NOVEL PEPTIDE AMPHIPHILES HAVING IMPROVED SOLUBILITY AND METHODS OF USING SAME

• US 20090042804A1
• Filed: 04/16/2008
• Published: 02/12/2009
• Est. Priority Date: 04/17/2007
• Status: Active Grant

• Current Assignee: Nanotope, Inc. (Arrowhead Pharmaceuticals, Inc.)
• Original Assignee: Nanotope, Inc. (Arrowhead Pharmaceuticals, Inc.)
• Inventors: Guler, Mustafa O., HULVAT, JAMES F.
• Granted Patent: US 8,076,295 B2
• Time in Patent Office: Days
• US Class Current: 514/1.100
• 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: A61K 47/542 Carboxylic acids, e.g. a fatty acid or an amino acid
• CPC Class Codes: C07K 7/08 having 12 to 20 amino acids gastrins C07K14/595; somatostatins C07K14/655; melanotropins C07K14/68

Abstract

Disclosed herein are novel peptide amphiphile molecules and compositions discovered to possess improved solubility in aqueous buffers which, in turn, facilitates purification required for pharmaceutical applications, particularly for in vivo administration to human patients. In addition, gels of such peptide amphiphile compositions are shown herein to possess unexpectedly superior gelation kinetics and rheological properties, including an increased mechanical stiffness, which better mimics the mechanical properties of natural central nervous system tissues.

Claims

1. A molecule having one of the following structures:

C₁₆H₃₁O-Ser-Leu-Ser-Leu-Ala-Ala-Ala-Glu-Glu-Ile-Lys-Val-Ala-Val-OH (SEQ ID NO;

2) or C₁₆H₃₁O-Ser-Leu-Ser-Leu-Ala-Ala-Ala-Asp-Ile-Lys-Val-Ala-Val-OH (SEQ ID NO;

4).

2. A method of treating nerve damage in a subject in need thereof comprising administering to said subject a composition comprising the compound of claim 1.

3. The method of claim 2, wherein the damaged nerve is a nerve in the spinal cord of said subject.

4. The method of claim 2, wherein the damaged nerve comprises a sensory neuron.

5. The method of claim 2, wherein the damage nerve comprises a motor neuron.

6. The method of claim 2, wherein said composition is administered intrathecally.

7. The method of claim 6, wherein said composition is an aqueous solution comprising said compound.

8. The method of claim 7, wherein said compound forms a nanofiber gel in the subject.

9. The method of claim 8, wherein the nanofiber gels forms upon contact with the damaged nerve.

10. The method of claim 2, wherein said composition further comprises one or more other agents selected from the group consisting of a neurotrophic factor, an inhibitor of a neuronal growth inhibitor, a neuronal growth attractant and a neuronal growth inhibitor.

20. A composition comprising one or more peptide amphiphile molecules of claim 1 self-assembled to form one or more fibrillar structures.

21. The composition of claim 20, wherein said fibrillar structures are cylindrical micelles.

22. A substrate having the composition of claim 20 coated thereon.

23. A biocompatible, biodegradable gel comprising the peptide amphiphile molecules of claim 1, said gel serving as a scaffold for tissue growth.

24. A biocompatible, biodegradable gel comprising the fibrillar structures of claim 20, said gel serving as a scaffold for tissue growth.

25. A matrix or scaffold comprising the composition of claim 20.

26. A pharmaceutical composition comprising one or more peptide amphiphile molecules of claim 1 in conjunction with a pharmaceutically acceptable carrier.

32. A method of promoting development of a neural progenitor cell comprising contacting said neural progenitor cell with a composition comprising the peptide amphiphile of claim 1.

33. The method of claim 32, wherein said development of a neuron comprises axonal growth.

34. The method of claim 33, wherein said axonal growth comprises descending motor fiber growth.

35. The method of claim 33, wherein said axonal growth comprises ascending sensory fiber growth.

36. The method of claim 32, wherein said composition further comprises a neurotrophic factor.

37. The method of claim 32, wherein said neural progenitor cell is accompanied by reduced astrogliosis in said subject.

38. The method of claim 32, wherein said contacting comprises intrathecal administration of an aqueous solution comprising said peptide amphiphile.

39. The method of claim 32, wherein said progenitor neural cell is a stem cell.

40. The method of claim 32, wherein said progenitor neural cell is a neurite or undeveloped neuron.

41. The method of claim 32, wherein said progenitor neural cell is immortalized.

42. The method of claim 32, wherein said altered development includes the growth of the progenitor neural cell.

43. The method of claim 32, wherein said altered development includes the differentiation and growth of the progenitor neural cell.

44. The method of claim 32, further comprising the inhibition of differentiation of the progenitor neural cells into astroglial cells.

11. A peptide amphiphile that is soluble in aqueous media and affords a gel under physiological conditions, said peptide amphiphile comprising:

(a) a lipophilic segment selected from the group consisting of a single, linear alkyl chain of the formula C_nH_2n-1O, wherein n=6−

22, linked to the N-terminus of the peptide via a peptidyl bond;

(b) a structural peptide segment intermediate to the molecule, comprising 3-8 amino acid residues with non-polar side chains, having a propensity for predominantly beta-sheet secondary structure formation; and

(c) a C-terminal, functional peptide segment selected from any one of SEQ ID NOs;

22-26, 31 and 32 or having a formula comprising (Xaa)_m-Ile-Lys-Val-Ala-Val-(Xbb)_p (SEQ ID NO;

27), or (Xaa)_m-Val-Ala-Val-Lys-Ile-(Xbb)_p (SEQ ID NO;

28), where m=0 to 5, p=0 to 3, Xaa is selected from one or more amino acid residues with acidic side chains, and Xbb is selected from any amino acid.

12. The peptide amphiphile molecule of claim 11, wherein Xaa is an amino acid residue selected the group consisting of aminomalonic acid (Ama), aspartic acid (Asp), glutamic acid (Glu), aminoadipic acid (Aib), aminoheptanedioic acid (Apm) or gammacarboxyglutamic acid (Gla).

13. The peptide amphiphile molecule of claim 11, wherein Xaa is an amino acid residue selected the group consisting of glutamic acid (Glu) and aspartic acid (Asp).

14. The peptide amphiphile molecule of claim 11, wherein n=16 and thus the lipophilic segment comprises palmitic acid.

15. The peptide amphiphile molecule of claim 11, wherein the structural peptide segment is selected from the group consisting of SEQ ID NOs:

8-21.

16. The peptide amphiphile molecule of claim 11, wherein Xaa is glutamic acid, m is 2 and p is 0.

17. The peptide amphiphile molecule of claim 11, wherein Xaa is aspartic acid, m is 1 and p is 0.

18. The peptide amphiphile molecule of claim 16, wherein the functional peptide segment is terminated in a free acid and comprises the sequence Glu-Glu-Ile-Lys-Val-Ala-Val-OH (SEQ ID NO:

29).

19. The peptide amphiphile molecule of claim 17, wherein the functional peptide segment is terminated in a free acid and comprises the sequence Asp-Ile-Lys-Val-Ala-Val-OH (SEQ ID NO:

30).

27. A method of treating a human patient suffering nerve damage comprising the step of administering a composition comprising the peptide amphiphile molecule of claim 11 to the subject under conditions that stimulate neuron regeneration.

28. The method of claim 27, wherein the damaged nerve has been damaged by traumatic spinal cord injury.

29. The method of claim 28, wherein said peptide amphiphile molecule has the

following structure:

C₁₆H₃₁O-Ser-Leu-Ser-Leu-Ala-Ala-Ala-Glu-Glu-Ile-Lys-Val-Ala-Val-OH (SEQ ID NO;

2).

30. The method of claim 28, wherein said peptide amphiphile molecule has the

following structure:

C₁₆H₃₁O-Ser-Leu-Ser-Leu-Ala-Ala-Ala-Asp-Ile-Lys-Val-Ala-Val-OH (SEQ ID NO;

4).

31. The method of claim 27, wherein the peptide amphiphile forms a gel upon contact with cerebrospinal fluid.

45. A method of making a peptide amphiphile, as described in claim 11, wherein solid phase peptide synthesis is performed using a polymeric resin support that is pre-loaded with a protected amino acid at a loading fraction of 0.1-0.4 mmole/g, said loading fraction selected to improve synthetic yield of the peptide.

46. A method of making a peptide amphiphile, as described in claim 11, wherein a serine amino acid residue is incorporated into the peptide sequence in the form of a pseudoproline (oxazolidine) dipeptide, said method selected to improve the synthetic yield of the peptide.