Co-delivery of stimulatory and inhibitory factors to create temporally stable and spatially restricted zones

• US 9,693,954 B2
• Filed: 06/27/2011
• Issued: 07/04/2017
• Est. Priority Date: 06/25/2010
• Status: Active Grant

• Current Assignee: President and Fellows of Harvard College (Harvard Corporation)
• Original Assignee: President and Fellows of Harvard College (Harvard Corporation)
• Inventors: Mooney, David J., Yuen, William W., Arany, Praveen
• Primary Examiner(s): Xie, Xiaozhen
• Application Number:
  US13/805,840
• Publication Number:
  US 20130302396A1
• Time in Patent Office: 2,199 Days
• Field of Search: None
• CPC Class Codes: A61F 2/30756 Cartilage endoprostheses A61F2/3603 takes precedence
• CPC Class Codes: A61F 2002/30766 Scaffolds for cartilage ingrowth and regeneration
• CPC Class Codes: A61K 2300/00 Mixtures or combinations of active ingredients, wherein at least one active ingredient is fully defined in groups A61K31/00 - A61K41/00
• CPC Class Codes: A61K 31/16 Amides, e.g. hydroxamic acids
• CPC Class Codes: A61K 31/335 having oxygen as the only ring hetero atom, e.g. fungichromin
• CPC Class Codes: A61K 31/381 having five-membered rings
• CPC Class Codes: A61K 31/4192 1,2,3-Triazoles
• CPC Class Codes: A61K 31/454 containing a five-membered ring with nitrogen as a ring hetero atom, e.g. pimozide, domperidone
• CPC Class Codes: A61K 35/12 Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells vaccines or medicinal preparations containing antigens or antibodies A61K39/00
• CPC Class Codes: A61K 35/28 Bone marrow; Haematopoietic stem cells; Mesenchymal stem cells of any origin, e.g. adipose-derived stem cells
• CPC Class Codes: A61K 35/545 Embryonic stem cells; Pluripotent stem cells; Induced pluripotent stem cells; Uncharacterised stem cells
• CPC Class Codes: A61K 38/1808 Epidermal growth factor [EGF] urogastrone
• CPC Class Codes: A61K 38/1841 Transforming growth factor [TGF]
• CPC Class Codes: A61K 38/185 Nerve growth factor [NGF]; Brain derived neurotrophic factor [BDNF]; Ciliary neurotrophic factor [CNTF]; Glial derived neurotrophic factor [GDNF]; Neurotrophins, e.g. NT-3
• CPC Class Codes: A61K 38/1866 Vascular endothelial growth factor [VEGF]
• CPC Class Codes: A61K 38/1875 Bone morphogenic factor; Osteogenins; Osteogenic factor; Bone-inducing factor
• CPC Class Codes: A61K 39/3955 against proteinaceous materials, e.g. enzymes, hormones, lymphokines
• CPC Class Codes: A61K 45/06 Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
• CPC Class Codes: A61K 9/0087 Galenical forms not covered by A61K9/02 - A61K9/7023
• CPC Class Codes: A61P 19/02 for joint disorders, e.g. arthritis, arthrosis
• CPC Class Codes: A61P 9/00 : Drugs for disorders of the cardiovascular system

Abstract

The invention provides methods and compositions for local manipulation of regenerative processes via exogenous factor delivery.

Claims

1. A method for promoting cartilage regeneration, comprising:

contacting a target site of a subject with a device comprising a polymeric scaffold comprising a first and a second spatially distinct restricted zones, wherein the first zone comprises a purified morphogen that promotes cartilage regeneration, and wherein the second zone comprises a purified inhibitor of the morphogen;

wherein the morphogen and the inhibitor are released from the device into the target site;

wherein the morphogen promotes cartilage regeneration over a sustained period of time at a spatially restricted first region of the target site contacted by the first zone, thereby creating a field of action; and

wherein the inhibitor inhibits the activity of the morphogen at a spatially restricted second region of the target site contacted by the second zone, thereby creating a boundary between the first region and the second region of the target site defining where cartilage regeneration is promoted.

2. The method of claim 1, wherein said morphogen is released at a first rate and said inhibitor is released at a second rate.

3. The method of claim 1, wherein the morphogen that promotes cartilage regeneration is a transforming growth factor-β

(TGF-β

).

4. The method of claim 3, wherein the TGF-β

is selected from the group consisting of TGF-β

1, TGF-β

2, TGF-β

3, and Latent TGF-β

1 (LTGF-β

1).

5. The method of claim 3, wherein the inhibitor is an anti-TGF-β

antibody.

6. The method of claim 5, wherein the anti-TGF-β

antibody is an anti-TGF-β

1 antibody.

7. The method of claim 3, wherein the inhibitor is the small molecule inhibitor SB431542.

8. The method of claim 1, wherein the purified morphogen that promotes cartilage regeneration is Latent TGF-β

1 and the purified inhibitor is the small molecule inhibitor SB431542.

9. The method of claim 1, wherein the morphogen is encapsulated within a microsphere.

10. The method of claim 1, wherein the inhibitor is encapsulated within a microsphere.

11. The method of claim 1, wherein the morphogen and the inhibitor are released from the device by diffusion.

12. The method of claim 1, wherein the polymeric scaffold comprises poly(lactic-co-glycolic) acid (PLGA).

13. The method of claim 1, wherein said device further comprises cells.

14. The method of claim 13, wherein said cells comprise mesenchymal stem cells, embryonic stem cells, or induced pluripotent stem cells.

15. The method of claim 1, wherein said device further comprises:

a) a second purified morphogen, wherein the second purified morphogen promotes bone regeneration, and wherein the second purified morphogen is present in a spatially distinct restricted zone from the purified morphogen that promotes cartilage regeneration; and

b) a second purified inhibitor, wherein the second purified inhibitor inhibits the activity of the second purified morphogen, and wherein the second purified inhibitor is present in a spatially distinct restricted zone from the second purified morphogen.

16. The method of claim 15, wherein the second purified morphogen is a bone morphogenetic protein (BMP).

17. The method of claim 16, wherein the bone morphogenetic protein is BMP4.

18. The method of claim 16, wherein the second purified inhibitor is an anti-BMP antibody.

19. The method of claim 18, wherein the anti-BMP antibody is an anti-BMP4 antibody.

20. The method of claim 1, wherein said method is performed in vivo.

21. A method of inducing cartilage regeneration in a spatially restricted region of a target site, comprising:

contacting the target site with a device comprising a polymeric scaffold comprising a first and a second spatially distinct restricted zones, wherein the first zone comprises a purified morphogen that promotes cartilage regeneration, and wherein the second zone comprises a purified inhibitor of the morphogen;

wherein the morphogen and the inhibitor are released from the device into the target site;

wherein the morphogen promotes cartilage regeneration at a spatially restricted first region of the target site contacted by the first zone, thereby creating a field of action; and

wherein the inhibitor inhibits the activity of the morphogen at a spatially restricted second region of the target site contacted by the second zone, thereby creating a boundary between the first region and the second region of the target site defining where cartilage regeneration is promoted, and inducing cartilage regeneration in a spatially restricted region of the target site.

22. The method of claim 21, wherein the morphogen that promotes cartilage regeneration is a transforming growth factor-β

(TGF-β

).

23. The method of claim 22, wherein the transforming growth factor-β

is selected from the group consisting of TGF-β

1, TGF-β

2, TGF-β

3, and Latent TGF-β

1 (LTGF-β

1).

24. The method of claim 22, wherein the inhibitor is an anti-TGF-β

antibody.

25. The method of claim 24, wherein the anti-TGF-β

antibody is an anti-TGF-β

1 antibody.

26. The method of claim 22, wherein the inhibitor is the small molecule inhibitor SB431542.

27. The method of claim 21, wherein the morphogen is encapsulated within a microsphere.

28. The method of claim 21, wherein the inhibitor is encapsulated within a microsphere.

29. The method of claim 21, wherein the morphogen and the inhibitor are released from the device by diffusion.

30. The method of claim 21, wherein the polymeric scaffold comprises PLGA.

31. The method of claim 21, wherein said device further comprises cells.

32. The method of claim 31, wherein said cells comprise mesenchymal stem cells, embryonic stem cells, or induced pluripotent stem cells.

33. The method of claim 21, wherein the device further comprises:

a) a second purified morphogen, wherein said second purified morphogen promotes bone regeneration, and wherein the second purified morphogen is present in a spatially distinct restricted zone from the purified morphogen that promotes cartilage regeneration; and

b) a second purified inhibitor, wherein said second purified inhibitor inhibits the activity of the second purified morphogen, and wherein the second purified inhibitor is present in a spatially distinct restricted zone from the second purified morphogen.

34. The method of claim 33, wherein the second purified morphogen is a bone morphogenetic protein (BMP).

35. The method of claim 34, wherein the bone morphogenetic protein is BMP4.

36. The method of claim 34, wherein the second purified inhibitor is an anti-BMP antibody.

37. The method of claim 36, wherein the anti-BMP antibody is an anti-BMP4 antibody.

38. The method of claim 21, wherein said method is performed in vivo.