethylamine, butylamine, hexylamine, 2-ethylhexylamine, 2-piperidin-1-yl-ethylamine, C Aziridin-1-yl-methylamine, 4-aminomethylpiperidine (AMPD), N-methylethylenediamine (MEDA), N-ethylethylenediamine (EEDA), N-hexylethylenediamine (HEDA) and Process for preparing at least one block copolymer selected from the group consisting of 1- (2-aminoethyl) piperazine (AEPZ) or mixtures thereof The method of claim 26, The secondary amine containing diamine compounds include piperazine, 4,4'-trimethylene dipiperidine, N, N'-dimethyl ethylene diamine, N, N'-diethyl ethylene diamine, imidazolidine and dia A method for producing at least one block copolymer selected from the group consisting of plate making. 24. The method of claim 23, The reaction molar ratio of the alkylene diacrylamide compound and the amine compound is 1: 0.5 to 4.0 method for producing a block copolymer. The method of claim 15, Method of producing a block copolymer of the molecular weight of the poly (amido amine) -based oligomer is 1,000 to 5,000. The method of claim 15, The poly (amido amine) -based oligomer is a method for producing a block copolymer comprising a tertiary amine group ionized at less than pH 7.0. The method of claim 15, The block copolymer is a method for producing a block copolymer consisting of the following formula.

Wherein k is a natural number in the range of 4 to 10, n is a natural number in the range of 11 to 45, and m is a natural number in the range of 10 to 100. The method of claim 15, The molecular weight of the block copolymer is 2,500 to 21,000 method for producing a block copolymer. The method of claim 15, A molecular weight ratio of the polyethylene glycol series compound and the poly (amido amine) series oligomer is 1: 1 to 4. The method of claim 15, The block copolymer forms a hydrogel at pH 7.0 to 7.4, and forms a sol state at pH 3.0 to less than 7.0. The method of claim 15, An amine group (-NH) and an alkylenediacrylamide group (CH ₂ ) are mixed with the imidazole polyethyleneglycol compound by mixing an amine-based compound, which is a precursor of a poly (amido amine) oligomer, which is a pH sensitive compound, and an alkylene diacrylamide. = CH-CONH-R-NHCO-CH = CH ₂ ) A method for producing a block copolymer prepared by coupling (coupling) with. The method of claim 15, Method for producing a block copolymer prepared in the imidazole polyethylene glycol compound as shown in the following scheme.

Wherein k is a natural number in the range of 4 to 10, n is a natural number in the range of 11 to 45, and m is a natural number in the range of 10 to 100. In the method of producing a temperature and pH sensitive block copolymer, Adding a carbonyldiimidazole to polyethylene glycol having a molecular weight of 500 to 8,000, and then forming a block copolymer having an imidazole bond; After the polyethylene glycol having an imidazole bond prepared above was put in a reactor at room temperature, chloroform was added to dissolve, and then trimethylenedipiperidine and 1,10 dekilene diol acrylamide were formed to form amidoamine blocks. By adding at room temperature to prepare a PAA-PEG-PAA triple block copolymer, Method for producing a temperature and pH sensitive block copolymer having excellent long-term drug release property of the polyethylene glycol block and poly (amido amine) block ratio of 1: 1 to 1: 4 and the total molecular weight of 2,500 to 21,000. A polymer hydrogel comprising a block copolymer according to any one of claims 1 to 14 or a block copolymer prepared by the method according to any one of claims 15 to 38. A drug carrier comprising a block copolymer according to any one of claims 1 to 14 or a block copolymer prepared by the method according to any one of claims 15 to 38. A block copolymer prepared by the method according to any one of claims 1 to 14 or a method according to any one of claims 15 to 38; And Drug delivery system comprising a bioactive material that can be encapsulated in the block copolymer.      The method of claim 41, wherein Bioactive substances that can be encapsulated in the block copolymer are insulin (insulin), leukocyte growth factor (G-SCF), human growth hormone (hGH), EPO, simlin (Exendin), somatokin ( Proteins such as Somatokine, Paclitaxal, Chlorambucil Interferon, Monoclonal antibodies, Vaccines, etc. Antiviral agents, anesthetics, antiemetics or antihistamines and the like, in addition to the aforementioned components, may include conventional additives known in the art, such as excipients, stabilizers, pH adjusting agents, antioxidants, preservatives, binders or disintegrants, etc. The composition may further comprise other additives, solvents and the like known in the art. The method of claim 41, wherein The carrier is an oral or parenteral drug, preferably in the form of intravenous, intramuscular or subcutaneous injection.

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