Materials and methods for making improved micelle compositions

• US 6,217,886 B1
• Filed: 01/27/1999
• Issued: 04/17/2001
• Est. Priority Date: 07/14/1997
• Status: Expired due to Term

• Current Assignee: Board of Trustees of The University of Illinois
• Original Assignee: Board of Trustees of The University of Illinois
• Inventors: Önyüksel, Hayat, Rubinstein, Israel
• Primary Examiner(s): Kishore, Gollamudi S.
• Application Number:
  US09/239,069
• Time in Patent Office: 811 Days
• Field of Search: 424/450, 424/1.21, 424/9.321, 424/9.51, 424/417, 424/96.3, 424/401, 514/2, 514/21, 514/937-943, 264/4.1, 264/4.3, 264/4.6
• US Class Current: 424/401
• CPC Class Codes: A61K 2800/57 Compounds covalently linked to a(n inert) carrier molecule, e.g. conjugates, pro-fragrances
• CPC Class Codes: A61K 38/095 Oxytocins; Vasopressins; Related peptides
• CPC Class Codes: A61K 38/1738 Calcium binding proteins, e.g. calmodulin
• CPC Class Codes: A61K 38/2278 Vasoactive intestinal peptide [VIP]; Related peptides (e.g. Exendin)
• CPC Class Codes: A61K 38/2285 Endothelin, vasoactive intestinal contractor [VIC]
• CPC Class Codes: A61K 47/6907 the form being a microemulsion, nanoemulsion or micelle
• CPC Class Codes: A61K 8/0291 Micelles
• CPC Class Codes: A61K 8/14 Liposomes; Vesicles
• CPC Class Codes: A61K 8/553 Phospholipids, e.g. lecithin
• CPC Class Codes: A61K 8/64 Proteins; Peptides; Derivatives or degradation products thereof
• CPC Class Codes: A61K 8/68 Sphingolipids, e.g. ceramides, cerebrosides, gangliosides
• CPC Class Codes: A61K 8/86 Polyethers
• CPC Class Codes: A61K 9/1075 Microemulsions or submicron emulsions; Preconcentrates or solids thereof; Micelles, e.g. made of phospholipids or block copolymers A61K9/0026 takes precedence
• CPC Class Codes: A61P 1/00 Drugs for disorders of the alimentary tract or the digestive system
• CPC Class Codes: A61Q 19/00 Preparations for care of the skin
• CPC Class Codes: A61Q 19/08 Anti-ageing preparations
• CPC Class Codes: G01N 33/58 involving labelled substances G01N33/53 takes precedence
• CPC Class Codes: G01N 33/92 involving lipids, e.g. cholesterol , lipoproteins, or their receptors steroid hormones G01N33/743

Abstract

Provided are methods for preparing improved biologically active micelle and crystalline products comprising a biologically active amphipathic compound in association with a micelle or crystalline product. Methods for producing the micelle or crystalline products as well as methods of using the micelle or crystalline products in therapeutic, diagnostic, and cosmetic, applications are also provided.

Claims

1. A method of preparing a biologically active micelle product comprising one or more biologically active amphipathic compounds in association with a micelle;

said method comprising the steps of;

a) mixing one or more lipids wherein at least one lipid component is covalently bonded to a water-soluble polymer;

b) forming sterically stabilized micelles from lipids;

c) incubating micelles from step (b) with one or more biologically active amphipathic compound(s) under conditions in which said compound(s) becomes associated with said micelles in a more biologically active conformation.

2. The method according to claim 1 wherein mixing in step (a) is carried out in an organic solvent, and forming sterically stabilized micelles in step (b) is carried out in steps comprising (i) removing the organic solvent to leave a dry film, and (ii) hydrating the dry film with an aqueous solution.

3. The method of claim 2 wherein the organic solvent is removed by evaporation or lyophilization.

4. The method according to claim 1 wherein mixing in step (a) is carried out in an aqueous solution.

10. The method according to any one of claims 1, 5, or 7 wherein said water soluble polymer is polyethylene glycol (PEG).

11. The method according to any one of claims 1, 5 or 7 wherein the amphipathic compound is characterized by having one or more α

- or π

-helical domains in its biologically active conformation.

12. The method according to claim 11 wherein the compound is a member of the vasoactive intestinal peptide (VIP)/growth hormone releasing factor (GRF) family of peptides.

13. The method according to claim 11 wherein the peptide is selected from the group consisting of VIP, galanin, endothelin, angiotensin II, calmodulin, and fragments, analogs, and modulators thereof.

14. The method according to any one of claims 1, 5, or 7 wherein the micelles have an average diameter of less than about 25 nm.

15. The method of any one of claims 1, 5, or 7 wherein the combination of lipids consists of distearoyl-phosphatidylethanolamine covalently bonded to PEG (PEG-DSPE).

16. A biologically active micelle product produced by the method of any of one claims 1, 5 or 7.

17. A composition comprising the biologically active micelle product of claim 16 wherein said biologically active amphipathic peptide, protein, fragment, analog, or modulators thereof has an activity selected from the group consisting of anti-oxidant activity, anti-pain, anti-inflammatory, wound healing activity, anti-microbial, antibronchospasm, metabolic activity, anti-cancer activity, cardiovascular activity, antiglaucoma activity, anti-apoptosis, anti-wrinkling activity, cryopreservation, and anti-aging activity.

18. The composition according to claim 17 wherein the composition is a cosmetic.

19. The composition according to claim 17 wherein the composition is a therapeutic.

20. A diagnostic composition comprising the micelle composition according to claim 17 and further comprising a detectable label.

21. The diagnostic composition according to claim 20 wherein the label is selected from the group consisting of a fluorescent label, a radioactive label, a dye, a gas, and a compound which enhances radiographic, magnetic resonance, and ultrasound imaging.

22. A diagnostic method comprising the steps of:

a) preparing a diagnostic composition according to claim 20;

b) administering a diagnostically effective amount of said composition to a target tissue or organ; and

c) detecting uptake of the composition at the target tissue or organ by detecting the presence of the label at the target tissue or organ.

23. The method according to claim 22 wherein the label is selected for the group consisting of a fluorescent label, a radioactive label, a dye, a gas, and a compound which enhances radiographic, magnetic resonance, and ultrasound imaging.

24. An oral controlled release preparation for the treatment of a gastrointestinal disorder produced according to the method of claim 12 wherein said method further comprises the step of encapsulating the biologically active micelle product.

25. The oral controlled release preparation of claim 24 wherein the gastrointestinal disorder is selected from the group consisting of inflammatory bowel disease, acute food impaction, chronic constipation, Hirschprung'"'"'s disease, achalasia, infantile hypertrophic pyloric stenosis, and ulcers.

26. A method of administering a biologically active amphipathic compound to a target tissue or organ comprising the steps of:

a) a biologically active micelle product comprising a biologically active amphipathic compound in association with a micelle according to the method of any one of claims 1, 5 or 7; and

b) administering a therapeutically effective amount of said micelle product to said target tissue or organ.

27. The method according to claim 26 wherein the amphipathic compound in a biologically active conformation is characterized by having one or more α

- or π

-helical domains.

28. The method of claim 27 wherein said peptide is a member of the vasoactive intestinal peptide (VIP)/growth hormone releasing factor (GRF) family of peptides.

29. The method of claim 27 wherein said peptide is selected from the group consisting of VIP, galanin, endothelin, and angiotensin II and fragments, analogs, and modulators thereof.

30. A method for preserving a bodily organ, tissue or cell type for transplantation or fertilization comprising the step of incubating said organ, tissue, or cell type in the micelle composition produced according to claim 12.

5. A method of preparing a biologically active micelle product comprising one or more biologically active amphipathic compounds in association with a micelle;

said method comprising the steps of;

a) mixing one or more lipids with one or more biologically active amphipathic compounds, wherein at least one lipid component is covalently bonded to a water-soluble polymer;

b) forming sterically stabilized micelles from the mixture of step (a) under conditions in which said compound(s) becomes associated with said micelles in a more biologically active conformation.

6. The method according to claim 5 wherein mixing in step (a) is carried out in an organic solvent and at least one lipid is conjugated to one or more targeting compound(s), and forming micelles in step (b) is carried out in a process comprising the steps of:

(i) removing the organic solvent to leave a dry film, and (ii) hydrating the dry film with an aqueous solution, said method further comprising step of;

(c) incubating said micelle products under conditions wherein the targeting compound(s) associates with said micelle products in an active conformation.

7. A method for preparing a biologically active sterically stabilized crystalline product comprising one or more biologically active compounds which are insoluble in aqueous solution;

said method comprising the steps of;

a) mixing the biologically active compound(s) with one or more lipids, wherein at least one of the lipids is conjugated to a water soluble polymer; and

b) forming sterically stabilized crystalline products.

8. The method according to claim 7 wherein mixing in step (a) is carried out in an organic solvent, and forming crystalline products in step (b) is carried out in a process comprising the steps of (i) removing the organic solvent to leave a dry film;

and (ii) hydrating the dry film with an aqueous solution, said method further comprising the steps of (c) contacting said crystalline products with one or more targeting compounds; and

(d) incubating said crystalline products under conditions wherein the targeting compound(s) associates with said crystalline products.

9. The method according to claim 7 wherein forming in step(b) is carried out in the steps comprising (i) removing the organic solvent to leave a dry film and (ii) hydrating the dry film with an aqueous solution.

31. The method of claim 7, wherein the soluble compound is selected from the group consisting of progesterone, estrogen, prednisolone, prednisone, 2,3 mercaptopropanol, testosterone, betulinic acid, doxorubicin, amphotericin B, diazepam, nystatin, propofol, and Taxol®

.

32. A pharmaceutical composition comprising a crystalline product produced by the method of claim 7.

33. The composition according to claim 32 which is a therapeutic, diagnostic, or cosmetic.

34. A method of administering an insoluble compound to a target tissue comprising the steps of:

a) preparing a biologically active micelle product comprising a biologically active amphipathic compound in association with a micelle prepared according to the method of claim 8; and

b) administering a therapeutically effective amount of said micelle product to said target tissue.