Functionalized derivatives of hyaluronic acid, formation of hydrogels in situ using same, and methods for making and using same

• US 6,630,457 B1
• Filed: 09/18/1998
• Issued: 10/07/2003
• Est. Priority Date: 09/18/1998
• Status: Expired due to Fees

• Current Assignee: Orthogene LLC
• Original Assignee: Orthogene LLC
• Inventors: Aeschlimann, Daniel, Bulpitt, Paul
• Primary Examiner(s): Fonda, Kathleen K.
• Application Number:
  US09/156,829
• Time in Patent Office: 1,845 Days
• Field of Search: 514/2, 514/8, 514/54, 536/53, 435/243, 435/253.6
• US Class Current: 514/54
• CPC Class Codes: A61K 31/728 Hyaluronic acid
• CPC Class Codes: A61K 47/36 Polysaccharides; Derivatives thereof, e.g. gums, starch, alginate, dextrin, hyaluronic acid, chitosan, inulin, agar or pectin
• CPC Class Codes: A61K 9/0024 Solid, semi-solid or solidifying implants, which are implanted or injected in body tissue compositions for intravenous administration, normal injectable solutions or dispersions for, e.g. subcutaneous administration A61K9/0019; brain implants A61K9/0085; (coated) prostheses, catheters or stents A61L
• CPC Class Codes: A61L 24/08 Polysaccharides A61L24/043 takes precedence
• CPC Class Codes: A61L 27/20 Polysaccharides
• CPC Class Codes: C08B 37/0072 Hyaluronic acid, i.e. HA or hyaluronan; Derivatives thereof, e.g. crosslinked hyaluronic acid (hylan) or hyaluronates
• CPC Class Codes: C08L 5/08 Chitin; Chondroitin sulfate; Hyaluronic acid; Derivatives thereof
• CPC Class Codes: C12N 2533/80 Hyaluronan
• CPC Class Codes: C12N 5/0068 General culture methods using substrates for specific animal cell type C12N5/06

Abstract

Methods for chemical modification of hyaluronic acid, formation of amine or aldehyde functionalized hyaluronic acid, and the cross-linking thereof to form hydrogels are provided. Functionalized hyaluronic acid hydrogels of this invention can be polymerized in situ, are biodegradable, and can serve as a tissue adhesive, a tissue separator, a drug delivery system, a matrix for cell cultures, and a temporary scaffold for tissue regeneration.

Claims

1. A composition comprising crosslinked derivatives of hyaluronic acid comprising disaccharide subunits, wherein at least one of said disaccharide subunits is a substituted disaccharide subunit having a substitution at a carboxyl group, such that the substituted disaccharide subunit is of the formula

wherein each of R′

and R″

is a side chain comprising one or more functional groups selected from the group consisting of hydrogen;

bioactive peptide;

alkyl;

aryl;

alkylaryl;

arylalkyl;

substituted alkylaryl containing an atom or atoms of oxygen, nitrogen, sulfur, or phosphorous;

substituted arylalkyl containing an atom or atoms of oxygen, nitrogen, sulfur;

phosphorous, halogen, or metal ion; and

substituted heterocycle containing an atom or atoms of oxygen, nitrogen, sulfur;

phosphorous, halogen or metal ion;

wherein said functional groups within or among each of said R′

or R″

side chains are either bound directly to each other or are separated by a member selected from the group consisting of ether, keto, amino, oxycarbonyl, sulfone, sulfoxide, carboxamide, alkyne and alkene; and

wherein each of said R′ and

R″

side chains terminates with a terminal functional group selected from the group consisting of hydrogen, peptide, aldehyde, amine, arylazide, hydrazide, maleimide, sulfhydryl, active ester, ester, carboxylate, imidoester, halogen and hydroxyl;

or

wherein said terminal functional groups of each of said R′ and

R″

side chains may be bound directly to each other, with the proviso that when one of R′

or R″

is hydrogen, halogen or univalent metal ion, then R′ and

R″

may not be bound directly to each other;

wherein said derivatives of hyaluronic acid are covalently crosslinked via one of said terminal functional groups; and

wherein said derivatives of hyaluronic acid are modified to an extent of 10-95%.

2. The composition of claim 1, wherein at least one of said terminal functional groups is selected from the group consisting of peptide, aldehyde, amine, arylazide, hydrazide, maleimide, sulfhydryl, and active ester, whereby said composition is amenable to crosslinking.

3. The composition of claim 1, wherein the molecular weight of said composition is at least 100,000 daltons.

4. The composition of claim 1, wherein the molecular weight of said composition is at most 100,000 daltons.

5. The composition of claim 1, wherein the molecular weight of said composition is at least 1,000,000 daltons.

6. The composition of claim 1, wherein said composition is water soluble.

7. A hydrogel of crosslinked HA derivatives, wherein said HA derivatives are compositions according to claim 1.

8. The hydrogel of crosslinked HA derivatives of claim 7, wherein said hydrogel is biodegradable.

9. A tissue adhesive comprising a hydrogel of claim 7, wherein the side chain is selected from the group consisting of activated ester, aldehyde, arylazide, and maleimide.

10. A tissue adhesive comprising a hydrogel of claim 7, wherein the crosslinked HA derivatives are formed using a cross-linker selected from the group consisting of polyvalent active ester, aldehyde, arylazide, and maleimide.

11. A tissue adhesive comprising a hydrogel of claim 7, wherein the cross-linked hydrogel is formed in the presence of at least one member selected from the group consisting of growth factors, cytokines, drugs, and bioactive peptides.

12. A tissue adhesive of claim 11, wherein the cross-linked hydrogel is formed in the presence of a growth factor and wherein the growth factor is TGF-β

or BMP-2.

13. A matrix for cell cultures comprising a hydrogel of claim 7, wherein the crosslinked HA-derivatives are formed using a cross-linker selected from the group consisting of polyvalent active ester, aldehyde, amine, arylazide, maleimide, and sulfhydryl.

14. A matrix for cell cultures comprising a hydrogel of claim 7, wherein the crosslinked hydrogel is formed in the presence of at least one member selected from the group consisting of growth factors, cytokines, drugs, and bioactive peptides.

15. A matrix for cell cultures according to claim 14, wherein the crosslinked hydrogel is formed in the presence of a growth factor and wherein the growth factor is TGF-β

or BMP-2.

16. A matrix for a scaffold comprising a hydrogel of claim 7, wherein the crosslinked HA-derivatives are formed using a cross-linker selected from the group consisting of polyvalent active ester, aldehyde, amine, arylazide, maleimide, and sulfhydryl.

17. A matrix for a scaffold comprising a hydrogel of claim 7, wherein the crosslinked hydrogel is formed in the presence of at least one member selected from the group consisting of growth factors, cytokines, drugs, and bioactive peptides.

18. A matrix for a scaffold according to claim 17, wherein the crosslinked hydrogel is formed in the presence of a growth factor and wherein the growth factor is TGF-β

or BMP-2.

19. The matrix of claim 17, wherein the matrix further comprises cells.