Hydrogels and use thereof in anastomosis procedures

Patent metadata

Field	Value
Publication number	US-2018000983-A1
Application number	US-201615545647-A
Country	US
Kind code	A1
Filing date	Jan 22, 2016
Priority date	Jan 22, 2015
Publication date	Jan 4, 2018
Grant date	—

Abstract

This disclosure provides novel hydrogels that can undergo multiple gel-sol transitions and methods of making and using such hydrogels, particularly in anastomosis procedures. The peptide hydrogels comprising a fibrillar network of peptides that are in an amphiphilic β-hairpin conformation. The peptides comprise photo-caged glutamate residues with a neutral photocage that can be photolytically selectively uncaged to disrupt the fibrillar network and trigger an irreversible gel-sol phase transition of the hydrogel. Isolated peptides for making the disclosed hydrogels are provided, as are methods of using the peptide hydrogels in anastomosis procedures.

First claim

1 . A peptide hydrogel comprising a fibrillar network of peptides, wherein:
the hydrogel undergoes a gel-sol phase transition upon application of shear stress, and a sol-gel phase transition upon removal of the shear stress; and the peptides are in an amphiphilic β-hairpin conformation and comprise photo-caged glutamate residues with a neutral photocage that can be photolytically selectively uncaged to disrupt the fibrillar network and trigger an irreversible gel-sol phase transition of the hydrogel.

2 . The peptide hydrogel of claim 1 , wherein:
the amphiphilic β-hairpin conformation comprises a β-turn, a first β-strand, a second β-strand, a hydrophobic face, and a hydrophilic face; the assembly of the peptides in the fibrillar network comprises hydrophobic interactions between the hydrophobic faces of the peptides; the first β-strand comprises the photocaged glutamate residue, the second β-strand comprises a glycine residue, and the sidechains of the photocaged glutamate residue and the glycine residue are proximal to each other on the hydrophobic faces of the peptides; and uncaging the photocaged glutamate residues disrupts the hydrophobic interactions between the peptides by exposing negative charges of the glutamate residues, thereby disrupting the fibrillar network and triggering the irreversible gel-sol phase transition of the hydrogel.

3 . The peptide hydrogel of claim 1 , comprising:
a storage modulus of greater than 40 Pascal in the absence of shear; from about 20 mM to about 400 mM NaCl; a pH of from about 7.0 to about 9.0; and/or from about 0.25% to about 4.0% w/v peptide.

4 - 5 . (canceled)

6 . The peptide hydrogel of claim 3 , wherein the pH is about 7.4

7 . (canceled)

8 . The peptide hydrogel of claim 3 , comprising from about 1% to about 2.0% w/v peptide.

9 . The peptide hydrogel of claim 1 , wherein the peptide is from 10 to 75 amino acids in length.

10 . The peptide hydrogel of claim 1 , wherein the peptide comprises or consists of a consensus peptide sequence selected from one of APCC1, APCC2, APCC3, APCC4, APCC4a, APCC5 APCC5a, APCC6, APCC6a, APCC7, APCC7a, APCC8, or APCC8a.

11 . (canceled)

12 . The peptide hydrogel of claim 1 , wherein the peptide the peptide comprises or consists of a peptide selected from one of APC1 APC1a, APC2, APC2a, APC3, APC3a, APC4, APC4a, APC5, APC5a, APC6, APC6a, APC7, APC7a, APC8, APC8a, APC9, ACP9a, APC10, APC10a, APC11, APC11a, APC12, APC12a, APC13, APC13a, APC14, APC14a, APC15, APC15a, APC16, APC16a, APC17, APC17a, APC18, APC18a, APC19, APC19a, APC20, APC20a, APC21, APC21a, APC22, APC22a, APC23, or APC23a.

13 . (canceled)

14 . The peptide hydrogel of claim 1 , wherein the photocaged glutamate residue is a 4-methoxy-7-nitroindolinyl-glutamate residue.

15 . A syringe, containing the peptide hydrogel of claim 1 .

16 . An isolated peptide that forms an amphiphilic β-hairpin conformation comprising a β-turn, a first β-strand, a second β-strand, a hydrophobic face, and a hydrophilic face when the peptide is dissolved in an aqueous solution comprising 150 mM NaCl and a pH of 7.4 at 25° C.; and
wherein the first β-strand comprises a photocaged glutamate residue with a neutral photocage, the second β-strand comprises a glycine residue, and the sidechains of the photocaged glutamate residue and the glycine residue are proximal to each other on the hydrophobic face of the peptide.

17 . The isolated peptide of claim 16 , wherein:
an aqueous solution containing 2% w/v of the peptide and 150 mM NaCl and a pH of 7.4 forms a peptide hydrogel comprising a fibrillar network of a plurality of the peptide when incubated at 25° C. in a container; the peptides assemble in the fibrillar network by hydrophobic interactions between the hydrophobic faces of the peptides; the hydrogel undergoes a gel-sol phase transition upon application of shear stress, and a sol-gel phase transition upon removal of the shear stress; and uncaging the photocaged glutamate residues disrupts the hydrophobic interactions between the peptides by exposing negative charges of the glutamate residues, thereby disrupting the fibrillar network and triggering an irreversible gel-sol phase transition of the hydrogel.

18 . (canceled)

19 . The isolated peptide of claim 16 , wherein the peptide is from 10 to 75 amino acids in length, particularly wherein the peptide is 18-22 amino acids in length, more particularly wherein the peptide is 20 amino acids in length.

20 . The isolated peptide of claim 16 , wherein the peptide comprises or consists of a peptide selected from one of APCC1, APCC2, APCC3, APCC4, APCC4a, APCC5, APCC5a, APCC6, APCC6a, APCC7, APCC7a, APCC8, or APCC8a.

21 . (canceled)

22 . The isolated peptide of claim 16 , wherein the peptide comprises or consists of a peptide selected from one of APC1, APC1a, APC2, APC2a, APC3, APC3a, APC4, APC4a, APC5, APC5a, APC6, APC6a, APC7, APC7a, APC8, APC8a, APC9, APC9a, APC10, APC10a, APC11, APC11a, APC12, APC12a, APC13, APC13a, APC14, APC14a, APC15, APC15a, APC16, APC16a, APC17, APC17a, APC18, APC18a, APC19, APC19a, APC20, APC20a, APC21, APC21a, APC22, APC22a, APC23, or APC23a.

23 . (canceled)

24 . The isolated peptide hydrogel of claim 16 , wherein the photocaged glutamate residue is a 4-methoxy-7-nitroindolinyl-glutamate residue.

25 . (canceled)

26 . A method of performing an anastomosis, comprising:
filling the lumen of each end of a severed vessel in a subject with the peptide hydrogel of claim 1 to support each end in an open configuration; apposing the two ends of the severed vessel and anastomosing the apposed ends; irradiating the hydrogel with a sufficient amount of light of a preselected wavelength to uncage the photocaged glutamate residues to disrupt the fibrillar network of the hydrogel and trigger the irreversible gel-sol phase transition of the hydrogel to a low viscosity gel capable of flow, wherein blood flow through the vessel disperses the disrupted hydrogel and restores patency to the vessel.

27 . The method of claim 26 , wherein the vessel is from about 50 μM to about 10 mM in diameter.

28 . The method of claim 26 , wherein:
filling the lumen of each end of the severed vessel comprises injecting the peptide hydrogel into the lumen of each end of the severed vessel with a syringe; securing the ends of the severed vessel to each other comprises connecting the ends of the severed vessel with one or more sutures; the anastomosis comprises end-to-end suturing of a severed blood vessel, a severed duct, or a severed lymphatic vessel; and/or the method further comprises applying the hydrogel to an interspace between the vessel ends to stabilize the positioning of vessel ends.

29 . The method of claim 26 , wherein the peptides in the peptide hydrogel comprise or consist of an APCC5 or APCC5a consensus peptide

30 . The method of claim 26 , wherein the peptides in the peptide hydrogel comprise or consist of APC1 or APC1a.

31 - 33 . (canceled)

Assignees

• The Us Secretary Department Of Health And Human
• Univ Johns Hopkins

Inventors

• Schneider Joel
• Brandacher Gerald
• Smith Daniel
• Brat Gabriel
• Grahammer Johanna

Classifications

• A61B2017/1107

  for blood vessels · CPC title

• A61B17/11

  for performing anastomosis; Buttons for anastomosis · CPC title

• A61L24/001

  Use of materials characterised by their function or physical properties · CPC title

• A61L2400/06

  Flowable or injectable implant compositions · CPC title

• A61L24/0031 Primary

  Hydrogels or hydrocolloids · CPC title