The invention features polymeric biomaterials formed by nucleophilic addition reactions to conjugated unsaturated groups. These biomaterials may be used for medical treatments.

Description of the Invention

SUMMARY OF THE INVENTION

The following new aspects feature compounds and methods that are useful in the coupling of a pharmaceutically active compound to a polymer, using a conjugate addition reaction, and the cross-linking of the polymers to form a biomaterial, in some embodiments using conjugate addition reactions. Alternatively, the cross-linking may be achieved through other mechanisms, such as free radical polymerization. A polymer coupled to a pharmaceutically active compound may also be cross-linked with another polymer to form a copolymer, such as a colloidal biomaterial. The compounds, precursor components, and biomaterials of the invention may be used in the treatment or prevention of a disease, disorder, or infection.

In a first aspect, the invention provides a compound having the formula: D-Y--C(O)--(CH.sub.2).sub.n--SH or D-Y--C(O)--(CH.sub.2).sub.n--NH.sub.2 wherein D is a pharmaceutically active moiety; n is 1 or 2; and Y is O, NH, or N.

In a second aspect, the invention features a compound the formula: D-Y--C(O)--(CH.sub.2).sub.n--S--(CH.sub.2).sub.2--COX--P, D-Y--C(O)--(CH.sub.2).sub.n--NH--(CH.sub.2).sub.2--COX--P, D-Y--C(O)--(CH.sub.2).sub.n--NH--U--P, or D-Y--C(O)--(CH.sub.2).sub.n--S--U--P wherein D is a pharmaceutically active moiety; n is 1 or 2; X is N or O; P is a water-soluble polymer or a water swellable polymer having one or more conjugated unsaturated groups; Y is O, NH, or N; and U is the product of the addition of a nucleophile to an electrophilic group that is attached to the polymer. It is also contemplated that the compound may have a hydrocarbon moiety in place of one or more hydrogens in one or more of the methylene (CH.sub.2) groups. The half-life of the ester or amide bond onto the pharmaceutically active moiety is between 1 hour and 1 year in an aqueous solution at pH 7.4 and 37.degree. C. Preferably, the half-life is between 1 day and 9 months, more preferably between 2 days and 6 months, and most preferably between 4 days and 3 weeks.

In a third aspect, the invention features a compound having the formula: D-Y--C(O)--CH.dbd.CH.sub.2 wherein D is a pharmaceutically active moiety, and Y is O, NH, or N. It is also contemplated that the compound may have a hydrocarbon moiety in place of one or more hydrogens in the alkene (--CH.dbd.CH.sub.2) group.

In a fourth aspect, the invention includes a compound having the formula: D-Y--C(O)--(CH.sub.2).sub.2--S-L-SH, D-Y--C(O)--(CH.sub.2).sub.2--NH-L-SH, D-Y--C(O)--(CH.sub.2).sub.2--S-L-NH.sub.2, or D-Y--C(O)--(CH.sub.2).sub.2--NH-L-NH.sub.2 wherein D is a pharmaceutically active moiety; Y is O, NH, or N; and L is a linear or branched linker.

In a fifth aspect, the invention features a compound having the formula: D-Y--C(O)--(CH.sub.2).sub.2--S-L-S--CH.sub.2--CH.sub.2--CO--X-P; D-Y--C(O)--(CH.sub.2).sub.2--S-L-S--U--P; D-Y--C(O)--(CH.sub.2).sub.2--NH-L-S--CH.sub.2--CH.sub.2--CO--X-P; D-Y--C(O)--(CH.sub.2).sub.2--NH-L-S--U--P; D-Y--C(O)--(CH.sub.2).sub.2--S-L-NH--CH.sub.2--CH.sub.2--CO--X-P; D-Y--C(O)--(CH.sub.2).sub.2--S-L-NH--U--P; D-Y--C(O)--(CH.sub.2).sub.2--NH-L-NH--CH.sub.2--CH.sub.2--CO--X-P; or D-Y--C(O)--(CH.sub.2).sub.2--NH-L-NH--U--P, wherein D is a pharmaceutically active moiety; L is a linear or branched linker; X is O or N; Y is O, NH, or N; P is a water-soluble polymer or a water-swellable polymer having one or more conjugated unsaturated groups; and U is the product of the addition of a nucleophile to an electrophilic group that is attached to the polymer. The half-life the ester or amide bond onto the pharmaceutically active moiety is between 1 hour and 1 year in an aqueous solution at pH 7.4 and 37.degree. C.

A sixth aspect of the invention features a biomaterial formed from the cross-linking of two or more precursor components having the formula: D-Y--C(O)--(CH.sub.2).sub.n--S--(CH.sub.2).sub.2--COX--P , D-Y--C(O)--(CH.sub.2).sub.n--NH--(CH.sub.2).sub.2--COX--P , D-Y--C(O)--(CH.sub.2).sub.n--NH--U--P, D-Y--C(O)--(CH.sub.2).sub.n--S--U--P, D-Y--C(O)--(CH.sub.2).sub.2--S-L-S--CH.sub.2--CH.sub.2--CO--X--P , D-Y--C(O)--(CH.sub.2).sub.2--S-L-S--U--P, D-Y--C(O)--(CH.sub.2).sub.2--NH-L-S--CH.sub.2-CH.sub.2--CO--X--P , D-Y--C(O)--(CH.sub.2).sub.2--NH-L-S--U--P, D-Y--C(O)--(CH.sub.2).sub.2--S-L-NH--CH.sub.2--CH.sub.2--CO--X--P , D-Y--C(O)--(CH.sub.2).sub.2--S-L-NH--U--P, D-Y--C(O)--(CH.sub.2).sub.2--NH-L-NH--CH.sub.2--CH.sub.2--CO--X--P , or D-Y--C(O)--(CH.sub.2).sub.2--NH-L-NH--U--P, wherein D is a pharmaceutically active moiety; Y is O, NH, or N; L is a linear or branched linker; X is O or N; P is a water-soluble polymer or a water-swellable polymer having one or more conjugated unsaturated groups; and U is the product of the addition of a nucleophile to an electrophilic group that is attached to the polymer. The half-life the ester or amide bond onto the pharmaceutically active moiety is between 1 year and 1 year in an aqueous solution at pH 7.4 and 37.degree. C. In one preferred embodiment, the cross-linking occurs through free radical polymerization or conjugate addition, possibly in the presence of an accelerator. In another preferred embodiment, the cross-linking forms a colloidal material, microsphere, or a nanosphere. The cross-linking may also occur in the presence of sensitive biological molecules or near or at a site in the body of a mammal, such as a human. Preferably, a pharmaceutically active compound is released and delivered to the site.

In a preferred embodiment of the first through sixth aspects of the invention, the pharmaceutically active moiety is derived from one of the group consisting of synthetic organic molecules, naturally occurring organic molecules, nucleic acid molecules, biosynthetic proteins or peptides, naturally occurring peptides or proteins, and modified naturally occurring peptides or proteins. Preferred organic molecules include paclitaxel, doxorubicin, 5-fluorodeoxyuridine, estradiol, 2-methoxyestradiol, and their derivatives.

In preferred embodiments of the second, fourth, fifth, and sixth aspects, the water-soluble or water-swellable polymer is selected from the group consisting of poly(ethylene glycol), poly(ethylene oxide), poly(vinyl alcohol), poly(acrylic acid), poly(ethylene-co-vinyl alcohol), poly(vinyl pyrrolidone), poly(acrylic acid), poly(ethyloxazoline), poly(ethylene oxide)-co-poly(propylene oxide) block copolymers, or water-soluble or water-swellable copolymers containing these polymers, and their derivatives having conjugated unsaturated groups. The unsaturated groups may be identical. One or more of the unsaturated groups may not be coupled to a pharmaceutically active moiety. Preferably, the unsaturated groups are not activated as to undergo nucleophilic substitution reactions. Preferred unsaturated groups include acrylates, methacrylates, acrylamides, methacrylamides, acrylonitiriles, quinones, and their derivatives. In another preferred embodiment, the hydrolysis of the compound results in the release of a pharmaceutically active compound having the formula D-OH, D-NH.sub.2, or D-NH.

In one preferred embodiment of the fourth through sixth aspects, the linker includes one ore more amino acids. Preferably, the linker comprises an adhesion site, growth factor binding site, or protease binding site. Preferred linker also include enzymatically degradable linkers.

If the linker of the fourth through sixth aspects is hydrophilic, it may increase the water solubility of the pharmaceutically active moiety and/or increase the rate of release of the a pharmaceutically active compound derived from D. If the linker is hydrophobic, it may decrease the water solubility of the pharmaceutically active moiety and/or decrease the rate of release of a pharmaceutically active compound derived from D. In other preferred embodiments, the linker includes a nucleophilic group that increases the rate of release of a pharmaceutically active compound having the formula D-OH, D-NH.sub.2, or D-NH by reacting with the ester or amide bond onto D. Preferred linkers also include hydrocarbon moieties containing between 1 and 4 carbon atoms, inclusive.

In a seventh aspect, the invention features a method for making a precursor component of a biomaterial. The method includes (a) attaching a pharmaceutically active compound to a linker molecule to produce a compound having the formula: D-Y--C(O)--(CH.sub.2).sub.n--SH or D-Y--C(O)--(CH.sub.2).sub.n--NH.sub.2 wherein D is a pharmaceutically active moiety; Y is O, NH, or N; and n is 1 or 2; and (b) coupling the product formed in step (a) to a water soluble polymer or a water swellable polymer having two or more conjugated unsaturated groups by a conjugate addition reaction.

A method for making a precursor component of a biomaterial is also provided by an eighth aspect of the invention. This method includes (a) attaching a pharmaceutically active compound to a linker molecule to produce a compound having the formula: D-Y--C(O)--CH.dbd.CH.sub.2 wherein D is a pharmaceutically active moiety, and Y is O, NH, or N; and (b) coupling the product formed in step (a) to a water soluble polymer or a water swellable polymer having two or more conjugated unsaturated groups by a conjugate addition reaction. Preferably, step (a) is performed by condensing an acrylic acid with an alcohol or amine on a pharmaceutically active compound to form an ester or amide bond and produce a modified pharmaceutically active compound.

The ninth aspect of the invention features a method for making a precursor component of a biomaterial which includes (a) attaching a pharmaceutically active compound to a linker to produce a compound having the formula: D-Y--C(O)--(CH.sub.2).sub.2--S-L-SH, D-Y--C(O)--(CH.sub.2).sub.2--NH-L-SH, D-Y--C(O)--(CH.sub.2).sub.2--S-L-NH.sub.2, or D-Y--C(O)--(CH.sub.2).sub.2--NH-L-NH.sub.2 wherein D is a pharmaceutically active moiety; Y is O, NH, or N; and L is a linear or branched linker; and (b) coupling the product formed in step (a) to a water soluble polymer or a water swellable polymer having two or more conjugated unsaturated groups by a conjugate addition reaction. Preferably step (a) is performed by condensing an acrylic acid with an alcohol or amine on a pharmaceutically active compound to form an ester or amide bond, reacting the product with a compound having one protected amine or thiol and one free amine or thiol, and removing the thiol- or amine-protecting group.

In a tenth aspect, the invention features a method for making a precursor component of a biomaterial that includes (a) condensing a linker consisting of one of the following: a thiol-protected mercaptopropionic acid, a thiol-protected mercaptoacetic acid, an amine-protected aminopropionic acid, or an amine-protected glycine; with an alcohol or amine on a pharmaceutically active compound to form an ester or amide bond and produce a modified pharmaceutically active compound; (b) removing the thiol- or amine-protecting group; and (c) coupling the product formed in step (b) to a water soluble polymer or a water swellable polymer having two or more conjugated unsaturated groups by a conjugate addition reaction.

In an eleventh aspect, the invention provides a method for making a precursor component of a biomaterial. This method includes (a) condensing an acrylic acid with an alcohol or amine on a pharmaceutically active compound to form an ester or amide bond and produce a modified pharmaceutically active compound; (b) reacting the modified pharmaceutically active compound with a linker containing one free thiol or amine and one protected thiol or amine through conjugate addition; (c) removing the thiol- or amine-protecting group; and (d) coupling the product formed in step (c) to a water soluble polymer or a water swellable polymer having two or more conjugated unsaturated groups by a conjugate addition reaction.

In a twelfth aspect, the invention features a method for making a precursor component of a biomaterial. This method includes (a) incorporating a nucleophilic amine or thiol into a pharmaceutically active compound and (b) coupling the product formed in step (a) to a water soluble polymer or a water swellable polymer having two or more conjugated unsaturated groups by a conjugate addition reaction. Preferably, the pharmaceutically active compound is DNA, RNA, peptide, or protein. In one preferred embodiment, the DNA or RNA has a base that is modified to contain a thiol.

In preferred embodiments of the seventh through twelfth aspects, the pharmaceutically active compound is selected from the group consisting of synthesized organic molecules, naturally occurring organic molecules, nucleic acid molecules, biosynthetic proteins or peptides, naturally occurring peptides or proteins, and modified naturally occurring peptides or proteins. Preferred organic molecules include paclitaxel, doxorubicin, 5-fluorodeoxyuridine, estradiol, 2-methoxyestradiol, and their derivatives. In one preferred embodiment, the amino acid sequence of the biosynthetic peptide or protein has a cysteine instead of another amino acid found in the corresponding location in a naturally occurring peptide or protein. The attachment of the pharmaceutically active compound to a linker or acrylic acid in step (a) can be performed in the presence of a condensing agent. In other preferred embodiments of these aspects, the water-soluble or water-swellable polymer is selected from the group consisting of poly(ethylene glycol), poly(ethylene oxide), poly(vinyl alcohol), poly(acrylic acid), poly(ethylene-co-vinyl alcohol), poly(vinyl pyrrolidone), poly(acrylic acid), poly(ethyloxazoline), poly(ethylene oxide)-co-poly(propylene oxide) block copolymers, or water-soluble or water-swellable copolymers containing these polymers, and their derivatives having conjugated unsaturated groups. In another preferred embodiment, the conjugated unsaturated groups are identical. Preferred conjugated unsaturated groups included acrylates, methacrylates, acrylamides, methacrylamides, acrylonitiriles, and quinones. In preferred embodiments of these aspects, one or more of the unsaturated groups is not coupled to the pharmaceutically active moiety. Preferably, the unsaturated groups are not activated as to undergo nucleophilic substitution reactions. The methods of these aspects of the invention may include a purification step that is performed prior to the last step. Preferably, the pharmaceutically active compound is released from the precursor component as the original unmodified pharmaceutically active compound. In one preferred embodiment, the number of conjugated unsaturated groups in the polymer is greater than the number of amine or thiol groups in the linker.

The linker molecule of the seventh through twelfth aspects of the invention can have the same embodiments as listed for the linker of the fourth through sixth aspects.

In a thirteenth aspect, the invention features a method of making a biomaterial. This method includes (a) attaching a pharmaceutically active compound to a linker molecule or incorporating a nucleophilic amine or thiol into a pharmaceutically active compound, (b) removing any thiol-or amine-protecting groups in the linker, (c) coupling a thiol, amine, or alkene group in the linker or incorporated into the pharmaceutically active compound to a water soluble polymer or a water swellable polymer having two or more conjugated unsaturated groups by a conjugate addition reaction to form a precursor component, and (d) cross-linking the uncoupled conjugated unsaturated groups in one or more of the precursor components. In one preferred embodiment, a polymer that has one or more conjugated unsaturated groups and that is not coupled to a pharmaceutically active moiety is incorporated into the biomaterial by performing the cross-linking in the presence of this polymer. In another preferred embodiment, the cross-linking is performed in the presence of a linker having two or more nucleophilic groups, and the linker is thereby incorporated into the biomaterial. Preferred linkers include a peptide with an amino acid sequence that is 80%, preferably 90%, or more preferably 100% identical to the sequence GCNNRGDNNCG (SEQ ID NO: 75). Other preferred linkers include those having an amino acid sequence or moiety that provides targeting to cells, tissues, organs, organ systems, or sites within a mammal. In one preferred embodiment, the cross-linking step and/or the formation of the precursor components of the biomaterial occurs within the body of a mammal, such as a human. In another preferred embodiment, the cross-linking occurs through free radical polymerization or conjugate addition reactions at or near a site within the body of a mammal. Preferably, the cross-linking occurs through a self-selective reaction between a thiol or an amine and a conjugated unsaturated group. In another preferred embodiment, the cross-linking forms a hydrogel, a colloidal material, a microsphere, or nanosphere that can be delivered to a mammal, such as a human. In yet another preferred embodiment, the pharmaceutically active compound or a derivative thereof is released from the biomaterial and delivered to a site within the body. Preferably, the half-life the ester or amide bond onto the pharmaceutically active moiety is between 1 hour and 1 year at the site within the body. Preferably, the half-life is between 1 hour and 1 year at pH 7.4 and 37.degree. C. in an aqueous solution. The conjugated unsaturated groups of this aspect may have the same embodiments as listed for the conjugated unsaturated groups of any of the previous aspects.

In a fourteenth aspect, the invention features a biomaterial having a pharmaceutically active moiety. The biomaterial includes an ester or amide bond onto the pharmaceutically active moiety, and this bond has a half-life of between 1 hour and 1 year in an aqueous solution at pH 7.4 and 37.degree. C. Preferably, the half-life of the ester or amide bond onto the pharmaceutically active moiety for this biomaterial, the biomaterial of the sixth aspect of the invention, and the biomaterials formed using the methods of the invention is between 1 day and 9 months, more preferably between 2 days and 6 months, and most preferably between 4 days and 3 weeks.

In preferred embodiments of the thirteenth and fourteenth aspects, the pharmaceutically active moiety has any of the preferred embodiments of the pharmaceutically active moiety of the previous aspects.

In a fifteenth aspect, the invention provides a method of treating or preventing a disease, disorder, or infection by administering to a mammal, such as a human, a compound having the formula: D-Y--C(O)--(CH.sub.2).sub.n--SH, D-Y--C(O)--(CH.sub.2).sub.n--NH.sub.2, D-Y--C(O)--(CH.sub.2).sub.n--S--(CH.sub.2).sub.2--COX--P , D-Y--C(O)--(CH.sub.2).sub.n--NH--(CH.sub.2).sub.2--COX--P , D-Y--C(O)--(CH.sub.2).sub.n--NH--U--P, D-Y--C(O)--(CH.sub.2).sub.n--S--U--P, D-Y--C(O)--CH.dbd.CH.sub.2, D-Y--C(O)--CH--CH.sub.2--P, D-Y--C(O)--(CH.sub.2).sub.2--S-L-SH, D-Y--C(O)--(CH.sub.2).sub.2--NH-L-SH, D-Y--C(O)--(CH.sub.2).sub.2--S-L-NH.sub.2, D-Y--C(O)--(CH.sub.2).sub.2--NH-L-NH.sub.2, D-Y--C(O)--(CH.sub.2).sub.2--S-L-S--CH.sub.2--CH.sub.2--CO--X--P , D-Y--C(O)--(CH.sub.2).sub.2--S-L-S--U--P, D-Y--C(O)--(CH.sub.2).sub.2--NH-L-S--CH.sub.2--CH.sub.2--CO--X--P , D-Y--C(O)--(CH.sub.2).sub.2--NH-L-S--U--P, D-Y--C(O)--(CH.sub.2).sub.2--S-L-NH--CH.sub.2--CH.sub.2--CO--X--P , D-Y--C(O)--(CH.sub.2).sub.2--S-L-NH--U--P, D-Y--C(O)--(CH.sub.2).sub.2--NH-L-NH--CH.sub.2--CH.sub.2--CO--X--P , D-Y--C(O)--(CH.sub.2).sub.2--NH-L-NH--U--P, or Z-P, wherein D is a pharmaceutically active moiety; Y is O, NH, or N; L is a linear or branched linker; X is O or N; Z is a pharmaceutically active moiety in which a nucleophilic amine or thiol has been incorporated; P is a water-soluble polymer or a water-swellable polymer having one or more conjugated unsaturated groups; and U is the product of the addition of a nucleophile to an electrophilic group that is attached to the polymer. The half-life of the ester or amide bond onto the pharmaceutically is between 1 hour and 1 year in an aqueous solution at pH 7.4 and 37.degree. C.

In a sixteenth aspect, the invention features a method of treating or preventing a disease, disorder, or infection in a mammal. This method includes administering to the mammal a biomaterial having a pharmaceutically active moiety. This biomaterial is formed from the cross-linking of one or more of the following precursor components: D-Y--C(O)--(CH.sub.2).sub.n--S--(CH.sub.2).sub.2--COX--P , D-Y--C(O)--(CH.sub.2).sub.n--NH--(CH.sub.2).sub.2--COX--P , D-Y--C(O)--(CH.sub.2).sub.n--NH--U--P, D-Y--C(O)--(CH.sub.2).sub.n--S--U--P, D-Y--C(O)--CH--CH.sub.2--P, D-Y--C(O)--(CH.sub.2).sub.2--S-L-S--CH.sub.2--CH.sub.2--CO--X--P , D-Y--C(O)--(CH.sub.2).sub.2--S-L-S--U--P, D-Y--C(O)--(CH.sub.2).sub.2--NH-L-S--CH.sub.2--CH.sub.2--CO--X--P , D-Y--C(O)--(CH.sub.2).sub.2--NH-L-S--U--P, D-Y--C(O)--(CH.sub.2).sub.2--S-L-NH--CH.sub.2--CH.sub.2--CO--X--P , D-Y--C(O)--(CH.sub.2).sub.2--S-L-NH--U--P, D-Y--C(O)--(CH.sub.2).sub.2--NH-L-NH--CH.sub.2--CH.sub.2--CO--X--P , D-Y--C(O)--(CH.sub.2).sub.2--NH-L-NH--U--P, or Z-P, wherein D is a pharmaceutically active moiety; Y is O, NH, or N; L is a linear or branched linker; X is O or N; Z is a pharmaceutically active moiety in which a nucleophilic amine or thiol has been incorporated; P is a water-soluble polymer or a water-swellable polymer having one or more conjugated unsaturated groups; and U is the product of the addition of a nucleophile to an electrophilic group that is attached to the polymer. The half-life of the ester or amide bond onto the pharmaceutically is between 1 hour and 1 year in an aqueous solution at pH 7.4 and 37.degree. C.

In a seventeenth aspect, the invention provides a method of treating or preventing a disease, disorder, or infection in a mammal. This method includes (a) attaching a pharmaceutically active compound to a linker molecule, (b) removing any thiol-or amine-protecting groups in the linker, (c) coupling a thiol, amine, or alkene group in the linker to a water soluble polymer or a water swellable polymer having two or more conjugated unsaturated groups by a conjugate addition reaction, and (d) cross-linking the uncoupled unsaturated groups in the polymer at a site within a mammal. In one embodiment of this aspect, one or more of steps (a) through (c) are also performed at a site within a mammal.

In an eighteenth aspect, the invention features a method of treating or preventing a disease, disorder, or infection in a mammal by administering to the mammal a biomaterial having a pharmaceutically active moiety. The biomaterial includes an ester or amide bond onto the pharmaceutically active moiety, and this bond has a half-life of between 1 hour and 1 year in an aqueous solution at pH 7.4 and 37.degree. C.

In a nineteenth aspect, the invention features a method for delivering a pharmaceutically active compound to a cell, tissue, organ, organ system, or body of a mammal. This method includes contacting the cell, tissue, organ, organ system or body with a biomaterial having an ester or amide bond onto a pharmaceutically active moiety. The bond has a half-life of between 1 hour and 1 year in an aqueous solution at pH 7.4 and 37.degree. C., and cleavage of the bond results in the release of a pharmaceutically active compound having the pharmaceutically active moiety.

In a twentieth aspect, the invention features a method for delivering a pharmaceutically active compound to a cell, tissue, organ, organ system, or body of a mammal. This method includes administering to the mammal a biomaterial having a pharmaceutically active moiety. The biomaterial is formed from the cross-linking of a precursor component in the presence of a linker having two or more nucleophilic groups. The precursor component includes a pharmaceutically active moiety coupled to a polymer having two or more conjugated unsaturated groups, and the linker provides targeting to a cell, tissue, organ, organ system, or site within the mammal. A pharmaceutically active compound having the pharmaceutically active moiety is released from the biomaterial at or near the cell, tissue, organ, organ system, or body of the mammal. In one preferred embodiment of this aspect, the biomaterial has an ester or amide bond onto the pharmaceutically active moiety, and the bond has a half-life of between 1 hour and 1 year in an aqueous solution at pH 7.4 and 37.degree. C.

In a twenty-first aspect, the invention provides a method of preventing adhesions, thrombosis, or restenosis in a mammal. This method includes contacting a site in the mammal with a precursor component and cross-linking the precursor component at the site. The precursor component has the formula: D-Y--C(O)--(CH.sub.2).sub.n--S--(CH.sub.2).sub.2--COX--P , D-Y--C(O)--(CH.sub.2).sub.n--NH--(CH.sub.2).sub.2--COX--P , D-Y--C(O)--(CH.sub.2).sub.n--NH--U--P, D-Y--C(O)--(CH.sub.2).sub.n--S--U--P, D-Y--C(O)--CH--CH.sub.2--P, D-Y--C(O)--(CH.sub.2).sub.2--S-L-S--CH.sub.2--CH.sub.2--CO--X--P , D-Y--C(O)--(CH.sub.2).sub.2--S-L-S--U--P, D-Y--C(O)--(CH.sub.2).sub.2--NH-L-S--CH.sub.2--CH.sub.2--CO--X--P , D-Y--C(O)--(CH.sub.2).sub.2--NH-L-S--U--P, D-Y--C(O)--(CH.sub.2).sub.2--S-L-NH--CH.sub.2--CH.sub.2--CO--X--P , D-Y--C(O)--(CH.sub.2).sub.2--S-L-NH--U--P, D-Y--C(O)--(CH.sub.2).sub.2--NH-L-NH--CH.sub.2--CH.sub.2--CO--X--P , D-Y--C(O)--(CH.sub.2).sub.2--NH-L-NH--U--P, or Z-P, wherein D is a pharmaceutically active moiety; Y is O, NH, or N; L is a linear or branched linker; X is O or N; Z is a pharmaceutically active moiety in which a nucleophilic amine or thiol has been incorporated; P is a water-soluble polymer or a water-swellable polymer having one or more conjugated unsaturated groups; and U is the product of the addition of a nucleophile to an electrophilic group that is attached to the polymer. The half-life of the ester or amide bond onto the pharmaceutically is between 1 hour and 1 year in an aqueous solution at pH 7.4 and 37.degree. C.

In a twenty-second aspect, the invention provides a method of preventing adhesions, thrombosis, or restenosis in a mammal. This method includes contacting a site within the mammal with a biomaterial having an ester or amide bond onto a pharmaceutically active moiety. The bond has a half-life of between 1 hour and 1 year in an aqueous solution at pH 7.4 and 37.degree. C., and cleavage of the bond results in the release of a pharmaceutically active compound having the pharmaceutically active moiety.

In one preferred embodiment of the fifteenth through twenty-second aspects, the compound, precursor component, or biomaterial is administered orally, intravenously, intramuscularly, subcutaneously, parenterally, or by any other route sufficient to provide an adequate dose for the prevention or treatment of a disease, disorder, or infection. In another preferred embodiment of these aspects, the ester or amide bond onto the pharmaceutically active moiety has a half-life of between 1 day and 9 months in an aqueous solution at pH 7.4 and 37.degree. C. More preferably, the half-life is between 2 days and 6 months, and most preferably it is between 4 days and 3 weeks in an aqueous solution at pH 7.4 and 37.degree. C. One disease that may be treated or prevented using the methods of these aspects is cancer. Preferably, the mammal is a human. The linker of these aspects may have the same embodiments as listed for the linker of the fourth through sixth aspects. The pharmaceutically active moiety or the conjugated unsaturated groups of these aspects may have the corresponding preferred embodiments listed for any of the previous aspects.

The aforementioned new aspects of the invention may include self-selective conjugate addition reactions between a strong nucleophile and a conjugated unsaturated group for cross-linking of precursor components to form a biomaterial, as we described in the U.S. patent application U.S. Ser. No. 09/496,231, which is incorporated herein by reference. For example, the novel precursor components of the present invention, which have a covalently bound pharmaceutically active moiety, may be cross-linked in the presence of a polymer having two or more nucleophilic groups to form a copolymer in methods that include self-selective conjugate addition reactions. In addition, the methods of the present invention may utilize a self-selective conjugate addition reaction for the coupling of a thiol or amine group, linked to or incorporated into a pharmaceutically active compound, to a conjugated unsaturated group on a polymer for the production of novel compounds.

We now describe the polymeric biomaterials that we previously developed, which are unique in their use of addition reactions between a strong nucleophile and a conjugated unsaturation for polymerizing or cross-linking two or more components in a manner that can be accomplished in the presence of sensitive biological materials. Applications of the process include formation of biomaterials in the presence of drugs, including proteins and DNA, formation of biomaterials in the presence of cells and cell aggregates, and also formation of biomaterials in vivo either within the body or upon the surface of the body. It is possible to form these biomaterials in the presence of sensitive biological materials because of the high self-selectivity of the addition reactions between strong nucleophiles and conjugated unsaturations, that are employed. The strong nucleophile of particular interest in the method described herein is the thiol.

In the formation of the biomaterial in the presence of the sensitive biological materials, two or more liquid components can be mixed together and react to form either an elastic solid, a viscoelastic solid (like a typical solid gel, for example, a gel like gelatin), a viscoelastic liquid (like a typical gel that can be induced to flow, for example, a gel like petroleum jelly), a viscoelastic liquid that is formed of gel microparticles (such as a Carbopol.TM. gel) or even a viscous liquid of a considerably higher viscosity than either of the two precursor components that are mixed together. The chemical conversion from the precursors to the final material is so selective that it can be carried out in the presence of the sensitive biological material, including the case when the biological material is the body itself.

A novel family of potentially highly biomimetic synthetic polymers has been developed. These polymers can: (i) be converted from liquid precursors to polymeric linear or cross-linked biomaterials either in the laboratory or in situ at a site of implantation; (ii) be hydrogels or more substantially non-swelling materials; (iii) present bioactive molecules that serve as adhesion sites, to provide traction for cell invasion; (iv) present bioactive molecules that serve as protease substrate sites, to make the material degrade in response to enzymes, such as collagenase or plasmin, which are produced by cells during cell migration; (v) present growth factor binding sites, to make the material interact with growth factors in a biomimetic manner, by binding them and then releasing them on cellular demand; and (vi) provide for the delivery of protein drugs by hydrolysis or enzymatic degradation of groups contained within the backbone of the polymers that form the gel.

Accordingly, in a twenty-third aspect the invention features a method for making a biomaterial, involving combining two or more precursor components of the biomaterial under conditions that allow polymerization of the two components, where polymerization occurs through self selective reaction between a strong nucleophile and a conjugated unsaturated bond or a conjugated unsaturated group, by nucleophilic addition. The functionality of each component is at least two, and the biomaterial does not comprise unprocessed albumin. In addition, the conjugated unsaturated bond or group is not a maleimide or a vinyl sulfone.

In one embodiment of the twenty-third aspect of the invention, the components are selected from the group consisting of oligomers, polymers, biosynthetic proteins or peptides, naturally occurring peptides or proteins, processed naturally occurring peptides or proteins, and polysaccharides. The polymer may be poly(ethylene glycol), poly(ethylene oxide), poly(vinyl alcohol), poly(ethylene-co-vinyl alcohol), poly(acrylic acid), poly(ethylene-co-acrylic acid), poly(ethyloxazoline), poly(vinyl pyrrolidone), poly(ethylene-co-vinyl pyrrolidone), poly(maleic acid), poly(ethylene-co-maleic acid), poly(acrylamide), or poly(ethylene oxide)-co-poly(propylene oxide) block copolymers. The peptide may comprise an adhesion site, growth factor binding site, or protease binding site.

In another embodiment, the components are functionalized to comprise a strong nucleophile or a conjugated unsaturated group or a conjugated unsaturated bond. Preferably the strong nucleophile is a thiol or a group containing a thiol. Preferably the conjugated unsaturated group is an acrylate, an acrylamide, a quinone, or a vinylpyridinium, for example, 2- or 4-vinylpyridinium. In another embodiment, one component has a functionality of at least three.

In yet other embodiments of the twenty-third aspect of the invention, the method further comprises combining the precursor components with a molecule that comprises an adhesion site, a growth factor binding site, or a heparin binding site and also comprises either a strong nucleophile or a conjugated unsaturated bond or a conjugated unsaturated group. Preferably the strong nucleophile is a thiol or the conjugated unsaturated bond or conjugated unsaturated group is an acrylate, an acrylamide, a quinone, or a vinyl pyridinium.

In still other embodiments of the twenty-third aspect of the invention, the biomaterial is a hydrogel. The biomaterial may also be degradable. The biomaterial may be made in the presence of sensitive biological molecules, or in the presence of cells or tissues. The biomaterial may also be made within or upon the body of an animal.

In still further embodiments of the twenty-third aspect of the invention, the method further comprises combining the precursor components with an accelerator prior to polymerization. The method may also further comprise mixing the precursor components with a component that comprises at least one conjugated unsaturated bond or conjugated unsaturated group and at least one amine reactive group. An additional component may also be applied to the cell or tissue surface site of polymerization, the additional component comprising at least one conjugated unsaturated bond or conjugated unsaturated group and at least one amine reactive group.

In a twenty-fourth aspect, the invention features a biomaterial formed by combining two or more precursor components of a biomaterial under conditions that allow polymerization of the two components, where polymerization occurs through self selective reaction between a strong nucleophile and a conjugated unsaturated bond or a conjugated unsaturated group, by nucleophilic addition. The functionality of each component is at least two, the biomaterial does not comprise unprocessed albumin, and the conjugated unsaturated bond or conjugated unsaturated group is not a maleimide or a vinyl sulfone.

In one embodiment of the twenty-fourth aspect of the invention, the components are selected from the group consisting of oligomers, polymers, biosynthetic proteins or peptides, naturally occurring peptides or proteins, processed naturally occurring peptides or proteins, and polysaccharides. The polymer may be poly(ethylene glycol), poly(ethylene oxide), poly(vinyl alcohol), poly(ethylene-co-vinyl alcohol), poly(acrylic acid), poly(ethylene-co-acrylic acid), poly(ethyloxazoline), poly(vinyl pyrrolidone), poly(ethylene-co-vinyl pyrrolidone), poly(maleic acid), poly(ethylene-co-maleic acid), poly(acrylamide), or poly(ethylene oxide)-co-poly(propylene oxide) block copolymers. The peptide may comprise an adhesion site, growth factor binding site, or protease binding site.

In another embodiment of the twenty-fourth aspect of the invention, the components are functionalized to comprise a strong nucleophile or a conjugated unsaturated group or a conjugated unsaturated bond. Preferably the strong nucleophile is a thiol or a group containing a thiol. Preferably the conjugated unsaturated group is an acrylate, an acrylamide, a quinone, or a vinylpyridinium, for example, 2- or 4-vinylpyridinium. In another embodiment, one component has a functionality of at least three.

In yet other embodiments of the twenty-fourth of the invention, the method further comprises combining the precursor components with a molecule that comprises an adhesion site, a growth factor binding site, or a heparin binding site and also comprises either a strong nucleophile or a conjugated unsaturated bond or a conjugated unsaturated group. Preferably the strong nucleophile is a thiol or the conjugated unsaturated bond or conjugated unsaturated group is an acrylate, an acrylamide, a quinone, or a vinyl pyridinium.

In still other embodiments of the twenty-fourth aspect of the invention, the biomaterial is a hydrogel. The biomaterial may also be degradable. The biomaterial may be made in the presence of sensitive biological molecules, or in the presence of cells or tissues. The biomaterial may also be made within or upon the body of an animal.

In still further embodiments of the twenty-fourth aspect of the invention, the method further comprises combining the precursor components with an accelerator prior to polymerization. The method may also further comprise mixing the precursor components with a component that comprises at least one conjugated unsaturated bond or conjugated unsaturated group and at least one amine reactive group. An additional component may also be applied to the cell or tissue surface site of polymerization, the additional component comprising at least one conjugated unsaturated bond or conjugated unsaturated group and at least one amine reactive group.

In a twenty-fifth aspect, the invention features a method for delivering a therapeutic substance to a cell, tissue, organ, organ system, or body of an animal said method involving contacting the cell, tissue, organ, organ system or body with the biomaterial of the twenty-fourth aspect of the invention, wherein the biomaterial contains a therapeutic substance, whereby the therapeutic substance is delivered to the cell, tissue, organ, organ system, or body of an animal.

In one embodiment, the therapeutic substance is selected from the group consisting of proteins, naturally occurring or synthetic organic molecules, nucleic acid molecules, for example DNA or RNA, and a viral particle. In another embodiment, the therapeutic substance is a prodrug. In still another embodiment, the nucleic acid molecule is an antisense nucleic acid molecule.

In a twenty-sixth aspect, the invention features a method of regenerating a tissue, involving introducing a scaffold to a site, under conditions which permit cell ingrowth. The scaffold may comprising the biomaterial of the twenty-fourth aspect of the invention.

In embodiments of the twenty-sixth aspect of the invention, the scaffold has been pre-seeded with cells. The tissue may be selected from the group consisting of bone, skin, nerve, blood vessel, and cartilage.

In a twenty-seventh aspect, the invention features a method of preventing adhesions, thrombosis, or restenosis, involving contacting a site with the biomaterial precursor components of the twenty-fourth aspect of the invention; and polymerizing the components at the site.

In a twenty-eighth aspect, the invention features a method of sealing a fluid or gas flow, said method comprising the steps of contacting a site within the body of an animal with the biomaterial precursor components of the twenty-fourth aspect of the invention, which may further comprise a component that includes at least one conjugated unsaturated bond or conjugated unsaturated group and a least one amine reactive group; and polymerizing the components at the site.

In preferred embodiments of the twenty-eighth aspect of the invention, the site is a lung, blood vessel, skin, dura barrier, or intestine.

In a twenty-ninth aspect, the invention features a method of encapsulating a cell or tissue, involving combining the precursor components of a biomaterial with a cell or tissue; and polymerizing the components, where polymerization occurs through self selected reaction between a strong nucleophile and a conjugated unsaturated bond or a conjugate unsaturated group, and where the cell or tissue is encapsulated by the polymerized biomaterial.

In an thirtieth aspect, the invention features a method for making a biomaterial, involving combining two or more precursor components of the biomaterial under conditions that allow polymerization of the two components, where the polymerization occurs through self selective reaction between an amine and a conjugated unsaturated bond or a conjugated unsaturated group, by nucleophilic addition, wherein the functionality of each component is at least two, and wherein the biomaterial does not comprise unprocessed albumin, and the unsaturated bond or group is not a maleimide or a vinyl sulfone.

In a thirty-first aspect, the invention features a biomaterial, formed by combining two or more precursor components of the biomaterial under conditions that allow polymerization of the two components, where the polymerization occurs through self selective reaction between an amine and a conjugated unsaturated bond or a conjugated unsaturated group, by nucleophilic addition, wherein the functionality of each component is at least two, and wherein the biomaterial does not comprise unprocessed albumin, and the unsaturated bond or group is not a maleimide or a vinyl sulfone.
 

Claim 1 of 42 Claims

1. A biomaterial formed by combining two or more precursor components under conditions that allow polymerization of the two components, wherein said polymerization occurs through self selective reaction between a strong nucleophile and a conjugated unsaturated bond or a conjugated unsaturated group, by nucleophilic addition, wherein each of said precursor components comprises at least two strong nucleophiles or at least two conjugated unsaturated bonds or conjugated unsaturated groups, said biomaterial does not comprise unprocessed albumin, and said unsaturated bond or group is not a maleimide or a vinyl sulfone.

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