Preparation and/or formulation of proteins cross-linked with polysaccharides
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 1-250 mg/mL of at least one protein residue derived from at least one amine-bearing protein;
ii) at least one saccharide-containing cross-linking residue derived from at least one saccharide-containing cross-linking molecule comprising one or more carboxyl groups; and
iii) at least one intermolecular cross-linkage comprising an amide bond between an amine of the at least one protein residue and a carboxyl group of the at least one saccharide-containing cross-linking molecule.
2. The composition of claim 1 , wherein the composition is an injectable composition.
3. The composition of claim 1 , wherein the composition is extrudable through needles of 18G to 31G to a length of between about 5 cm to about 30 cm and the extruded composition substantially holds together without surface support.
4. 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.
5. 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.
6. The composition of claim 1 , wherein the at least one cross-linked protein matrix comprises a 50-99.99% full length protein residue or a full length protein residue.
7. The composition of claim 1 , wherein the composition enables the at least one protein residue to retain its full length, and wherein the composition is needle injectable, retains a coherent structure, and is sufficiently cross-linked to slow the composition's resorption in vivo.
8. The composition of claim 1 , wherein the at least one protein residue is 50-99.99% full length or full length.
9. The composition of claim 1 , wherein the composition is tissue compatible, enhances tissue in-growth, enhances tissue re-growth, or combinations thereof.
10. The composition of claim 1 , wherein the at least one saccharide-containing cross-linking residue has one or more of the following properties: substantially bioavailable, substantially biodegradeable, substantially bioabsorbable, or substantially bioresorbable.
11. The composition of claim 1 , wherein the at least one saccharide-containing cross-linking residue is derived from or comprises the residue of hyaluronic acid, a cellulose derivative, carboxy cellulose, carboxymethyl cellulose, hydroxymethyl cellulose, hydroxypropyl cellulose, hydroxypropyl methylcellulose, hydroxy-propylcellulosecarboxymethyl amylose, xanthan gum, guar gum, α-glucan, β-glucan, β-1,4-glucan, β-1,3-glucan, alginates, carboxymethyl dextran, a glycosaminoglycan derivative, chondroitin-6-sulfate, dermatin sulfate, heparin, heparin sulfate, polylactic acid, polyglycolic acid, poly(lactic-co-glycolic) acid, tricalcium phosphate, 1-hydroxyapatite, pharmaceutically acceptable salts of, derivatives of, or combinations thereof.
12. The composition of claim 1 , wherein the at least one protein residue is derived from or comprises the residue of tropoelastin, elastin, albumin, collagen, collagen monomers, immunoglobulins, insulin, derivatives of, or combinations thereof.
13. The composition of claim 1 , wherein the concentration of the at least one protein residue is in the range of between 1-200 mg/mL.
14. The composition of claim 1 , wherein the at least one cross-linked protein matrix is derived from or comprises the reside of a synthetic protein substantially identical to a naturally occurring human protein.
15. The composition of claim 1 , wherein the at least one cross-linked protein matrix is prepared from at least one saccharide-containing cross-linking molecule comprising at least 5% of activated carboxyl groups, modified carboxyl groups, or combinations thereof.
16. The composition of claim 1 , wherein the number of protein units not incorporated into the at least one cross-linked protein matrix and left unbound is at least 1%, 3%, 5%, or 7%.
17. The composition of claim 1 , wherein the at least one cross-linkage further comprises a spacer group.
18. The composition of claim 17 , wherein the spacer group comprises a glycol moiety, an ethylene oxide moiety, a polyamine, a polyol, or combinations thereof.
19. The composition of claim 1 , wherein the concentration of the at least one protein residue is in the range of between 5-30 mg/mL.
20. The composition of claim 1 , wherein the concentration of the at least one saccharide-containing cross-linking residue is in the range of between 1-30 mg/mL.
21. The composition of claim 20 , wherein the concentration of the at least one protein residue is in the range of between 25-50 mg/mL.
22. The composition of claim 21 , wherein:
i) the at least one protein residue is derived from or comprises the residue of tropoelastin; and
ii) the at least one saccharide-containing cross-linking residue is derived from or comprises the residue of hyaluronic acid.
23. The composition of claim 8 , wherein said at least one protein residue is devoid of intramolecular cross-links.
24. A method of preparing a cross-linked protein composition, comprising:
cross-linking at least one amine-bearing protein with at least one saccharide-containing cross-linking molecule comprising one or more carboxyl groups to form at least one cross-linked protein matrix;
wherein the at least one cross-linked matrix comprises:
i) at least one protein residue; and
ii) at least one saccharide-containing cross-linking residue; and
iii) at least one intermolecular cross-linkage comprising an amide bond between the amine of the at least one amine-bearing protein and the carboxyl group of the at least one saccharide-containing cross-linking molecule.
25. The method of claim 24 , wherein the prepared cross-linked protein composition is an injectable composition.
26. The method of claim 24 , wherein the prepared cross-linked protein composition is extrudable through needles of 18G to 31G to a length of between 5-30 cm and the extruded composition substantially holds together without surface support.
27. The method of claim 24 , wherein the prepared cross-linked protein composition retains sufficient cohesiveness after needle extrusion such that strings of extruded composition do not break during extrusion.
28. The method of claim 24 , wherein the at least one protein residue is 50-99.99% full length, or full length.
29. The method of claim 28 , wherein said at least one protein residue is devoid of intramolecular cross-links.
30. The method of claim 24 , wherein the at least one saccharide-containing cross-linking residue has one or more of the following properties: substantially bioavailable, substantially biodegradeable, substantially bioabsorbable, or substantially bioresorbable.
31. The method of claim 24 , wherein the at least one saccharide-containing cross-linking residue is derived from or comprises the residue of hyaluronic acid, a cellulose derivative, carboxy cellulose, carboxymethyl cellulose, hydroxymethyl cellulose, hydroxypropyl cellulose, hydroxypropyl methylcellulose, hydroxy-propylcellulosecarboxymethyl amylose, xanthan gum, guar gum, α-glucan, β-glucan, β-1,4-glucan, β-1,3-glucan, alginates, carboxymethyl dextran, a glycosaminoglycan derivative, chondroitin-6-sulfate, dermatin sulfate, heparin, heparin sulfate, polylactic acid, polyglycolic acid, poly(lactic-co-glycolic) acid, tricalcium phosphate, 1-hydroxyapatite, pharmaceutically acceptable salts of, derivatives of, or combinations thereof.
32. The method of claim 24 , wherein the at least one protein residue is derived from or comprises the residue of tropoelastin, elastin, albumin, collagen, collagen monomers, immunoglobulins, insulin, derivatives of, or combinations thereof.
33. The method of claim 24 , wherein the prepared cross-linked protein comprises a reside of a synthetic protein substantially identical to a naturally occurring human protein.
34. The method of claim 24 , wherein the number of protein units not incorporated into the at least one cross-linked protein matrix and left unbound in the prepared cross-linked protein composition is at least 1%, 3%, 5%, or 7%.
35. The method of claim 24 , wherein the at least one saccharide-containing cross-linking molecule comprises at least 5% of activated carboxyl groups, modified carboxyl groups, or combinations thereof.
36. The method of claim 24 , wherein the prepared cross-linked protein composition comprises a concentration in the range of between 1-250 mg/mL of the at least one protein residue derived from the at least one amine-bearing protein.
37. The method of claim 24 , wherein the prepared cross-linked protein composition comprises a concentration in the range of between 5-30 mg/mL of the at least one protein residue.
38. The method of claim 24 , wherein the prepared cross-linked protein composition comprises a concentration in the range of between 1-30 mg/mL of the at least one saccharide-containing cross-linking residue.
39. The method of claim 38 , wherein the prepared cross-linked protein composition comprises a concentration in the range of between 25-50 mg/mL of the at least one protein residue.
40. The method of claim 39 , wherein:
i) the at least one protein residue is derived from or comprises the residue of tropoelastin; and
ii) the at least one saccharide-containing cross-linking residue is derived from or comprises the residue of hyaluronic acid.