Injectable cryogel vaccine devices and methods of use thereof

• US 9,675,561 B2
• Filed: 01/28/2014
• Issued: 06/13/2017
• Est. Priority Date: 04/28/2011
• Status: Active Grant

• Current Assignee: President and Fellows of Harvard College (Harvard Corporation)
• Original Assignee: President and Fellows of Harvard College (Harvard Corporation)
• Inventors: Bencherif, Sidi A., Mooney, David J., Sands, Roger Warren, Koshy, Sandeep T.
• Primary Examiner(s): ZISKA, SUZANNE E
• Application Number:
  US14/166,689
• Publication Number:
  US 20140227327A1
• Time in Patent Office: 1,232 Days
• CPC Class Codes: A61K 2039/5152 Tumor cells
• CPC Class Codes: A61K 2039/55522 Cytokines; Lymphokines; Interferons
• CPC Class Codes: A61K 2039/55561 CpG containing adjuvants; Oligonucleotide containing adjuvants
• CPC Class Codes: A61K 39/0011 Cancer antigens
• CPC Class Codes: A61K 39/001106 Her-2/neu/ErbB2, Her-3/ErbB3 or Her 4/ErbB4
• CPC Class Codes: A61K 39/39 characterised by the immunostimulating additives, e.g. chemical adjuvants
• CPC Class Codes: A61K 39/4611 T-cells, e.g. tumor infiltrating lymphocytes [TIL], lymphokine-activated killer cells [LAK] or regulatory T cells [Treg]
• CPC Class Codes: A61K 39/4644 Cancer antigens
• 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 47/42 Proteins; Polypeptides; Degradation products thereof; Derivatives thereof, e.g. albumin, gelatin or zein oligopeptides having up to five amino acids A61K47/183; polyamino acids A61K47/34
• 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: A61K 9/7007 Drug-containing films, membranes or sheets A61K9/0041, A61K9/0043, A61K9/006, A61K9/0063 take precedence
• CPC Class Codes: A61L 2300/252 Polypeptides, proteins, e.g. glycoproteins, lipoproteins, cytokines
• CPC Class Codes: A61L 2300/258 Genetic materials, DNA, RNA, genes, vectors, e.g. plasmids
• CPC Class Codes: A61L 2400/06 Flowable or injectable implant compositions
• CPC Class Codes: A61L 27/26 Mixtures of macromolecular compounds
• CPC Class Codes: A61L 27/50 Materials characterised by their function or physical properties , e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
• CPC Class Codes: A61L 27/54 Biologically active materials, e.g. therapeutic substances A61L27/227 takes precedence
• CPC Class Codes: A61L 27/56 Porous materials, e.g. foams or sponges
• CPC Class Codes: C08L 5/04 Alginic acid; Derivatives thereof
• CPC Class Codes: C08L 89/06 : derived from leather or skin , e.g. gelatin

Abstract

The invention provides polymer compositions for cell and drug delivery.

Claims

1. An injectable device comprising a cell-adhesive, highly crosslinked cryogel composition comprising open interconnected macropores,

wherein said cryogel composition comprises at least 75% pores,

wherein said cryogel composition is characterized by shape memory following deformation by compression through a needle,

wherein said cryogel composition comprises a crosslinked gelatin polymer, wherein said gelatin is methacrylated,

wherein said cryogel composition comprises a methacrylated gelatin concentration of 0.5% to 1.4% (w/v), and

wherein said cryogel composition comprises a live attenuated breast cancer cell in one or more of said open interconnected pores.

2. The device of claim 1, wherein the device recruits cells into the cryogel composition upon injection into a subject.

3. The device of claim 2, wherein the cryogel composition is degraded by the recruited cells.

4. The device of claim 1, wherein the cryogel composition is formed by cryopolymerization of methacrylated gelatin.

5. The device of claim 4, wherein the cryogel composition further comprises a methacrylated alginate macromonomer.

6. The device of claim 1, wherein said cryogel composition comprises a biomolecule in one or more of said open interconnected pores.

7. The device of claim 6, wherein said biomolecule comprises a small molecule, nucleic acid, or protein.

8. The device of claim 7, wherein said protein comprises granulocyte-macrophage colony-stimulating factor (GM-CSF).

9. The device of claim 7, wherein said nucleic acid comprises a CpG nucleic acid oligonucleotide (CpG-ODN).

10. The device of claim 1, wherein the device is between 100 um³ to 100 mm³ in size.

11. A method for eliciting an immune response, comprising injecting the device of claim 1 into a subject, wherein an immune response is elicited against the live attenuated breast cancer cell.

12. The method of claim 11, wherein the device is injected into the subject one to 5 times in the lifetime of the subject.

13. The method of claim 11, wherein the injected device comprises at least 0.5×

10⁶ immune cells at least 1 day after injection into the subject.

14. The method of claim 11, wherein the injected device comprises 10⁷ or fewer cells 15 days or more after injection.

15. The method of claim 11, wherein the injected device, a tissue within 10 cm of the injected device, or both comprises an elevated level of a cytokine compared to the level of the cytokine at a site in the subject more than 10 cm away from the injected device.

16. The method of claim 11, wherein the device increases the survival time of at least 80% of subjects diagnosed with a cancer by at least 1 month compared to the survival time of untreated subjects, wherein increased survival time is determined by comparing the prognosis for survival in the subjects from a time period prior to administration of the device to the prognosis for survival in the subjects following administration of the device, wherein an increase in predicted survival time indicates that the treatment increased survival of the subjects following administration of the device.

17. The method of claim 11, wherein the device reduces the rate of tumor growth in the subject compared to the rate of tumor growth in an untreated subject.

18. The device of claim 7, wherein said nucleic acid comprises deoxyribonucleic acid (DNA).

19. The device of claim 1, wherein the device is in the shape of a disc, cylinder, square, rectangle, or string.

20. The device of claim 1, wherein said highly crosslinked cryogel composition comprises a crosslinking density of at least 50% polymer crosslinking.

21. The device of claim 1, wherein said highly crosslinked cryogel composition comprises a crosslinking density of at least 50-100% polymer crosslinking.

22. The device of claim 1, wherein said device comprises live attenuated HER-2/neu expressing breast cancer cells.

23. The device of claim 1, wherein said live attenuated breast cancer cell is an irradiated breast cancer cell.

24. The device of claim 1, wherein said cryogel composition comprises macropores having a diameter of 20 μ

m to 300 μ

m.

32. The method of claim 11, wherein the device

(i) comprises live attenuated HER-2/neu expressing breast cancer cells, and

(ii) increases the level of HER-2/neu-specific IgG antibody in subjects diagnosed with a cancer by approximately 70-fold compared to untreated subjects.

33. The device of claim 1, wherein said cryogel composition is characterized by shape memory following deformation by compression through a needle such that said cryogel composition returns to its original undeformed three-dimensional shape less than one second after compression through the needle.

34. The device of claim 6, wherein said biomolecule comprises a pathogen-associated molecular pattern (PAMP).

25. A syringe comprising an injectable device comprising a cell-adhesive, highly crosslinked cryogel composition comprising open interconnected pores,

wherein said cryogel composition comprises at least 75% pores,

wherein said cryogel composition is characterized by shape memory following deformation by compression,

wherein said cryogel composition comprises a crosslinked gelatin polymer, wherein said gelatin is methacrylated,

wherein said cryogel composition comprises a methacrylated gelatin concentration of 0.5% to 1.4% (w/v), and

wherein said cryogel composition comprises live attenuated breast cancer cells in one or more of said open interconnected pores.

26. The syringe of claim 25, comprising a 16-gauge, an 18-gauge, a 22-gauge, a 24-gauge, a 26-gauge, a 28-gauge, a 30-gauge, a 32-gauge, or a 34-gauge needle.

27. The syringe of claim 25, comprising an 18 to 30-gauge needle.

28. The syringe of claim 26, wherein the device is between 1 mm³ to 50 mm³ in size.

29. The syringe of claim 26, wherein 90% or more of the cancer cells survive passage of the device through the bore of the needle.

30. The syringe of claim 25, wherein said cryogel composition is characterized by shape memory following deformation by compression through a needle, such that said cryogel returns to its original undeformed three-dimensional shape less than one second after compression through the needle.

31. The syringe of claim 25, wherein said cryogel composition comprises macropores having a diameter of 20 μ

m to 300 μ

m.

35. An injectable device comprising a cell-adhesive, highly crosslinked cryogel composition comprising open interconnected macropores,

wherein said cryogel composition comprises at least 75% pores,

wherein said cryogel composition is characterized by shape memory following deformation by compression through a needle,

wherein said cryogel composition comprises a crosslinked gelatin polymer, wherein said gelatin is methacrylated, and

wherein said cryogel composition comprises a methacrylated gelatin concentration of 0.5% to 1.4% (w/v).

36. The device of claim 35, wherein said cryogel composition comprises macropores having a diameter of 20 μ

m to 300 μ

m.

37. The device of claim 35, wherein said cryogel composition is characterized by shape memory following deformation by compression through a needle, such that said cryogel returns to its original undeformed three-dimensional shape less than one second after compression through the needle.

38. The device of claim 35, wherein said highly crosslinked cryogel composition comprises a crosslinking density of at least 50-100% polymer crosslinking.

39. A syringe comprising the device of claim 35.