Metal-binding compounds and uses therefor
View Patent ↗The invention provides a method of reducing the damage done by reactive oxygen species (ROS) in an animal. The invention also provides a method of reducing the concentration of a metal in an animal. These methods comprise administering to the animal an effective amount of a metal-binding compound as further described in the application. The invention further provides a method of reducing the damage done by ROS to a cell, a tissue or an organ that has been removed from an animal. This method comprising contacting the cell, tissue or organ with a solution or medium containing an effective amount of a metal-binding compound of the invention. The invention further provides novel metal-binding compounds, pharmaceutical compositions comprising the metal-binding compounds, and kits comprising a container holding a metal-binding compound of the invention.
1. A method of treating ischemia in a mammal comprising administering to the animal an effective amount of a linear metal-binding peptide which does not have a metal ion bound to it, or a physiologically-acceptable salt of the peptide, the sequence of the peptide being:
P 1 -P 2 ,
wherein:
P 1 is:
Xaa 1 Xaa 2 His or
Xaa 1 Xaa 2 His Xaa 3 ;
Xaa 1 is glycine, alanine, valine, leucine, isoleucine, serine, threonine, aspartic acid, isoaspartic acid, asparagine, glutamic acid, isoglutamic acid, glutamine, lysine, hydroxylysine, histidine, arginine, ornithine, phenylalanine, tyrosine, tryptophan, cysteine, methionine, or α-hydroxymethylserine, and the only substituents on the α-amino group of Xaa 1 are hydrogen;
Xaa 2 is glycine, alanine, β-alanine, valine, leucine, isoleucine, serine, threonine, aspartic acid, asparagine, glutamic acid, glutamine, lysine, hydroxylysine, histidine, arginine, ornithine, phenylalanine, tyrosine, tryptophan, cysteine, methionine, or α-hydroxymethylserine; and
Xaa 3 is glycine, alanine, valine, leucine, isoleucine, lysine, histidine, arginine, ornithine, aspartic acid, glutamic acid, asparagine, glutamine or tryptophan;
P 2 is a peptide sequence which comprises a sequence of a Cu(I) binding site, and P 2 contains no more than 10 amino acids; and
optionally, the terminal —COOH of the peptide is substituted to produce —COR 2 , wherein R 2 is —NH 2 , —NHR 1 , —N(R 1 ) 2 , —OR 1 or —R 1 , wherein R 1 is an alkyl, aryl or heteroaryl.
2. The method of claim 1 wherein:
Xaa 1 is glycine, alanine, valine, leucine, isoleucine, serine, threonine, aspartic acid, glutamic acid, lysine, hydroxylysine, histidine, arginine or α-hydroxymethylserine,
Xaa 2 is glycine, alanine, valine, leucine, isoleucine, threonine, serine, asparagine, glutamine, methionine, lysine, hydroxylysine, histidine, arginine or α-hydroxymethylserine, and
Xaa 3 , when present, is alanine, aspartic acid, histidine or lysine.
3. The method of claim 2 wherein:
Xaa 1 is alanine, serine, threonine, lysine, histidine, aspartic acid or glutamic acid, and
Xaa 2 is glycine, alanine, valine, leucine, isoleucine, threonine, serine or histidine.
4. The method of claim 3 wherein Xaa 1 is aspartic acid or glutamic acid and Xaa 2 is glycine, alanine, valine, leucine or isoleucine.
5. The method of claim 4 wherein P 1 is Asp Ala His.
6. The method of claim 4 wherein P 1 is Asp Ala His Lys [SEQ ID NO:1].
7. The method of claim 3 wherein P 1 is Ser Gly His, Thr Leu His, His Ala His or Ala Ala His.
8. The method of claim 3 wherein P 1 is Lys His His Lys [SEQ ID NO:11], Asp His His Ala [SEQ ID NO:12] or Asp His His Asp [SEQ ID NO:15].
9. The method of claim 3 wherein P 1 is His Ala His Ala [SEQ ID NO:13], Ala His His Ala [SEQ ID NO:14] or Ala Ala His His [SEQ ID NO:16].
10. The method of any one of claims 1 - 9 wherein the sequence of the Cu(I) binding site of P 2 is one of the following sequences:
Met Xaa 4 Met,
Met Xaa 4 Xaa 4 Met,
Cys Cys,
Cys Xaa 4 Cys,
Cys Xaa 4 Xaa 4 Cys,
Met Xaa 4 Cys Xaa 4 Xaa 4 Cys,
Gly Met Xaa 4 Cys Xaa 4 Xaa 4 Cys,
[SEQ ID NO: 7]
Gly Met Thr Cys Xaa 4 Xaa 4 Cys,
[SEQ ID NO: 8]
Gly Met Thr Cys Ala Asn Cys,
[SEQ ID NO: 9]
or
γ-Glu Cys Gly,
wherein Xaa 4 is any amino acid.
11. The method of claim 10 wherein the sequence of the Cu(I) binding site of P 2 is one of the following sequences:
Gly Met Xaa 4 Cys Xaa 4 Xaa 4 Cys,
[SEQ ID NO: 7]
Gly Met Thr Cys Xaa 4 Xaa 4 Cys
[SEQ ID NO: 8]
or
Gly Met Thr Cys Ala Asn Cys.
[SEQ ID NO: 9]
12. The method of claim 11 wherein the sequence of the Cu(I) binding site of P 2 is Gly Met Thr Cys Ala Asn Cys [SEQ ID NO:9].
13. The method of claim 1 wherein at least one amino acid of P 1 other than β-alanine or glycine, when present, is a D-amino acid.
14. The method of claim 1 or 13 wherein at least one amino acid of P 2 other than β-alanine or glycine, when present, is a D-amino acid.
15. The method of claim 1 wherein the terminal —COOH of the peptide is substituted to produce —COR 2 , wherein R 2 is —NH 2 , —NHR 1 , —N(R 1 ) 2 , —-OR 1 or —R 1 , wherein R 1 is an alkyl, aryl or heteroaryl.
16. The method of claim 1 wherein the ischemia is located in the central nervous system.
17. The method of claim 16 wherein the ischemia is located in the brain.
18. The method of claim 17 wherein the ischemia is due to a stroke.
19. The method of claim 17 wherein the ischemia is due to surgery, an injury or a trauma.
20. The method of claim 17 wherein the ischemia is due to a transient ischemic attack, a seizure or hypoxia.
21. The method of claim 1 wherein the ischemia is located in the cardiovascular system.
22. The method of claim 20 wherein the ischemia is cardiac ischemia.
23. The method of claim 22 wherein the cardiac ischemia is due to acute myocardial infarction or angina pectoris.
24. The method of claim 22 wherein the cardiac ischemia is due to surgery.
25. The method of claim 24 wherein the surgery is angioplasty.
26. The method of claim 24 wherein the surgery is cardiac bypass surgery.
27. The method of claim 1 wherein the ischemia is located in the bowel.
28. The method of claim 1 wherein the ischemia is placental ischemia.
29. The method of any one of claims 16 - 28 wherein:
Xaa 1 is alanine, serine, threonine, lysine, histidine, aspartic acid or glutamic acid;
Xaa 2 is glycine, alanine, valine, leucine, isoleucine, threonine, serine or histidine;
Xaa 3 , when present, is alanine, aspartic acid, histidine or lysine; and
the sequence of the Cu(I) binding site of P 2 is one of the following sequences:
Met Xaa 4 Met,
Met Xaa 4 Xaa 4 Met,
Cys Cys,
Cys Xaa 4 Cys,
Cys Xaa 4 Xaa 4 Cys,
Met Xaa 4 Cys Xaa 4 Xaa 4 Cys,
Gly Met Xaa 4 Cys Xaa 4 Xaa 4 Cys,
[SEQ ID NO: 7]
Gly Met Thr Cys Xaa 4 Xaa 4 Cys,
[SEQ ID NO: 8]
Gly Met Thr Cys Ala Asn Cys,
[SEQ ID NO: 9]
or
γ-Glu Cys Gly,
wherein Xaa 4 is any amino acid.
30. The method of claim 29 wherein:
P 1 is Asp Ala His Lys [SEQ ID NO:1]; and
the sequence of the Cu(I) binding site of P 2 is one of the following sequences:
Gly Met Xaa 4 Cys Xaa 4 Xaa 4 Cys,
[SEQ ID NO: 7]
Gly Met Thr Cys Xaa 4 Xaa 4 Cys
[SEQ ID NO: 8]
or
Gly Met Thr Cys Ala Asn Cys.
[SEQ ID NO: 9]
31. The method of claim 30 wherein the sequence of the Cu(I) binding site of P 2 is Gly Met Thr Cys Ala Asn Cys [SEQ ID NO:9].