Peptide amphiphiles having improved solubility and methods of using same

• US 8,076,295 B2
• Filed: 04/16/2008
• Issued: 12/13/2011
• Est. Priority Date: 04/17/2007
• Status: Expired due to Fees

• Current Assignee: Nanotope, Inc. (Arrowhead Pharmaceuticals, Inc.)
• Original Assignee: Nanotope, Inc. (Arrowhead Pharmaceuticals, Inc.)
• Inventors: Hulvat, James F., Guler, Mustafa O.
• Primary Examiner(s): Russel, Jeffrey E
• Application Number:
  US12/104,407
• Publication Number:
  US 20090042804A1
• Time in Patent Office: 1,336 Days
• Field of Search: None
• US Class Current: 514/17.7
• 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:

C₁₅H₃₁C(O)-Ser-Leu-Ser-Leu-Ala-Ala-Ala-Glu-Glu-Ile-Lys-Val-Ala-Val-OH (SEQ ID NO;

2) or C₁₅H₃₁C(O)-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 molecule 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 damaged 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 molecule.

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

9. The method of claim 8, wherein the nanofiber gel 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.

18. A composition comprising one or more molecules of claim 1 or peptide amphiphile molecules of claim 11 self-assembled to form one or more fibrillar structures.

19. The composition of claim 18, wherein said fibrillar structures are cylindrical micelles.

20. A substrate having the composition of claim 18 coated thereon.

21. A biocompatible, biodegradable gel comprising the molecules of claim 1 or peptide amphiphile molecules of claim 11, said gel serving as a scaffold for tissue growth.

22. A biocompatible, biodegradable gel comprising the fibrillar structures of claim 18, said gel serving as a scaffold for tissue growth.

23. A matrix or scaffold comprising the composition of claim 18.

24. A pharmaceutical composition comprising one or more claim 1 or peptide amphiphile molecules of claim 11 in conjunction with a pharmaceutically acceptable carrier.

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

(a) a lipophilic segment selected from the group consisting of a single, linear moiety of the formula C<sub>n−

1</sub>H<sub>2n−

1</sub>C(O)—

, wherein n=6-22, linked to the N-terminus of the peptide via a peptidyl bond;

(b) a structural peptide segment intermediate to the peptide amphiphile 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;

24-26, and (Xaa)_m-Val-Ala-Val-Lys-Ile-(Xbb)_p (SEQ ID NO;

28), where m=1 to 5, p=0 to 3, Xaa is selected from the group consisting of aminomalonic acid (Ama), aspartic acid (Asp), aminoadipic acid (Aib), aminoheptanedioic acid (Apm) and gammacarboxyglutamic acid (Gla), and Xbb is selected from any amino acid.

12. The peptide amphiphile molecule of claim 11, wherein the lipophilic segment comprises palmitic acid.

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

8-21.

14. The peptide amphiphile molecule of claim 11, wherein the C-terminal functional peptide segment is SEQ ID NO:

24.

15. The peptide amphiphile molecule of claim 11, wherein the C-terminal functional peptide segment is SEQ ID NO:

25.

16. The peptide amphiphile molecule of claim 11, wherein the C-terminal functional peptide segment is SEQ ID NO:

26.

17. The peptide amphiphile molecule of claim 11, wherein the C-terminal functional peptide segment is (Xaa)_m-Val-Ala-Val-Lys-Ile-(Xbb)_p (SEQ ID NO:

28), where m=1 to 5, p=0 to 3, Xaa is selected from the group consisting of aminomalonic acid (Ama), aspartic acid (Asp), aminoadipic acid (Aib), aminoheptanedioic acid (Apm) and gammacarboxyglutamic acid (Gla), and Xbb is selected from any amino acid.

27. A method of making a peptide amphiphile molecule, 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 for use in the peptide amphiphile molecule.

28. A method of making a peptide amphiphile molecule, 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 for use in the peptide amphiphile molecule.

25. A method of treating a human subject suffering nerve damage comprising the step of administering a composition comprising the peptide amphiphile molecule having the following structure:

C₁₅H₃₁C(O)-Ser-Leu-Ser-Leu-Ala-Ala-Ala-Glu-Glu-Ile-Lys-Val-Ala-Val-OH (SEQ ID NO;

2) under conditions that stimulate neuron regeneration.

26. A method of treating a human subject suffering nerve damage comprising the step of administering a composition comprising the peptide amphiphile molecule having the following structure:

C₁₅H₃₁C(O)-Ser-Leu-Ser-Leu-Ala-Ala-Ala-Asp-Ile-Lys-Val-Ala-Val-OH (SEQ ID NO;

4) under conditions that stimulate neuron regeneration.