Direct cellular energy delivery system

• US 9,066,866 B2
• Filed: 06/17/2011
• Issued: 06/30/2015
• Est. Priority Date: 05/14/2002
• Status: Active Grant

• Current Assignee: University of Louisville
• Original Assignee: University of Louisville
• Inventors: Ehringer, William D., Chien, Sufan
• Primary Examiner(s): Shomer, Isaac
• Application Number:
  US13/163,546
• Publication Number:
  US 20110311442A1
• Time in Patent Office: 1,474 Days
• Field of Search: None
• US Class Current: 1/1
• CPC Class Codes: A61K 31/7076 containing purines, e.g. adenosine, adenylic acid
• CPC Class Codes: A61K 38/28 Insulins
• CPC Class Codes: A61K 9/0019 Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner non-active ingredients are additionally classified in A61K47/00
• CPC Class Codes: A61K 9/127 Liposomes
• CPC Class Codes: A61P 11/00 Drugs for disorders of the respiratory system
• CPC Class Codes: A61P 17/02 for treating wounds, ulcers, burns, scars, keloids, or the like
• CPC Class Codes: A61P 23/00 Anaesthetics
• CPC Class Codes: A61P 3/02 Nutrients, e.g. vitamins, minerals
• CPC Class Codes: A61P 31/00 Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 39/00 General protective or antinoxious agents
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 9/00 Drugs for disorders of the cardiovascular system
• CPC Class Codes: Y02A 50/30 Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
• CPC Class Codes: Y10S 977/726 using chemical reaction/biological energy, e.g. ATP
• CPC Class Codes: Y10S 977/907 Liposome

Abstract

A lipid vesicle comprising a phospholipid which is a stable vesicle former and at least one unstable vesicle forming member, wherein the unstable vesicle forming member is selected from the group consisting of a polar lipid which is not a stable vesicle former, a PEG, a raft former and a fusion protein is provided. The vesicle can further comprise a biomolecule, such as for example ATP. Methods of using the vesicle for delivery of biomolecules are also provided.

Claims

1. Vesicles, comprising:

(a) a first lipid that is a phosphatidylcholine; and

(b) a second lipid having a structure selected from the group consisting of formulas

(XXIV), (XXV), (XXVI), (XXVII), (XXIX), (XXXI), (XXXII), (XXXIII), and (XXXIV);

2. The vesicles of claim 1, wherein the high-energy phosphate is ATP.

3. The vesicles of claim 2, wherein the ATP is Mg-ATP.

4. The vesicles of claim 2, wherein the ATP is present at a concentration of about 50 mM or less when the vesicle is in solution.

5. The vesicles of claim 4, wherein the ATP is present at a concentration of from about 0.001 mM to about 50 mM when the vesicles are in solution.

6. The vesicles of claim 5, wherein the ATP is present at a concentration of from about 1 mM to about 50 mM when the vesicles are in solution.

7. The vesicles of claim 1, wherein the vesicles an absorption rate of at least 10³ vesicle absorptions per second per cell when the vesicles are contacted with target cells.

8. The vesicles of claim 7, wherein the vesicle has an absorption rate of at least 10⁶ vesicle absorptions per second per cell when the vesicles are contacted with target cells.

9. The vesicles of claim 1, wherein the vesicles have a mole:

mole ratio of the first lipid to the second lipid of 1;

9 to 100,000;

1.

10. The vesicles of claim 9, wherein the vesicles have a mole:

mole ratio of the first lipid to the second lipid of 1;

1 to 1,000;

1.

11. The vesicles of claim 1, wherein the vesicles have hydrodynamic radii of from about 20 nm to about 600 nm.

12. The vesicles of claim 11, wherein the vesicles have hydrodynamic radii of from about 100 nm to about 300 nm.

13. The vesicles of claim 1, wherein the ATP is selected from the group consisting of Mg-ATP or Na-ATP.

14. The vesicles of claim 1, wherein the first lipid is selected from the group consisting of soy phosphatidylcholine and DOPC;

and wherein the second lipid has the structure of formula (XXXIII).

15. The vesicles of claim 1, wherein the vesicles are freeze-dried.

16. The vesicles of claim 1, wherein the vesicles are formulated in a solution.

17. The vesicles of claim 16, wherein the vesicles are provided in the solution at a concentration of about 5 mg/ml.

18. The vesicles of claim 1, wherein the vesicles are formulated for topical delivery.

19. The vesicles of claim 1, wherein the vesicles are formulated for transdermal delivery.

20. The vesicles of claim 1, wherein the vesicles are provided in an ointment, salve, gel, or cream.

21. The vesicles of claim 16, wherein the vesicles are provided in the solution at a concentration of about 0.5 mg/ml-20 mg/ml.

22. The vesicles of claim 1, wherein the phosphatidylcholine is selected from the group consisting of soy phosphatidylcholine, egg phosphatidylcholine, E. coli extract 5 phosphatidylcholine, 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), 1-palmitoyl-2-docosahexaenoyl-sn-glycero-3-phosphocholine (PDPC), dimyristoyl phosphatidylcholine (DMPC), dipalmitoyl phosphatidylcholine (DPPC), distearoyl phosphatidylcholine (DSPC) and mixtures thereof.

23. The vesicles of claim 1, wherein the second lipid has the structure of formula XXXII.

45. A method of delivering a high energy phosphate to a cell, comprising contacting the cell with the vesicles of claim 1,

wherein the high energy phosphate is selected from the group consisting of adenosine triphosphate (ATP), adenosine diphosphate (ADP), adenosine monophosphate (AMP), cytosine triphosphate (CTP), cytosine diphosphate (CDP), cytosine monophosphate (CMP), uracil triphosphate (UTP), uracil diphosphate (UDP), uracil monophosphate (UMP), guanosine triphosphate (GTP), guanosine diphosphate (GDP), guanosine monophosphate (GMP), thymine triphosphate (TTP), thymine diphosphate (TDP), thymine monophosphate (TMP), inosine triphosphate (ITP), inosine diphosphate (IDP), inosine monophosphate (IMP), and phosphocreatine.

46. The method of claim 45, wherein the biomolecule is ATP.

47. The method of claim 46, wherein an amount of ATP delivered to the cell is sufficient to meet metabolic demand of the cell.

48. A method for treating a wound, the method comprising contacting the wound with a composition comprising the vesicles of claim 1.

49. The method of claim 48, wherein the composition further comprises becaplermin, fibroblast growth factor, vascular endothelial growth factor, an antibiotic, silver containing compositions, a skin graft composition or combinations thereof.

50. The method of claim 48, the method further comprising contacting the wound with a skin graft composition.

51. A method of preserving tissue, comprising contacting tissue with the vesicles of claim 1.

24. A vesicle, comprising:

adenosine triphosphate (ATP);

soy phosphatidylcholine; and

1,2-dioleoyl-3-trimethylammonium-propane (DOTAP),

wherein the vesicle is unilamellar.

25. The vesicle of claim 24, wherein the ATP is present at a concentration of from about 0.01 mM to about 50 mM when the vesicle is in solution.

26. The vesicle of claim 24, wherein the vesicle has a mole:

mole ratio of soy phosphatidylcholine to DOTAP of 50;

1.

27. The vesicle of claim 24, wherein the vesicle has a hydrodynamic radius of from about 100 nm to about 300 nm.

28. The vesicle of claim 24, wherein the vesicle has a hydrodynamic radius of from about 10 nm to about 600 nm.

29. The vesicle of claim 24, wherein the ATP is selected from Mg-ATP or Na-ATP.

30. The vesicle of claim 24, further comprising a lipid having the structure of formula XXXII.

31. A vesicle, comprising:

adenosine triphosphate (ATP);

1,2-dioleoyl-sn-glycero-3-phosphatidylcholine (DOPC) and

1,2-dioleoyl-3-trimethylammonium-propane (DOTAP),

wherein the vesicle is unilamellar.

32. The vesicle of claim 31, wherein the ATP is present at a concentration of from about 0.01 mM to about 50 mM when the vesicle is in solution.

33. The vesicle of claim 31, wherein the vesicle has a mole:

mole ratio of DOPC to DOTAP of 50;

1.

34. The vesicle of claim 31, wherein the vesicle has a hydrodynamic radius of from about 100 nm to about 300 nm.

35. The vesicle of claim 31, wherein the vesicle has a hydrodynamic radius of from about 10 nm to about 600 nm.

36. The vesicle of claim 31, further comprising a lipid having the structure of formula XXXII.

37. A vesicle, comprising:

A first lipid that is a phosphatidylcholine;

A second lipid having the structure of formula XXXII

38. A vesicle, comprising:

a high energy phosphate selected from the group consisting of adenosine triphosphate (ATP), adenosine diphosphate (ADP), adenosine monophosphate (AMP), cytosine triphosphate (CTP), cytosine diphosphate (CDP), cytosine monophosphate (CMP), uracil triphosphate (UTP), uracil diphosphate (UDP), uracil monophosphate (UMP), guanosine triphosphate (GTP), guanosine diphosphate (GDP), guanosine monophosphate (GMP), thymine triphosphate (TTP), thymine diphosphate (TDP), thymine monophosphate (TMP), inosine triphosphate (ITP), inosine diphosphate (IDP), inosine monophosphate (IMP), and phosphocreatinea phosphatidylcholine; and

1,2-dioleoyl-3-trimethylammonium-propane (DOTAP),

wherein the vesicle is unilamellar.

39. The vesicle of claim 38, wherein the high energy phosphate is ATP.

40. The vesicle of claim 38, wherein the phosphatidylcholine is soy phosphatidylcholine, egg phosphatidylcholine, E. coli extract 5 phosphatidylcholine, 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), 1-palmitoyl-2-docosahexaenoyl-sn-glycero-3-phosphocholine (PDPC), dimyristoyl phosphatidylcholine (DMPC), dipalmitoyl phosphatidylcholine (DPPC), distearoyl phosphatidylcholine (DSPC) or a mixture thereof.

41. The vesicle of claim 38, wherein the vesicle is freeze-dried.

42. The vesicle of claim 38, wherein the vesicle is formulated in a solution.

43. The vesicle of claim 42, wherein the vesicle is provided in the solution at a concentration of about 0.5 mg/ml-20 mg/ml.

44. The vesicle of claim 38, wherein the vesicle is provided in an ointment, salve, gel, or cream.