Hybrid matrix implants and explants

• US 5,965,125 A
• Filed: 10/25/1995
• Issued: 10/12/1999
• Est. Priority Date: 10/25/1995
• Status: Expired due to Fees

• Current Assignee: Shire Human Genetic Therapies, Inc. (Shire plc)
• Original Assignee: Transkaryotic Therapies Incorporated (Shire plc)
• Inventors: Mineau-Hanschke, Rochelle
• Primary Examiner(s): Campell, Bruce R.
• Application Number:
  US08/548,002
• Time in Patent Office: 1,448 Days
• Field of Search: 435/240.22, 435/240.2, 435/172.3, 435/320.1, 435/240.23, 435/382, 935/6, 935/9, 935/10, 935/11, 935/12, 935/13, 935/14, 935/15, 935/66, 935/70, 935/59, 935/62, 424/422, 424/423, 424/424, 424/93.1, 424/93.2, 424/93.21
• US Class Current: 424/93.21
• CPC Class Codes: A61F 2310/00365 Proteins; Polypeptides; Degradation products thereof
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: A61L 2300/414 Growth factors
• CPC Class Codes: A61L 2300/426 Immunomodulating agents, i.e. cytokines, interleukins, interferons
• CPC Class Codes: A61L 2300/428 Vitamins, e.g. tocopherol, riboflavin
• CPC Class Codes: A61L 2300/64 Animal cells
• CPC Class Codes: A61L 27/24 Collagen
• CPC Class Codes: A61L 27/38 containing added animal cells organs or tissue containing native cells A61L27/36
• CPC Class Codes: A61L 27/3839 characterised by the site of application in the body materials for artificial blood vessels A61L27/507; materials for use in artificial skin A61L27/60
• CPC Class Codes: A61L 27/44 having a macromolecular matrix
• CPC Class Codes: A61L 27/54 Biologically active materials, e.g. therapeutic substances A61L27/227 takes precedence
• CPC Class Codes: A61P 17/02 for treating wounds, ulcers, burns, scars, keloids, or the like
• CPC Class Codes: C07K 14/61 Growth hormone [GH], i.e. somatotropin

Abstract

An implantable device having a body of matrix material made up of insoluble collagen fibrils, and disposed therewithin

(a) a plurality of vertebrate cells; and

(b) a plurality of microspheres each of which consists primarily of one or more of the following materials: collagen, polystyrene, dextran, polyacrylamide, cellulose, calcium alginate, latex, polysulfone, or glass.

Claims

1. A composition comprising a body of matrix material comprising insoluble collagen fibrils, there being embedded within the body of matrix material

(a) a plurality of cultured vertebrate cells genetically engineered to express a medically useful polyeptide;

and

(b) a plurality of microspheres.

2. The composition of claim 1, wherein the cultured vertebrate cells are selected from the group consisting of adipocytes, astrocytes, cardiac muscle cells, chondrocytes, endothelial cells, epithelial cells, fibroblasts, gangliocytes, glandular cells, glial cells, hematopoietic cells, hepatocytes, keratinocytes, myoblasts, neural cells, osteoblasts, pancreatic beta cells, renal cells, smooth muscle cells, striated muscle cells, and precursors of any of the above.

3. The composition of claim 1, wherein the cultured vertebrate cells are human cells.

4. The composition of claim 1, wherein the cultured vertebrate cells are transfected cells containing exogenous DNA encoding the medically useful polypeptide.

6. The composition of claim 4, wherein the polypeptide is selected from the group consisting of enzymes, hormones, cytokines, colony stimulating factors, vaccine antigens, antibodies, clotting factors, regulatory proteins, transcription factors, receptors, and structural proteins.

7. The composition of claim 4, wherein the polypeptide is an angiogenesis factor.

8. The composition of claim 4 wherein the polypeptide is selected from the group consisting of human growth hormone, Factor VIII, Factor IX, erythropoietin, and insulin.

10. The composition of claim 1, wherein the polypeptide is selected from the group consisting of alpha-1 antitrypsin, calcitonin, glucocerebrosidase, low density lipoprotein (LDL) receptor, IL-2 receptor, globin, immunoglobulin, catalytic antibodies, the interleukins, insulin-like growth factor 1 (IGF-1), parathyroid hormone (PTH), leptin, the interferons, the nerve growth factors, basic fibroblast growth factor (bFGF), acidic FGF (aFGF), epidermal growth factor (EGF), endothelial cell growth factor, platelet derived growth factor (PDGF), transforming growth factors, endothelial cell stimulating angiogenesis factor (ESAF), angiogenin, tissue plasminogen activator (t-PA), granulocyte colony stimulating factor (G-CSF), and granulocyte-macrophage colony stimulating factor (GM-CSF).

11. The composition of claim 1, wherein the microspheres are beads of type I collagen.

12. The composition of claim 1, wherein the majority of the microspheres have a diameter between approximately 0.1 and approximately 2 mm.

13. The composition of claim 1, wherein the collagen in the matrix material is type I.

14. The composition of claim 13, wherein the matrix material additionally comprises a substance selected from the group consisting of a second type of collagen, agarose, alginate, fibronectin, laminin, hyaluronic acid, heparan sulfate, dermatan sulfate, sulfated proteoglycans, fibrin, elastin, and tenascin.

15. A method of making the composition of claim 1, comprising

forming a mixture comprising (a) a plurality of cultured vertebrate cells genetically engineered to express a medically useful polypeptide;

(b) a plurality of microspheres; and

(c) a solution comprising soluble collagen;

subjecting the soluble collagen in the mixture to conditions effective to form a gel; and

exposing the gel to culture conditions which cause the gel to contract, thereby forming the body of the composition.

16. The method of claim 15, wherein the microspheres are porous collagen beads.

17. The method of claim 15, wherein the solution additionally comprises a substance selected from the group consisting of a second type of collagen, agarose, alginate, fibronectin, laminin, hyaluronic acid, heparan sulfate, dermatan sulfate, sulfated proteoglycans, fibrin, elastin, and tenascin.

18. The method of claim 15, wherein the solution is an acidic aqueous solution of soluble collagen, and gelation is accomplished by raising the pH of the solution.

19. The method of claim 15, wherein the gelation step takes place in a mold, so that, prior to the contracting step, the gel is in the shape of the mold.

20. The method of claim 15, wherein the cultured vertebrate cells are cultured in the presence of the microspheres prior to being mixed with the solution.

21. The composition of claim 1, additionally comprising noncollagen fibers dispersed within the body of matrix material.

22. The composition of claim 21, wherein the noncollagen fibers comprise a material selected from the group consisting of nylon, dacron, polytetrafluoroethylene, polyglycolic acid, polylactic/polyglycolic acid mixture, polystyrene, polyvinylchloride copolymer, cat gut, cotton, linen, polyester and silk.

52. The composition of claim 1, wherein the cultured vertebrate cells are fibroblasts.

54. The composition of claim 1, wherein the microspheres have an approximately spherical shape.

59. The composition of claim 4 wherein the polypeptide is human growth hormone.

60. The composition of claim 4 wherein the polypeptide is erythropoietin.

61. The composition of claim 4 wherein the polypeptide is Factor VIII.

62. The composition of claim 4 wherein the polypeptide is Factor IX.

68. The composition of claim 3, wherein non-collagen fibers are disposed within the body of matrix material.

69. The composition of claim 68, wherein the non-collagen fibers comprise a material selected from the group consisting of nylon, dacron, polytetrafluoroethylene, polyglycolic acid, polylactic/polyglycolic acid mixture, polystyrene, polyvinylchloride co-polymer, cat gut, cotton, linen, polyester, and silk.

70. The composition of claim 1, configured to be implanted into a patient.

71. The composition of claim 70, wherein the cultured vertebrate cells are derived from one or more cells removed from the patient.

72. The composition of claim 70, wherein the cultured vertebrate cells consist of a clonal population.

73. The composition of claim 70, wherein the cultured vertebrate cells are transfected cells containing exogenous DNA which includes a regulatory sequence that activates expression of an endogenous gene encoding a medically useful polypeptide.

74. The composition of claim 11, wherein the cultured vertebrate cells are transfected human cells and the collagen in the matrix material is type 1.

75. A method of administering a medically useful polypeptide to a patient in need thereof, comprising providing the composition of claim 70, wherein the cultured vertebrate cells secrete the medically useful polypeptide;

and

implanting the composition in the patient.

76. The method of claim 75, wherein the cultured vertebrate cells are derived from one or more cells removed from the patient, and have been transfected in vitro with exogenous DNA encoding the medically useful polypeptide.

77. The method of claim 75, wherein the implanting is carried out at a subcutaneous site in the patient.

78. The method of claim 75, wherein the implanting is carried out at an intraperitoneal, sub-renal capsular, inguinal, intramuscular, intraventricular, or intrathecal site in the patient.

79. The method of claim 75, wherein the medically useful polypeptide is one which promotes wound healing, and the implanting is carried out at the site of a preexisting wound of the patient.

80. The composition of claim 70, wherein the cultured vertebrate cells are transfected cells containing exogenous DNA encoding a medically useful polypeptide.

5. The composition of claim l, wherein the cultured vertebrate cells are transfected cells containing exogenous DNA which includes a regulatory sequence that activates expression of a gene encoding the medically useful polypeptide, said gene being endogenous to said vertebrate cells both prior to and after they are transfected.

9. The composition of claim 5 wherein the polypeptide is selected from the group consisting of human growth hormone, Factor VIII, Factor IX, erythropoietin, and insulin.

63. The composition of claim 5 wherein the polypeptide is an angiogenesis factor.

64. The composition of claim 5 wherein the polypeptide is human growth hormone.

65. The composition of claim 5 wherein the polypeptide is erythropoietin.

66. The composition of claim 5 wherein the polypeptide is Factor VIII.

67. The composition of claim 5 wherein the polypeptide is Factor IX.

23. A composition comprising a body of matrix material comprising insoluble collagen fibrils, there being embedded within the body of matrix material

(a) a plurality of cultured vertebrate cells genetically engineered to express a medically useful polypeptide;

and

(b) a plurality of microspheres, each of which consists primarily of one or more substances selected from the list consisting of collagen, polystyrene, dextran, polyacrylamide, cellulose, calcium alginate, latex, polysulfone, and glass.

24. The composition of claim 23, wherein the cultured, genetically engineered vertebrate cells are selected from the group consisting of adipocytes, astrocytes, cardiac muscle cells, chondrocytes, endothelial cells, epithelial cells, fibroblasts, gangliocytes, glandular cells, glial cells, hematopoietic cells, hepatocytes, keratinocytes, myoblasts, neural cells, osteoblasts, pancreatic beta cells, renal cells, smooth muscle cells, striated muscle cells, and precursors of any of the above.

25. The composition of claim 23, wherein the cultured, genetically engineered vertebrate cells are human cells.

26. The composition of claim 23, wherein the cultured, genetically engineered vertebrate cells are transfected cells containing exogenous DNA encoding the polypeptide.

27. The composition of claim 23, wherein the cultured, genetically engineered vertebrate cells are transfected cells containing exogenous DNA which includes a regulatory sequence that activates expression of a gene encoding the polypeptide, said gene being endogenous to said vertebrate cells both prior to and after they are transfected.

28. The composition of claim 23, wherein the polypeptide is selected from the group consisting of enzymes, hormones, cytokines, colony stimulating factors, vaccine antigens, antibodies, clotting factors, regulatory proteins, transcription factors, receptors, and structural proteins.

29. The composition of claim 23, wherein the polypeptide is an angiogenesis factor.

30. The composition of claim 23 wherein the polypeptide is human growth hormone.

31. The composition of claim 23 wherein the polypeptide is erythropoietin.

32. The composition of claim 26, wherein the polypeptide is selected from the group consisting of enzymes, hormones, cytokines, colony stimulating factors, vaccine antigens, antibodies, clotting factors, regulatory proteins, transcription factors, receptors, and structural proteins.

33. The composition of claim 26, wherein the polypeptide is an angiogenesis factor.

34. The composition of claim 26 wherein the polypeptide is human growth hormone.

35. The composition of claim 26 wherein the polypeptide is erythropoietin.

36. The composition of claim 27, wherein the polypeptide is selected from the group consisting of enzymes, hormones, cytokines, colony stimulating factors, vaccine antigens, antibodies, clotting factors, regulatory proteins, transcription factors, receptors, and structural proteins.

37. The composition of claim 27, wherein the polypeptide is an angiogenesis factor.

38. The composition of claim 27 wherein the polypeptide is human growth hormone.

39. The composition of claim 27 wherein the polypeptide is erythropoietin.

40. The composition of claim 23, wherein the polypeptide is selected from the group consisting of alpha-1 antitrypsin, calcitonin, glucocerebrosidase, low density lipoprotein (LDL) receptor, IL-2 receptor, globin, immunoglobulin, catalytic antibodies, the interleukins, insulin-like growth factor 1 (IGF-1), parathyroid hormone (PTH), leptin, the interferons, the nerve growth factors, basic fibroblast growth factor (bFGF), acidic FGF (aFGF), epidermal growth factor (EGF), endothelial cell growth factor, platelet derived growth factor (PDGF), transforming growth factors, endothelial cell stimulating angiogenesis factor (ESAF), angiogenin, tissue plasminogen activator (t-PA), granulocyte colony stimulating factor (G-CSF), and granulocyte-macrophage colony stimulating factor (GM-CSF).

41. The composition of claim 23, wherein the majority of the microspheres have a diameter between approximately 0.1 and approximately 2 mm.

42. The composition of claim 23, wherein the collagen in the matrix material is type I.

43. The composition of claim 42, wherein the matrix material additionally comprises a substance selected from the group consisting of a second type of collagen, agarose, alginate, fibronectin, laminin, hyaluronic acid, heparin sulfate, dermatan sulfate, sulfated proteoglycans, fibrin, elastin, and tenascin.

44. A method of making the composition of claim 23, comprising

forming a mixture comprising (a) a plurality of cultured vertebrate cells genetically engineered to express a medically useful polypeptide;

(b) a plurality of microspheres, each of which consists primarily of one or more substances selected from the list consisting of collagen, polystyrene, dextran, polyacrylamide, cellulose, calcium alginate, latex, polysulfone, and glass; and

(c) a solution comprising soluble collagen;

subjecting the soluble collagen in the mixture to conditions effective to form a gel; and

exposing the gel to culture conditions which cause the gel to contract, thereby forming the body of the composition.

45. The method of claim 44, wherein the microspheres are porous collagen beads.

46. The method of claim 44, wherein the solution additionally comprises a substance selected from the group consisting of a second type of collagen, agarose, alginate, fibronectin, laminin, hyaluronic acid, heparin sulfate, dermatan sulfate, sulfated proteoglycans, fibrin, elastin, and tenascin.

47. The method of claim 44, wherein the solution is an acidic aqueous solution of soluble collagen, and gelation is accomplished by raising the pH of the solution.

48. The method of claim 44, wherein the gelation step takes place in a mold, so that, prior to the contracting step, the gel is in the shape of the mold.

49. The method of claim 44, wherein the cultured vertebrate cells are cultured in the presence of the microspheres prior to being mixed with the solution.

50. The composition of claim 23, additionally comprising noncollagen fibers dispersed within the body of matrix material.

51. The composition of claim 50, wherein the noncollagen fibers comprise a material selected from the group consisting of nylon, dacron, polytetrafluoroethylene, polyglycolic acid, polylactic/polyglycolic acid mixture, polystyrene, polyvinylchloride copolymer, cat gut, cotton, linen, polyester and silk.

53. The composition of claim 23, wherein the cultured vertebrate cells are fibroblasts.

55. The composition of claim 23, wherein the microspheres have an approximately spherical shape.

56. The composition of claim 23 wherein the polypeptide is Factor VIII.

57. The composition of claim 23 wherein the polypeptide is Factor IX.

58. The composition of claim 23 wherein the polypeptide is insulin.