Membrane-permeant peptide complexes for medical imaging, diagnostics, and pharmaceutical therapy

• US 6,589,503 B1
• Filed: 04/25/2000
• Issued: 07/08/2003
• Est. Priority Date: 06/20/1998
• Status: Expired due to Term

• Current Assignee: Washington University In St Louis
• Original Assignee: Washington University In St Louis
• Inventors: Piwnica-Worms, David
• Primary Examiner(s): JONES, DAMERON LEVEST
• Application Number:
  US09/557,465
• Time in Patent Office: 1,169 Days
• Field of Search: 530/300, 530/324-330, 530/331, 530/333, 530/334, 530/338, 530/350, 530/388.3, 530/388.35, 424/1.11, 424/1.65, 424/1.69, 424/9.1, 424/9.3, 424/9.4, 424/9.5, 424/9.36, 424/9.7, 424/9.8, 206/222, 206/560, 206/570
• US Class Current: 424/1.69
• CPC Class Codes: A61K 2123/00 Preparations for testing in vivo
• 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 47/65 Peptidic linkers, binders or spacers, e.g. peptidic enzyme-labile linkers
• CPC Class Codes: A61K 51/088 conjugates with carriers being peptides, polyamino acids or proteins antibodies A61K51/10
• CPC Class Codes: A61P 17/00 Drugs for dermatological disorders
• CPC Class Codes: A61P 19/02 for joint disorders, e.g. arthritis, arthrosis
• CPC Class Codes: A61P 21/04 for myasthenia gravis
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: A61P 25/02 for peripheral neuropathies
• 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 29/00 Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• CPC Class Codes: A61P 31/00 Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
• 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 37/00 Drugs for immunological or allergic disorders
• CPC Class Codes: A61P 37/04 Immunostimulants
• 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: C07K 14/005 : from viruses
• CPC Class Codes: C12N 2740/16322 : New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
• CPC Class Codes: C12N 9/6475 : Interleukin 1-beta convertase-like enzymes (3.4.22.10; 3.4.22.36; 3.4.22.63)

Abstract

Methods and compositions for medical imaging, evaluating intracellular processes and components, radiotherapy of intracellular targets, and drug delivery by the use of novel cell membrane-permeant peptide conjugate coordination and covalent complexes having target cell specificity are provided. Kits for conjugating radionuclides and other metals to peptide coordination complexes are also provided.

Claims

1. A compound, comprising:

a cell membrane-permeant peptide;

a diagnostic or pharmaceutically active substance; and

a functional linker moiety linking said peptide and said diagnostic or pharmaceutically active substance;

wherein said compound further comprises at least one D-amino acid and said functional linker moiety confers target cell specificity to said compound; and

wherein said functional linker moiety interacts with intracellular components;

or a pharmaceutically acceptable salt of said compound.

2. The compound of claim 1, wherein the majority of the amino acids comprising said compound are D-amino acids.

3. The compound of claim 2, wherein said cell membrane-permeant peptide is comprised of D-amino acids.

4. The compound of claim 1, wherein said cell membrane-permeant peptide is selected from the group consisting of SEQ ID NO:

1, SEQ ID NO;

2, SEQ ID NO;

3, SEQ ID NO;

4, SEQ ID NO;

5, and a peptide derivable from HIV-1 Tat protein.

5. The compound of claim 4, wherein said cell membrane-permeant peptide is a peptide derivable from HIV-1 Tat protein.

6. The compound of claim 5, wherein said peptide derivable from HIV-1 Tat protein comprises at least about four to about nine amino acids.

7. The compound of claim 5, wherein said peptide derivable from HIV-1 Tat protein comprises Tat amino acids 48-57, SEQ ID NO:

7.

8. The compound of claim 1, wherein said functional linker moiety is selected from the group consisting of a peptide, a protein, an oligonucleotide, a peptide nucleic acid, an oligosaccharide, and a hydrocarbon chain.

9. The compound of claim 1, wherein said functional linker moiety comprises a sequence selected from the group consisting of a peptide or protein binding motif, a protein kinase consensus sequence, a protein phosphatase consensus sequence, a protease-reactive sequence, an α

-, β

-, or γ

-secretase-reactive sequence, a peptidase-reactive sequence, a transferase-reactive sequence, a hydrolase-reactive sequence, an isomerase-reactive sequence, a ligase-reactive sequence, an HIV protease-reactive sequence, an extracellular metalloprotease-reactive sequence, a lysosomal protease-reactive sequence, a β

-lactamase-reactive sequence, an oxidoreductase-reactive sequence, an esterase-reactive sequence, a glycosidase-reactive sequence, and a nuclease-reactive sequence.

10. The compound of claim 9, wherein said sequence is a protease-reactive sequence.

11. The compound of claim 10, wherein said protease-reactive sequence is a caspase protease-reactive sequence.

12. The compound of claim 11, wherein said caspase protease-reactive sequence is cleaved by a caspase protease selected from the group consisting of caspase 1, caspase 2, caspase 3, caspase 4, caspase 5, caspase 6, caspase 7, caspase 8, caspase 9, caspase 10, caspase 1 1, caspase 12, and caspase 13.

13. The compound of claim 1, wherein said diagnostic substance is selected from the group consisting of a radionuclide, a relaxivity metal, a fluorochrome, a dye, and an enzyme substrate.

14. The compound of claim 13, wherein said radionuclide or relaxivity metal is coordinated to a chelation ligand linked to said functional linker moiety.

15. The compound of claim 14, wherein said chelation ligand is selected from the group consisting of modified peptide chelators, or DTPA, EDTA, and DOTA.

16. The compound of claim 13, wherein said radionuclide is a radioactive isotope of a metal selected from the group consisting Tc, Ru, In, Ga, Co, Pt, Fe, Os, Ir, W, Re, Cr, Mo, Mn, Ni, Rh, Pd, Nb, Cu, and Ta.

17. The compound of claim 13, wherein said relaxivity metal is a paramagnetic isotope of a metal selected from the group consisting of Mn, Cr, Fe, Gd, Eu, Dy, Ho, Cu, Co, Ni, Sm, Tb, Er, Tm, and Yb.

23. The compound of claim 1, wherein said cell membrane-permeant peptide comprises amino acids selected from the group consisting of proteogenic D-amino acids and non-proteogenic amino acids.

24. The compound of claim 23, wherein said amino acids comprise modified D-amino acids having substitutions wherein a first plurality of amino acids is substituted by a second plurality of amino acids having a hydropathic index within about 1 of the first plurality of amino acids.

25. The compound of claim 23, wherein said amino acids comprise modified D-amino acids having substitutions wherein a first plurality of amino acids is substituted by a second plurality of amino acids having a hydropathic index within about 0.5 of the first amino acid.

26. The compound of claim 5, wherein said peptide derived from HIV-1 Tat protein comprises SEQ ID NO:

7.

27. The compound of claim 9, wherein said sequence is selected from the group consisting of SEQ ID NO:

8, SEQ ID NO;

9, SEQ ID NO;

10, SEQ ID NO;

11, SEQ ID NO;

12, SEQ ID NO;

22, SEQ ID NO;

23, xR(R/K)x(S/T)x for Protein Kinase A, x(R/K)_2-3x(S/T)x for Protein Kinase G, x(R/K_1-3,x_0-2)(S/T)(x_0-2,R/K_1-3)x for Protein Kinase C, xRxx(S/T)x for Calmodulin Kinase 1I, SEQ ID NO;

24, SEQ ID NO;

25, SEQ ID NO;

26, SEQ ID NO;

27, SEQ ID NO;

28, and SEQ ID NO;

29 wherein x is any amino acid.

28. The compound of claim 13, wherein said radionuclide is a radioactive isotope of a metal selected from the group consisting of Tc-m99, In-111, Ga-67, Ga-68, Cu-64, Ru-97, Cr-51, Co-57, Re-188, and Re-186.

31. The compound of claim 4, wherein said membrane-permeant peptide comprises at least one D-amino acid.

32. The compound of claim 4, wherein the majority of the amino acids comprising said membrane-permeant peptide are D-amino acids.

33. The compound of claim 4, wherein said membrane-permeant peptide is comprised of D-amino acids.

34. The compound of claim 5, wherein said membrane-permeant peptide comprises at least one D-amino acid.

35. The compound of claim 5, wherein the majority of the amino acids comprising said membrane-permeant peptide are D-amino acids.

36. The compound of claim 5, wherein said membrane-permeant peptide is comprised of D-amino acids.

37. The compound of claim 6, wherein said membrane-permeant peptide comprises at least one D-amino acid.

38. The compound of claim 6, wherein the majority of the amino acids comprising said membrane-permeant peptide are D-amino acids.

39. The compound of claim 6, wherein said membrane-permeant peptide is comprised of D-amino acids.

40. The compound of claim 7, wherein said membrane-permeant peptide comprises at least one D-amino acid.

41. The compound of claim 7, wherein the majority of the amino acids comprising said membrane-permeant peptide are D-amino acids.

42. The compound of claim 7, wherein said membrane-permeant peptide is comprised of D-amino acids.

18. A composition, comprising a compound comprising:

a cell membrane-permeant peptide;

a diagnostic or pharmaceutically active substance; and

a functional linker moiety linking said peptide and said diagnostic or pharmaceutically active substance;

wherein said compound further comprises at least one D-amino acid and said functional linker moiety confers target cell specificity to said compound; and

wherein said functional linker moiety interacts with intracellular components.

19. The composition of claim 18, further comprising a pharmaceutically acceptable carrier, excipient, or diluent.

20. The composition of claim 18, wherein the majority of amino acids comprising said compound are D-amino acids.

21. The composition of claim 20, wherein said membrane permeant peptide comprises D-amino acids.

29. The composition of claim 18, wherein said composition comprises from about 1 μ

Ci to about 100 mCi of said compound when said compound is conjugated to a radionuclide.

30. The composition of claim 18, wherein said composition comprises about from about 1 mCi to about 50 mCi when said compound is conjugated to a radionuclide.

22. A kit, comprising a compound comprising:

a cell membrane-permeant peptide;

a metal chelation ligand; and

a functional linker moiety linking said peptide and said metal chelation ligand;

wherein said compound further comprises at least one D-amino acid and said functional linker moiety confers target cell specificity to said compound;

wherein said functional linker moiety interacts with intracellular components; and

a reducing agent capable of reducing a metal that can be coordinately incorporated into said metal chelation ligand.