Methods for preparing purified polypeptide compositions

• US 9,394,336 B2
• Filed: 01/13/2012
• Issued: 07/19/2016
• Est. Priority Date: 09/22/2008
• Status: Active Grant

• Current Assignee: Aileron Therapeutics Inc.
• Original Assignee: Aileron Therapeutics Inc.
• Inventors: Nash, Huw M., Iadanza, Matthew, Leitheiser, Christopher, Kawahata, Noriyuki
• Primary Examiner(s): Skowronek, Karlheinz R
• Assistant Examiner(s): Lee, Li
• Application Number:
  US13/350,644
• Publication Number:
  US 20120264674A1
• Time in Patent Office: 1,649 Days
• Field of Search: None
• US Class Current: 1/1
• 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: A61P 1/00 Drugs for disorders of the alimentary tract or the digestive system
• CPC Class Codes: A61P 1/04 for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
• CPC Class Codes: A61P 11/00 Drugs for disorders of the respiratory system
• CPC Class Codes: A61P 11/06 Antiasthmatics
• CPC Class Codes: A61P 13/12 of the kidneys
• CPC Class Codes: A61P 17/00 Drugs for dermatological disorders
• CPC Class Codes: A61P 17/02 for treating wounds, ulcers, burns, scars, keloids, or the like
• CPC Class Codes: A61P 19/00 Drugs for skeletal disorders
• CPC Class Codes: A61P 19/02 for joint disorders, e.g. arthritis, arthrosis
• CPC Class Codes: A61P 21/02 Muscle relaxants, e.g. for tetanus or cramps
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: A61P 25/14 for treating abnormal movements, e.g. chorea, dyskinesia
• CPC Class Codes: A61P 25/16 Anti-Parkinson drugs
• CPC Class Codes: A61P 25/28 for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
• CPC Class Codes: A61P 27/02 Ophthalmic agents
• CPC Class Codes: A61P 3/00 Drugs for disorders of the metabolism of the blood or the extracellular fluid A61P7/00
• CPC Class Codes: A61P 3/06 Antihyperlipidemics
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 31/12 Antivirals
• CPC Class Codes: A61P 35/00 : Antineoplastic agents
• CPC Class Codes: A61P 35/02 : specific for leukemia
• CPC Class Codes: A61P 35/04 : specific for metastasis
• CPC Class Codes: A61P 37/06 : Immunosuppressants, e.g. drugs for graft rejection
• CPC Class Codes: A61P 43/00 : Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 5/00 : Drugs for disorders of the endocrine system
• CPC Class Codes: A61P 7/00 : Drugs for disorders of the blood or the extracellular fluid
• CPC Class Codes: A61P 7/02 : Antithrombotic agents; Anticoagulants; Platelet aggregation inhibitors
• CPC Class Codes: A61P 7/06 : Antianaemics
• CPC Class Codes: A61P 9/00 : Drugs for disorders of the cardiovascular system
• CPC Class Codes: A61P 9/04 : Inotropic agents, i.e. stimulants of cardiac contraction; Drugs for heart failure
• CPC Class Codes: A61P 9/10 : for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
• CPC Class Codes: A61P 9/12 : Antihypertensives
• CPC Class Codes: C07K 1/107 : by chemical modification of precursor peptides
• CPC Class Codes: C07K 14/00 : Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
• CPC Class Codes: C07K 9/00 : Peptides having up to 20 amino acids, containing saccharide radicals and having a fully defined sequence; Derivatives thereof

Abstract

The present invention relates to purified peptidomimetic macrocycles. The invention additionally provides methods of preparing and using such macrocycles, for example in therapeutic applications.

Claims

1. A pharmaceutically acceptable composition comprising:

a peptidomimetic macrocycle or a pharmaceutically acceptable salt thereof comprising an alpha-helix and a cross-linker connecting a first amino acid and a second amino acid, the cross-linker spanning from 1 turn to 5 turns of the alpha-helix; and

a metal residue, from a metathesis catalyst, at a concentration of from about 0.5 ppm to about 10 ppm by weight as measured by nuclear inductively coupled plasma analysis (ICP) or nuclear inductively coupled plasma mass spectrometry analysis (ICP-ms);

wherein the pharmaceutically acceptable composition is suitable for administration to a human subject,

and wherein the peptidomimetic macrocycle or the pharmaceutically acceptable salt is of Formula (I);

2. The pharmaceutically acceptable composition of claim 1, wherein the metal residue comprises ruthenium or osmium.

3. The pharmaceutically acceptable composition according to claim 1, wherein at least one of the first and second amino acids is an α

,α

-disubstituted amino acid.

4. The pharmaceutically acceptable composition of claim 3, wherein the first or second amino acid is α

,α

-disubstituted.

5. The pharmaceutically acceptable composition of claim 1, wherein the first amino acid and the second amino acid are separated by three amino acids.

6. The pharmaceutically acceptable composition of claim 5, wherein the cross-linker comprises from 6 to 14 consecutive bonds.

7. The pharmaceutically acceptable composition of claim 6, wherein the cross-linker comprises from 8 to 12 consecutive bonds.

8. The pharmaceutically acceptable composition of claim 1, wherein the first amino acid and the second amino acid are separated by six amino acids.

9. The pharmaceutically acceptable composition of claim 8, wherein the cross-linker comprises from 8 to 16 consecutive bonds.

10. The pharmaceutically acceptable composition of claim 9, wherein the cross-linker comprises from 10 to 13 consecutive bonds.

11. The pharmaceutically acceptable composition of claim 1, wherein the cross-linker spans 1 or 2 turns of the alpha helix.

12. The pharmaceutically acceptable composition of claim 1, wherein the length of the cross-linker is about 5 Å

to about 9 Å

per turn of the alpha-helix.

13. The pharmaceutically acceptable composition of claim 1, wherein the peptidomimetic macrocycle or the pharmaceutically acceptable salt thereof carries a net positive charge at pH 7.4.

14. The pharmaceutically acceptable composition of claim 1, wherein the concentration of the metal residue is about 0.5 ppm by weight.

15. The pharmaceutically acceptable composition of claim 1, wherein the concentration of the metal residue is about 1 ppm by weight.

16. The pharmaceutically acceptable composition of claim 1, wherein the concentration of the metal residue is about 1 to about 5 ppm by weight.

17. The pharmaceutically acceptable composition of claim 1, wherein the concentration of the metal residue is about 1 to about 10 ppm by weight.

18. The pharmaceutically acceptable composition of claim 1, wherein the first amino acid and the second amino acid are separated by two amino acids.

19. The pharmaceutically acceptable composition of claim 1, wherein the concentration of the metal residue is about 0.5 ppm to about 5 ppm by weight.

20. The pharmaceutically acceptable composition of claim 1, wherein the concentration of the metal residue is about 0.5 ppm to about 1 ppm by weight.

37. The pharmaceutically acceptable composition of claim 2, wherein the metal residue comprises ruthenium.

38. The pharmaceutically acceptable composition of claim 1, wherein the peptidomimetic macrocycle is a therapeutic agent.

21. A pharmaceutically acceptable composition comprising:

a peptidomimetic macrocycle or a pharmaceutically acceptable salt thereof comprising an alpha-helix and a cross-linker connecting a first amino acid and a second amino acid, the cross-linker spanning from 1 turn to 5 turns of the alpha-helix; and

a metal residue comprising ruthenium, from a metathesis catalyst, at a concentration of below 10 ppm by weight as measured by nuclear inductively coupled plasma analysis (ICP) or nuclear inductively coupled plasma mass spectrometry analysis (ICP-ms);

wherein the pharmaceutically acceptable composition is suitable for administration to a human subject,

and wherein the peptidomimetic macrocycle or the pharmaceutically acceptable salt is of Formula (I);

22. The pharmaceutically acceptable composition according to claim 21, wherein at least one of the first and second amino acids is an α

,α

-disubstituted amino acid.

23. The pharmaceutically acceptable composition of claim 21, wherein the first amino acid and the second amino acid are separated by two amino acids.

24. The pharmaceutically acceptable composition of claim 21, wherein the first amino acid and the second amino acid are separated by three amino acids.

25. The pharmaceutically acceptable composition of claim 24, wherein the cross-linker comprises from 6 to 14 consecutive bonds.

26. The pharmaceutically acceptable composition of claim 25, wherein the cross-linker comprises from 8 to 12 consecutive bonds.

27. The pharmaceutically acceptable composition of claim 21, wherein the first amino acid and the second amino acid are separated by six amino acids.

28. The pharmaceutically acceptable composition of claim 27, wherein the cross-linker comprises from 8 to 16 consecutive bonds.

29. The pharmaceutically acceptable composition of claim 28, wherein the cross-linker comprises from 10 to 13 consecutive bonds.

30. The pharmaceutically acceptable composition of claim 21, wherein the cross-linker spans 1 or 2 turns of the alpha helix.

31. The pharmaceutically acceptable composition of claim 21, wherein the length of the cross-linker is about 5 Å

to about 9 Å

per turn of the alpha-helix.

32. The pharmaceutically acceptable composition of claim 21, wherein the peptidomimetic macrocycle or the pharmaceutically acceptable salt thereof carries a net positive charge at pH 7.4.

33. The pharmaceutically acceptable composition of claim 21, wherein the concentration of the metal residue is less than about 5 ppm by weight.

34. The pharmaceutically acceptable composition of claim 33, wherein the concentration of the metal residue is less than about 2.5 ppm by weight.

35. The pharmaceutically acceptable composition of claim 34, wherein the concentration of the metal residue is less than about 2 ppm by weight.

36. The pharmaceutically acceptable composition of claim 35, wherein the concentration of the metal residue is less than about 1 ppm by weight.

39. The pharmaceutically acceptable composition of claim 21, wherein the peptidomimetic macrocycle is a therapeutic agent.