# Patent

## Title
Biodegradable copolymer and thermosensitive material

## Abstract

The disclosed is a biodegradable copolymer, an amphiphilic diblock copolymer, composed of a hydrophilic segment and a hydrophobic segment. The hydrophilic segment is an endcapped PEG or derivatives thereof. The hydrophilic segment is a random polymer polymerized of lactone or cyclic C3-C6 molecule and lactic acid/glycolic acid. There is no coupling agent between the hydrophilic and hydrophobic segments, and the biodegradable copolymer is formed by one-pot ring-opening polymerization. The biodegradable copolymer can be dissolved in water to form a thermosensitive material having a phase transfer temperature of 25 to 50° C., thereby being applied to biological activity factor delivery, tissue engineering, cell culture and biological glue.

## Background
CROSS REFERENCE TO RELATED APPLICATIONS
This Application claims priority of Taiwan Patent Application No. 097111275, filed on Mar. 28, 2008, the entirety of which is incorporated by reference herein. This Application claims priority of Taiwan Patent Application No. 097141336, filed on Oct. 28, 2008, the entirety of which is incorporated by reference herein
BACKGROUND OF THE INVENTION
1. Field of the Invention
The present invention relates to a biodegradable copolymer, and in particular relates to the thermosensitive property of a solution thereof.
2. Description of the Related Art
For hard tissue recovery/surgery, fixation of bone fragments in a comminuted or joint fracture is one of the most challenging procedures. For fixation, a K-pin, a bone nail, or a steel wire may be utilized, however, ablation of soft tissue adhered on the bone fragments is necessary before using the fixation materials. Therefore, blood circulation and bone healing are degraded, and the bone fragments increase risk of destruction into a more powdery form. Commercially available bone cement (e.g. polymethylmethacrylate), serves as an adhesion for bone fragments or a filler for bone cavities. The biological inert bone cement rarely causes allergic reactions and cannot be absorbed by the human body, such that complete bone healing of the bone fragments fixed by the bone cement is difficult. As such, the bone cement is not suitable for fracture caused by trauma. For orthopedist, an absorbable bioadhesion material for bone fragment adhesion to improve bone healing is demanded.
For soft tissue recovery/surgery, an endoscope is utilized for minimal invasive surgery, specially in shoulder reconstructive surgery. Conventional techniques utilize drills to drill a hole through the shoulder bone to seam bone and tendons, thereby consuming time. Recently, sutureless anchors and knotless anchors have been popularly utilized to become mainstream techniques for saving operating time due to inconvenience, and further prevent abrasion and tissue reaction caused from residue knots. If the sutureless anchor is utilized for an ablated tissue, the stress of the anchor will be concentrated on the ablated tissue to form a bulge, such that the bioadhesion is utilized to adhere to the ablated tissues to accelerate tissue regeneration. If the knotless anchor is utilized for the ablated tissue, the healing effect will be dependant upon the operating skill of the surgeon, anchor strength, and degree of tissue regeneration. Meanwhile, the bioadhesion is utilized to adhere to the ablated tissues to improve the healing effect.
For a hip joint, avascular necrosis (AVN) is mainly found in youths and elderly. Although an artificial hip joint can replace a hip joint with AVN, early treatment can also be utilized to cure 75% AVN patients. The present treatment for AVN is core decompression, wherein a hole is drilled from the edge to the front end of the thighbone, and autogenic bone grafts full of veins are implanted into the drilled hole, such that blood is able to flow to the necrosis zone for bone regeneration. However, the described treatment needs an additional operation to collect the autogenic bone grafts from the patient's body, thereby extending the healing period. Note also that a combination of bioadhesion and medicine can be implanted into the thighbone to stimulate veins and bone regeneration following degradation of the bioadhesion to immediately release the medicine.
Meanwhile, delivery of biological activity factors such as medicines, cells, growth factors, and genes are important in biological medicine applications such as medicine therapy, gene therapy, and tissue engineering. The materials which serve as a delivery carrier must possess bio-compatibility and biodegrability for an implanting in vivo. In addition, the material should easily flow in vitro to evenly mix with medicine. The material which is subsequently injected into a body by a catheter or an endoscope, should transform to a gel after injection for fixing activity factors in the predetermined tissue regions, and slowly releasing the activity factors to complete treatment. Presently, a suitable delivery material is rare. Some materials form a gel through chemical reaction, thereby influencing the activity of the biological activity factors or damaging the implanted tissue region. Some materials have excellent thermosensitivity and gel formability but poor biodegradability, thereby preventing applicability for an implantation in vivo.
In U.S. Pat. No. 5,514,380, Song discloses a biodegradable copolymer gel which serves as a medicine delivery matrix. The copolymer is composed of hydrophilic and hydrophobic segments, the hydrophilic segment is primarily polyethyleneoxide (PEO), and the hydrophobic segment is polylactide (PLA), polyglycolide (PGA), polylactide-glycolide (PLGA), or polycaprolactone (PCL). The multi block copolymer is applied to a drug releasing carrier. However, the patent discloses a multi block copolymer such as ABABAB, wherein A means hydrophilic segment and B means hydrophobic segment, without disclosing the thermosensitive property. Furthermore, the patent also fails to disclose that the hydrophobic segment is a random copolymer of lactone or cyclic C3-C6 molecule and lactic acid/glycolic acid.
MacroMed Corporation has four U.S. Pat. No. 5,702,717, No. 6,004,573, U.S. Pat. No. 6,117,949, and U.S. Pat. No. 6,201,072. The patents disclose biodegradable thermosensitive triblock copolymer, such as ABA or BAB. A hydrophilic segment A is a polyethylene glycol (PEG), and a hydrophobic segment B is a polyester. The biodegradable triblock copolymer has a molecular weight of 2000 to 4990 g/mol, and a reverse thermal gelation. The hydrogel, prepared from the copolymer, can be mixed with a medicine at room temperature. The mixture will transform to a gel after being injected into homoiothermic animals, and the medicine release rate is determined by the hydrolysis rate of the gel in vivo. The hydrolysis product of the gel is free of bio-toxicity. However, the patents do not disclose the thermosensitive diblock copolymer AB, and that the hydrophobic segment is a random copolymer of lactone or cyclic C3-C6 molecule and lactic acid/glycolic acid.
In U.S. Pat. Nos. 6,451,346 and 6,004,573, Amgen Corporation discloses ABA and AB block copolymers with pH and thermo sensitivities. The hydrophilic segment A of the copolymer is PEG, and the hydrophilic segment B is PLA or PLGA. However, the patent does not disclose that the hydrophobic segment is a random copolymer of lactone or cyclic C3-C6 molecule and lactic acid/glycolic acid. Furthermore, succinic anhydride is necessary to be a crosslinking agent between the hydrophobic and hydrophilic segments, and is different from the direct ring-opening product of the invention.
In U.S. Pat. No. 7,087,244, Byeongmoon Jeong discloses triblock copolymers such as ABA and BAB with thermosensitivity. The copolymers are applied as a biological activity factor release carrier. However, the patent does not disclose that the hydrophobic segment is a random copolymer of lactone or cyclic C3-C6 molecule and lactic acid/glycolic acid, and the copolymer thereof is triblock not diblock.
In U.S. Pat. Nos. 5,410,016, 5,567,435, and 5,986,043, Hubbell discloses a diblock copolymer AB system. The initiator thereof is a photo initiator or a thermo initiator. In U.S. Pat. No. 5,410,016, the hydrophobic segment B is poly(α-hydroxy acid) (PHA), poly(glycolic acid) (PGA), PLA, or polylactone such as poly(ε-caprolactone) (PCL), poly(δ-valerolactone) (PVL), or poly(λ-butyrolactone) (PBL). However, the patent does not disclose that the hydrophobic segment is a random copolymer of lactone or cyclic C3-C6 molecule and lactic acid/glycolic acid. Alternatively, in the U.S. Pat. Nos. 5,567,435, and 5,986,043, the diblock copolymer has a PEG center with extension such as PHA, PGA, PLA, polylactone, poly(amino acid), polyanhydride, poly(orthoester), poly(orthocarbonate), or poly(phosphoester). However, similarly, the patents also fail to disclose that the hydrophobic segment is a random copolymer of lactone or cyclic C3-C6 molecule and lactic acid/glycolic acid. In addition, U.S. Pat. No. 6,060,582 discloses a photopolymerized diblock copolymer with three segments B, L, and P. P is an ethylene segment for photo initiation, B is PEG, and L is PHA, acrylic ester monomer or oligomer. The P segment and L segment are first coupled to form a hydrophobic segment, and the hydrophobic segment is then coupled with the B segment. Although the diblock copolymer in U.S. Pat. No. 6,060,582 can be applied for anti-adhesion after surgery, releasing medicine, tissue adhesion, and preventing cell adhesion to tissue, it still fails to disclose that the hydrophobic segment is a random copolymer of lactone or cyclic C3-C6 molecule and lactic acid/glycolic acid.
In U.S. Pat. No. 5,514,380, Samyang Corporation discloses a copolymer with hydrophilic and hydrophobic segments. The hydrophilic segment is PEO and the hydrophobic segment is PLA, PGA, PLGA, or PCL. The copolymer is a thermoplastic biodegradable gel. However, the patent fails to disclose thermosensitivity, and also fails to disclose that the hydrophobic segment is a random copolymer of lactone or cyclic C3-C6 molecule and lactic acid/glycolic acid.
In U.S. Pat. No. 6,136,333, Life Medical Sciences Corporation discloses block copolymers AB and ABA. Segment A is polyester, segment B is polyoxyalkylene polymer unit, segments A and B have a ratio of 1:0.1 to 1:100, and the segments A and B have hexamethylene diisocyanate (HMDI) as crosslinking agent therebetween. The block copolymers serve as anti-adhesion material. However, the patent does not disclose the thermosensitivity, and fails to disclose that the hydrophobic segment is a random copolymer of lactone or cyclic C3-C6 molecule and lactic acid/glycolic acid.
In U.S. Pat. No. 6,841,617, Battle Memorial Institute discloses an An(B) block copolymer that is a thermosensitive and biodegradable gel, wherein A is PEG and B is polyester. However, the polymerization thereof is through grafting, and it also fails to disclose that the hydrophobic segment is a random copolymer of lactone or cyclic C3-C6 molecule and lactic acid/glycolic acid.
In U.S. Pat. No. 7,094,810, Lapopharm Corporation discloses a block copolymer AB. The hydrophilic segment A is PEO, and an the hydrophobic segment B is poly(butyl(alkyl)acrylate-co-(alkyl)acrylic acid. However, the patent fails to disclose that the hydrophobic segment is a random copolymer of lactone or cyclic C3-C6 molecule and lactic acid/glycolic acid.
In U.S. Pat. No. 7,153,520, Samyang Corporation discloses an amphiphilic diblock copolymer that is a slow-degradable drug delivery material, wherein the hydrophilic segment is PEG, and hydrophobic is PLA, PCL, PLGA, PLDO, poly(lactide-co-p-dioxanone), poly(orthoester), polyanhydride, poly(amino acid), and polycarbonate. However, the patent does not disclose that the copolymer is thermosensitive.
In U.S. Pat. No. 4,188,373, Krezanoski discloses an aqueous solution PEO-PPO polymer with a phase transfer temperature of 25° C. to 40° C., wherein the hydrogel system can be applied as a drug delivery carrier. However, the polymer material thereof has no hydrophobic segment.
In U.S. Pat. No. 4,474,752, Haslam discloses a medicine delivery system, wherein the medicine is liquid at room temperature and transforms to a semisolid or gel at body temperature. The hydrogel has 40-80% PEO and 20-60% PPO and a molecular weight of 7,000 to 50,000. However, the hydrogel does not have a hydrophobic segment.
In the Chem. Commun. 2001, 1516-1517, Jeong discloses a thermosensitive copolymer PLGA-g-PEG. The backbone of the polymer is PLGA and the side chains PEG are grafted on the backbone, wherein the lactide, glycolic acid, and PEG have a molar ratio of 3.2:1:2.8. The copolymer has a molecular weight of 9,300, a weight percent in water of 25 wt %, and a phase transfer temperature of 30° C. However, the paper fails to disclose that the hydrophobic segment is a random copolymer of lactone or cyclic C3-C6 molecule and lactic acid/glycolic acid.
In the Polymer 2000, 41, 7091-7097, Lee discloses an amphiphilic polymer material with thermosensitivity. The polymer material is copolymerized of 2-ethyl-2-oxazoline and ε-caprolactone. However, the paper does not disclose that the hydrophobic segment is a random copolymer of lactone or cyclic C3-C6 molecule and lactic acid/glycolic acid.
In the Journal of Controlled Release 2002, 80, 69-77, Kim discloses a thermosensitive composite polymer hydrogel, wherein the hydrogel is photopolymerized of polyethylene oxide-polypropylene oxide and hyaluronic acid. The hydrogel serves as a human growth hormone delivery carrier to study drug release kinetics of related medicines and proteins. However, the polymer hydrogel does not have a hydrophobic segment.
In 1999, You Han Bae utilized condensation of PEG and PLA with carboxylic acid terminals to obtain a thermosensitive block copolymer PLA-PEG-PLA. The PEG has a molecular weight (Mn) of 1,000 to 2,000, and the PLA has a molecular weight of 820 to 3,150. The 1 wt % PLA-PEG-PLA aqueous solution has a low critical solution temperature (LCST) of 27° C. to 45° C., the aqueous solution does not transform into a gel or a semisolid, and the LCST is modified by tuning the length of the hydrophobic segment PLA. However, the thermosensitive polymer is not a diblock copolymer, and the hydrophobic segments thereof were not random copolymers of lactone or cyclic C3-C6 molecule and lactic acid/glycolic acid.
BRIEF SUMMARY OF THE INVENTION
The invention discloses a biodegradable copolymer, having a general formula: A-B; wherein A is a hydrophilic segment, having a general formula:




R1 is C1-8 alkyl; R2 is ethyl, C1-8 alkyl ethyl, C1-8 alkoxyl ethyl, or combinations thereof; and n is an integral of 5-20; and B is a hydrophobic segment, having a general formula:





R3 is hydrogen or methyl; R4 is a linear, branched or cyclic, saturated or unsaturated C2-C5 alkanediyl group optionally comprising one or more hetero atoms including O, S or N; J is C═O, C═S, C—CHO, C═NH, or C═NR5, wherein R5 is C1-C3 alkyl group; K is O, S, C—CHO, NH, or NR6, wherein R6 is C1-C3 alkyl group; x is an integral of 5-20; and y is an integral of 5-20.

The invention also provides a thermosensitive material, comprising water and the biodegradable copolymer as described above dissolved in the water to form a solution, wherein the hydrophobic segment B aggregates to form micelles.
A detailed description is given in the following embodiments with reference to the accompanying drawings.



BRIEF DESCRIPTION OF THE DRAWINGS
The present invention can be more fully understood by reading the subsequent detailed description and examples with references made to the accompanying drawings, wherein:
FIG. 1 is an NMR spectrum of mPEG-PPLA in one example of the invention;
FIG. 2 is a micelle diameter distribution diagram of mPEG-PPLA in one example of the invention;
FIG. 3 is a critical micelle concentration diagram of mPEG-PPLA in one example of the invention;
FIG. 4 is a UV transmittance versus temperature diagram of mPEG-PPLA, mPEG-PVLA, and mPEG-PCLA in examples of the invention;
FIG. 5 is a viscosity versus temperature diagram of mPEG-PPLA, mPEG-PVLA, and mPEG-PCLA in examples of the invention;
FIG. 6 is an NMR spectrum of mPEG-PVLA in one example of the invention;
FIG. 7 is a micelle diameter distribution diagram of mPEG-PVLA in one example of the invention;
FIG. 8 is a critical micelle concentration diagram of mPEG-PVLA in one example of the invention;
FIG. 9 is a medicine release curve of cyclosporine corresponding to different concentrations of mPEG-PVLA solution in one example of the invention;
FIG. 10 is an NMR spectrum of mPEG-PCLA in one example of the invention;
FIG. 11 is a micelle diameter distribution diagram of mPEG-PCLA in one example of the invention;
FIG. 12 is a critical micelle concentration diagram of mPEG-PCLA in one example of the invention;
FIG. 13 is a medicine release curve of cyclosporine corresponding to different concentrations of mPEG-PCLA solution in one example of the invention;
FIG. 14 is an NMR spectrum of mPEG-PSLA in one example of the invention;
FIG. 15 is a micelle diameter distribution diagram of mPEG-PSLA in one example of the invention;
FIG. 16 is a critical micelle concentration diagram of mPEG-PSLA in one example of the invention;
FIG. 17 is a UV transmittance versus temperature diagram of mPEG-PSLA in examples of the invention;
FIG. 18 is a viscosity versus temperature diagram of mPEG-PSLA in examples of the invention;
FIG. 19 is a Sol-Gel-Sol related to temperature and concentration of mPEG-PSLA in examples of the invention;
FIG. 20 is an NMR spectrum of mPEG-PITLA in one example of the invention;
FIG. 21 is a micelle diameter distribution diagram of mPEG-PITLA in one example of the invention;
FIG. 22 is a critical micelle concentration diagram of mPEG-PITLA in one example of the invention;
FIG. 23 is a UV transmittance versus temperature diagram of mPEG-PITLA in examples of the invention;
FIG. 24 is a viscosity versus temperature diagram of mPEG-PITLA in examples of the invention;
FIG. 25 is a UV transmittance versus temperature diagram of mPEG-PNLA, mPEG-PPDLA, mPEG-PTTLA, mPEG-POTLA, mPEG-PACTLA, mPEG-PTCALA in examples of the invention; and
FIG. 26 is a viscosity versus temperature diagram of mPEG-PLA in one comparative example of the invention.



DETAILED DESCRIPTION OF THE INVENTION
The following description is of the best-contemplated mode of carrying out the invention. This description is made for the purpose of illustrating the general principles of the invention and should not be taken in a limiting sense. The scope of the invention is best determined by reference to the appended claims.
The invention discloses a biodegradable copolymer having a general formula as shown in Formula 1. The hydrophilic segment A has a general formula as shown in Formula 2. In Formula 2, R1 is C1-8 alkyl. R2 is ethyl, C1-8 alkyl ethyl, C1-8 alkoxyl ethyl, or combinations thereof. n is an integral of 5-20. The hydrophobic segment B has a general formula as shown in Formula 3. In Formula 3, R3 is hydrogen or methyl. R4 is a linear, branched or cyclic, saturated or unsaturated C2-C5 alkanediyl group; this alkanediyl group can optionally be interrupted by one or more hetero atoms such as O, S or N. J is C═O, C═S, C—CHO, C═NH, or C═NR5, wherein R5 is C1-C3 alkyl group. K is O, S, C—CHO, NH, or NR6, wherein R6 is C1-C3 alkyl group. x is an integral of 5-20. y is an integral of 5-20.

A-B  (Formula 1)





Manufacturing of the biodegradable copolymer is described as below. First, since an equivalent ratio of alcohol and ethylene glycol or derivative thereof is reacted under an acid condition, therefore one terminal of the ethylene glycol is endcapped by an alkoxy group. In one embodiment, the ethylene glycol derivative can be a C1-8 alkyl ethylene glycol such as methyl ethylene glycol, or a C1-8 alkoxy ethylene glycol such as methoxy ethylene glycol. The alcohol is C1-8 alcohol. In one embodiment, the alcohol is methanol. In one embodiment, the methoxy endcapped poly ethylene glycol, the hydrophilic segment of the biodegradable copolymer, has a molecular weight of 300 to 1000.
Next, lactic acid, glycolic acid, or the precursors thereof are selected as a necessary monomer of the hydrophobic segment. The lactic acid can be any type of enantiomers or mixtures thereof, such as D-lactic acid, L-lactic acid, or DL-lactic acid. The lactic acid precursor can be lactide of any type of enantiomers or mixtures thereof, such as D-lactide, L-lactide, or DL-lactide. The lactide has a general formula as shown in Formula 4. The glycolic acid precursor includes glycolide as shown in Formula 5.




In addition, the lactone or the cyclic C3-C6 molecule can be selected as one necessary monomer of the hydrophobic segment. Suitable lactones includes β-propiolactone, γ-thiobutyrolactone, δ-valerolactone, or ε-caprolactone. Suitable cyclic C3-C6 molecule includes 2-Iminothiolane hydrochloride(2-IT), (4R)-(−)-2-Thioxo-4-thiazolidinecarboxylic acid, (−)-2-Oxo-4-thiazolidinecarboxylic acid, DL-N-Acetylhomocysteine thiolactone, 5-Thiazolecarboxaldehyde, γ-butyrolactam, or 1-Methyl-2-pyrrolidinone (NMP).
Subsequently, the endcapped polyethylene glycol or derivatives thereof, lactic acid and/or glycolic acid and precursor thereof, lactone, or cyclic C3-C6 molecule are mixed and reacted to form a diblock copolymer by ring-opening polymerization. The copolymer is a biodegradable material, the hydrophilic segment is endcapped polyethylene glycol or derivatives thereof, and the hydrophobic segment is copolymerized of lactic acid/glycolic acid (or derivatives thereof), lactone, or cyclic C3-C6 molecule. In one embodiment, the lactic acid/glycolic acid molar ratio (x in Formula 3) is greater than or equal to the lactone or cyclic C3-C6 molecule molar ratio (y in formula 3). In one embodiment, the lactic acid/glycolic acid and the lactone or cyclic C3-C6 molecule have a molar ratio (x:y) between 50:50 to 90:10.
Because the manufacturing of the biodegradable polymer of the invention is by one-pot reaction without any additional crosslinking agent or coupling agent between the hydrophilic and hydrophobic segments, synthesis periods and steps are reduced.
The described biodegradable copolymer is further dissolved in water to form a thermosensitive material, so that properties may be measured, e.g. critical micelle concentration, micelle diameter, gelling time, hemolysis, adhesion strength, and drug release. In one embodiment, the biodegradable copolymer and water have a weight ratio of 2:98 to 40:60. In another embodiment, the biodegradable copolymer and water have a weight ratio of 15:85 to 30:70. If the weight ratio is less than 2:98, the solution cannot be gelled. On the other hand, the biodegradable copolymer cannot be totally dissolved in water if the weight ratio is greater than 40:60 or 30:70. As known from invention experiments, the critical micelle concentration of the solution is about 0.003 wt % to 0.07 wt % with a micelle diameter of 5 nm to 500 nm. In one embodiment, the described thermosensitive material has a phase transfer temperature of 25° C. to 50° C. In one embodiment, the described thermosensitive material has a phase transfer temperature of 30° C. to 40° C. The thermosensitive material is liquid when the temperature is lower than the phase transfer temperature, and forms a hydrogel when the temperature is higher than the phase transfer temperature. According to the measurement, the gelling time of the thermosensitive material is less than 30 seconds. The hydrogel has excellent adhesion strength, non-hemolysis, and high biocompatibility. The phase transfer is reversible, meaning that the hydrogel will return to a liquid phase if the temperature is reduced to lower than the phase transfer temperature. In addition, the hydrophobic segment of the copolymer can be hydrolysis to a harmless compound for a human body. The hydrophilic segment of the copolymer can be dissolved in water and then egested by human body. In drug release experiments, the thermosensitive material of the invention releases drug in different ratios corresponding to the concentration thereof, and it can be applied as a long-acting medicine carrier. Accordingly, the thermosensitive material can be applied to biological activity factor delivery, tissue engineering, cell culture, or biological glue, wherein the biological activity factor includes medicine, cell, growth factor, inorganic salt, ceramic material, poly-(amino acid), peptide, protein, gene, DNA sequence or RNA sequence. The biological glue is applied in a cell organism, tissue, implant surface adhesion, soft and hard tissue recovery, or an implant carrier filler, or a hemostatic glue. The thermosensitive material type may be applied through injection, pastille, powder, gel, solution, or oral liquid
EXAMPLES AND COMPARATIVE EXAMPLES
Example 1
Copolymer of Methoxy Endcapped Polyethylene Glycol and Lactide/β-Propiolactone Random Copolymer, Hereinafter mPEG-PPLA
21.14 g of methoxy endcapped polyethylene glycol (mPEG, Molecular weight is 550 g/mol), 50 g of lactide, and 18.77 g of β-propiolactone were subsequently charged in an anhydrous reactor, and the reactor temperature was slowly increased until the substances were totally dissolved. The temperature was continuously increased to 160° C., and 42.0 μL stannous catalyst (stannous 2-ethylhexanoate) was added in the reactor to process polymerization at 160° C. for 8 hours. The reaction is shown in Formula 6. The reaction result was precipitated in ethyl ether/hexane (volume ratio is 1:9) as a semitransparent gel. The semitransparent gel was washed three times to remove monomers and then dried in a vacuum at 40° C. for 24 hours to obtain product mPEG-PPLA.




The copolymer mPEG-PPLA had an NMR spectrum as shown in FIG. 1, and a diameter analysis with instrument Zetasizer 1000H as shown in FIG. 2. The mPEG-PPLA dissolved in water is the thermosensitive material of the invention, and the critical micelle concentration analysis is shown in FIG. 3, the UV transmittance related to temperature is shown in FIG. 4, and the viscosity related to temperature is shown in FIG. 5. When the mPEG-PPLA liquid transforms to hydrogel, its UV transmittance was reduced. The thermosensitive material was a flowable transparent solution at a low temperature, a semitransparent solution with higher viscosity at 25° C., and a non-flowable opaque hydrogel at 40° C. The gelling time of the mPEG-PPLA at 37° C. measured in a water sink was about 27.2 seconds. The hydrogel had adhesion strength of 52 gf/mm2, which is better than that of a commercially available fibrin glue (about 49 gf/mm2).
Example 2
Copolymer of Methoxy Endcapped Polyethylene Glycol and Lactide/δ-Valerolactone Random Copolymer, Hereinafter mPEG-PVLA
17.03 g of methoxy endcapped polyethylene glycol (mPEG, Molecular weight is 550 g/mol), 33.6 g of lactide, and 10.0 g of δ-valerolactone were subsequently charged in an anhydrous reactor, and the reactor temperature was slowly increased until the substances were totally dissolved. The temperature was continuously increased to 160° C., and 34.0 μL stannous catalyst (stannous 2-ethylhexanoate) was added in the reactor to process polymerization at 160° C. for 8 hours. The reaction is shown in Formula 7. The reaction result was precipitated in ethyl ether/hexane (volume ratio is 1:9) as a semitransparent gel. The semitransparent gel was washed three times to remove monomers and then dried in vacuum at 40° C. for 24 hours to obtain product mPEG-PVLA.




The copolymer mPEG-PVLA has an NMR spectrum as shown in FIG. 6, and a diameter analysis with instrument Zetasizer 1000H as shown in FIG. 7. The mPEG-PVLA dissolved in water is the thermosensitive material of the invention, the critical micelle concentration analysis is shown in FIG. 8, the UV transmittance related to temperature is shown in FIG. 4, and the viscosity related to temperature is shown in FIG. 5. When the mPEG-PVLA liquid transformed to hydrogel, its UV transmittance was reduced. The thermosensitive material was a flowable transparent solution at a low temperature, a semi-transparent solution with higher viscosity at 25° C., and a non-flowable opaque hydrogel at 40° C. The gelling time of the mPEG-PVLA at 37° C. measured in a water sink was about 27.5 seconds. The hydrogel had adhesion strength of 58 gf/mm2, which is better than that of commercially available fibrin glue (about 49 gf/mm2). The medicine release experiment of cyclosporine corresponding to different concentrations of mPEG-PVLA solution is shown in FIG. 9. The thermosensitive material mPEG-PVLA solution is suitable for biological activity factor delivery such as medicine release.
Example 3
Copolymer of Methoxy Endcapped Polyethylene Glycol and Lactide/ε-Caprolactone Random Copolymer, Hereinafter mPEG-PCLA
17.62 g of methoxy endcapped polyethylene glycol (mPEG, Molecular weight is 550 g/mol), 33.6 g of lactide, and 11.4 g of ε-caprolactone were subsequently charged in an anhydrous reactor, and the reactor temperature was slowly increased until the substances were totally dissolved. The temperature was continuously increased to 160° C., and 35.0 μL stannous catalyst (stannous 2-ethylhexanoate) was added in the reactor to process polymerization at 160° C. for 8 hours. The reaction is shown in Formula 8. The reaction result was precipitated in ethyl ether/hexane (volume ratio is 1:9) as a semitransparent gel. The semitransparent gel was washed three times to remove monomers and then dried in vacuum at 40° C. for 24 hours to obtain product mPEG-PCLA.




The copolymer mPEG-PCLA had an NMR spectrum as shown in FIG. 10, and a diameter analysis with instrument Zetasizer 1000H as shown in FIG. 11. The mPEG-PCLA dissolved in water is the thermosensitive material of the invention, the critical micelle concentration analysis is shown in FIG. 12, the UV transmittance related to temperature is shown in FIG. 4, and the viscosity related to temperature is shown in FIG. 5. When the mPEG-PCLA liquid transformed to hydrogel, its UV transmittance was reduced. The thermosensitive material was a flowable transparent solution at a low temperature, a semi-transparent solution with higher viscosity at 25° C., and a non-flowable opaque hydrogel at 40° C. The gelling time of the mPEG-PCLA at 37° C. measured in a water sink was about 29.2 seconds. The hydrogel had adhesion strength of 100 gf/mm2, which is better than that of commercially available fibrin glue (about 49 gf/mm2). The medicine release experiment of cyclosporine corresponding to different concentrations of mPEG-PCLA solution is shown in FIG. 13. The thermosensitive material mPEG-PCLA solution is suitable for biological activity factor delivery such as medicine release.
Example 4
Copolymer of Methoxy Endcapped Polyethylene Glycol and Lactide/γ-Thiobutyrolactone Random Copolymer, Hereinafter mPEG-PSLA
18.43 g of methoxy endcapped polyethylene glycol (mPEG, Molecular weight is 550 g/mol), 40 g of lactide, and 7.083 g of γ-thiobutyrolactone were subsequently charged in an anhydrous reactor, and the reactor temperature was slowly increased until the substances were totally dissolved. The temperature was continuously increased to 160° C., and 26.026 μL stannous catalyst (stannous 2-ethylhexanoate) was added in the reactor to process polymerization at 160° C. for 8 hours. The reaction is shown in Formula 9. The reaction result was precipitated in ethyl ether/hexane (volume ratio is 1:9) as a semitransparent gel. The semitransparent gel was washed three times to remove monomers and then dried in vacuum at 40° C. for 24 hours to obtain product mPEG-PSLA.




The copolymer mPEG-PSLA had an NMR spectrum as shown in FIG. 14, and a diameter analysis with instrument Zetasizer 1000H as shown in FIG. 15. The mPEG-PSLA dissolved in water is the thermosensitive material of the invention, the critical micelle concentration analysis is shown in FIG. 16, the UV transmittance related to temperature is shown in FIG. 17, and the viscosity related to temperature is shown in FIG. 18. Sol-Gel-Sol related to temperature and concentration is shown in FIG. 19. When the mPEG-PSLA liquid transformed to hydrogel, its UV transmittance was reduced. The thermosensitive material was a flowable transparent solution at a low temperature, a semi-transparent solution with higher viscosity at 25° C., and a non-flowable opaque hydrogel at 46° C. The gelling time of the mPEG-PSLA at 37° C. measured in a water sink was about 11.83 seconds.
Example 5
Copolymer of Methoxy Endcapped Polyethylene Glycol and Lactide/2-Iminothiolane Hydrochloride Random Copolymer, Hereinafter mPEG-PITLA
2.03 g of methoxy endcapped polyethylene glycol (mPEG, Molecular weight is 550 g/mol), 4.185 g of lactide, and 1 g of 2-Iminothiolane hydrochloride were subsequently charged in an anhydrous reactor, and the reactor temperature was slowly increased until the substances were totally dissolved. The temperature was continuously increased to 200° C., and 2.886 μL stannous catalyst (stannous 2-ethylhexanoate) was added in the reactor to process polymerization at 200° C. for 4 hours. The reaction is shown in Formula 10. The reaction result was precipitated in ethyl ether/hexane (volume ratio is 1:9) as a semitransparent brown gel. The semitransparent gel was washed three times to remove monomers and then dried in vacuum at 40° C. for 24 hours to obtain product mPEG-PITLA.




The copolymer mPEG-PITLA had an NMR spectrum as shown in FIG. 20, and a diameter analysis with instrument Zetasizer 1000H as shown in FIG. 21. The mPEG-PITLA dissolved in water is the thermosensitive material of the invention, the critical micelle concentration analysis is shown in FIG. 22, the UV transmittance related to temperature is shown in FIG. 23, and the viscosity related to temperature is shown in FIG. 24. When the mPEG-PITLA liquid transformed to hydrogel, its UV transmittance was reduced. The thermosensitive material was a flowable transparent solution at a low temperature, a semi-transparent solution with higher viscosity at 25° C., and a non-flowable opaque hydrogel at 40° C.
Example 6
Copolymer of Methoxy Endcapped Polyethylene Glycol and Lactide/N-Methyl-2-Pyrrolidone Random Copolymer, Hereinafter mPEG-PNLA
7.99 g of methoxy endcapped polyethylene glycol (mPEG, Molecular weight is 550 g/mol), 17.43 g of lactide, and 3 g of N-Methyl-2-Pyrrolidone were subsequently charged in an anhydrous reactor, and the reactor temperature was slowly increased until the substances were totally dissolved. The temperature was continuously increased to 160° C., and 11.37 μL stannous catalyst (stannous 2-ethylhexanoate) was added in the reactor to process polymerization at 160° C. for 8 hours. The reaction is shown in Formula 11. The reaction result was precipitated in ethyl ether/hexane (volume ratio is 1:9) as a semitransparent brown gel. The semitransparent gel was washed three times to remove monomers and then dried in vacuum at 40° C. for 24 hours to obtain product mPEG-PNLA. The UV transmittance related to temperature is shown in FIG. 25.




Example 7
Copolymer of Methoxy Endcapped Polyethylene Glycol and Lactide/γ-Butyrolactam Random Copolymer, Hereinafter mPEG-PPDLA
9.12 g of methoxy endcapped polyethylene glycol (mPEG, Molecular weight is 550 g/mol), 20.31 g of lactide, and 3 g of γ-butyrolactam were subsequently charged in an anhydrous reactor, and the reactor temperature was slowly increased until the substances were totally dissolved. The temperature was continuously increased to 160° C., and 12.97 μL stannous catalyst (stannous 2-ethylhexanoate) was added in the reactor to process polymerization at 160° C. for 8 hours. The reaction is shown in Formula 12. The reaction result was precipitated in ethyl ether/hexane (volume ratio is 1:9) as a semitransparent brown gel. The semitransparent yellow gel was washed three times to remove monomers and then dried in vacuum at 40° C. for 24 hours to obtain product mPEG-PPDLA. The UV transmittance related to temperature is shown in FIG. 25




Example 8
Copolymer of Methoxy Endcapped Polyethylene Glycol and Lactide/(−)-2-Oxo-4-Thiazolidinecarboxylic Acid Random Copolymer, Hereinafter mPEG-POTLA
4.81 g of methoxy endcapped polyethylene glycol (mPEG, Molecular weight is 550 g/mol), 9.79 g of lactide, and 2.5 g of (−)-2-Oxo-4-thiazolidinecarboxylic acid were subsequently charged in an anhydrous reactor, and the reactor temperature was slowly increased until the substances were totally dissolved. The temperature was continuously increased to 180° C., and 6.838 μL stannous catalyst (stannous 2-ethylhexanoate) was added in the reactor to process polymerization at 180° C. for 8 hours. The reaction is shown in Formula 13. The reaction result was precipitated in ethyl ether/hexane (volume ratio is 1:9) as a semitransparent brown gel. The semitransparent yellow gel was washed three times to remove monomers and then dried in vacuum at 40° C. for 24 hours to obtain product mPEG-POTLA. The UV transmittance related to temperature is shown in FIG. 25.




Example 9
Copolymer of Methoxy Endcapped Polyethylene Glycol and Lactide/(4R)-(−)-2-Thioxo-4-Thiazolidinecarboxylic Acid Random Copolymer, Hereinafter mPEG-PTTLA
2.31 g of methoxy endcapped polyethylene glycol (mPEG, Molecular weight is 550 g/mol), 4.59 g of lactide, and 1.3 g of (4R)-(−)-2-Thioxo-4-thiazolidinecarboxylic acid were subsequently charged in an anhydrous reactor, and the reactor temperature was slowly increased until the substances were totally dissolved. The temperature was continuously increased to 180° C., and 3.277 μL stannous catalyst (stannous 2-ethylhexanoate) was added in the reactor to process polymerization at 180° C. for 8 hours. The reaction is shown in Formula 14. The reaction result was precipitated in ethyl ether/hexane (volume ratio is 1:9) as a semitransparent brown gel. The semitransparent brown gel was washed three times to remove monomers and then dried in vacuum at 40° C. for 24 hours to obtain product mPEG-PTTLA. The UV transmittance related to temperature is shown in FIG. 25.




Example 10
Copolymer of Methoxy Endcapped Polyethylene Glycol and Lactide/DL-N-Acetylhomocysteine Thiolactone Random Copolymer, Hereinafter mPEG-PACTLA
9.04 g of methoxy endcapped polyethylene glycol (mPEG, Molecular weight is 550 g/mol), 18.09 g of lactide, and 5 g of DL-N-Acetylhomocysteine thiolactone were subsequently charged in an anhydrous reactor, and the reactor temperature was slowly increased until the substances were totally dissolved. The temperature was continuously increased to 160° C., and 12.86 μL stannous catalyst (stannous 2-ethylhexanoate) was added in the reactor to process polymerization at 160° C. for 8 hours. The reaction is shown in Formula 15. The reaction result was precipitated in ethyl ether/hexane (volume ratio is 1:9) as a semitransparent yellow gel. The semitransparent brown gel was washed three times to remove monomers and then dried in vacuum at 40° C. for 24 hours to obtain product mPEG-PACTLA. The UV transmittance related to temperature is shown in FIG. 25.




Example 11
Copolymer of Methoxy Endcapped Polyethylene Glycol and Lactide/5-Thiazolecarboxaldehyde Random Copolymer, Hereinafter mPEG-PTCALA
2.384 g of methoxy endcapped polyethylene glycol (mPEG, Molecular weight is 550 g/mol), 5.09 g of lactide, and 1 g of 5-Thiazolecarboxaldehyde were subsequently charged in an anhydrous reactor, and the reactor temperature was slowly increased until the substances were totally dissolved. The temperature was continuously increased to 160° C., and 3.39 μL stannous catalyst (stannous 2-ethylhexanoate) was added in the reactor to process polymerization at 160° C. for 8 hours. The reaction is shown in Formula 16. The reaction result was precipitated in ethyl ether/hexane (volume ratio is 1:9) as a semitransparent yellow gel. The semitransparent yellow gel was washed three times to remove monomers and then dried in vacuum at 40° C. for 24 hours to obtain product mPEG-PTCALA. The UV transmittance related to temperature is shown in FIG. 25.




Comparative Example 1
Copolymer of Methoxy Endcapped Polyethylene Glycol and Lactide, Hereinafter mPEG-PLA
17.36 g of methoxy endcapped polyethylene glycol (mPEG, Molecular weight is 550 g/mol) and 50 g of lactide were subsequently charged in an anhydrous reactor, and the reactor temperature was slowly increased until the substances were totally dissolved. The temperature was continuously increased to 160° C., and 34.0 μL stannous catalyst (stannous 2-ethylhexanoate) was added in the reactor to process polymerization at 160° C. for 8 hours. The reaction is shown in Formula 17. The reaction result was precipitated in ethyl ether/hexane (volume ratio is 1:9) as a semitransparent gel. The semitransparent gel was washed three times to remove monomers and then dried in vacuum at 40° C. for 24 hours to obtain product mPEG-PLA.




The mPEG-PLA solution viscosity was measured as shown in FIG. 26. Even if the solution was heated to 45° C., the solution viscosity does not remarkably change. Accordingly, the lactone is necessary to copolymerize with the lactide for completing the hydrophobic segment. If the hydrophobic segment is only polymerized by lactide, the diblock copolymer composed of methoxy endcapped polyethylene glycol and polylactide will not have thermosensitivity.
While the invention has been described by way of example and in terms of the preferred embodiments, it is to be understood that the invention is not limited to the disclosed embodiments. To the contrary, it is intended to cover various modifications and similar arrangements (as would be apparent to those skilled in the art). Therefore, the scope of the appended claims should be accorded the broadest interpretation so as to encompass all such modifications and similar arrangements.

## Claims

1. A biodegradable copolymer, having a general formula



in which:
A- is a hydrophilic segment selected from those having a formula:




—B is a hydrophobic segment selected from segments such that the biodegradable copolymer is selected from those having a formula:




n is an integer of 5-20;
x is an integer of 5-20; and
y is an integer of 5-20.

2. The biodegradable copolymer as claimed in claim 1, wherein the hydrophilic segment A- of the biodegradable copolymer has a molecular weight of 300 to 1000.

3. The biodegradable copolymer as claimed in claim 1, wherein the hydrophobic segment —B of the biodegradable copolymer has a molecular weight of 800 to 2000.

4. The biodegradable copolymer as claimed in claim 1, wherein x is greater than or equal to y.

5. The biodegradable copolymer as claimed in claim 1, wherein x and y has a molar ratio of 50:50 to 90:10.

6. A thermosensitive material, comprising:
water; and
the biodegradable copolymer as claimed in claim 1 dissolved in the water to form a solution,
wherein the hydrophobic segment —B aggregates to form micelles.

7. The thermosensitive material as claimed in claim 6, wherein the biodegradable material and the water have a weight ratio of 2:98 to 40:60.

8. The thermosensitive material as claimed in claim 6, wherein the biodegradable material and the water have a weight ratio of 15:85 to 30:70.

9. The thermosensitive material as claimed in claim 6, wherein the micelles have a diameter of 5 nm to 500 nm.

10. The thermosensitive material as claimed in claim 6, further having a phase transfer temperature of 25 to 50° C.

11. The thermosensitive material as claimed in claim 6, further having a phase transfer temperature of 30 to 40° C.

12. The thermosensitive material as claimed in claim 6 is applied through injection, pastille, powder, gel, solution, or oral liquid.

13. The biodegradable copolymer as claimed in claim 1, wherein the hydrophobic segment —B has a formula:



in which:
L is synthesized from a lactone or a C3-C6 molecule selected from, γ-thiobutyrolactone, 2 iminothiolane hydrochloride, (4R)-(−)-2-thioxo-4-thiazolidinecarboxylic acid, (−)-2-oxo-4-thiazolidinecarboxylic acid, DL-N-acetylhomocysteine thiolactone, 5-thiazolecarboxaldehyde, γ-butyrolactam, and 1-methyl-2-pyrrolidinone;
x is an integer of 5-20; and
y is an integer of 5-20.

14. The biodegradable copolymer as claimed in claim 1, wherein the biodegradable copolymer is selected from those having a formula:



in which:
n is an integer of 5-20;
x is an integer of 5-20; and
y is an integer of 5-20.

15. The biodegradable copolymer as claimed in claim 1, wherein the biodegradable copolymer is selected from those having a formula:



in which:
n is an integer of 5-20;
x is an integer of 5-20; and
y is an integer of 5-20.

16. The biodegradable copolymer as claimed in claim 1, wherein the biodegradable copolymer is selected from those having a formula:



in which:
n is an integer of 5-20;
x is an integer of 5-20; and
y is an integer of 5-20.

17. The biodegradable copolymer as claimed in claim 1, wherein the biodegradable copolymer is selected from those having a formula:



in which:
n is an integer of 5-20;
x is an integer of 5-20; and
y is an integer of 5-20.

18. The biodegradable copolymer as claimed in claim 1, wherein the biodegradable copolymer is selected from those having a formula:



in which:
n is an integer of 5-20;
x is an integer of 5-20; and
y is an integer of 5-20.

19. The biodegradable copolymer as claimed in claim 1, wherein the biodegradable copolymer is selected from those having a formula:



in which:
n is an integer of 5-20;
x is an integer of 5-20; and
y is an integer of 5-20.

20. The biodegradable copolymer as claimed in claim 1, wherein the biodegradable copolymer is selected from those having a formula:



in which:
n is an integer of 5-20;
x is an integer of 5-20; and
y is an integer of 5-20.

21. The biodegradable copolymer as claimed in claim 1, wherein the biodegradable copolymer is selected from those having a formula:



in which:
n is an integer of 5-20;
x is an integer of 5-20; and
y is an integer of 5-20.