WO2006109945A1 - Copolymere bloc sensible a la temperature et au ph, et hydrogels polymere l'utilisant - Google Patents

Copolymere bloc sensible a la temperature et au ph, et hydrogels polymere l'utilisant Download PDF

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Publication number
WO2006109945A1
WO2006109945A1 PCT/KR2006/001185 KR2006001185W WO2006109945A1 WO 2006109945 A1 WO2006109945 A1 WO 2006109945A1 KR 2006001185 W KR2006001185 W KR 2006001185W WO 2006109945 A1 WO2006109945 A1 WO 2006109945A1
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Prior art keywords
block copolymer
poly
compound
group
copolymer according
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PCT/KR2006/001185
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English (en)
Inventor
Doo-Sung Lee
Min-Sang Kim
Je-Sun You
Huynh Dai Phu
Bong-Sup Kim
Minh Khanh Nguyen
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Sungkyunkwan University Foundation For Corporate Collaboration
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Priority claimed from KR1020050097674A external-priority patent/KR100665672B1/ko
Application filed by Sungkyunkwan University Foundation For Corporate Collaboration filed Critical Sungkyunkwan University Foundation For Corporate Collaboration
Priority to US11/815,960 priority Critical patent/US8835492B2/en
Priority to DE112006000685.1T priority patent/DE112006000685B4/de
Priority to JP2008503960A priority patent/JP4753992B2/ja
Publication of WO2006109945A1 publication Critical patent/WO2006109945A1/fr

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    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08G—MACROMOLECULAR COMPOUNDS OBTAINED OTHERWISE THAN BY REACTIONS ONLY INVOLVING UNSATURATED CARBON-TO-CARBON BONDS
    • C08G81/00—Macromolecular compounds obtained by interreacting polymers in the absence of monomers, e.g. block polymers
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08L—COMPOSITIONS OF MACROMOLECULAR COMPOUNDS
    • C08L7/00—Compositions of natural rubber
    • C08L7/02—Latex
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08L—COMPOSITIONS OF MACROMOLECULAR COMPOUNDS
    • C08L71/00—Compositions of polyethers obtained by reactions forming an ether link in the main chain; Compositions of derivatives of such polymers
    • C08L71/02—Polyalkylene oxides
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08L—COMPOSITIONS OF MACROMOLECULAR COMPOUNDS
    • C08L2205/00—Polymer mixtures characterised by other features
    • C08L2205/05—Polymer mixtures characterised by other features containing polymer components which can react with one another
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08L—COMPOSITIONS OF MACROMOLECULAR COMPOUNDS
    • C08L67/00—Compositions of polyesters obtained by reactions forming a carboxylic ester link in the main chain; Compositions of derivatives of such polymers
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08L—COMPOSITIONS OF MACROMOLECULAR COMPOUNDS
    • C08L77/00—Compositions of polyamides obtained by reactions forming a carboxylic amide link in the main chain; Compositions of derivatives of such polymers

Definitions

  • the present invention relates to a biodegradable block copolymer useful for temperature and pH sensitive drug carriers, a method for preparing the same, and a polymeric hydrogel type drug composition comprising the above block copolymer. More particularly, the present invention relates to a multiblock copolymer derived from a temperature-sensitive block copolymer comprising a polyethylene glycol type compound and a biodegradable polyester polymer compound, and a poly( ⁇ -anino ester) compound and/or a poly(arrido amine) compound that shows ionization characteristics depending on pH values.
  • the multiblock copolymer according to the present invention is capable of target-directed drug delivery depending on temperature and pH variations in the body.
  • the present invention also relates to a polymeric hydrogel comprising the above multiblock copolymer.
  • US Patent No. 4,942,035 discloses the use of a copolymer of polyalkylene glycol, as a hydrophilic polymer, with polylactide, polyglycolide or polycaprolactone, as a biodegradable polyester polymer, for improving the problem of non-degradability of a so-called pluronic gel (polyethylene glycol and polyethylene oxide-polypropylene oxide-polyethylene oxide block copolymer) in the body.
  • pluronic gel polyethylene glycol and polyethylene oxide-polypropylene oxide-polyethylene oxide block copolymer
  • US Patent No. 5,476,909 discloses an A-B-A type triblock copolymer as a biodegradable polyester polymer, wherein the hydrophobic block (A) is limited to polylactide (PLA), polyglycolide (PGA) and copolymers thereof, and the hydrophilic block (B) is limited to polyethylene glycol (PEQ and derivatives thereof.
  • PLA polylactide
  • PGA polyglycolide
  • PEQ polyethylene glycol
  • Korean Laid-Open Patent No. 2000-0012970 discloses a pH-sensitive polymer containing a sulfoneanide group and a method for preparing the same. More particularly, the Korean Patent discloses variations in the solubility of a linear polymer formed by random copolymerization of a sulfoneanide monomer with dimethylacryl anide or isopropylacryl anlde, or the swelling degree of the crosslinked polymer thereof.
  • the aforementioned prior art utilizes a block copolymer of a hydrophobic biodegradable polymer with a hydrophilic polymer, which shows a sol-gel transition phenomenon depending on temperatures. More particularly, when the block copolymer is injected into the body in the form of an aqueous solution present in a sol state, it undergoes a transition into a gel state, so that it can be used as a release-controlled drug carrier that carries a drug stably in the body and releases the drug gradually.
  • the block copolymer hydrogel according to the present invention shows sol-gel transition behavior sensitive to pH values as well as temperatures.
  • the hydrogel according to the present invention shows sol-gel transition behavior, wherein it undergoes gelling at a pH range of 7-7.4 similar to pH values in the body, while it is converted into sol at a pH value lower than the above range.
  • the block copolymer hydrogel according to the present invention forms gel stably with no problem of occlusion of an injection needle, occurring in the conventional temperature-sensitive hydrogel.
  • the resultant block can be applied as a carrier for use in release-controlled drug delivery, which carries a drug stably at a specific temperature and a specific pH value in the body and releases the drug gradually.
  • a block copolymer formed by coupling the following components with each other: (a) a copolymer (A) of a polyethylene glycol (PEQ compound with a biodegradable polymer; and (b) at least one oligomer (B) selected from the group consisting of poly( ⁇ -anino ester) and poly(anido amine). There is also provided a method for preparing the same block copolymer.
  • A polyethylene glycol
  • B at least one oligomer selected from the group consisting of poly( ⁇ -anino ester) and poly(anido amine).
  • a polymeric hydrogel type drug composition comprising the above temperature and pH-sensitive block copolymer and a physiologically active substance that can be encapsulated with the block copolymer.
  • the present invention is characterized in that a temperature-sensitive block copolymer formed by the copolymerization of a polyethylene glycol type compound and a biodegradable polymer compound is coupled with poly( ⁇ -arrino ester), poly(amido amine) or a combination (PAEA) thereof, which experiences a change in the ionization degree depending on pH values, to provide a temperature and pH sensitive multiblock copolymer, which is sensitive to pH variations as well as temperature variations in the body, and to provide polymeric hydrogel using the same multiblock copolymer.
  • a temperature-sensitive block copolymer formed by the copolymerization of a polyethylene glycol type compound and a biodegradable polymer compound is coupled with poly( ⁇ -arrino ester), poly(amido amine) or a combination (PAEA) thereof, which experiences a change in the ionization degree depending on pH values, to provide a temperature and pH sensitive multiblock copolymer, which is sensitive to pH variations as well as temperature
  • the conventional block copolymer comprising a hydrophilic polymer such as polyethylene glycol and a biodegradable hydrophobic polymer shows sol-gel transition behavior due to a change in the physical properties of the hydrophilic block and the hydrophobic block.
  • a hydrophilic polymer such as polyethylene glycol
  • a biodegradable hydrophobic polymer shows sol-gel transition behavior due to a change in the physical properties of the hydrophilic block and the hydrophobic block.
  • it was difficult to apply such conventional block copolymers to an actual drug delivery system due to their poor sensitivity to temperature variations and side effects in the body, caused by the thermal equilibrium of a transfer medium.
  • the temperature-sensitive block copolymer comprising the hydrophilic polymer and the biodegradable polymer is coupled with a compound such as poly( ⁇ -arrino ester) (PAE), poly(arrido amine) (PAA) or a combination (PAEA) thereof, which shows an ionization degree varied with pH values. Therefore, the resultant block copolymer has pH sensitivity as well as temperature sensitivity, and thus can solve the aforementioned problem related to the conventional temperature-sensitive hydrogel and can form more stable hydrogel.
  • a compound such as poly( ⁇ -arrino ester) (PAE), poly(arrido amine) (PAA) or a combination (PAEA) thereof, which shows an ionization degree varied with pH values. Therefore, the resultant block copolymer has pH sensitivity as well as temperature sensitivity, and thus can solve the aforementioned problem related to the conventional temperature-sensitive hydrogel and can form more stable hydrogel.
  • the temperature and pH sensitive block copolymer according to the present invention forms a physically and chemically stable hydrogel in a specific pH range, and undergoes a transition into a sol state in the other pH ranges.
  • the block copolymer according to the present invention can show reversible sol-gel transition behavior.
  • a low pH range e.g. pH ⁇ 7.0
  • PAE poly( ⁇ -anino ester)
  • PAE PAE
  • the temperature and pH sensitive block copolymer is safe in the human body, and thus can be used as a release-controlled drug carrier in the medical field and in the gene transfer and drug delivery systems, in particular, as a drug carrier and drug releasing substance. Also, the block copolymer can be applied to a carrier for cell transfer, used as an injectable scaffold.
  • the block copolymer according to the present invention it is possible to modify the constitutional elements forming the block copolymer and physical properties thereof, for example, the composition, molar ratio, molecular weight and/or functional groups in the blocks, so as to design the reversible sol-gel transition behavior of the block copolymer in various manners.
  • the block copolymer according to the present invention can be used for various applications, including cancer cell mutation, gene mutation and others.
  • One constitutional element forming the temperature- and pH-sensitive block copolymer according to the present invention is a copolymer (A) of a PEG type compound with a biodegradable compound.
  • the copolymer (A) allows a sol-gel transition depending on temperature variations, because the hydropbilic PEG type compound and the hydrophobic biodegradable polymer are present in the same molecule.
  • the PEG type compound forming the copolymer (A) is a general PEG compound known to one skilled in the art. Although there is no particular limitation in selection of the PEG compound, it is preferable to use a PEG type compound represented by the following formula 1.
  • R is a hydrogen atom or a C1 ⁇ C5 alkyl group
  • n is a natural number ranging from 11 to 45.
  • the polyethylene glycol type compound preferably has a molecular weight of 500 to 5000.
  • the polyethylene glycol preferably has a molecular weight of 1000 to 2000.
  • the methoxypolyethylene glycol preferably has a molecular weight of 500 to 5000.
  • the polyethylene glycol type compound has a molecular weight (Mn) away from the above range (for example, a molecular weight of less than 500 or greater than 5000), it is difficult to form gel. Even if gel is formed from the block copolymer, the resultant gel shows poor gel strength, so that it cannot be used as a carrier for drug delivery.
  • Mn molecular weight
  • the biodegradable polymer forming the copolymer (A) is a conventional biodegradable polymer known to one skilled in the art.
  • the biodegradable polymer is a biodegradable aliphatic polyester polymer, and non-liniting examples thereof include caprolactone (CL), glycolide (GA), lactide (LA) or a copolymer thereof.
  • CL caprolactone
  • G glycolide
  • LA lactide
  • any biodegradable polymers capable of forming a copolymer with the PEG type compound may be used in the scope of the present invention.
  • copolymer (A) formed by the polymerization of the polyethylene type compound and the biodegradable polymer there is no particular limitation in the copolymer (A) formed by the polymerization of the polyethylene type compound and the biodegradable polymer, as long as the copolymer comprises the above components.
  • the copolymer has at least one substituent selected from the group consisting of a primary amine group, a secondary amine group and a double bond, which are capable of reacting with a poly( ⁇ -anino ester) and/or poly(anido anine) block.
  • Non-liniting examples of the copolymer (A) include polylactide (PLA), polyglycolide (PGA), poly- caprolactone (PCL), poly(caprolactone-lactide) random copolymer (PCLA), poly(caprolactone-glycolide) random copolymer (PCGA), poly(lactide-glycolide) random copolymer (PLGA), or the like.
  • the ratio is preferably 1:1-3. If the ratio is less than 1:1, it is not possible to form gel. If the ratio is greater than 1:3, hydrophobicity increases undesirably, so that the resultant block copolymer may not be dissolved in water.
  • PCGA or PLGA it is possible to improve the temperature sensitivity and pH sensitivity by controlling the molar ratio thereof.
  • Another constitutional element forming the temperature- and pH-sensitive block copolymer according to the present invention may be any compound that shows an ionization degree varied with pH values with no particular limitation.
  • the compound is an oligomer (B) formed from a poly( ⁇ -anino ester) and/or poly(amido anine) type compound having hydrophobicity and pH-sensitivity at the same time.
  • the poly( ⁇ -anino ester)- and poly(anido anine)-based oligomer (PAEA) comprising poly( ⁇ -anino ester) (PAEj, poly(arrido anine) (PAA) or a mixture containing them in an adequate ratio have ionization characteristics characterized by water solubility varied with pH values due to the presence of a tertiary anine group ionized at a pH of 7.2 or less. Therefore, the oligomer can show pH sensitivity by forming hydrogel or by maintaining a sol state depending on pH variations in the body.
  • the above compounds may be prepared by a process known to one skilled in the art.
  • a bisacrylate compound and/or a bisacrylanide compound having a double bond is polymerized with an amine compound via the Michael reaction mechanism to produce a poly( ⁇ -amino ester) (PAE), poly(a ⁇ ido anine) (PAA) or a nixed oligomer containing them in an adequate ratio.
  • PAE poly( ⁇ -amino ester)
  • PAA poly(a ⁇ ido anine)
  • nixed oligomer containing them in an adequate ratio.
  • R is a C1-C30 alkyl group.
  • the bisacrylanide compound used in the above process may be represented by the following formula 3, and non-liniting examples of such bisacrylanide compounds include N,N'-methylene bisacrylanide (MDA), N,N'-ethylene bisacrylanide and mixtures thereof.
  • MDA N,N'-methylene bisacrylanide
  • the bisacrylanide compound is allowed to react with an anine compound such as 4-aninomethylpiperidine (AMPD), N-methyl ethylenedianine (MEDA) or l-(2-aninoethyl)piperidine (AEPZ) through a conventional reaction mechanism such as the Michael reaction mechanism.
  • AMPD 4-aninomethylpiperidine
  • MEDA N-methyl ethylenedianine
  • AEPZ l-(2-aninoethyl)piperidine
  • R is a C 1 ⁇ C20 alkyl group.
  • the bisacrylate compound and/or the bisacrylanide compound may be used alone or in combination.
  • the weight ratio of the compounds forming the mixture may be controlled freely in a range of 100:0-0:100.
  • the amine compound there is no particular limitation in selection of the amine compound as long as the amine compound has an amine group.
  • a primary amine represented by the following formula 4 a secondary anine-containing diamine compound represented by the following formula 5, or a mixture thereof are used.
  • Non-liniting examples of such primary amine compounds include
  • Non- liniting examples of such secondary anine-containing diamine compounds include piperazine, piperidine, pyrrolidine, 3,3-dimethylpiperidine, 4,4'-trimethylene dip- iperidine, N,N'-dimethyl ethylene diamine, N,N'-diethyl ethylene diamine, ini- dazolidine, diazepine, etc.
  • the hisacrylate or bisacrylanide compound is allowed to react with the anine compound preferably in a molar ratio of 1 :0.5 ⁇ 2.0.
  • the molar ratio of the anine compound is less than 0.5 or greater than 2.0, the resultant polymer has a broad molecular weight distribution and shows poor pH sensitivity, and it is difficult to control the length of a block in the resultant block copolymer.
  • the oligomer preferably has a molecular weight of 500-20,000. If the molecular weight is less than 500, the resultant block copolymer cannot show sol-gel transition behavior depending on pH variations. On the other hand, if the molecular weight is greater than 20,000, the block copolymer cannot show temperature sensitivity.
  • the block copolymer according to the present invention formed by coupling the copolymer (A) of a PEG type compound and a biodegradable polymer with the poly( ⁇ -anino ester) and/or poly(anido anine) oligomer (B) is preferably a tri- or higher block copolymer, and is more preferably a triblock or pentablock copolymer. More particularly, the block copolymer may be represented by any one of the following formulae 6-11 :
  • the block copolymer represented by any one of the above formulae 6-11 can form hydrogel or can maintain a sol state depending on pH variations due to its amphiphilic property and pH sensitivity.
  • the block copolymer according to the present invention can be used satisfactorily in various applications requiring sensitivity depending on pH variations in the body (for example, carriers for release-controlled drug delivery).
  • the block copolymer represented by formula 9 has a hydroxy group only at one end of the copolymer (MPEGPGLA) of a PEG type compound and a biodegradable polyester polymer. Therefore, the hydroxy group may be substituted with an acrylate group, and thus the resultant block copolymer has a block structure coupled with a ⁇ -amino ester at one side.
  • the molecular weight of the block copolymer it is preferable that the block copolymer has a molecular weight of 5,000 to 30,000. When the block copolymer has a molecular weight away from the above range, it is difficult to form gel due to a failure in hydrophilicity/hydrophobicity balance.
  • the temperature- and pH-sensitive block copolymer according to the present invention may further comprise other components or additives, currently used in the art.
  • the method comprises the steps of: (a) polymerizing a PEG type compound with a biodegradable polymer to form a copolymer (A); (b) introducing an acrylate group into the copolymer (A) of PEG with the biodegradable polymer; (c) and coupling the resultant copolymer (A) with at least one oligomer selected from the group consisting of poly( ⁇ -anino ester) and poly(ai ⁇ ido amine).
  • a PEG type compound is polymerized with a biodegradable polyester polymer to form a copolymer, and the reaction may be represented by the following Reaction Scheme 1:
  • Copolymerization of a PEG type compound with a biodegradable polyester polymer is preferably carried out via a ring-opening polymerization reaction.
  • polymerization temperature and time may be controlled in a range known to one skilled in the art, and the polymerization is carried out preferably at a temperature of 130 ⁇ 150°C for 12-48 hours.
  • a catalyst may be used for accelerating the reaction, and particular examples of the catalyst that may be used include stannous octoate, stannous chloride, metal oxides (GeO , Sb O , SnO , etc.), aluminum triiso-
  • the step of introducing an acrylate group into the copolymer formed by the ring- opening polymerization of PEG is preferably carried out by way of the reaction between the terminal hydroxy group (-OH) of the polyethylene glycol-biodegradable polyester copolymer and the halogen of an acryloyl chloride.
  • the reaction may be represented by the following Reaction Scheme 2: [80] [Reaction Scheme 2]
  • the coupling reaction may be represented by the following Reaction Scheme 3:
  • a primary amine compound and a secondary anine-containing diamine compound, used for the preparation of a poly( ⁇ -arrino ester) oligomer, as well as a diol diacrylate type compound that reacts with the above amine compound to form a ⁇ -anino ester block may be subjected to the same process as described above.
  • the multiblock copolymer obtained as described above has a combination of a hy- drophilic block, a hydrophobic block and a poly( ⁇ -a ⁇ no ester) and/or poly(arrido amine) oligomer that shows an ionization degree varied with pH values, and thus the multiblock copolymer can show temperature sensitivity and pH sensitivity at the same time.
  • the block copolymer according to the present invention for example, methoxypolyethylene glycol-polycaprolactone-beta amino ester (MPEGPCL- ⁇ -amino ester) for the introduction of each functional group and for the reaction of terminal groups by using FT-IR and H-NMR. It was also possible to determine that the block copolymer according to the present invention has a structure formed by coupling of a copolymer of a PEG type compound and a biodegradable polymer with a ⁇ -anino ester oligomer, through an increase in the molecular weight of the block copolymer, as observed by GPC (gel permeation chromatography).
  • GPC gel permeation chromatography
  • sol-gel transition characteristics of the block copolymer were measured by varying pH at a certain temperature. After the measurement, it was shown that the multiblock copolymer according to the present invention has pH sensitivity.
  • a polymeric hydrogel type drug composition which comprises: (a) the temperature and pH sensitive block copolymer; and (b) a physiologically active substance that can be encapsulated with the block copolymer.
  • any physiologically active substances can be used and can be encapsulated with the polymeric hydrogel type block copolymer according to the present invention with no particular limitation.
  • active substances include anti- cancer agents, antibacterial agents, steroids, antiphlogistic analgesic agents, sexual hormones, immunosuppressants, antiviral agents, anesthetic agents, antiemetic agents, antihistamine agents, etc.
  • the drug composition according to the present invention may further comprise conventional additives such as vehicles, stabilizers, pH adjusting agents, antioxidants, preservatives, binders and disintegrating agents.
  • the composition may further comprise other conventional additives, solvents, or the like.
  • the polymeric hydrogel type drug composition may be provided as oral formulation or parenteral formulation. Particularly, the polymeric hydrogel type drug composition may be provided for intravenous, intramuscular or subcutaneous injection.
  • a carrier for drug delivery or medical diagnosis which comprises the temperature and pH sensitive block copolymer.
  • any materials are encapsulated with the block copolymer, as long as they are for treatment, prevention or diagnosis of diseases.
  • a copolymer comprising: (a) at least one hydrophilic block; (b) at least one biodegradable block having hydrophobicity varied with temperatures; and (c) at least one unit having an ionization degree varied with pH values, as a carrier for drug delivery or medical diagnosis.
  • the unit (c) having an ionization degree varied with pH values may be at least one oligomer (B) selected from the group consisting of poly ( ⁇ -anino ester) and poly(anido amine), but is not United thereto.
  • the hydrophilic block (a) and the biodegradable block (b) having different hydrophobicity depending on temperature variations are the same as defined above. Also, it is possible to use other hydrophilic and/or hydrophobic materials known to one skilled in the art. Brief Description of the Drawings
  • FIG. 1 is a graph showing the sol-gel transition behavior of the triblock copolymer according to Example 1, depending on temperature and pH variations, the triblock copolymer comprising a pH-sensitive polyethylene glycol type compound, a biodegradable polycaprolactone compound and a pH-sensitive biodegradable ⁇ oly( ⁇ -anino ester) compound;
  • FIG. 2 is a graph showing the sol-gel transition behavior of the pentablock copolymer according to Example 2, depending on temperature and pH variations, the pentablock copolymer comprising a polyethylene glycol type compound, poly- caprolactone compound and polylactic acid compound, and a poly ⁇ -arrino ester) compound;
  • FIG. 3 is a graph showing the sol-gel transition behavior of the pentablock copolymer according to Example 4, depending on pH variations, the pentablock copolymer comprising a polyethylene glycol type compound, polycaprolactone compound and polylactic acid compound, and poly( ⁇ -a ⁇ ino ester) and poly(anido amine) nixed in a weight ratio of 60:40; and
  • FIG. 4 is a graph showing variations in the molecular weight of each of the block copolymers as a function of time at pH 7.4 according to Examples 2 to 7. Mode for the Invention
  • MPEGPCL block copolymer having a desired molecular weight by controlling the amount of MPEG and the addition amount of ⁇ -caprolactone so as to control the molecular weight.
  • Stannous octoate was used in an amount of 0.5 wt% based on the amount of MPEG.
  • the reaction mixture was cooled to room temperature, and a small amount of methylene chloride was added thereto to dissolve the reactants.
  • the reaction mixture was added to an excessive amount of ethyl ether and precipitated therein to remove unreacted materials.
  • the product, from which unreacted materials were removed was dried under vacuum at 40 0 C for 48 hours. By doing so, a block copolymer (MPEGPCL) of polyethylene glycol with ⁇ -caprolactone (biodegradable polyester polymer compound) was obtained with a yield of 85% or more.
  • the above reaction was carried out in an ice-bath under nitrogen atmosphere for 24 hours. After the completion of the reaction, the reaction mixture was precipitated in ethyl ether to remove unreacted materials, filtered and dried under vacuum at room temperature to obtain a polyethylene glycol-polycaprolactone-acrylate block copolymer (MPEGPCL-A) having a double bond at its terminal group, with a yield of 80% or more.
  • MPEGPCL-A polyethylene glycol-polycaprolactone-acrylate block copolymer having a double bond at its terminal group, with a yield of 80% or more.
  • PCLA-PEGPCLA triblock copolymers
  • the block copolymer (acrylated PCLA-PEGPCLA) having a double bond, obtained as described above, was introduced into a reactor at room temperature, and chloroform was added thereto to dissolve the block copolymer.
  • chloroform was added thereto to dissolve the block copolymer.
  • 4,4 / -trimethylene dipiperazine and 1,4-butanediol diacrylate were added thereto at room temperature and dissolved. Then, the reaction mixture was allowed to react at 50 0 C for 48 hours.
  • molecular weight of MPEG molecular weight of the biodegradable polymer (PCLA), molar ratio of MPEG to the biodegradable polymer, and molecular weight of poly( ⁇ -anino ester) were varied as shown in the following Table 1 to provide triblock copolymers (PCLA-PEGPCLA) and pentablock copolymers (poly( ⁇ -anino ester)-PCLA-PEGPCLA-poly( ⁇ -amino ester)) having various molecular weights. Molecular weight of each block copolymer is also shown in Table 1.
  • PDI refers to a polydispersity index as measured by GPC, and is used to determine whether the block copolymer has a uniform molecular distribution or not. As PDI of a block copolymer decreases, the block copolymer has a more uniform molecular weight distribution. On the other hand, as PDI of a block copolymer increases, the block copolymer has a less uniform molecular weight distribution. [112] Table 1
  • Example 3 [114] Example 2 was repeated to provide a pentablock copolymer of poly( ⁇ -a ⁇ ino ester- a ⁇ ido a ⁇ ine)-PCLA-PEGPCLA-poly( ⁇ -anino ester-anido anine) having a molecular weight of 6800, except that 1,6-hexanediol diacrylate and N,N'-methylene bisacrylanide were used in a weight ratio of 80:20 instead of 1 ,4-butanediol diacrylate.
  • Example 4 [116] Example 2 was repeated to provide a pentablock copolymer of poly( ⁇ -anino ester- anido amine)-PCLA-PEGPCLA ⁇ poly( ⁇ -anino ester-anido anine) having a m olecular weight of 6500, except that 1,6-hexanediol diacrylate and N,N'-methylene bisacrylanide were used in a weight ratio of 60:40 instead of 1,4-butanediol diacrylate.
  • Example 5 [118] Example 2 was repeated to provide a pentablock copolymer of poly( ⁇ -anino ester- anido anine)-PCLA-PEGPCLA-poly( ⁇ -anino ester-anido anine) having a molecular weight of 6500, except that 1,6-hexanediol diacrylate and N,N'-methylene bisacrylanide were used in a weight ratio of 40:60 instead of 1,4-butanediol diacrylate.
  • Example 6 [120] Example 2 was repeated to provide a pentablock copolymer of poly( ⁇ -anino ester- anido a ⁇ ine)-PCLA-PEGPCLA-poly( ⁇ -anino ester-anido anine) having a molecular weight of 6500, except that 1,6-hexanediol diacrylate and N,N'-methylene bisacrylanide were used in a weight ratio of 20:80 instead of 1,4-butanediol diacrylate. [121] Example 7
  • Example 2 was repeated to provide a pentablock copolymer of poly(amido anine)-PCLA-PEGPCLA-poly(anido amine) having a molecular weight of 6500, except that 1,6-hexanediol diacrylate and N,N'-methylene hisacrylanide were used in a weight ratio of 0:100 instead of 1 ,4-bitanediol diacrylate.
  • Example 2 was repeated to provide a pentablock copolymer of poly(anido arrine)-PCLA-PEG-PCLA-poly(amido amine) having a molecular weight of 5000, except that the oligomer formed from poly(anido amine) had a molecular weight of less than 500. However, the copolymer showed no sol-gel transition behavior under the body temperature conditions (37°C, pH 7.4).
  • Example 2 was repeated to provide a pentablock copolymer of poly(amido arrine)-PCLA-PEG-PCLA-poly(anido amine) having a molecular weight of 25500, except that the oligomer formed from poly(arrido amine) had a molecular weight of 21000. However, the copolymer showed no sol-gel transition behavior under the body temperature conditions (37°C, pH 7.4).
  • Example 2 was repeated to provide a pentablock copolymer of poly( ⁇ -anino ester- airido airine)-PCLA-PEGPCLA-poly( ⁇ -anino ester-a ⁇ ido amine) having a molecular weight of 5000, except that MPEG4000 was used instead of MPEG2000. However, the copolymer showed no sol-gel transition behavior under the body temperature conditions (37°C, pH 7.4).
  • Example 2 was repeated to provide a pentablock copolymer of poly( ⁇ -anino ester- a ⁇ ido anine)-PCLA-PE&PCLA-poly( ⁇ -anino ester-arrido anine) having a molecular weight of 7700, except that MPEG6000 was used instead of MPEG5000.
  • MPEG6000 was used instead of MPEG5000.
  • it was difficult to form a gel from the copolymer because the copolymer had unbalanced hydrophilic/hydrophobic blocks and could not allow a sol-gel transition due- to the use of MPEG having a high molecular weight.
  • FIGs. 1-3 show the sol-gel transition behavior of each block copolymer depending on temperature and pH variations.
  • the block copolymer according to the present invention allowed reversible sol-gel transition behavior depending on pH variations as well as temperature variations, due to a change in the ionization degree of the poly( ⁇ -anino ester) type oligomer depending on pH variations and a change in the hydrophobicity of the biodegradable polymer depending on temperature variations (see FIGs. 1-3).
  • the block copolymer according to the present invention shows reversible sol-gel transition behavior under the same conditions as the human body (37°C, pH 7.4). Therefore, it is expected that the block copolymer according to the present invention has industrial applicability as a drug carrier.
  • the block copolymer according to the present invention was prepared under various conditions to provide various kinds of block copolymers. Then, the block copolymers were evaluated for their sol-gel transition behavior depending on temperature and pH variations.
  • thermosensitive block and the molecular weight of a hydrophilic MPEG block should be considered in addition to the molecular weight ratio and the block ratio of MPEGPCL, in order to provide a temperature- and pH-sensitive block copolymer that shows reversible sol-gel transition behavior under the body temperature and pH conditions.
  • the temperature- and pH- sensitive block copolymer hydrogel according to the present invention can be obtained not by a simple combination of a hydrophilic block, a hydrophobic block and a pH- sensitive block but by controlling the molecular weight of each constitutional element, molar ratio of constitutional elements and molar ratio of blocks to optimal conditions. Additionally, it is possible to design a medical drug carrier that requires various conditions by such controlling technique and to obtain a commercially available drug carrier.
  • the block copolymer according to the present invention shows pH sensitivity as well as temperature sensitivity, and thus can solve the problems occurring in conventional temperature-sensitive block copolymers according to the prior art. Additionally, the block copolymer according to the present invention can form more stable hydrogel at an adequate temperature and pH, and can solve the problem related to the in vivo stability. Therefore, the block copolymer according to the present invention can be used for various applications in the medical field, in particular, as a drug carrier for drug delivery.

Abstract

La présente invention concerne un copolymère bloc formé par couplage des composants suivants les uns avec les autres: (a) un copolymère (A) d'un type de polyéthylène-glycol (PEG) en composition avec un polymère biodégradable; et (b) au moins un oligomère (B) choisi dans le groupe constitué de poly (ß-amino ester) et de poly(amido amine). L'invention concerne également un procédé d'élaboration de ce copolymère bloc, et une composition médicamenteuse de type hydrogel polymère comprenant le copolymère bloc sensible à la température et au pH, et une substance physiologiquement active pouvant être encapsulée avec le copolymère bloc. Le copolymère multibloc s'obtient par copolymérisation d'un oligomère sensible au pH de type poly(ß-amine ester) et/ou poly(amide amine) avec un composé hydrophile et thermosensible de type polyéthylène glycol et un polymère hydrophobe et biodégradable. Ainsi, ce copolymère bloc qui convient à la formation d'une structure hydrogel polymère en raison d'une amphiphilicité résultant de la combinaison d'un groupe hydrophile et d'un groupe hydrophobe dans le copolymère et de caractéristiques d'ionisation liées à des variations de pH, convient par conséquent comme vecteur de médication pour l'administration médicamenteuse ciblée en fonction de variations de pH anatomique.
PCT/KR2006/001185 2005-04-13 2006-03-31 Copolymere bloc sensible a la temperature et au ph, et hydrogels polymere l'utilisant WO2006109945A1 (fr)

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US11/815,960 US8835492B2 (en) 2005-04-13 2006-03-31 Temperature and pH sensitive block copolymer and polymeric hydrogels using the same
DE112006000685.1T DE112006000685B4 (de) 2005-04-13 2006-03-31 Temperatur- und pH-empfindliches Blockcopolymer und daraus hergestellte Polymerhydrogele
JP2008503960A JP4753992B2 (ja) 2005-04-13 2006-03-31 新規な温度及びｐＨ感受性のブロック共重合体及びこれを用いた高分子ヒドロゲル

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Cited By (19)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2008222745A (ja) * 2007-03-08 2008-09-25 Sungkyunkwan Univ Foundation For Corporate Collaboration Ｐｈ及び温度敏感性のブロック共重合体ヒドロゲル
WO2009031861A2 (fr) * 2007-09-06 2009-03-12 Sungkyunkwan University Foundation For Corporate Collaboration Copolymère séquencé sensible à la température et au ph extrêmement sûr in vivo, hydrogel et système d'administration de médicament utilisant ceux-ci
JP2009132825A (ja) * 2007-11-30 2009-06-18 Agency For Science Technology & Research 刺激応答型生分解性高分子及びその製造方法
WO2010030728A2 (fr) * 2008-09-12 2010-03-18 Boston Scientific Scimed, Inc. Dispositifs et systèmes pour distribuer des agents thérapeutiques à des lumières corporelles
JP2010530906A (ja) * 2007-06-11 2010-09-16 ルレデーラ フンダシオン パラ エル デサローヨ テクノロジコ イ ソシアル 不溶性薬剤の制御放出のための、シクロデキストリンおよびポリ（アミドアミン）に基づく超分岐ポリマー
US8163861B2 (en) 2009-01-15 2012-04-24 Living Proof, Inc. Beta-amino ester compounds and uses thereof
WO2011111067A3 (fr) * 2010-03-09 2012-05-31 Council Of Scientific & Industrial Research Composition d'hydrogel polymère biodégradable
WO2012126005A2 (fr) 2011-03-17 2012-09-20 Georgia Tech Research Corporation Hydrogels de polymère pour applications in vivo et leurs procédés d'utilisation et de préparation
US8703194B2 (en) 2007-11-02 2014-04-22 Agency For Science, Technology And Research Stimulus-responsive biodegradable polymers and methods of preparation
US8796234B2 (en) 2009-11-24 2014-08-05 Agency For Science, Technology And Research Crosslinking branched molecule through thiol-disulfide exchange to form hydrogel
CN104710625A (zh) * 2015-03-19 2015-06-17 南京邮电大学 一种近红外响应可降解水溶性共聚物及其高效合成方法
EP2766407A4 (fr) * 2011-10-12 2015-12-30 Univ Missouri Polymères pentablocs
US9271929B2 (en) 2008-11-25 2016-03-01 École Polytechnique Fédérale De Lausanne (Epfl) Block copolymers and uses thereof
CN105754107A (zh) * 2016-04-25 2016-07-13 苏州大学 一种亲水性聚（ω-己内酯）及其制备方法
US9505867B2 (en) 2005-05-31 2016-11-29 Ecole Polytechmique Fédérale De Lausanne Triblock copolymers for cytoplasmic delivery of gene-based drugs
US9611353B2 (en) 2013-05-16 2017-04-04 The Curators Of The University Of Missouri Drug loading pentablock polymers
CN108102117A (zh) * 2017-12-28 2018-06-01 长春工业大学 一种共价键交联的碱基仿生粘韧水凝胶体系的制备方法
US10711106B2 (en) 2013-07-25 2020-07-14 The University Of Chicago High aspect ratio nanofibril materials
CN114573808A (zh) * 2022-03-17 2022-06-03 浙江大学杭州国际科创中心 一种端双键离子响应型超支化聚合物及其制备方法和应用

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS62218456A (ja) * 1986-03-19 1987-09-25 Sakata Shokai Ltd 被覆用樹脂組成物
US4882168A (en) * 1986-09-05 1989-11-21 American Cyanamid Company Polyesters containing alkylene oxide blocks as drug delivery systems
JPH08143651A (ja) * 1994-11-22 1996-06-04 Daicel Chem Ind Ltd アミノ基含有ポリラクトン類およびその製造方法
US6476156B1 (en) * 2000-02-14 2002-11-05 Korea Institute Of Science And Technology Biodegradable triblock copolymers and process for their preparation

Family Cites Families (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
GB9713979D0 (en) * 1997-07-03 1997-09-10 Univ Nottingham Method for attaching peg to macromolecules
US6998115B2 (en) * 2000-10-10 2006-02-14 Massachusetts Institute Of Technology Biodegradable poly(β-amino esters) and uses thereof
MXPA03004112A (es) * 2000-11-09 2003-08-19 Astrazeneca Ab Composicion farmaceutica oral que contiene un copolimero de bloque.

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS62218456A (ja) * 1986-03-19 1987-09-25 Sakata Shokai Ltd 被覆用樹脂組成物
US4882168A (en) * 1986-09-05 1989-11-21 American Cyanamid Company Polyesters containing alkylene oxide blocks as drug delivery systems
JPH08143651A (ja) * 1994-11-22 1996-06-04 Daicel Chem Ind Ltd アミノ基含有ポリラクトン類およびその製造方法
US6476156B1 (en) * 2000-02-14 2002-11-05 Korea Institute Of Science And Technology Biodegradable triblock copolymers and process for their preparation

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US9505867B2 (en) 2005-05-31 2016-11-29 Ecole Polytechmique Fédérale De Lausanne Triblock copolymers for cytoplasmic delivery of gene-based drugs
JP2008222745A (ja) * 2007-03-08 2008-09-25 Sungkyunkwan Univ Foundation For Corporate Collaboration Ｐｈ及び温度敏感性のブロック共重合体ヒドロゲル
JP2010530906A (ja) * 2007-06-11 2010-09-16 ルレデーラ フンダシオン パラ エル デサローヨ テクノロジコ イ ソシアル 不溶性薬剤の制御放出のための、シクロデキストリンおよびポリ（アミドアミン）に基づく超分岐ポリマー
WO2009031861A2 (fr) * 2007-09-06 2009-03-12 Sungkyunkwan University Foundation For Corporate Collaboration Copolymère séquencé sensible à la température et au ph extrêmement sûr in vivo, hydrogel et système d'administration de médicament utilisant ceux-ci
WO2009031861A3 (fr) * 2007-09-06 2009-05-07 Univ Sungkyunkwan Found Copolymère séquencé sensible à la température et au ph extrêmement sûr in vivo, hydrogel et système d'administration de médicament utilisant ceux-ci
US9469728B2 (en) 2007-09-06 2016-10-18 Sungkyunkwan University Foundation For Corporate Temperature and pH-sensitive block copolymer having excellent safty in vivo and hydrogel and drug delivery system using thereof
US8703194B2 (en) 2007-11-02 2014-04-22 Agency For Science, Technology And Research Stimulus-responsive biodegradable polymers and methods of preparation
JP2009132825A (ja) * 2007-11-30 2009-06-18 Agency For Science Technology & Research 刺激応答型生分解性高分子及びその製造方法
US8187222B2 (en) 2008-09-12 2012-05-29 Boston Scientific Scimed, Inc. Devices and systems for delivery of therapeutic agents to body lumens
WO2010030728A2 (fr) * 2008-09-12 2010-03-18 Boston Scientific Scimed, Inc. Dispositifs et systèmes pour distribuer des agents thérapeutiques à des lumières corporelles
WO2010030728A3 (fr) * 2008-09-12 2010-06-03 Boston Scientific Scimed, Inc. Dispositifs et systèmes pour distribuer des agents thérapeutiques à des lumières corporelles
US10335499B2 (en) 2008-11-25 2019-07-02 École Polytechnique Fédérale De Lausanne (Epfl) Block copolymers and uses thereof
US9271929B2 (en) 2008-11-25 2016-03-01 École Polytechnique Fédérale De Lausanne (Epfl) Block copolymers and uses thereof
US9175114B2 (en) 2009-01-15 2015-11-03 Living Proof, Inc. Beta-amino ester compounds and uses thereof
US8362175B2 (en) 2009-01-15 2013-01-29 Living Proof, Inc. Beta-amino ester compounds and uses thereof
US8748551B2 (en) 2009-01-15 2014-06-10 Living Proof, Inc. β-amino ester compounds and uses thereof
US8163861B2 (en) 2009-01-15 2012-04-24 Living Proof, Inc. Beta-amino ester compounds and uses thereof
US8796234B2 (en) 2009-11-24 2014-08-05 Agency For Science, Technology And Research Crosslinking branched molecule through thiol-disulfide exchange to form hydrogel
US9034930B2 (en) 2010-03-09 2015-05-19 Council Of Scientific & Industrial Research Biodegradable polymeric hydrogel composition
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US10039831B2 (en) 2011-03-17 2018-08-07 Georgia Tech Research Corporation Polymer hydrogels for in vivo applications and methods for using and preparing same
WO2012126005A2 (fr) 2011-03-17 2012-09-20 Georgia Tech Research Corporation Hydrogels de polymère pour applications in vivo et leurs procédés d'utilisation et de préparation
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