Gels for encapsulation of biological materials

• US 5,858,746 A
• Filed: 01/25/1995
• Issued: 01/12/1999
• Est. Priority Date: 04/20/1992
• Status: Expired due to Term

• Current Assignee: Board of Regents of the University of Texas System (University of Texas System)
• Original Assignee: Board of Regents of the University of Texas System (University of Texas System)
• Inventors: Pathak, Chandrashekhar P., Sawhney, Amarpreet S., Desai, Neil P., Hossainy, Syed F. A., Hubbell, Jeffrey A., Hill, Jennifer L.
• Primary Examiner(s): Lilling, Herbert J.
• Application Number:
  US08/377,911
• Time in Patent Office: 1,448 Days
• Field of Search: 524/702, 524/704, 524/849, 524/733, 524/734, 524/850, 524/852, 524/856, 524/56, 435/240.4, 435/182, 435/177, 435/178, 424/499, 424/450, 424/489, 424/483, 424/487, 514/2, 514/772.1, 514/773, 514/777, 525/54.1, 525/54.2, 525/408, 525/413, 528/361
• US Class Current: 435/177
• CPC Class Codes: A61K 2035/128 capsules, e.g. microcapsules
• CPC Class Codes: A61K 38/42 Haemoglobins; Myoglobins
• CPC Class Codes: A61K 38/44 Oxidoreductases (1)
• CPC Class Codes: A61K 47/645 Polycationic or polyanionic oligopeptides, polypeptides or polyamino acids, e.g. polylysine, polyarginine, polyglutamic acid or peptide TAT
• CPC Class Codes: A61K 47/6903 the form being semi-solid, e.g. an ointment, a gel, a hydrogel or a solidifying gel
• CPC Class Codes: A61K 47/6927 the form being a solid microparticle having no hollow or gas-filled cores
• CPC Class Codes: A61K 9/1635 obtained by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyvinyl pyrrolidone, poly(meth)acrylates
• CPC Class Codes: A61K 9/1647 Polyesters, e.g. poly(lactide-co-glycolide)
• CPC Class Codes: A61K 9/5026 obtained by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyvinyl pyrrolidone, poly(meth)acrylates
• CPC Class Codes: A61K 9/5031 obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyethylene glycol, poly(lactide-co-glycolide)
• CPC Class Codes: A61K 9/5073 having two or more different coatings optionally including drug-containing subcoatings
• CPC Class Codes: A61L 24/0031 Hydrogels or hydrocolloids
• CPC Class Codes: A61L 24/0042 Materials resorbable by the body
• CPC Class Codes: A61L 24/046 obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds A61L24/043 takes precedence
• CPC Class Codes: A61L 26/0019 obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds
• CPC Class Codes: A61L 27/34 Macromolecular materials
• CPC Class Codes: A61L 27/3683 subjected to a specific treatment prior to implantation, e.g. decellularising, demineralising, grinding, cellular disruption/non-collagenous protein removal, anti-calcification, crosslinking, supercritical fluid extraction, enzyme treatment
• CPC Class Codes: A61L 27/38 containing added animal cells organs or tissue containing native cells A61L27/36
• CPC Class Codes: A61L 27/52 Hydrogels or hydrocolloids
• CPC Class Codes: A61L 29/085 Macromolecular materials
• CPC Class Codes: A61L 31/10 : Macromolecular materials
• CPC Class Codes: A61L 31/145 : Hydrogels or hydrocolloids
• CPC Class Codes: A61L 31/148 : Materials at least partially resorbable by the body
• CPC Class Codes: B82Y 5/00 : Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery
• CPC Class Codes: C08B 37/003 : Chitin, i.e. 2-acetamido-2-deoxy-(beta-1,4)-D-glucan or N-acetyl-beta-1,4-D-glucosamine; Chitosan, i.e. deacetylated product of chitin or (beta-1,4)-D-glucosamine; Derivatives thereof
• 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: C08B 37/0084 : 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 Class Codes: C08F 290/00 : Macromolecular compounds obtained by polymerising monomers on to polymers modified by introduction of aliphatic unsaturated end or side groups
• CPC Class Codes: C08F 290/02 : on to polymers modified by introduction of unsaturated end groups
• CPC Class Codes: C08F 290/06 : Polymers provided for in subclass C08G
• CPC Class Codes: C08F 290/062 : Polyethers
• CPC Class Codes: C08F 290/08 : on to polymers modified by introduction of unsaturated side groups
• CPC Class Codes: C08F 290/10 : Polymers provided for in subclass C08B
• CPC Class Codes: C08F 291/00 : Macromolecular compounds obtained by polymerising monomers on to macromolecular compounds according to more than one of the groups C08F251/00 - C08F289/00
• CPC Class Codes: C08L 71/02 : Polyalkylene oxides
• CPC Class Codes: C09D 153/00 : Coating compositions based on block copolymers containing at least one sequence of a polymer obtained by reactions only involving carbon-to-carbon unsaturated bonds; Coating compositions based on derivatives of such polymers
• CPC Class Codes: C12N 11/04 : entrapped within the carrier, e.g. gel or hollow fibres
• CPC Class Codes: C12N 2533/30 : Synthetic polymers
• CPC Class Codes: C12N 2533/32 : Polylysine, polyornithine
• CPC Class Codes: C12N 2533/40 : Polyhydroxyacids, e.g. polymers of glycolic or lactic acid (PGA, PLA, PLGA); Bioresorbable polymers
• CPC Class Codes: C12N 2533/74 : Alginate
• CPC Class Codes: C12N 5/0012 : Cell encapsulation
• CPC Class Codes: C12N 5/0677 : Three-dimensional culture, tissue culture or organ culture; Encapsulated cells
• CPC Class Codes: Y10S 530/812 : Peptides or proteins is immobilized on, or in, an organic carrier
• CPC Class Codes: Y10S 530/813 : Carrier is a saccharide
• CPC Class Codes: Y10S 530/815 : Carrier is a synthetic polymer
• CPC Class Codes: Y10S 530/817 : Entrapped within the carrier, e.g. gel, hollow fibre
• CPC Class Codes: Y10T 428/2987 : Addition polymer from unsaturated monomers only
• CPC Class Codes: Y10T 428/2989 : Microcapsule with solid core [includes liposome]

Abstract

Water soluble macromers are modified by addition of free radical polymerizable groups, such as those containing a carbon-carbon double or triple bond, which can be polymerized under mild conditions to encapsulate tissues, cells, or biologically active materials. The polymeric materials are particularly useful as tissue adhesives, coatings for tissue lumens including blood vessels, coatings for cells such as islets of Langerhans, coatings, plugs, supports or substrates for contact with biological materials such as the body, and as drug delivery devices for biologically active molecules.

Claims

1. A method for encapsulating, sealing, plugging, or supporting of mammalian cells, cell aggregates, or cell tissue comprising:

a) applying (i) a water soluble biocompatible macromer comprising at least two free radical-polymerizable substituents, wherein the macromer is non-toxic and has a molecular weight of at least 400, and (ii) a non-toxic free radical polymerization initiator selected from the group consisting of visible light or long wavelength ultraviolet light-activatable free radical initiators, thermal activatable free radical initiators, benzoyl peroxide, potassium persulfate and ammonium persulfate;

to (iii) mammalian cells, cell aggregates, or cell tissue; and

b) exposing the mixture to an activating agent to cause polymerization of the macromers.

2. The method of claim 1 wherein the biological material is cell tissue.

3. The method of claim 1 further comprising biologically active molecules selected from the group consisting of peptides of less than one hundred amino acids, proteins of one hundred or more amino acids, polysaccharides, nucleic acids, organic drugs, and inorganic drugs.

4. The method of claim 1 wherein the free-radical polymerizable substituents contain carbon-carbon double or triple bonds.

5. The method of claim 1 where a non-toxic catalyst or accelerator is added to the mixing step.

6. The method of claim 1 wherein the water soluble macromer is selected from the group consisting of poly(ethylene glycol), poly(ethylene oxide), poly(vinyl alcohol), poly(vinylpyrrolidone), poly(ethyloxazoline), poly(amino acid), polysaccharides, proteins, or a block or random copolymer thereof comprising two or more polymerizable substituents.

7. The method of claim 6 wherein the polysaccharide is selected from the group consisting of alginate, hyaluronic acid, chondroitin sulfate, dextran, dextran sulfate, heparin, heparin sulfate, heparan sulfate, chitosan, gellan gum, xanthan gum, guar gum, and K-carrageenan.

8. The method of claim 6, wherein the protein is selected from the group consisting of gelatin, collagen and albumin.

9. The method of claim 1 wherein the initiator is a thermal initiator and the initiating agent is a change in temperature.

10. The method of claim 1 wherein the free-radical polymerizable substituents are selected from the group of macromers containing two or more acrylate groups.

11. The method of claim 1 wherein the gel is prepared from macromers comprising an acrylate terminated poly(ethylene glycol).

12. The method of claim 1 wherein the polymerization initiator is selected from the group consisting of an eosin dye, a substituted eosin dye, riboflavin, acetophenone, a substituted acetophenone, a fluoroscein dye, a substituted fluoroscein dye, camphorquinone, rose bengal, methylene green, methylene blue, eosin Y, ethyl eosin, acridine orange, xanthine dye, and thioxanthine dyes.

13. The method of claim 1 wherein the polymerization initiator is selected from the group consisting of erythrosin, phloxime, and thionine.

14. The method of claim 5 wherein the catalyst or accelerator is an amine.

15. The method of claim 14 wherein the amine is selected from the group consisting of triethanolamine, triethylamine, ethanolamine, N-methyl diethanolamine, N,N-dimethyl benzylamine, dibenzyl amine, N-benzyl ethanolamine, N-isopropyl benzylamine, tetramethyl ethylene-diamine, lysine, ornithine, histidine and arginine.

16. The method of claim 1 wherein polymerization is initiation by light having a wavelength of between 320 and 800 nm.

17. The method of claim 16 or 2 wherein the light has a wavelength of 514 nm or 365 nm.

18. The method of claim 2, wherein the mammalian cell, mammalian cell aggregate, or mammalian tissue is contacted with a solution of a light sensitive photoinitiator to allow binding of the photoinitiator to the mammalian cell, mammalian cell aggregate, or mammalian tissue and the unbound initiator is removed prior to contacting it with the polymer or oligomer.

19. The method of claim 18 wherein the unbound initiator is removed by dilution with the macromer solution such that polymerization occurs only at the surface of the mammalian cell, mammalian cell aggregate, or mammalian tissue.

20. The method of claim 1, wherein the macromer solution is shaped and then polymerized.

21. The method of claim 1 wherein the gel contains a supporting structure.

22. The method of claim 1 wherein the polymer is selected and the polymerization is controlled to produce a desired permeability around the encapsulated material.

23. The method of claim 1 wherein polymerization of the macromer solution adheres tissue to other tissue or cells.

24. The method of claim 2 wherein biologically active material is encapsulated with the cells or tissue.

25. The method of claim 1, wherein a photopolymerizable polycation is preadsorbed to the molecule or material being encapsulated to increase attachment of the gel to the molecule or material.

26. The method of claim 1 wherein the macromer solution is applied to a tissue lumen and then polymerized to form a coating or support on the surface of the tissue lumen.

27. A method for the preparation of a biocompatible substrate comprising:

a) mixing (i) a water soluble biocompatible macromer comprising at least two free radical-polymerizable substituents, wherein the macromer is non-toxic and has a molecular weight of at least 400, (ii) a biocompatible substrate for mammalian cells, cell aggregates and cell tissue and (iii) a non-toxic free radical polymerization initiator selected from the group consisting of visible light or long wavelength ultraviolet light-activatable free radical initiators, thermal activatable free radical initiators, benzoyl peroxide, potassium persulfate and ammonium persulfate; and

b) exposing the mixture to an activating agent to cause polymerization of the macromers.

28. The method of claim 27 wherein a catalyst or accelerator is added in the mixing step.

29. The method of claim 27 wherein the substrate is selected from the group consisting of microspheres, membranes, woven matrices, porous matrices and prosthetic implants.

30. The method of claim 27, wherein the substrate is treated with an initiator, the unbound initiator is removed, the macromer is applied to the substrate and polymerized.

31. A method for applying a polymeric coating to mammalian tissue or cells comprising applying a photoinitiator to the cells or tissue, removing unbound photoinitiator, applying a solution of water soluble macromer comprising at least two free radical-polymerizable substituents to the cells or tissue, and exposing the solution to visible or long wavelength ultraviolet light.