Preparation and/or formulation of proteins cross-linked with polysaccharides

Abstract

Therapeutic compositions and/or formulations are provided, comprising: at least one cross-linked protein matrix, wherein the at least one cross-linked protein matrix comprises at least one protein residue and at least one saccharide-containing residue, and methods of producing the same. The cross-linked protein matrix may be derived from cross-linking a full length or substantially full length protein, such as tropoelastin, elastin, albumin, collagen, collagen monomers, immunoglobulins, insulin, and/or derivatives or combinations thereof, with a saccharide containing cross-linking agent, such as a polysaccharide cross-linking agent derived from, for example, hyaluronic acid or a cellulose derivative. The therapeutic compositions may be administered topically or by injection. The present disclosure also provides methods, systems, and/or kits for the preparation and/or formulation of the compositions disclosed herein.

1 . A composition, comprising at least one cross-linked protein matrix, wherein the at least one cross-linked protein matrix comprises:
 i) between 25-50 mg/ml of a protein, wherein the protein is tropoelastin;   ii) about 0.25% to about 5% (w/v) of a saccharide-containing residue wherein the saccharide containing residue is hyaluronic acid; and   iii) at least one intermolecular cross-linkage comprising an amide bond between an amine on the tropoelastin and a carboxyl group on the hyaluronic acid.   
     
     
         2 . The composition of  claim 1 , comprising 30 mg/ml tropoelastin and 5% (w/v) hyaluronic acid. 
     
     
         3 . The composition of  claim 1 , wherein the composition is an injectable composition. 
     
     
         4 . The composition of  claim 1 , wherein the composition is extrudable through needles of 18 G to 31 G to a length of between about 5 cm to about 30 cm and the extruded composition substantially holds together without surface support. 
     
     
         5 . The composition of  claim 1 , wherein the composition is extrudable to a length of between 5 cm to 30 cm, 10 cm to 20 cm; or 15 cm to 30 cm without further surface support when extruded through a fine gauge needle and wherein the extruded composition is substantially coherent and substantially holds together. 
     
     
         6 . The composition of  claim 1 , wherein the composition retains sufficient cohesiveness after needle extrusion such that strings of extruded composition do not break during extrusion. 
     
     
         7 . The composition of  claim 1 , wherein the tropoelastin is full length tropoelastin. 
     
     
         8 . The composition of  claim 1 , wherein the composition is tissue compatible, enhances tissue in-growth, enhances tissue re-growth, or combinations thereof. 
     
     
         9 . The composition of  claim 1 , wherein at least 1%, 3%, 5%, or 7% of the tropoelastin is not intermolecularly crosslinked to hyaluronic acid. 
     
     
         10 . The composition of  claim 1 , wherein the at least one cross-linkage further comprises a spacer group. 
     
     
         11 . The composition of  claim 10 , wherein the spacer group comprises a glycol moiety, an ethylene oxide moiety, a polyamine, a polyol, or combinations thereof. 
     
     
         12 . The composition of  claim 1 , wherein the tropoelastin is devoid of intramolecular cross-links. 
     
     
         13 . A prefilled syringe, wherein the prefilled syringe is filled with a composition comprising at least one cross-linked protein matrix, wherein the at least one cross-linked protein matrix comprises:
 i) between 25-50 mg/ml of a protein, wherein the protein is tropoelastin;   ii) about 0.25% to about 5% (w/v) of a saccharide-containing residue wherein the saccharide containing residue is hyaluronic acid; and   iii) at least one intermolecular cross-linkage comprising an amide bond between an amine on the tropoelastin and a carboxyl group on the hyaluronic acid.   
     
     
         14 . The prefilled syringe of  claim 13 , further comprising a needle, wherein the needle comprises a gauge range from about 18 gauge to about 31 gauge. 
     
     
         15 . The prefilled syringe of  claim 14 , wherein the needle comprises a gauge range from about 25 gauge to about 31 gauge. 
     
     
         16 . The prefilled syringe of  claim 13 , further comprising a needle delivery system. 
     
     
         17 . The prefilled syringe of  claim 16 , wherein the needle delivery system is selected from a needle roller ball type system, an automatic injection pen type system, and a mesotherapy injection gun type system. 
     
     
         18 . A composition, comprising at least one cross-linked protein matrix, wherein the at least one cross-linked protein matrix comprises:
 i) about 30 mg/ml of a protein, wherein the protein is tropoelastin;   ii) about 5% (w/v) of a saccharide-containing residue, wherein the saccharide containing residue is hyaluronic acid; and   iii) at least one intermolecular cross-linkage comprising an amide bond between an amine on the tropoelastin and a carboxyl group on the hyaluronic acid.   
     
     
         19 . The composition of  claim 18 , wherein the composition is injectable, and wherein the composition is extrudable through needles of 18 G to 31 G to a length of between about 5 cm to about 30 cm. 
     
     
         20 . The composition of  claim 18 , wherein the composition is tissue compatible, enhances tissue in-growth, enhances tissue re-growth, or combinations thereof.