COMPOSITIONS AND METHODS FOR NUCLEIC ACID DELIVERY

• US 20140302116A1
• Filed: 03/07/2014
• Published: 10/09/2014
• Est. Priority Date: 03/15/2013
• Status: Active Grant

• Current Assignee: Massachusetts Institute of Technology
• Original Assignee: Massachusetts Institute of Technology
• Inventors: Hammond, Paula T., Castleberry, Steven A.
• Granted Patent: US 9,463,244 B2
• Time in Patent Office: Days
• US Class Current: 424/443
• CPC Class Codes: A61K 31/7105 Natural ribonucleic acids, i.e. containing only riboses attached to adenine, guanine, cytosine or uracil and having 3'-5' phosphodiester links
• CPC Class Codes: A61K 45/06 Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
• CPC Class Codes: A61L 2400/12 Nanosized materials, e.g. nanofibres, nanoparticles, nanowires, nanotubes; Nanostructured surfaces
• CPC Class Codes: A61L 2420/08 Coatings comprising two or more layers
• CPC Class Codes: A61L 27/12 Phosphorus-containing materials, e.g. apatite
• CPC Class Codes: A61L 27/22 Polypeptides or derivatives thereof , e.g. degradation products
• CPC Class Codes: A61L 27/46 with phosphorus-containing inorganic fillers
• CPC Class Codes: A61L 31/026 Ceramic or ceramic-like structures, e.g. glasses
• CPC Class Codes: A61L 31/047 Other specific proteins or polypeptides not covered by A61L31/044 - A61L31/046
• CPC Class Codes: A61L 31/127 containing fillers of phosphorus-containing inorganic materials
• CPC Class Codes: A61L 31/16 Biologically active materials, e.g. therapeutic substances A61L31/047 takes precedence
• CPC Class Codes: A61P 17/02 for treating wounds, ulcers, burns, scars, keloids, or the like

Abstract

The present invention provides, among other things, multilayer film coating compositions, coated substrates and methods thereof. In some embodiments, a structure includes a substrate; and a multilayer film coated on the substrate, wherein adjacent layers of the multilayer film are associated with one another via one or more non-covalent interactions, wherein the multilayer film comprises a first nucleic acid agent present at a loading density, and further wherein the multilayer film is characterized in that, when the structure is placed on a subject so that the multilayer film contacts cells, the first nucleic acid agent is released with a profile characterized by a feature selected from the group consisting of 1) being a burst-free release; 2) being a sustained release; and 3) exhibiting in vitro and/or in vivo biological effectiveness.

Claims

1. A structure for localized and controlled release of nucleic acid agents comprising:

a substrate;

a multilayer film coated on the substrate, wherein adjacent layers of the multilayer film are associated with one another via one or more non-covalent interactions,

wherein the multilayer film comprises a first nucleic acid agent present at a loading density of at least about 1 μ

g/cm², and

further wherein the multilayer film is characterized in that, when the structure is placed in contact with cells of a subject so that the multilayer film contacts the cells, the first nucleic acid agent is released with a profile characterized by a feature selected from the group consisting of

a) being a burst-free release;

b) being a sustained release; and

c) exhibiting in vitro and/or in vivo biological effectiveness.

2. The structure of claim 1, wherein the loading density of the first nucleic acid agent is at least about 1 μ

g/cm², about 2 μ

g/cm², about 5 μ

g/cm², about 8 μ

g/cm², about 10 μ

g/cm², about 12 μ

g/cm², about 15 μ

g/cm², about 18 μ

g/cm², about 20 μ

g/cm², about 25 μ

g/cm², about 30 μ

g/cm², about 50 μ

g/cm², or about 100 μ

g/cm².

3. The structure of claim 1, wherein the burst-free release is characterized by releasing less than about 10%, about 20%, about 30%, about 40%, or about 50% of the first nucleic acid agent in the first 24 hours after placement on the subject.

4. The structure of claim 1, wherein the sustained release is characterized in that the first nucleic acid agent is released from the multilayer film over an extended period of time of at least about 3 days, about 5 days, about 7 days, about 10 days, about 15 days, about 30 days, about 1 month, about 2 months, about 3 months, about 6 months, or about 1 year.

5. The structure of claim 1, wherein the first nucleic acid agent is or comprises one or more members selected from the group consisting of deoxyribonucleic acid (DNA), ribonucleic acid (RNA), peptide nucleic acid (PNA), morpholino nucleic acid, locked nucleic acid (LNA), glycol nucleic acid (GNA), and threose nucleic acid (TNA).

6. The structure of claim 1, wherein the first nucleic acid agent is or comprises an siRNA agent.

7. The structure of claim 6, wherein the siRNA agent inhibits expression of a polypeptide.

8. The structure of claim 7, wherein the polypeptide is selected from the group consisting of matrix metallopeptidase 9 (MMP-9), neutral endopeptidase (NEP) and protein tyrosine phosphatase 1B (PTP 1B).

9. The structure of claim 1, wherein at least one layer of the multilayer film comprises or consists of the first nucleic acid agent.

10. The structure of claim 9, wherein the at least one layer comprises the first nucleic acid agent and further a ceramic material.

11. The structure of claim 10, wherein the ceramic material comprises one or more members selected from the group consisting of hydroxyapatite, floroapatite, carbonate apatide, tricalcium phosphate, octacalcium phosphate, calcium pyrophosphate, tetracalcium phosphate, and dicalcium phosphate dehydrate.

12. The structure of claim 10, wherein the ceramic material is or comprises a calcium phosphate particle.

13. The structure of claim 10, wherein the first nucleic acid agent is associated with the ceramic material.

14. The structure of claim 1, wherein the multilayer film comprises one or more architectures selected from the group consisting of bilayer, trilayer, and tetralayer.

15. The structure of claim 14, wherein the multilayer film comprises or consists of a number of each of the one or more architectures at the number being selected from 1, 2, 5, 10, 20, 30, 40, 50, 60, 80, 100, 150 or 200.

16. The structure of claim 1, wherein the multilayer film comprises a layer comprising or consisting of silicate.

17. The structure of claim 16, wherein the silicate is Laponite.

18. The structure of claim 1, wherein the multilayer film comprises a layer comprising or consisting of polysaccharide.

19. The structure of claim 18, wherein the polysaccharide is selected from the group consisting of sodium alginate, chitosan, agar, agarose, carragenaan or any combination thereof.

20. The structure of claim 1, wherein the multilayer film comprises a layer comprising or consisting of a degradable polyelectrolyte.

21. The structure of claim 1, wherein the multilayer film comprises at least one base layer.

22. The structure of claim 21, wherein the at least one base layer comprises a degradable polyelectrolyte.

23. The structure of claim 20, wherein the degradable polyelectrolyte is a hydrolytically degradable polyelectrolyte.

24. The structure of claim 23, wherein the hydrolytically degradable polyelectrolyte comprises a polymer selected from the group consisting of polyesters, polyanhydrides, polyorthoesters, polyphosphazenes, polyphosphoesters, and combinations thereof.

25. The structure of claim 24, wherein the polyesters are selected from the group consisting of poly(β

-amino ester)s, poly(L-lactide-co-L-lysine), poly(serine ester), poly(4-hydroxy-L-proline ester), poly[α

-(4-aminobutyl)-L-glycolic acid], PLGA, and combinations thereof.

26. The structure of claim 25, wherein the poly(β

-amino ester) is selected from the group consisting of those whose chemical structure is represented by;

27. The structure of claim 25, wherein the poly(β

-amino ester) is selected from the group consisting of those whose chemical structure is represented by;

28. The structure of claim 25, wherein the poly(β

-amino ester) is selected from the group consisting of those whose chemical structure is represented by;

29. The structure of claim 1, further comprising an agent.

30. The structure of claim 29, wherein the agent is selected from the group consisting of an antibiotic, an anti-inflammatory agent or any combination thereof.

31. The structure of claim 1, wherein the substrate is or comprises at least a portion of a medical device.

32. The structure of claim 1, wherein the substrate is a wound dressing.

33. The structure of claim 1, wherein the multilayer film further comprises a second nucleic acid agent present at a loading density of at least about 1 μ

g/cm².

34. The structure of claim 33, wherein the first and second nucleic acid agents are presented at a predetermined ratio.

35. A method of making a structure comprising steps of:

depositing layer-by-layer a multilayer film onto a substrate, wherein adjacent layers of the multilayer film are associated with one another via one or more non-covalent interactions,

wherein the multilayer film comprises a first nucleic acid agent presents at a loading density of at least about 1 μ

g/cm², and

further wherein the multilayer film is characterized in that, when the structure is placed in contact with cells of a subject so that the multilayer film contacts cells, the first nucleic acid agent is released with a profile characterized by a feature selected from the group consisting of

a) being a burst-free release;

b) being a sustained release; and

c) exhibiting in vitro and/or in vivo biological effectiveness.

36. The method of claim 35, wherein the step of depositing comprises incorporating the first nucleic acid agent in the multilayer film.

37. The method of claim 36, wherein the step of incorporating comprises associating the first nucleic acid agent with an adjacent layer of the multilayer film.

38. The method of claim 36, wherein the step of incorporating comprises depositing a layer comprising or consisting of a ceramic material.

39. The method of claim 38, wherein the step of incorporating further comprises associating the first nucleic acid agent with the ceramic material.

40. The method of claim 35, wherein the multilayer film further comprises a second nucleic acid agent present at a loading density of at least about 1 μ

g/cm².

41. The structure of claim 40, wherein the first and second nucleic acid agents are presented in the same layer of the multilayer film.

42. The structure of claim 41, wherein the step of depositing comprises contacting a cocktail comprising the first and second nucleic acid agents at a predetermined ratio.

43. A method of using a structure comprising steps of:

providing a structure comprising a multilayer film on a substrate, wherein adjacent layers of the multilayer film are associated with one another via one or more non-covalent interactions,

wherein the multilayer film comprises a first nucleic acid agent presents at a loading density of at least about 1 μ

g/cm², and

releasing the first nucleic acid, when the structure is placed in contact with cells of a subject so that the multilayer film contacts cells, with a profile characterized by a feature selected from the group consisting of

a) being a burst-free release;

b) being a sustained release; and

c) exhibiting in vitro and/or in vivo biological effectiveness.

44. The method of claim 43, wherein the substrate comprises at least a portion of a medical device.

45. The method of claim 44, further comprising a step of applying or implanting the medical device to the subject.

46. The method of claim 43, wherein the multilayer film further comprises a second nucleic acid agent present at a loading density of at least about 1 μ

g/cm².

47. The structure of claim 46, wherein the first and second nucleic acid agents are presented at a predetermined ratio.

48. The method of claim 43, further comprising a step of simultaneously or sequentially releasing the first and second nucleic acid agents.

49. A method of treating a chronic diabetic wound of a patient, which method comprises:

topically applying a wound dressing to a chronic diabetic wound, wherein the wound dressing is coated with a multilayer film,

wherein adjacent layers of the multilayer film are associated with one another via one or more non-covalent interactions,

wherein the multilayer film comprises a first siRNA agent present at a loading density of at least about 1 μ

g/cm², and

further wherein the first siRNA agent inhibits expression of MMP-9;

and

releasing the first siRNA agent with a profile characterized by a feature selected from the group consisting of

a) being a burst-free release;

b) being a sustained release; and

c) exhibiting in vitro and/or in vivo biological effectiveness.

50. The method of claim 49, wherein the chronic diabetic wound is a diabetic foot ulcer.

51. The method of claim 49, wherein the wound dressing is a bandage.