Photodegradable Polycaprolactone Fumarate Block Copolymers

Abstract

A polycaprolactone fumarate copolymer useful as a material for a biocompatible scaffold for tissue engineering applications is disclosed. The copolymer includes at least one caprolactone unit, at least one fumarate unit, and at least one third unit selected from the group consisting of acrylate units and styrenic units. A linking moiety forms a link between the third unit and at least one caprolactone unit or at least one fumarate unit. The linking moiety can be photodegradable. In one form, the third unit includes at least one methyl methacrylate unit. The copolymer can be used to form the wall of a nerve conduit.

Claims

1. A method for forming a nanoporous structure, the method comprising: (a) preparing a copolymer comprising (i) at least one caprolactone unit, (ii) at least one fumarate unit, (iii) at least one third unit selected from the group consisting of acrylate units and styrenic units, and (iv) a linking moiety that forms a link between the third unit and at least one caprolactone unit or at least one fumarate unit; and (b) exposing the linking moiety to photons thereby breaking the link and forming the nanoporous structure.

2. The method of claim 1 wherein: the nanoporous structure includes pores having a size in a range from 1 to 500 nanometers.

3. The method of claim 1 wherein: the nanoporous structure includes pores having a size in a range from 10 to 100 nanometers.

4. The method of claim 1 wherein: the nanoporous structure has a porosity in a range of about 10% to about 50%.

5. The method of claim 1 wherein: the nanoporous structure has a porosity in a range of about 20% to about 40%.

6. The method of claim 1 wherein: at least one caprolactone unit and at least one fumarate unit form a main chain of the copolymer, and at least one third unit forms a side chain of the copolymer.

7. The method of claim 1 wherein: the third unit is selected from the group consisting of methyl methacrylate units.

8. The method of claim 1 wherein: the copolymer includes a first block in the main chain, the first block having at least one caprolactone unit or at least one fumarate unit, and the copolymer includes a second block in the main chain, the second block having at least one caprolactone unit or at least one fumarate unit, and the linking moiety links the first block and the second block.

9. The method of claim 1 wherein: the linking moiety comprises an alkanoyl group or an alkanedioyl group.

10. The method of claim 1 wherein: the linking moiety comprises a dioxyalkyl group that links the first block and the second block.

11. The method of claim 1 wherein: the linking moiety comprises a trivalent radical of a [(halo-alkylalkanoyl)oxy]-nitrobenzyl dihydroxyalkyl alkanedioate.

12. The method of claim 1 wherein: the copolymer comprises a polycaprolactone fumarate block and a poly(methyl methacrylate) block linked via a covalent bond.

13. The method of claim 12 wherein: the method comprises exposing the copolymer to ultraviolet light thereby breaking the covalent bond.

14. The method of claim 13 wherein: exposing the copolymer to ultraviolet light does not degrade the polycaprolactone fumarate block.

15. The method of claim 13 wherein: breaking the covalent bond allows for selective removal of the poly(methyl methacrylate) block.

16. The method of claim 12 wherein: the copolymer assembles into an ordered nanomaterial.

17. A copolymer comprising: at least one caprolactone unit; at least one fumarate unit; and a linking moiety that links (i) at least one caprolactone unit and at least one fumarate unit, or (ii) at least one caprolactone unit and at least one caprolactone unit, or (iii) at least one fumarate unit and at least one fumarate unit, wherein the linking moiety comprises a carboxylic acid group.

18. The copolymer of claim 17 wherein: the linking moiety is a bivalent radical of a dihydroxyalkyl ester of alkane-dicarboxylic acid.

19. A photodegradable initiator comprising: a [(halo-alkylalkanoyl)oxy]-nitrobenzyl dihydroxyalkyl alkanedioate.

20. The photodegradable initiator of claim 19 wherein: the [(halo-alkylalkanoyl)oxy]-nitrobenzyl dihydroxyalkyl alkanedioate is (5-[(2-bromo-2-methylpropanoyl)oxy]-2-nitrobenzyl dihydroxypropyl butanedioate).

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0018] FIG. 1 shows one embodiment of a nerve conduit according to the invention.

[0019] FIG. 2 shows example nanostructures formed from block copolymers of the invention.

[0020] FIG. 3 shows a reaction scheme for synthesizing an example PCLF-b-PMMA copolymer according to the invention.

[0021] FIG. 4 shows a reaction scheme for synthesizing a UV degradable initiator according to the invention and for synthesizing another example PCLF-b-PMMA copolymer according to the invention.

[0022] FIG. 5 shows a reaction scheme for synthesis of an example biodegradable nanoporous structure according to the invention.

[0023] FIG. 6 shows differential scanning calorimetry results for various polymers, including an example PCLF-b-PMMA copolymer according to the invention.

[0024] FIG. 7 shows transmission electron microscopy images of a PCL-b-PMMA film.

[0025] FIG. 8 shows transmission electron microscopy images of a PCLF-b-PMMA film according to the invention.

DETAILED DESCRIPTION OF THE INVENTION

[0026] Looking at FIG. 1, there is shown a nerve conduit 10 having a tubular biodegradable wall 12 with hollow channels 14. The channels 14 can have an average transverse diameter in a range from 1 to 500 nanometers, or in a range from 5 to 250 nanometers, or in a range from 10 to 100 nanometers. The wall 12 can have a porosity in a range of about 1% to about 50%, in a range of about 20% to about 40%, or in a range of about 20% to about 35%.

[0027] The nanoporosity provided by the channels 14 has many advantages. For example, nanoporosity allows nutrient and waste exchange between a nerve repair site and the adjacent outer environment. The nanoporosity prevents fibrous tissue infiltration through conduit wall 12. The nanoporosity may provide a route to future control of diffusion by differences in molecules 16 hydrodynamic radii. Optionally, functionalizing the pores with proteins can further control diffusion.

[0028] Referring now to FIG. 2, accessing ten to one hundred nanometer structures can be difficult and numerous tools are not available that allow physical manipulation at a nanometer size scale. Block copolymer assembly is one method that can be used for nanomaterial fabrication. Various nanoporous structures (such as wall 12 in FIG. 1) are shown in FIG. 2 with porosities in the ranges of 0-21%, 21-33% (cylindrical), 33-37% (gyroidal), and 37-50%. Nanostructure block copolymers of the invention can assemble into the structures of FIG. 2.

[0029] The invention provides a biocompatible, biodegradable copolymer suitable for use in wall 12 of the nerve conduit 10 to achieve a nanoporous structure with desired porosity levels. The biocompatible, biodegradable copolymer includes at least one caprolactone unit as follows:

##STR00001##

and at least one fumarate unit as follows:

##STR00002##

and at least one third unit selected from the group consisting of acrylate units and styrenic units. In one embodiment, the third unit is a methyl methacrylate unit as follows:

##STR00003##

[0030] In one form of the copolymer, at least one caprolactone unit and at least one fumarate unit form a main chain of the copolymer, and the third unit forms a side chain of the copolymer. In another form, the copolymer includes a first block in the main chain, and the first block has at least one caprolactone unit and/or at least one fumarate unit; and the copolymer includes a second block in the main chain, and the second block has at least one caprolactone unit and/or at least one fumarate unit.

[0031] A photodegradable linking moiety forms a link between the third unit and at least one caprolactone unit or at least one fumarate unit. In one form, the linking moiety links the first block and the second block in the main chain. In one non-limiting example, the linking moiety comprises an alkanoyl group or an alkanedioyl group. In another non-limiting example, the linking moiety comprises a dioxyalkyl group that links the first block and the second block. In another non-limiting example, the linking moiety comprises a benzyl group. In another non-limiting example, the linking moiety comprises a carbonyl group. In another non-limiting example, the linking moiety comprises a trivalent radical of a [(halo-alkylalkanoyl)oxy]-nitrobenzyl dihydroxyalkyl alkanedioate. In another non-limiting example, the linking moiety comprises a trivalent radical of (5-[(2-bromo-2-methylpropanoyl)oxy]-2-nitrobenzyl dihydroxypropyl butanedioate).

[0032] In one form, the copolymer has a number average molecular weight in the range of 3000 to 60,000 g mol.sup.1, or in the range of 5000 to 25,000 g mol.sup.1, or in the range of 10,000 to 20,000 g mol.sup.1. The copolymer may have a polydispersity index in the range of 1 to 6.

[0033] The invention provides another biocompatible, biodegradable copolymer suitable for use in wall 12 of the nerve conduit 10 to achieve a nanoporous structure with desired porosity levels. The biocompatible, biodegradable copolymer includes at least one caprolactone unit, at least one fumarate unit, and a linking moiety that links (i) at least one caprolactone unit and at least one fumarate unit, or (ii) at least one caprolactone unit and at least one caprolactone unit, or (iii) at least one fumarate unit and at least one fumarate unit. The linking moiety can include a carboxylic acid group. The linking moiety can be a bivalent radical of a dihydroxyalkyl ester of alkane-dicarboxylic acid. The linking moiety can be a bivalent radical of a dihydroxypropyl ester of butane-dicarboxylic acid. In one form, the copolymer has a number average molecular weight in the range of 3000 to 60,000 g mol.sup.1, or in the range of 5000 to 25,000 g mol.sup.1, or in the range of 10,000 to 20,000 g mol.sup.1. The copolymer may have a polydispersity index in the range of 1 to 6.

[0034] Any copolymer according to the invention can be used to form a biodegradable, nanoporous structure. In one form, the structure is a nerve conduit, and a wall of the nerve conduit comprises the copolymer. The structure can include pores having an average diameter in a range from 1 to 500 nanometers, or in a range from 5 to 250 nanometers, or in a range from 10 to 100 nanometers. The structure can have a porosity in a range of about 10% to about 50% or in a range of about 20% to about 40%. The structure can be a scaffold for tissue regeneration.

[0035] The invention also provides a method for forming a nanoporous structure. In the method, one prepares a copolymer including (i) at least one caprolactone unit, (ii) at least one fumarate unit, (iii) at least one third unit selected from the group consisting of acrylate units and styrenic units, and (iv) a linking moiety that forms a link between the third unit and at least one caprolactone unit or at least one fumarate unit. The linking moiety is exposed to photons thereby breaking the link and forming the nanoporous structure. The third unit can be selected from methyl methacrylate units. The nanoporous structure can include pores having a size in a range from 10 to 100 nanometers. The linking moiety can include an alkanoyl group or an alkanedioyl group. The linking moiety can include a carbonyl group. The linking moiety can include a benzyl group. In one version of the invention, the third unit is washed away after exposing the linking moiety to photons, which may have a wavelength in the ultraviolet range. The formed nanoporous structure can have a porosity in a range of about 20% to about 40% or in a range of about 10% to about 50%.

[0036] The invention also provides a photodegradable initiator comprising a [(halo-alkylalkanoyl)oxy]-nitrobenzyl dihydroxyalkyl alkanedioate. In one non-limiting example, the [(halo-alkylalkanoyl)oxy]-nitrobenzyl dihydroxyalkyl alkanedioate is (5-[(2-bromo-2-methylpropanoyl)oxy]-2-nitrobenzyl dihydroxypropyl butanedioate).

[0037] As used herein, a biocompatible material is one which stimulates only a mild, often transient, implantation response, as opposed to a severe or escalating response. As used herein, a biodegradable material is one which decomposes under normal in vivo physiological conditions into components which can be metabolized or excreted. The term number average molecular weight (M.sub.n) refers to the total weight of all the molecules in a polymer sample divided by the total number of moles present (M.sub.n=.sub.iN.sub.iM.sub.i/.sub.IN.sub.i). Although number average molecular weight can be determined in a variety of ways, with some differences in result depending upon the method employed, it is convenient to employ gel permeation chromatography or endgroup analysis. As used herein, weight average molecular weight is defined as M.sub.w=.sub.iN.sub.iM.sub.i.sup.2/.sub.iN.sub.iM.sub.i. Although weight average molecular weight (M.sub.w) can be determined in a variety of ways, with some differences in result depending upon the method employed, it is convenient to employ gel permeation chromatography. As used herein, the term polydispersity or polydispersity index (PDI) refers to the ratio of a materials' weight average molecular weight divided by its number average molecular weight (M.sub.w/M.sub.n).

EXAMPLES

[0038] The following Examples have been presented in order to further illustrate the invention and are not intended to limit the invention in any way.

A. Synthesis of Poly(Caprolactone Fumarate)

[0039] A poly(caprolactone fumarate) macromer can be synthesized using the method described in Example A of U.S. Patent Application Publication No. 2007/0043202.

B. Synthesis of a PCLF-b-PMMA Copolymer

[0040] Looking at FIG. 3, compound 3a (2,3-dihydroxypropyl 2-bromo-2-methylpropanoate) is reacted with poly(caprolactone fumarate) and methyl methacrylate using ring opening polymerization (ROP) and/or atom transfer radical polymerization (ATRP) and/or condensation polymerization to form compound 3b (a PCLF-b-PMMA copolymer) wherein n, f, and E are integers preferably in the range of 1 to 50.

C. Synthesis of a UV Degradable Initiator

[0041] Looking at FIG. 4, compound 4a ((2,2-dimethyl-1,3-dioxolan-4-yl)methanol) is reacted with compound 4b (dihydrofuran-2,5-dione) in the presence of dichloromethane and ethanolamine to produce compound 4c (4-[(2,2-dimethyl-1,3-dioxolan-4-yl)methoxy]-4-oxobutanoic acid). Compound 4c is reacted with compound 4d (3-(hydroxymethyl)-4-nitrophenyl 2-bromo-2-methylpropanoate) in the presence of sulfur oxychloride, dichloromethane and ethanolamine and washed with acetic acid and water to produce compound 4e (5-[(2-bromo-2-methylpropanoyl)oxy]-2-nitrobenzyl dihydroxypropyl butanedioate), a UV degradable initiator.

D. Synthesis of a PCLF-b-PMMA Copolymer

[0042] Still referring to FIG. 4, compound 4e is reacted with poly(caprolactone fumarate) and methyl methacrylate using ring opening polymerization (ROP) and/or atom transfer radical polymerization (ATRP) and/or condensation polymerization to form compound 4f (a PCLF-b-PMMA copolymer) wherein n, f, and E are integers preferably in the range of 1 to 50.

E. Scheme for Synthesis of a Biodegradable Nanoporous Structure

[0043] Looking at FIG. 5, we have developed a block copolymer approach whereby an A-B block copolymer is connected via a photodegradable linkage (see compound 5a). After assembly into a nanostructure, the individual polymer chains can be separated by exposure to UV light. At this point, the unwanted block (compound 5b) can be washed away, and the biodegradable nanoporous structure remains (compound 5c). Compound 5c possesses a single carboxylic acid per polymer. The intramolecular rearrangement is advantageous for solid state reactions.

F. Characterization of a PCLF-b-PMMA Copolymer

[0044] The number average molecular weight (M.sub.n) of a polycaprolactone (PCL) sample, a polycaprolactone-b-poly(methyl methacrylate) (PCL-b-PMMA) sample, and compound 4f (PCLF-b-PMMA) were determined by gel permeation chromatography. The composition percentage was determined by .sup.1H NMR methyl vs. methylene. Table 1 shows the results below.

TABLE-US-00001 TABLE 1 M.sub.n PCL PMMA Polymer (g mol.sup.1) % % PCL 7500 PCL-b-PMMA 12500 67 PCLF-b-PMMA 16000 33

G. Characterization of Thermal Transitions and Crystalline Properties

[0045] In order investigate the material properties of a polycaprolactone (PCL) sample, a (PCL-b-PMMA) sample, and a PCLF-b-PMMA sample after exposure to UV light, differential scanning calorimetry experiments were performed. FIG. 6 shows that PCL is approximately 32% crystalline and has a H of 66 J/g. PCL-b-PMMA shows 19% crystalline and a H of 40 J/g. PCLF-b-PMMA after UV shows that the material is back to 33% crystalline and has a H of 68 J/g. This indicates the material after UV degradation is mainly PCLF.

H. Transmission Electron Microscopy (TEM)

[0046] Films of PCL-b-PMMA and PCLF-b-PMMA were cast from a chloroform solvent. The nanostructure block copolymer films were cut into 90 nanometer sections, stained with OsO.sub.4, and imaged using TEM. Referring to FIGS. 7 and 8, three dimensional structures were viewed in two dimensions for the 90 nanometer sections. Results were dependent on the film/section orientation. In FIG. 7, PCL-b-PMMA showed a porosity in the range 33-37%. In FIG. 8, the PCLF-b-PMMA showed a porosity in the range 21-33%.

I. UV Degradation Tests

[0047] Films of PCLF-b-PMMA were cast from a chloroform solvent. The films were irradiated with 365 nanometer light. Submersion in methanol during irradiation aided removal of poly(methyl methacrylate) (PMMA). The films turned orange in color as degradation occurred after roughly thirty minutes. Irradiation times ranged from 30 minutes to 36 hours. .sup.1H NMR characterization after UV exposure showed that 80-90% of the PMMA was removed from the entire sample.

[0048] Thus, we have synthesized a UV degradable initiator that can be used to synthesize PCLF block copolymers. We have also synthesized a UV degradable poly(caprolactone fumarate)-b-poly(methyl methacrylate) block copolymer. Polycaprolactone fumarate (PCLF) was chosen as the degradable polymer of choice to create a nanoporous scaffold for nerve conduit applications. We have assembled this block copolymer into nanostructured materials which were imaged using transmission electron microscopy. We have further showed that we can degrade this material and selectively remove 90% of the poly(methyl methacrylate) block as shown by .sup.1H NMR. We have also shown that poly(methyl methacrylate) (PMMA) is mostly removed by differential scanning calorimetry. Although we have made a PCLF-b-PMMA block copolymer, PMMA can be substituted with any acrylate or styrene with no changes. Additionally, the initiator and block copolymer design can be altered to incorporate different initiators for other polymer systems.

[0049] Although the present invention has been described in detail with reference to certain embodiments, one skilled in the art will appreciate that the present invention can be practiced by other than the described embodiments, which have been presented for purposes of illustration and not of limitation. Therefore, the scope of the appended claims should not be limited to the description of the embodiments contained herein.