WO2013089491A1

WO2013089491A1 - Multiblock-copolymer aqueous solution composition with improved-stability

Info

Publication number: WO2013089491A1
Application number: PCT/KR2012/010923
Authority: WO
Inventors: 김봉오; 서민효
Original assignee: 주식회사 삼양바이오팜
Priority date: 2011-12-16
Filing date: 2012-12-14
Publication date: 2013-06-20
Also published as: KR20130069196A; KR101466893B1

Abstract

The present invention relates to: an aqueous solution composition wherein the stability of the sol state of a multiblock-copolymer having sol-gel transition characteristics can be improved; and a stabilisation method therefor. More specifically, the present invention relates to: a multiblock-copolymer aqueous solution composition for refrigerated storage, which comprises a multiblock copolymer dissolved in distilled water or a buffer solution, and in which the multiblock copolymer has a molecular weight reduced by no more than 30% when stored for 2 years; and a stabilisation method therefor.

Description

【Specification】

[Name of invention]

Aqueous Composition of Multiblock Copolymers with Improved Stability

The present invention relates to an aqueous solution composition in which the sol state of a multiblock copolymer having sol-gel transition properties can have improved stability and to a stabilization method thereof. More specifically, the present invention relates to a multi-block copolymer aqueous solution composition for cold storage containing a distilled water or a complete solution in which the multi-block copolymer is dissolved and having a reduced amount of the molecular weight of the multi-block copolymer in two years or less and a stabilization method thereof. It is about.

Background Art

Block copolymers composed of polyethylene oxide and polypropylene oxide form gels when the polyethylene oxide and polypropylene oxide polymers absorb water at certain concentrations and temperatures (US patent). 4,188, 373, 4,478,822 and 4,474,751. Poloxamer is well known as an example of the polymer. Commonly known poloxamers have the structure of triblock copolymers of polyethylene oxide (PE0) -polypropylene oxide (PP0) and / or polybutylene oxide (PB0) -polyethylene oxide (PE0) and are in solution at low temperatures. One is a thermosensitive material that gels when the temperature rises. Poloxamers have already been commercialized by BASF and used as drug carriers and anti-adhesion agents.

However, poloxamers form a polymer gel in an aqueous solution, but have a disadvantage in that they do not stay in one place for a sufficient time to prevent drug delivery and adhesion due to weak gel strength.

In order to solve this problem, the present inventors increase the molecular weight while maintaining the constituent ratio of poloxamer and sol-gel transition phenomenon. As a method, a method of synthesizing a multiblock co-polymer comprising a poloxamer ABA-type triblock as a unit block using a chain extender was developed and obtained a patent (Domestic Patent Registration No. 10-0835738). The patent provides multiblock copolymers comprising two or more ABA-type triple blocks covalently bonded with a biodegradable dicarboxyl linker. A is a polyethylene oxide block, B represents a polypropylene oxide block, a polybutylene oxide block or a complex thereof, and the multiblock copolymer includes MX at both ends, and M is H or a cationic group. And X is an anionic group. For example, the patent provides a multiblock copolymer represented by the following formula:

M-X-0- [PE0-Y-PE0-C (= 0) -R-C (O) -0] η-ΡΕΟ-Υ-ΡΕΟ-0-Χ-Μ

In the above, PE0 is a polyethylene oxide block,

Y is PP0 or PB0, or a complex of PP0 and PB0,

PP0 is a polypropylene oxide block,

PB0 is a polybutylene oxide block,

^' X is H or an anion group,

n is an integer between 1 and 100,

R is aryl containing-(CH ₂ ) m- or Cm ',

m is an integer from 0 to 20,

tn 'is an integer between 6 and 12, and

M is H or a cationic group, provided that both M and X are not H, and M is absent when X is H.

The multiblock copolymers thus synthesized can be used in many applications such as drug delivery systems and anti-adhesion agents. However, since the ester bond formed by the reaction of the chain extender and the multiblock copolymer applied to increase the molecular weight is hydrolyzed in the aqueous solution, the ester bond is hydrolyzed when the multiblock copolymer is stored in the aqueous solution for a long time. As the molecular weight decreases, the strength of the gel formed by the micelle or body temperature of the micelle is reduced, so the desired physical properties are expected. It is difficult. Therefore, the final product requiring product stability by long-term storage is released in the solid state, not the aqueous state.

Accordingly, in using the multiblock copolymer as a drug carrier and an anti-adhesion agent, the multiblock copolymer and the drug are dissolved in distilled water, lyophilized and stored in a solid state, and then dissolved again in distilled water immediately before use. There is a feeling. In addition, since the multiblock copolymer has a large molecular weight, it takes a long time to dissolve in distilled water, which is inconvenient to be dissolved in an aqueous solution at least 24 hours before. In addition, the film-type adhesion inhibitors do not adhere well to the biological surface when applied to the internal organs, there is a problem that the effect of preventing the adhesion of organs is insufficient due to agglomeration with each other in the tissue itself.

Therefore, if the final product can be provided in an aqueous solution state by improving the stability of the multiblock copolymer in an aqueous solution state, it is expected to be more easily applicable as a drug carrier and an anti-adhesion agent, and further expansion of the field of application is expected.

[Detailed Description of the Invention]

[Technical problem]

Therefore, the present inventors have studied to develop a method for improving the stability of a multi-block copolymer aqueous solution and to develop a stabilized multi-block copolymer aqueous solution composition. By confirming that the stability of the aqueous solution state can be improved even during long-term storage, the present invention was completed.

Technical Solution

It is an object of the present invention to provide a multiblock copolymer aqueous solution composition which ensures stability during long-term storage in aqueous solution.

Specifically, one object of the present invention is a multi-block copolymer aqueous solution comprising the step of dissolving the multi-block copolymer in distilled water or complete solution, and the step of storing the dissolved multi-block copolymer in storage conditions It is to provide a method of stabilizing the composition.

It is another object of the present invention to provide a multiblock copolymer aqueous solution composition stabilized by the above method.

It is another object of the present invention to provide a multiblock copolymer aqueous solution composition for storage of distilled water containing distilled water or buffer solution in which the multiblock copolymer molecular weight is reduced by 30% or less when stored for 2 years. will be.

It is another object of the present invention to provide a pharmaceutical composition comprising the multiblock copolymer aqueous solution composition and a bioactive agent.

[Brief Description of Drawings]

Figure 1 shows the sol-gel transition under the conditions of dissolving the multiblock copolymer in distilled water (10 w / v%) at room temperature (Comparative Example 1) (M means month).

Figure 2 shows the sol-gel transition under the conditions of dissolving the multiblock copolymer in a buffer solution (10 w / v%) at room temperature (Comparative Example 2) (M means month).

Figure 3 shows the sol-gel transition under conditions (Example 1) in which the multiblock copolymer is dissolved (10 w / v%) in distilled water and stored for a long time (M means month).

FIG. 4 shows the sol-gel transition under the conditions of dissolving the multiblock copolymer in a complete solution (10 w / v%) and refrigerated (Example 2) (M means month).

FIG. 5 shows the sol-gel transition under the conditions in which the multiblock copolymer is dissolved (15 w / v%) in a complete solution and stored for a long time (Example 3) (M means month).

[Best form for implementation of the invention]

In order to achieve the above object, the present invention as one aspect (a) dissolving a multiblock copolymer comprising two or more ABA-type triblocks covalently bonded with a biodegradable dicarboxyl linker in distilled water or a complete solution,

A is a polyethylene oxide block,

B is a polypropylene oxide block, a polybutylene oxide block or a composite thereof, and

The multiblock copolymer comprises M-X at both ends, M is H or a cationic group, and X is an anionic group; And

(b) storing the dissolved multiblock copolymer under storage conditions;

A method for stabilizing a multiblock copolymer aqueous solution composition. In a preferred embodiment, the multiblock copolymer can be represented by the following formula.

M-X-0- [PE0-Y-PE0-C (= 0) -R-C (= 0) -0] η-ΡΕΟ-Υ-ΡΕΟ-0-Χ-Μ

In the above, PE0 is a polyethylene oxide block,

Y is PP0 or PB or a complex of PP0 and PB0,

PP0 is polypropylene oxide block,

PB0 is a polybutylene oxide block,

X is H or an anion group,

n is an integer between 1 and 100,

Is aryl containing-(C¾) m- or Cm ',

m is an integer from 0 to 20,

m 'is an integer between 6 and 12, and

Also, more preferably, wherein X may be a negative group selected from the group consisting of -S0 ₃ —, -P0 ₃ ² —, and -C (= 0) -RC (= 0) —0—, wherein M May be a cationic group selected from the group consisting of Li, Na, K, Ag, Au, Ca, Mg, Zn, Fe, Cu, Co, and Ni, The unit ratio of the PEO and Y may be 0.2: 1 to 40: 1, the weight average molecular weight of the Υ may be in the range of 1,000 to 20,000 Daltons, more preferably may be poloxamer,

R is selected from the group consisting of oxalic acid, malonic acid, malic acid, succinic acid, glutaric acid, adipic acid, pimelic acid, sebacolic acid, suberic acid, dodecanoic acid, fumaric acid, maleic acid, phthalic acid, and terephthalic acid. selected from those may be a biodegradable di-carboxyl groups derived linker, ^"

The weight average molecular weight of the multiblock copolymer may range from 40,000 to 1,000,000 Daltons (Dalton),

The content of the multiblock copolymer may be 0.1 to 50 w / v, more preferably 5 to 25 w / v%.

In a preferred embodiment, the buffer solution in the present invention is glycine buffer (Clycine buffer), citrate buffer (Citrate buffer), succinate buffer (Succinate buffer), malate complete solution (Mai eat e buffer), acetate complete layer Acetate buffer, MOPS buffer (3- (N-morphol ino) propanesul fonic acid buffer), MES buffer solution (2- (N-morpholino) ethanesulfonic acid buffer), PIPES complete solution (2- [4- ( 2-sul foethyl) piperazin-l-yl lethanesul fonic acid buffer), TES complete solution (N-Tris (hydroxymethyl) methyl-2-aminoethanesul fonic acid buffer), bicine complete layer solution (Ν, Ν-bis (2-Hydroxyhethyl glycine), phosphate buffer, HEPES buffer

(4- (2-hydroxyoxyt hy l) -l-piperazineet hane su 1 f on ic acid buffer), Tricine buffer, Tris (hydroxymethyl) am inome thane buffer), TABS It may be selected from the group consisting of a mild solution (N ᅳ tris (hydroxymethyl) -4-amino-butanesul fonic acid buffer) and a cacodylate buffer (Cacodylate buffer).

As a preferred mode, refrigeration conditions in the present invention can range from 0 to 10 ° C, more preferably from 2 to 5 ⁰ C. In addition, the pH of the aqueous solution of the multiblock copolymer may be 3 to 8. As another aspect, the present invention relates to a multi-block copolymer aqueous solution composition stabilized by the above method.

As another aspect, the present invention comprises a multi-block copolymer solubilized distilled water or buffer solution containing two or more ABA-type triblock covalently bonded with a biodegradable dicarboxyl group linker,

A is a polyethylene oxide block,

The multiblock copolymer includes M-X at both ends, M is H or a cationic group, and X is an anionic group,

The present invention relates to a multi-block copolymer aqueous solution composition for storage of storage, wherein the amount of reduction of the molecular weight of the multiblock co-polymer when stored for two years under refrigeration is 30% or less. Preferably, the reduced amount of the multiblock copolymer molecular weight in the aqueous solution is 20% or less for 6 months storage, 25% or less for 1 year storage and / or 30% or less for 2 years storage under the storage conditions.

At this time, a preferred embodiment of the multiblock copolymer is as described above.

As another aspect, the present invention relates to a pharmaceutical composition comprising the multi-block copolymer aqueous solution composition for cold storage and a bioactive agent.

Preferably the bioactive agent may be in the concentration range of 0.01 to 50 / \ ^.

Also preferably, the bioactive agent may be a protein or a peptide,

More preferably, the bioactive agent is a growth hormone (growth hormone,

GH), interferon (IFN), granulocyte colony stimulation factor (G-CSF), granulocyte macrophage colony stimulation factor (GMCSF), erythropoietin (EPO), interleukin (interleukin) , ID, follicle-promoting hormone (fibroblast growth factor, follicle stimulating hormone (FSH), nerve growth factor (NGF), octreotide, insulin, insulin-like growth factor (IGF), calcitonin (calcitonin), tumor necrosis factor ( tumor necrosis factor (TNF), vascular endothel ial growth factor (VEGF), epidermal growth factor (EGF), platelet growth factor (pi ate let-derived growth factor (PDGF)), bone formation factor (bone morphogenetic protein (BMP)), thrombolytics (tissue lasminogen activator (TPA), thrombopoietin (TPO), tissue growth factor (TGF), and tumor necrosis factor (TNF) It may be a protein selected from the group consisting of.

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

The present inventors confirmed that the multiblock copolymer was dissolved in distilled water or a complete solution, and then stored at the storage conditions to improve stability of the aqueous solution even during long-term storage.

The multiblock copolymer provided by the present inventors in Korean Patent No. 10-0835738 is in the form of a multiblock copolymer comprising two or more ABA-type triblocks covalently bonded with a biodegradable dicarboxyl linker ( Wherein A is a polyethylene oxide block, B is a polypropylene oxide block, a polybutylene oxide block or a complex thereof, and the multiblock copolymer includes MX at both ends, and M is H or a cationic group. , And X is an anionic group), when the multiblock copolymer is hydrolyzed in an aqueous solution, the pH is reduced by the carboxyl group formed at the polymer terminal, and in such a low pH condition, hydrolysis can be accelerated to improve stability. Lowers.

Therefore, in step (a) of the stabilization method of the present invention, the multiblock copolymer may be dissolved in an aqueous solvent to minimize the pH decrease caused by the carboxyl group generated by the hydrolysis reaction, thereby increasing the stability of the multiblock copolymer. have. As the aqueous solvent used in the present invention, distilled water or a buffer solution can be used. Complete layer of glycine as a buffer solution, citrate Buffer solution, succinate buffer solution, maleate complete solution, acetate complete solution,

Select from among MOPS complete solution, MES complete solution, PIPES buffer solution, TES complete solution, bicine complete solution, phosphate buffer solution, HEPES buffer solution, tricine buffer solution, Tris buffer solution, TABS buffer solution, and cacodylate complete solution Can be used, but is not limited thereto.

In addition, the content of the multiblock copolymer in the aqueous solution may be 0.1 to 50 w / v%, specifically 5 to 25 wMo. When the content of the polymer is less than 0.1 w / v%, the sol-gel transition phenomenon is not observed in the aqueous solution of the polymer. Instead, it is a condition for forming a polymer micelle and is applicable to a drug delivery system that solubilizes poorly soluble drugs. At 50 w / v%, sol-gel transition is observed and can be applied to anti-adhesion agents and drug delivery systems that delay drug release. If the content of the polymer is more than 50w / v%, the viscosity of the sol is high even in the storage conditions of the store, and because the gel strength increases immediately before the final use has a disadvantage inconvenient to use.

In step (b) of the stabilization method of the present invention, the multi-block copolymer aqueous solution is stored in a storage state, the storage conditions are 0 ~ 10 ^o C, specifically 2 ~ 5 ⁰ C. If it is less than 0 ^o C, the polymer solution freezes, which is inconvenient to be dissolved immediately before use, and if it is above 10 ^o C, decomposition of the multiblock copolymer may occur under long term storage conditions.

The stabilization method of the present invention can be applied to multiblock copolymers, preferably multiblock poloxamer aqueous solutions.

As used herein, the term "poloxamer" is a triblock structure compound of the ABA-type, wherein A is a polyethylene oxide block (PE0), B is a polypropylene oxide block (PP0), a polybutylene oxide block (PB0) Or a complex thereof, each block is connected by an ether bond, and in general, the weight average molecular weight of the poloxamer is 1,000 Daltons to 20,000 Daltons or less, and the terminal group is a hydroxyl group. Poloxamer 188 (Phironic® F-68), poloxamer 407 (Pluronic® F-127), and the like. As used herein, the term "multiblock copolymer" refers to a polymer in which the poloxamer is bonded to two or more molecules by a biodegradable dicarboxyl linker, and has a weight average molecular weight in the range of 40,000 to 1,000 Daltons. Say. In other words, in the present invention, the multiblock copolymer refers to a multiblock copolymer including two or more ABA-type triblocks covalently bonded with a biodegradable dicarboxyl linker, wherein A is a polyethylene oxide block, and B Means a polypropylene oxide block, a polybutylene oxide block or a composite thereof.

As used herein, the term "decarboxyl linker '" refers to the terminal hydroxyl group (-0H group) of the PEO—PP0 or PB0-PE0 block, oxalic acid, malonic acid, succinic acid, An ester bond formed by the reaction of an alkyl or aryl compound having two carboxyl groups in a molecule such as dipic acid, etc. The dicarboxyl linker is an oxalic acid, malonic acid, malic acid, succinic acid, glutaric acid, It may be provided by an alkyl dicarboxylic acid selected from the group consisting of adipic acid, pimelic acid, sebacolic acid, suberic acid, and dodecanoic acid, and the dicarboxyl linker may be an unsaturated di such as fumaric acid or maleic acid. Aryl dicarboxylic acids such as carboxylic acid, or phthalic acid, and terephthalic acid.

As mentioned above, the dicarboxyl linker may be linked to the PE by ester bonds of hydroxyl groups present at both terminal groups of PP0 (and / or PB0) -PE0, which ester bonds are hydrolyzed or Enzymes can be broken down into carboxylic acids and PE0-PP0 (and / or PB0) -PE0 units. As used herein, the term "sol-gel phase transition" is present in a fluid state, i.e., in a sol state, below a certain temperature, then changes to a gel state when the temperature rises above a certain temperature, and the temperature is changed to a specific temperature. Lowering below means reversibly changing back to the fluid state. The specific temperature varies depending on the type and molecular weight of the polymer, the concentration of the copolymer aqueous solution, the presence of salt, the proton concentration, etc., but is usually in the range of 5 to 37 ⁰ C, preferably in the range of 20 to 35 ° C. The multiblock copolymers of the present invention form hydrogels at sufficient concentrations and at certain temperatures, exhibit zolzogel phase transitions and have biodegradable characteristics. In the multiblock copolymers of the present invention, the PE0-PP0 (and / or The PBC-PE0 block is linked with a high molecular weight biodegradable dicarboxyl linker to show enhanced gel persistence, while the ionic end groups of the copolymer provide the effect of delayed drug release from the gel.

One embodiment of the invention is a multiblock copolymer, which can be represented by the formula:

M-X-0- [PE0-Y-PE0-C (= 0) -R-C (= 0) -0] n-PE0-Y-PE0-0-X-M

In the above, PE0 is a polyethylene oxide block,

Y is PP0 or PB0, or a complex of PP0 and PB0,

PP0 is a polypropylene oxide block,

PB0 is a polybutylene oxide block,

X is H or an anion group,

n is an integer between 1 and 100,

R is aryl comprising-(C¾) m 'or Cm',

m is an integer from 0 to 20,

m 'is an integer between 6 and 12, and

M is H or a cationic group, provided that both M and X are not 1 ^ and M is absent when X is H.

The polyethylene oxide block in the multiblock copolymer may be composed of ethylene oxide units having a number of about 2 to 2000, preferably about 5 to 500, and more preferably about 80 to 120 units. In the above formula f, the number of units of each ethylene oxide constituting two PE0 blocks may be the same or different. The number of units of propylene oxide or butylene oxide in the polypropylene oxide or polybutylene oxide block may be present in the range of 2 to 2000, preferably about 20 to 500, and more preferably about 30 to 250.

The multiblock copolymer of the present invention is 40, 000 Daltons to 1,000,000 Daltons, It may preferably have a weight average molecular weight in the range of 40,000 daltons to 500,000 daltons, more preferably 80,000 daltons to 130,000 daltons.

The unit ratio of ethylene oxide to propylene oxide or butylene oxide units in the PE0-PP0 (and / or PB0) -PE0 block can be adjusted for diversity of the polymer. For example, the unit ratio of PE0 to PP0 or PB0 of the multiblock copolymer may vary within the range of maintaining the water solubility of the triblock copolymer itself, about 0.2: 1 to 40: 1, preferably 1: 1 to 7.5: 1, and more preferably 1: 1 to 5: 1, wherein the PE0 block is 10 to 85 weight ¾ of the PE0-PP0 (and / or PE0) -PB0 unit, preferably 40 To 85% by weight.

Both ends of the multiblock block copolymer of the present invention are a hydroxyl group or an ionic group. As the ionic group at the end of the copolymer, anionic groups such as -S0 ₃ ᅳ, -P0 ₃ ^{2 "} , -C (= 0) -RC (= 0) -0 ^" are preferred, and the salt corresponding to the anionic group is Li, Monovalent metal cations such as Na, K, Ag, or Au, or divalent metal cations such as Ca, Mg, Zn, Fe, Cu, Co or Ni.

In particular, one or more multiblock copolymers of the present invention having anionic groups at both ends form a complex with a divalent divalent metal, thereby maintaining a more stable form of gel to sustain release of the drug from the gel. have. When the multiblock copolymer of the present invention having an anionic group is mixed with a cationic drug in an aqueous solution, an ionic salt is formed, which can reduce the initial release rate of the drug from the multiblock copolymer gel and thus release the drug. It can improve the persistence of. When a divalent cationic metal salt such as calcium chloride, zinc chloride, or magnesium chloride is added to the mixture of the multiblock copolymer of the present invention having an anionic group and a drug having an anionic group at the terminal, the divalent metal cation is bound to the drug. Complexes can be formed to sustain release of the drug from the gel. Thus, the multiblock copolymers of the present invention can be used as nonionic and heterozygous drug carriers for controlled drug release.

In addition, the present invention includes distilled water or a buffer solution in which the multiblock copolymer is dissolved, and when stored under refrigerated conditions for 2 years, the molecular weight of the multiblock copolymer It provides a multi-block copolymer aqueous solution composition for cold storage having a reduction amount of 30% or less.

More specifically, the present invention includes distilled water or a complete solution in which a multiblock copolymer comprising two or more ABA-type triblocks covalently bonded with a biodegradable dicarboxyl group linker is dissolved.

A is a polyethylene oxide block,

The multiblock copolymer includes M—X at both ends, M is H or a cationic group, and X is an anionic group,

Provided is a multi-block copolymer aqueous solution composition for cold storage, wherein the amount of reduction of the molecular weight of the multiblock copolymer when stored for two years under storage conditions is 30% or less. Preferably, the aqueous solution of the multiblock copolymer may be 20% or less for 6 months and / or 25% or less for 1 year when stored in refrigerated conditions.

According to a specific embodiment of the present invention, when the multi-block copolymer is stored for a long time at room temperature conditions, the strength of the gel is significantly reduced, it can be seen that a significant difference between the initial physical properties (Fig. 1 and 2) . In addition, the molecular weight rapidly decreased with time, and it can be seen that the hydrolysis rate of the multi-block copolymer in distilled water at room temperature was faster than that of the complete solution (Table 1).

However, when the multiblock copolymer was dissolved in distilled water or buffer solution and stored under long-term storage conditions, there was little change in gel strength even after long-term storage for 6 months, 12 months and 24 months. It can be seen that the difference is not large (FIGS. 3 to 5). In addition, it can be confirmed that the molecular weight change of the multiblock copolymer in the aqueous solution under cold storage conditions is well maintained without any significant influence on the content of the multiblock copolymer or the type of the complete solution (Table 1).

Therefore, the aqueous composition of the multi-block copolymer for cold storage of the present invention Long-term storage stability is maintained, and when used in the final use, it has the advantage of skipping the time to solubilize the polymer can be used immediately, using this property increases the ease of use of the aqueous solution composition can be developed in various applications do.

Accordingly, the present invention also provides a pharmaceutical composition comprising the aqueous composition of the multi-block copolymer for cold storage and a bioactive agent.

As used herein, the term “bioactive agent” or “drug” refers to a chemical or biological material that exhibits the desired biological or pharmaceutical efficacy and is injectable by the method used in the present invention or by methods already known in the art or Means a compound. Such efficacy may include the prevention of unwanted biological effects such as the prevention of disease in living organisms and the prevention of infection, the reduction of disease states, for example, the reduction of pain or inflammation caused by disease, or the reduction of disease states from organisms, Including but not limited to reduction or cure. The efficacy can be local or systemic, providing local anesthetic efficacy.

The bioactive agent may be any kind of drug, and may be, for example, nonionic and bivalent drugs. The drug may include, but is not limited to, microparticles, peptides, proteins, polysaccharides, nucleotides, and the like. Preferably, the drug may be an ionic drug, in particular a peptide or protein having a large number of carboxyl groups and aminoion groups in its molecule. Examples of the peptide or protein include growth hormone, interferon, leukocyte growth factor, giant leukocyte proliferation factor, erythroid-producing protein, interleukin, follicle-producing hormone, nerve growth factor, octreotide, insulin, insulin-like growth factor, calcitonin, tumor Necrosis factor, angiogenesis factor, epithelial growth factor, platelet growth factor, bone formation factor, thrombolytic agent, thrombopoietin, tissue growth factor, tumor necrosis factor, etc. The peptides and proteins may be natural, processed, Natural, glycosylated forms, and modified forms with polymers such as PEG, fragments with biological activity, and analogs thereof.

The drug may comprise within the range of 0.01 w / v% to 50 w / v%, wherein The multiblock copolymer aqueous solution may be used as a drug carrier as long as it exhibits phase transition, and may be preferably used at a concentration of 0.5 to 50 w / v%.

[Form for implementation of invention]

Hereinafter, the present invention will be described in detail by way of examples. However, the following examples are merely to illustrate the invention, the present invention is not limited by the following examples. Preparation Example 1 Preparation of Multiblock Copolymer

A multiblock copolymer was prepared by a slightly modified method based on the preparation method of Example 1 of Korean Patent Registration No. 10-0835738. Specifically, 10 g of purified poloxamer [Pluronic® F127 (BASF; poloxamer 407)] was placed in a flask reactor (100 ml) with a magnetic rod and heated and depressurized in an oil bath heated to 120 ° C (1 torr or less). ) While the water contained in the polymer was removed for 2 hours. The pressure was released and the reaction temperature was set to 120 ° C. while flowing nitrogen, and then 50 ml of dehydrated acetonitrile was added to completely dissolve the poloxamer. 192 ul of succinyl chloride (2 equivalents to poloxamer) was dilute in 10 ml of dehydrated acetonitrile and slowly added for 20 hours using a syringe pump. When all of the succinyl chloride was added, the reaction was further reacted at the same temperature for 4 hours to give a total reaction time of 24 hours. The synthesized multiblock poloxamer was precipitated in 1 L of diethyl ether, filtered and dried in vacuo to give the product as a white solid (8.4 g). The multiblock copolymer thus prepared was used in the following Comparative Examples and Examples. Comparative Example 1. Conditions for storing the multi-block copolymer in distilled water (10 w / v ¾) at room temperature

5 g of the multiblock copolymer was added to 50 ml of distilled water, and dissolved in a washroom of 5 ° C or less. Once the polymer was completely dissolved, the aqueous polymer solution was stored in a thermostat controlled at 25 ° C. Comparative Example 2 Conditions of Storing the Phases by Dissolving the Multiblock Copolymer in a Complete Solution (10w / v%)

A phosphate buffered saline (Phosphate Buffer Saline, hereinafter PBS) (10 1,!) 117.4 was used in place of distilled water, and the same method as in Comparative Example 1 was performed. Example 1. Conditions for storing the multi-block co-polymer in distilled water (10 w / v%) for storage

The polymer solution was carried out in the same manner as in Comparative Example 1 except for long-term storage in a thermostat controlled at 4 ° C. Example 2 Conditions for Refrigerating by Dissolving Multiblock Copolymer in Buffer Solution (10 w / v ¾)

Except for long-term storage of the polymer solution in a thermostat controlled at 4 ° C, it was carried out in the same manner as in Comparative Example 2. Example 3 Conditions for Refrigerating by Dissolving a Multiblock Copolymer in a Buffer Solution (15 w / v%)

The same procedure as in Example 2 was performed except that 7.5 g of the multiblock copolymer was added to 50 ml of PBSdOmM, pH7.4). Example 4 Conditions for Storage by Dissolving Multiblock Copolymer in Complete Solution (1 w / v ¾)

Except for using 0.5 g of a multiblock copolymer, the above example

It was carried out in the same manner as 2. Example 5 Conditions for Refrigerating by Dissolving (10 w / v%) the Multiple Blocky Copolymer in a Complete Solution The same procedure was followed as in Example 2, except that the complete acetate solution (10 mM, pH 5.0) was used as the complete solution. Example 6 Conditions for Storage of the Multiblock Copolymer by Dissolving in Complete Solution (10w / v%)

The same procedure was followed as in Example 2, except that HEPES complete solution (10 mM, pH 7.4) was used as the complete solution. The change in molecular weight decrease is shown in Table 1. Experimental Example 1. Measurement of sol-gel transition phenomenon

The sol-gel transition phenomenon of the polymer aqueous solution was measured according to a certain storage period, while storing for two years under the conditions of Comparative Examples 1-2 and Example 1-3, respectively. In order to observe the sol-gel transition phenomenon, the distilled water circulator and viscometer (BR00KFIELD, DV-II +) with temperature control were used. By measuring the temperature at which the gel starts to form and the strength of the gel varies with storage period, it was confirmed whether there was a decrease in gel strength due to decomposition of the multiblock copolymer. The results of the sol-gel transition phenomenon are shown in FIGS. 1 (Comparative Example 1), 2 (Comparative Example 2), 3 (Example 1), 4 (Example 2) and 5 (Example 3). It was.

3 to 5, which correspond to Examples 1 to 3, when the multi-block copolymer is dissolved in distilled water or a complete solution and stored under refrigerated storage conditions, gel strength is maintained even when stored for 6 months, 12 months and 24 months. It can be seen that there is little change and the gel strength is not significantly different from the initial value. However, in FIGS. 1 and 2 corresponding to Comparative Example 1 and Comparative Example 2, when the multiblock copolymer is stored at room temperature for a long time, the strength of the gel is remarkably decreased, resulting in a significant difference in initial physical properties. Therefore, it can be seen that it is not suitable to apply to the expected long-term field because the decrease in physical properties by long-term storage at room temperature. Experimental Example 2 Measurement of Molecular Weight Change of Multiblock Copolymer

After storage for a period of time, the multiblock obtained by freeze-drying the aqueous solution The copolymer was dissolved in chloroform, and the precipitated salt was removed by filtration, and then the change in molecular weight was observed during storage using GPC (gel permeation chromatography). The change in molecular weight decrease is shown in Table 1.

Table 1

* Molecular weight change (%) = (molecular weight after initial period / initial molecular weight) x 100 In Examples 1 to 6, the molecular weight change of the multiblock copolymer under the storage conditions is shown in the multiblock copolymer content or complete solution in the aqueous solution. It was shown that the initial molecular weight was well maintained without significant influence on the type of. However, looking at Comparative Example 1 and Comparative Example 2, when the long-term storage of the multi-block copolymer at room temperature conditions, the molecular weight was drastically decreased with time, the hydrolysis rate of the multi-block copolymer in distilled water at room temperature conditions to the complete solution It was faster than the phenomenon.

Based on the experimental results, refrigeration of the aqueous solution composition in which the multiblock copolymer is dissolved can minimize the deterioration of physical properties during long-term storage, and can be used immediately after eliminating the time required to solubilize the polymer in the final use. There is an advantage. Using these properties, the aqueous solution composition Ease of use will be increased and development will be possible for various applications. Having described the specific part of the present invention in detail, it is apparent to those skilled in the art that such a specific technology is only a preferred embodiment, and the scope of the present invention is not limited thereto. Accordingly, the substantial scope of the invention will be defined by the appended claims and equivalents thereof.

Industrial Applicability

According to the present invention, by storing the aqueous solution composition in which the multiblock copolymer is dissolved in distilled water or a complete solution, the storage stability can be improved, can be used immediately, and the ease of use is increased, thereby expanding the field of use.

Claims

[Range of request]

[Claim 1]

(a) dissolving a multiblock copolymer comprising two or more ABA-type triblocks covalently bonded with a biodegradable dicarboxyl linker in distilled water or a complete solution,

A is a polyethylene oxide block,

B is a polypropylene oxide block, a polybutylene oxide block or a complex thereof, and

(b) storing the dissolved multiblock copolymer under storage conditions;

Method of stabilizing a multiblock copolymer aqueous solution composition.

[Claim 2]

The method of claim 1, wherein the multiblock copolymer is represented by the following formula:

M-X-0- [PE0-Y-PE0-C (= 0) -R-C (= 0) -0] η-ΡΕΟ-Υ-ΡΕΟ-0-Χ-Μ

In the above, PE0 is a polyethylene oxide block,

Y is PP0 or PB0, or a complex of PP0 and PB0,

PP0 is a polypropylene oxide block

PB0 is a polybutylene oxide block,

X is H or an anionic group,

n is an integer from 1 to 100

R is aryl containing-(C¾) m- or Cm ',

m is an integer from 0 to 20,

m 'is an integer between 6 and 12, and

[Claim 3]

3. The method of claim 2, wherein X is an anionic group selected from the group consisting of -S (, -P0 ₃ ^{2 "} , and -C (= 0) -RC (= 0)-(T.

[Claim 4]

The method of claim 12, wherein M is a cationic group selected from the group consisting of Li, Na, K, Ag, An, Ca, Mg, Zn, Fe, Cu, Co, and Ni.

[Claim 5]

The method of claim 2 wherein the weight average molecular weight of the multiblock copolymer is in the range of 40,000 to 1,000,000 Daltons.

[Claim 6]

The method of claim 2, wherein the unit ratio of PE0 and Y is 0.2: 1 to 40: 1.

[Claim 7]

The method of claim 2, wherein the weight average molecular weight of Y is in the range of 1,000 to 20,000 Daltons.

[Claim 8]

The method of claim 2, wherein Y is poloxamer.

[Claim 9]

The method of claim 2, wherein R is oxalic acid, malonic acid, malic acid, succinic acid, glutaric acid, adipic acid, pimelic acid, sebacicolic acid, suberic acid, dodecanoic acid, fumaric acid, maleic acid phthalic acid, And a biodegradable dicarboxyl linker derived from one selected from the group consisting of terephthalic acid.

[Claim 10]

The method of claim 1, wherein the content of the multiblock copolymer is 0.1 to 50 w / v.

[Claim 11]

The method of claim 1, wherein the content of the multiblock copolymer is from 5 to 25 w / v%.

[Claim 12]

According to claim 1, wherein the complete solution is a glycine buffer (Glycine buffer), citrate buffer (Citrate buffer), succinate buffer (Succinate buffer), maleate buffer (Maleate buffer), acetate complete solution (Acetate buffer), MOPS complete solution (3- (N-morphol ino) propanesul fonic acid buffer), MES complete solution (2- (N-mor pho 1 i no) et hane su 1 f on ic acid buffer), PIPES buffer solution (2- [4- (2-sul foethyl) piperazin-l-yl Jethanesul fonic acid buffer), TES complete solution (N-Tris (hydroxymethyl) methyl-2-aminoethanesul fonic acid buffer), bicine complete layer solution (Ν, Ν-bis (2-Hydroxyhethyl) glycine, Phosphate buffer, HEPES buffer

(4- (2-hydroxyethyl) -l-piperazineethanesul fonic acid buffer), Tricin buffer, Tris buffer (tri hydr oxyme t hy 1) mi nome t hane buffer, TABS complete solution (N at least one selected from the group consisting of -tris (hydroxymethy 1) -4-am i no-but anesu 1 f on ic acid buffer) and cacodylate buffer.

[Claim 13]

The method of claim 1, wherein the storage condition is 0 to 10 ^° C.

[Claim 14]

The method of claim 13, wherein the refrigeration conditions are 2 to 5 ° C.

[Claim 15]

The method of claim 1, wherein the pH of the aqueous solution is 3 to 8.

[Claim 16]

Aqueous composition of the multi-blox copolymer stabilized by the method of any one of claims 1 to 15.

[Claim 17]

It includes distilled water or a complete solution in which a multi-blocky copolymer comprising two or more ABA 'type triblocks covalently bonded with a biodegradable dicarboxyl linker is dissolved.

A is a polyethylene oxide block,

Wherein the multiblock copolymer comprises M-X at both ends, wherein M is H or a cationic group, and X is an anionic group,

A multi-block copolymer aqueous solution composition for cold storage, wherein the reduced amount of the multiblock copolymer molecular weight is 30% or less when stored under refrigerated conditions for 2 years.

[Claim 18]

A pharmaceutical composition comprising the aqueous solution composition of claim 16 and a bioactive agent.

[Claim 19]

The pharmaceutical composition of claim 18, wherein the bioactive agent is in a concentration range of 0.01 to 50 / ^.

[Claim 20]

The pharmaceutical composition of claim 19, wherein the bioactive agent is a protein or peptide.

[Claim 21]

The method of claim 20, wherein the protein is a growth hormone (GH), interferon (IFN), granulocyte colony stimulation factor (G-CSF), granulocyte macrophage colony stimulation factor, GMCSF), erythropoietin (EPO) interleukin (IL), fibroblast growth factor fol icle stimulating hormone (FSH), nerve growth factor (NGF), octreotide octreotide, insulin, insulin-like growth factor (IGF), calcitonin, tumor necrosis factor (TNF), vascular endothelial growth factor (VEGF), epithelium Epidermal growth factor (EGF), platelet-derived growth factor (PDGF), bone morphogenet ic protein (BMP), tissue lasminogen activator (TPA), thrombopoietin ( thrombopoietin (TP0), tissue growth factor (TGF), and tumor necrosis factor (TNF).