Hydrogels formed from polypeptide micelles and methods of use thereof

• US 9,956,300 B2
• Filed: 03/18/2016
• Issued: 05/01/2018
• Est. Priority Date: 03/18/2015
• Status: Active Grant

• Current Assignee: Duke University, UNM Rainforest Innovations (f/k/a STC.UNM) (The University of New Mexico)
• Original Assignee: Duke University
• Inventors: Lopez, Gabriel P., Chilkoti, Ashutosh, Ghoorchian, Ali, Simon, Joseph R.
• Primary Examiner(s): Lieb, Jeanette M
• Application Number:
  US15/074,132
• Publication Number:
  US 20160271262A1
• Time in Patent Office: 774 Days
• Field of Search: None
• CPC Class Codes: A61K 47/64 Drug-peptide, drug-protein or drug-polyamino acid conjugates, i.e. the modifying agent being a peptide, protein or polyamino acid which is covalently bonded or complexed to a therapeutically active agent peptidic linkers A61K47/65
• CPC Class Codes: A61K 47/6907 the form being a microemulsion, nanoemulsion or micelle
• CPC Class Codes: A61K 9/06 Ointments; Bases therefor; Other semi-solid forms, e.g. creams, sticks, gels composition of ointments, creams or gels A61K47/00
• CPC Class Codes: A61L 2300/252 Polypeptides, proteins, e.g. glycoproteins, lipoproteins, cytokines
• CPC Class Codes: A61L 2300/626 Liposomes, micelles, vesicles
• CPC Class Codes: A61L 27/227 Other specific proteins or polypeptides not covered by A61L27/222, A61L27/225 or A61L27/24
• CPC Class Codes: A61L 27/52 Hydrogels or hydrocolloids
• CPC Class Codes: C07K 14/78 Connective tissue peptides, e.g. collagen, elastin, laminin, fibronectin, vitronectin or cold insoluble globulin [CIG]
• CPC Class Codes: C08J 2389/00 Characterised by the use of proteins; Derivatives thereof
• CPC Class Codes: C08J 3/075 Macromolecular gels

Abstract

The presently disclosed subject matter is directed to compositions, methods, and systems (e.g., platforms) comprising the same, the systems comprising, consisting of, or consisting essentially of micro- and macro-hydrogels that are formed from polypeptide micelles. The systems have enhanced mechanical properties that can be useful in a wide variety of applications, can be used for controlled release of drug-loaded micelles, and can be designed to reversibly assemble and disassemble on demand.

Claims

1. A hydrogel comprised of reversibly crosslinked micelles, the hydrogel comprising:

a. a plurality of a peptide polymer consisting essentially of a hydrophobic portion of 40 to 80 repeats of a VPGVG (SEQ ID NO;

     1) peptide motif, a hydrophilic portion of 10 to 50 repeats of a VPGAGVPGGG (SEQ ID NO;

     2) peptide motif, and a separate peptide metal binding sequence at a terminus of the hydrophilic portion, wherein the peptide polymer is in the form of a plurality of micelles having the hydrophobic portion at a core of the micelles and the metal binding sequence at a surface of the micelles; and

b. a plurality of metal ions bound at the metal binding sequences, wherein the plurality of micelles are reversibly crosslinked through the bound metal ions.

2. The hydrogel of claim 1, wherein the peptide metal binding sequence comprises a motif HEDGHWDGSEHGY (SEQ ID NO:

3) and the metal comprises one or more of transition metals Zn2+, Ni2+, or Cu2+.

3. The hydrogel of claim 2, wherein the metal is Zn2+.

4. The hydrogel of claim 1, wherein the peptide polymer further comprises a hydrophobic therapeutic agent attached to the hydrophobic portion at the core of the micelles.

5. The hydrogel of claim 4, wherein the hydrophobic therapeutic agent is attached to the hydrophobic portion through a linker, and wherein the linker includes an enzymatic cleavage site to allow for release of the hydrophobic therapeutic agent from the micelle.

6. The hydrogel of claim 4, wherein the hydrophobic therapeutic agent is attached to the hydrophilic portion through a linker, and wherein the linker includes a pH sensitive cleavage site to allow for release of the hydrophobic therapeutic agent from the micelle.

7. The hydrogel of claim 1, further comprising an interpenetrating mesh consisting essentially of covalently crosslinked polymers, wherein the mesh surrounds the reversibly crosslinked micelles without being covalently attached to the micelles.

8. The hydrogel of claim 7, wherein the covalently crosslinked polymers comprise polypeptides, elastin-like polypeptides (ELP), biopolymers, synthetic polymers, or branched polymers, and combinations thereof.

9. A method of making a hydrogel comprised of reversibly crosslinked micelles, the method comprising:

heating a plurality of a peptide polymer consisting essentially of a hydrophobic portion of 40 to 80 repeats of a VPGVG (SEQ ID NO;

     1) peptide motif, a hydrophilic portion of 10 to 50 repeats of a VPGAGVPGGG (SEQ ID NO;

     2) peptide motif, and a separate peptide metal binding sequence at near a terminus of the hydrophilic portion, wherein the peptide polymer takes the form of a plurality of micelles having the hydrophobic portion at a core of the micelles and the metal binding sequences at a surface of the micelles; and

adding a plurality of metal ions to the micelles, wherein the plurality of micelles are reversibly crosslinked through the bound metal ions at the metal binding sequences.

10. The method of claim 9, wherein the peptide polymer further comprises a hydrophobic therapeutic agent attached to the hydrophobic portion, wherein the hydrophobic therapeutic agent is at the core of the micelles along with the hydrophobic portion.

11. The method of claim 9, further comprising adding a plurality of a polymer and a crosslinking agent, wherein the plurality of polymers are covalently crosslinked to each other to form an interpenetrating mesh consisting essentially of the covalently crosslinked polymers, wherein the interpenetrating mesh surrounds the reversibly crosslinked micelles without being covalently attached to the micelles.

12. A hydrogel loaded with a therapeutic molecule, the hydrogel comprising:

a. a plurality of a peptide polymer consisting essentially of a hydrophilic portion of 60 to 100 repeats of a VPGAG (SEQ ID NO;

     6) peptide motif and a separate peptide metal binding sequence at a terminus of the hydrophilic portion; and

having a hydrophobic therapeutic agent attached near an opposing terminus of the hydrophilic portion, wherein the peptide polymer is in the form of a plurality of micelles having the hydrophobic therapeutic agent at a core of the micelles and the metal binding sequences at a surface of the micelles; and

b. a plurality of metal ions bound at the metal binding sequences, wherein the plurality of micelles are reversibly crosslinked through the bound metal ions.

13. The hydrogel of claim 12, wherein the peptide metal binding site comprises a motif HEDGHWDGSEHGY (SEQ ID NO:

3) and the metal comprises one or more of transition metals Zn2+, Ni2+, or Cu2+.

14. The hydrogel of claim 13, wherein the metal is Zn2+.

15. The hydrogel of claim 12, wherein the hydrophobic therapeutic agent is attached to the hydrophilic portion through a linker, and wherein the linker includes an enzymatic cleavage site to allow for release of the hydrophobic therapeutic agent from the micelle.

16. The hydrogel of claim 12, wherein the hydrophobic therapeutic agent is attached to the hydrophilic portion through a linker, and wherein the linker includes a pH sensitive cleavage site to allow for release of the hydrophobic therapeutic agent from the micelle.

17. The hydrogel of claim 12, further comprising an interpenetrating mesh consisting essentially of covalently crosslinked polymers, wherein the mesh surrounds the reversibly crosslinked micelles without being covalently attached to the micelles.

18. The hydrogel of claim 17, wherein the covalently crosslinked polymers comprise polypeptides, elastin-like polypeptides (ELP), biopolymers, synthetic polymers, or branched polymers, and combinations thereof.

19. A method of making a hydrogel loaded with a therapeutic agent, the method comprising:

attaching a hydrophobic therapeutic agent at a terminus of a plurality of a peptide polymer consisting essentially of 60 to 100 repeats of a VPGAG (SEQ ID NO;

     6) peptide motif and a separate peptide metal binding sequence at an opposing terminus, wherein the peptide polymer takes the form of a plurality of micelles having the attached hydrophobic therapeutic agent a core of the micelles and the metal binding sequences at a surface of the micelles; and

adding a plurality of metal ions to the micelles, wherein the plurality of micelles are reversibly crosslinked through the bound metal ions at the metal binding sequences.

20. The method of claim 19, further comprising adding a plurality of a polymer and a crosslinking agent, wherein the plurality of polymers are covalently crosslinked to each other to form an interpenetrating mesh consisting essentially of the covalently crosslinked polymers, wherein the interpenetrating mesh surrounds the reversibly crosslinked micelles without being covalently attached to the micelles.