Click-crosslinked hydrogels and methods of use

Patent metadata

Field	Value
Publication number	US-2025161536-A1
Application number	US-202418752542-A
Country	US
Kind code	A1
Filing date	Jun 24, 2024
Priority date	Apr 4, 2014
Publication date	May 22, 2025
Grant date	—

Abstract

The present disclosure provides click-crosslinked hydrogels and methods of use.

First claim

1 . A hydrogel comprising a first polymer and a second polymer, wherein the first polymer is alginate and the second polymer is collagen, wherein the first polymer is connected to the second polymer by linkers of formula (A):

wherein
bond is a single or a double bond;
R 1 is —C 0 -C 6 alkyl-NH—, —C 0 -C 6 alkyl-O—, or —C 0 -C 3 alkyl-C(O)—;
R 2 is a bond, aryl, or heteroaryl, wherein aryl and heteroaryl are optionally substituted with halogen, hydroxy, C 1 -C 6 alkyl, C 1 -C 6 alkoxy, (C 1 -C 6 alkyl)amino, or di(C 1 -C 6 alkyl)amino;
R 3 is —C 0 -C 6 alkyl-NH—, —C 0 -C 6 alkyl-O—, or —C 0 -C 3 alkyl-C(O)—; and
R 4 is hydrogen, C 1 -C 6 alkyl, aryl, or heteroaryl, wherein aryl and heteroaryl are optionally substituted with halogen, hydroxy, C 1 -C 6 alkyl, C 1 -C 6 alkoxy, (C 1 -C 6 alkyl)amino, or di(C 1 -C 6 alkyl)amino.

2 . The hydrogel according to claim 1 , wherein:
bond is a single bond; R 1 is —C 0 -C 6 alkyl-NH—, or —C 0 -C 3 alkyl-C(O)—; R 2 is a bond or aryl optionally substituted with halogen, hydroxy, C 1 -C 6 alkyl, C 1 -C 6 alkoxy, (C 1 -C 6 alkyl)amino, or di(C 1 -C 6 alkyl)amino; R 1 is —C 0 -C 6 alkyl-NH—, or —C 0 -C 3 alkyl-C(O)—; and R 4 is hydrogen, C 1 -C 6 alkyl, or heteroaryl, wherein heteroaryl is optionally substituted with halogen, hydroxy, C 1 -C 6 alkyl, C 1 -C 6 alkoxy, (C 1 -C 6 alkyl)amino, or di(C 1 -C 6 alkyl)amino.

3 . The hydrogel according to claim 2 , wherein R 1 and R 3 are both -methyl-NH—; or R 1 and R 3 are both —C(O)—.

4 . The hydrogel according to claim 1 , wherein the linkers of formula (A) are selected from the group consisting of formula (I):

or formula (II):

or formula (III):

5 - 10 . (canceled)

11 . The hydrogel of claim 1 , wherein the first polymer or the second polymer further comprises a cell adhesive peptide.

12 . The hydrogel of claim 11 , wherein the cell adhesive peptide comprises the amino acid sequence RGD.

13 . The hydrogel of claim 12 , wherein the cell adhesive peptide comprises the amino acid sequence RGDC.

14 . (canceled)

15 . The hydrogel of claim 1 , wherein the Young's modulus of the hydrogel is 50 to 50,000 Pa.

16 . The hydrogel of claim 1 , wherein the hydrogel further comprises a cell, a biological factor, and/or a small molecule.

17 . The hydrogel of claim 16 , wherein the cell is a mammalian cell.

18 . The hydrogel of claim 16 , wherein the biological factor is a protein, nucleic acid, lipid, or carbohydrate.

19 . The hydrogel of claim 18 , wherein the protein is a growth factor or fragment thereof or an antibody or fragment thereof.

20 . A method for preparing a hydrogel of claim 1 , comprising,
a) providing a first polymer comprising a tetrazine moiety and a second polymer comprising a norbornene moiety; and b) contacting the second polymer with the first polymer to form the hydrogel of claim 1 .

21 . The method of claim 20 , wherein: (i) each molecule of the first polymer comprises 1-50,000 tetrazine moieties; (ii) each molecule of the second polymer comprises 1-50,000 norbornene moieties; and/or (iii) step b) comprises contacting a second polymer with a first polymer at a ratio of about 1:10 to about 10:1.

22 - 23 . (canceled)

24 . The method of claim 20 , further comprising reacting the first polymer with a) benzyl amine tetrazine, benzyl alcohol tetrazine, or benzoic acid tetrazine, and a b) coupling agent to provide the first polymer comprising a tetrazine moiety.

25 . The method of claim 24 , wherein the coupling agent is 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC).

26 . The method of claim 20 , further comprising reacting the second polymer with a) norbornene methanamine, norbornene methanol, or norbornene carboxylic acid, and b) a coupling agent to provide the second polymer comprising a norbornene moiety.

27 . The method of claim 26 , wherein the coupling agent comprises 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) and optionally N-Hydroxysuccinimidc (NHS).

28 . A method of regenerating a tissue in a subject in need thereof, the method comprising contacting the tissue with the hydrogel of claim 1 .

29 - 33 . (canceled)

34 . A method of delivering a cell, a biological factor and/or a small molecule to a subject, the method comprising administering to the subject the hydrogel of claim 16 , wherein after administration of the hydrogel, the cell, the biological factor and/or the small molecule is released from the hydrogel into a surrounding tissue of the subject, thereby delivering the cell, the biological factor and/or the small molecule to the subject.

Assignees

• Harvard College

Inventors

• Desai Rajiv
• Joshi Neel Satish
• Mooney David J
• Koshy Sandeep T
• Stafford Alexander

Classifications

• C08B37/0084

  Guluromannuronans, e.g. alginic acid, i.e. D-mannuronic acid and D-guluronic acid units linked with alternating alpha- and beta-1,4-glycosidic bonds; Derivatives thereof, e.g. alginates · CPC title

• C07K14/78

  Connective tissue peptides, e.g. collagen, elastin, laminin, fibronectin, vitronectin or cold insoluble globulin [CIG] · CPC title

• A61L2400/18

  Modification of implant surfaces in order to improve biocompatibility, cell growth, fixation of biomolecules, e.g. plasma treatment · CPC title

• A61L2400/06

  Flowable or injectable implant compositions · CPC title

• A61L2300/64

  Animal cells · CPC title