AROMATIC-CATIONIC PEPTIDES AND USES OF SAME

Abstract

The disclosure provides compositions and methods relating to aromatic-cationic peptides. The methods comprise administering to the subject an effective amount of an aromatic-cationic peptide to subjects in need thereof. For example, the peptides may be administered to subjects in need of a mitochondrial-targeted antioxidant.

Claims

1. An aromatic-cationic peptide selected from the group consisting of: TABLE-US-00010 6-Butyric acid CoQ0-Phe-D-Arg-Phe-Lys-NH.sub.2 6-Decanoic acid CoQ0-Phe-D-Arg-Phe-Lys-NH.sub.2 Arg-Arg-Dmt-Phe Arg-Cha-Lys Arg-Dmt Arg-Dmt-Arg Arg-Dmt-Lys Arg-Dmt-Lys-Phe Arg-Dmt-Lys-Phe-Cys Arg-Dmt-Phe Arg-Dmt-Phe-Lys Arg-Lys-Dmt-Phe Arg-Lys-Phe-Dmt Arg-Phe-Dmt-Lys Arg-Phe-Lys Arg-Trp-Lys Arg-Tyr-Lys Arg-Tyr-Lys-Phe D-Arg-D-Dmt-D-Lys-D-Phe-NH.sub.2 D-Arg-D-Dmt-D-Lys-L-Phe-NH.sub.2 D-Arg-D-Dmt-L-Lys-D-Phe-NH.sub.2 D-Arg-D-Dmt-L-Lys-L-Phe-NH.sub.2 D-Arg-Dmt-D-Lys-NH.sub.2 D-Arg-Dmt-D-Lys-Phe-NH.sub.2 D-Arg-Dmt-Lys-D-Phe-NH.sub.2 D-Arg-Dmt-Lys-NH.sub.2 D-Arg-Dmt-Lys-Phe-Cys D-Arg-Dmt-NH.sub.2 D-Arg-L-Dmt-D-Lys-D-Phe-NH.sub.2 D-Arg-L-Dmt-D-Lys-L-Phe-NH.sub.2 D-Arg-L-Dmt-L-Lys-D-Phe-NH.sub.2 D-Arg-Phe-Lys-NH.sub.2 D-Arg-Trp-Lys-NH.sub.2 D-Arg-Tyr-Lys-NH.sub.2 Dmt-Arg Dmt-Lys Dmt-Lys-D-Phe-NH.sub.2 Dmt-Lys-NH.sub.2 Dmt-Lys-Phe Dmt-Lys-Phe Dmt-Lys-Phe-NH.sub.2 Dmt-Phe-Arg-Lys H-Arg-D-Dmt-Arg-NH.sub.2 H-Arg-D-Dmt-Lys-NH.sub.2 H-Arg-D-Dmt-Lys-Phe-NH.sub.2 H-Arg-D-Dmt-NH.sub.2 H-Arg-Dmt-Lys-Phe-NH.sub.2 H-D-Arg-2,6-dichloro-L-tyrosine-L-Lys-L-Phe-NH.sub.2 H-D-Arg-2,6-dichlorotyrosine-Lys-Phe-NH.sub.2 H-D-Arg-2,6-difluoro-L-tyrosine-L-Lys-L-Phe-NH.sub.2 H-D-Arg-2,6-difluorotyrosine-Lys-Phe-NH.sub.2 H-D-Arg-2,6-dimethyl-L-phenylalanine-L-Lys-L-Phe-NH.sub.2 H-D-Arg-2,6-dimethylphenylalanine-Lys-Phe-NH.sub.2 H-D-Arg-4-methoxy-2,6-dimethyl-L-tyrosine-L-Lys-L-Phe-NH.sub.2 H-D-Arg-4-methoxy-2,6-dimethyltyrosine-Lys-Phe-NH.sub.2 H-D-Arg-Arg-Dmt-Phe-NH.sub.2 H-D-Arg-Cha-Lys-NH.sub.2 H-D-Arg-D-Dmt-D-Lys-D-Phe-NH.sub.2 H-D-Arg-D-Dmt-Lys-Phe-NH.sub.2 H-D-Arg-D-Dmt-NH.sub.2 H-D-Arg-Dmt-D-Lys-D-Phe-NH.sub.2 H-D-Arg-Dmt-Lys-2,6-dimethylphenylalanine-NH.sub.2 H-D-Arg-Dmt-Lys-3-hydroxyphenylalanine-NH.sub.2 H-D-Arg-Dmt-Lys-NH.sub.2 H-D-Arg-Dmt-Lys-OH H-D-Arg-Dmt-Lys-Phe-OH H-D-Arg-Dmt-N6-acetyllysine-Phe-NH.sub.2 H-D-Arg-Dmt-OH H-D-Arg-Dmt-Phe-Lys-NH.sub.2 H-D-Arg-Dmt-Phe-NH.sub.2 H-D-Arg-D-Phe-L-Lys-L-Phe-NH.sub.2 H-D-Arg-D-Trp-L-Lys-L-Phe-NH.sub.2 H-D-Arg-D-Tyr-L-Lys-L-Phe-NH.sub.2 H-D-Arg-L-Dmt-L-Lys-2,6-dimethyl-L-phenylalanine-NH.sub.2 H-D-Arg-L-Dmt-L-Lys-3-hydroxy-L-phenylalanine-NH.sub.2 H-D-Arg-L-Dmt-L-Lys-D-Dmt-NH.sub.2 H-D-Arg-L-Dmt-L-Lys-D-Trp-NH.sub.2 H-D-Arg-L-Dmt-L-Lys-D-Tyr-NH.sub.2 H-D-Arg-L-Dmt-L-Lys-L-Dmt-NH.sub.2 H-D-Arg-L-Dmt-L-Lys-L-Dmt-NH.sub.2 H-D-Arg-L-Dmt-L-Lys-L-Trp-NH.sub.2 H-D-Arg-L-Dmt-L-Lys-L-Tyr-NH.sub.2 H-D-Arg-L-Dmt-L-Phe-L-Lys-NH.sub.2 H-D-Arg-L-Dmt-N6-acetyl-L-lysine-L-Phe-NH.sub.2 H-D-Arg-L-Lys-L-Dmt-L-Phe-NH.sub.2 H-D-Arg-L-Lys-L-Phe-L-Dmt-NH.sub.2 H-D-Arg-L-Phe-L-Dmt-L-Lys-NH.sub.2 H-D-Arg-L-Phe-L-Lys-L-Dmt-NH.sub.2 H-D-Arg-L-Phe-L-Lys-L-Phe-NH.sub.2 H-D-Arg-L-Trp-L-Lys-L-Phe-NH.sub.2 H-D-Arg-L-Tyr-L-Lys-L-Phe-NH.sub.2 H-D-Arg-Lys-Dmt-Phe-NH.sub.2 H-D-Arg-Lys-Phe-Dmt-NH.sub.2 H-D-Arg-Phe-Dmt-Lys-NH.sub.2 H-D-Arg-Phe-Lys-Dmt-NH.sub.2 H-D-Arg-Tyr-Lys-Phe-NH.sub.2 H-D-Dmt-Arg-NH.sub.2 H-D-His-L-Dmt-L-Lys-L-Phe-NH.sub.2 H-D-Lys-L-Dmt-L-Lys-L-Phe-NH.sub.2 H-Dmt-D-Arg-Lys-Phe-NH.sub.2 H-Dmt-D-Arg-NH.sub.2 H-Dmt-D-Arg-Phe-Lys-NH.sub.2 H-Dmt-D-Phe-Arg-Lys-NH.sub.2 H-Dmt-Lys-D-Arg-Phe-NH.sub.2 H-Dmt-Lys-Phe-D-Arg-NH.sub.2 H-Dmt-Phe-D-Arg-Lys-NH.sub.2 H-Dmt-Phe-Lys-D-Arg-NH.sub.2 H-D-N2-acetylarginine-Dmt-Lys-Phe-NH.sub.2 H-D-N8-acetylarginine-Dmt-Lys-Phe-NH.sub.2 H-D-Phe-D-Arg-D-Phe-D-Lys-NH.sub.2 H-L-Dmt-D-Arg-L-Lys-L-Phe-NH.sub.2 H-L-Dmt-D-Arg-L-Phe-L-Lys-NH.sub.2 H-L-Dmt-L-Lys-D-Arg-L-Phe-NH.sub.2 H-L-Dmt-L-Lys-L-Phe-D-Arg-NH.sub.2 H-L-Dmt-L-Phe-D-Arg-L-Lys-NH.sub.2 H-L-Dmt-L-Phe-L-Lys-D-Arg-NH.sub.2 H-L-His-L-Dmt-L-Lys-L-Phe-NH.sub.2 H-L-Lys-D-Arg-L-Dmt-L-Phe-NH.sub.2 H-L-Lys-D-Arg-L-Phe-L-Dmt-NH.sub.2 H-L-Lys-L-Dmt-D-Arg-L-Phe-NH.sub.2 H-L-Lys-L-Dmt-L-Lys-L-Phe-NH.sub.2 H-L-Lys-L-Dmt-L-Phe-D-Arg-NH.sub.2 H-L-Lys-L-Phe-D-Arg-L-Dmt-NH.sub.2 H-L-Lys-L-Phe-L-Dmt-D-Arg-NH.sub.2 H-L-Phe-D-Arg-L-Dmt-L-Lys-NH.sub.2 H-L-Phe-D-Arg-L-Lys-L-Dmt-NH.sub.2 H-L-Phe-L-Dmt-D-Arg-L-Lys-NH.sub.2 H-L-Phe-L-Dmt-L-Lys-D-Arg-NH.sub.2 H-L-Phe-L-Lys-D-Arg-L-Dmt-NH.sub.2 H-L-Phe-L-Lys-L-Dmt-D-Arg-NH.sub.2 H-Lys-D-Arg-Dmt-Phe-NH.sub.2 H-Lys-D-Arg-Phe-Dmt-NH.sub.2 H-Lys-Dmt-D-Arg-Phe-NH.sub.2 H-Lys-Dmt-Phe-D-Arg-NH.sub.2 H-Lys-D-Phe-Arg-Dmt-NH.sub.2 H-Lys-Phe-D-Arg-Dmt-NH.sub.2 H-Lys-Phe-Dmt-D-Arg-NH.sub.2 H-N2-acetyl-D-arginine-L-Dmt-L-Lys-L-Phe-NH.sub.2 H-N7-acetyl-D-arginine-Dmt-Lys-Phe-NH.sub.2 H-Phe(d5)-D-Arg-Phe(d5)-Lys-NH.sub.2 H-Phe-Arg-Phe-Lys-NH.sub.2 H-Phe-D-Arg-Dmt-Lys-NH.sub.2 H-Phe-D-Arg-Dmt-Lys-NH.sub.2 H-Phe-D-Arg-D-Phe-Lys-NH.sub.2 H-Phe-D-Arg-Lys-Dmt-NH.sub.2 H-Phe-D-Arg-Phe-D-Lys-NH.sub.2 H-Phe-D-Arg-Phe-Lys-Glu-Cys-Gly-NH.sub.2 H-Phe-D-Dmt-Arg-Lys-NH.sub.2 H-Phe-Dmt-D-Arg-Lys-NH.sub.2 H-Phe-Dmt-Lys-D-Arg-NH.sub.2 H-Phe-Lys-D-Arg-Dmt-NH.sub.2 H-Phe-Lys-Dmt-D-Arg-NH.sub.2 L-Arg-D-Dmt-D-Lys-D-Phe-NH.sub.2 L-Arg-D-Dmt-D-Lys-D-Phe-NH.sub.2 L-Arg-D-Dmt-D-Lys-L-Phe-NH.sub.2 L-Arg-D-Dmt-D-Lys-L-Phe-NH.sub.2 L-Arg-D-Dmt-L-Lys-D-Phe-NH.sub.2 L-Arg-D-Dmt-L-Lys-D-Phe-NH.sub.2 L-Arg-D-Dmt-L-Lys-L-Phe-NH.sub.2 L-Arg-D-Dmt-L-Lys-L-Phe-NH.sub.2 L-Arg-L-Dmt-D-Lys-D-Phe-NH.sub.2 L-Arg-L-Dmt-D-Lys-D-Phe-NH.sub.2 L-Arg-L-Dmt-D-Lys-L-Phe-NH.sub.2 L-Arg-L-Dmt-D-Lys-L-Phe-NH.sub.2 L-Arg-L-Dmt-L-Lys-D-Phe-NH.sub.2 L-Arg-L-Dmt-L-Lys-D-Phe-NH.sub.2 L-Arg-L-Dmt-L-Lys-L-Phe-NH.sub.2 L-Arg-L-Dmt-L-Lys-L-Phe-NH.sub.2 Lys-Dmt-Arf Lys-Dmt-D-Arg-NH.sub.2 Lys-Phe Lys-Phe-Arg-Dmt Lys-Phe-NH.sub.2 Lys-Trp-Arg Lys-Trp-D-Arg-NH.sub.2 Phe-Arg-Dmt-Lys Phe-Arg-Phe-Lys Phe-Arg-Phe-Lys Phe-Arg-Phe-Lys Phe-Arg-Phe-Lys Phe-Arg-Phe-Lys Phe-Arg-Phe-Lys-Glu-Cys-Gly Phe-Dmt-Arg-Lys Phe-Lys-Dmt Phe-Lys-Dmt-NH.sub.2 Succinic monoester CoQ0-Phe-D-Arg-Phe-Lys-NH.sub.2

2. A pharmaceutical composition comprising one or more aromatic-cationic peptides of claim 1 and pharmaceutically acceptable salts thereof.

3. The pharmaceutical composition of claim 2 further comprising a pharmaceutically acceptable carrier.

4. A method of reducing the number of mitochondria undergoing mitochondrial permeability transition (MPT), or preventing mitochondrial permeability transitioning in a mammal in need thereof, the method comprising administering to the mammal an effective amount of one or more aromatic-cationic peptides of claim 1.

5. A method for reducing oxidative damage in a mammal in need thereof, the method comprising administering to the mammal an effective amount of one or more aromatic-cationic peptides of claim 1.

6. A method for increasing the ATP synthesis rate in a mammal in need thereof, the method comprising administering to the mammal an effective amount of one or more aromatic-cationic peptides of claim 1.

7. A method for determining the presence or amount of an administered aromatic-cationic peptide in a subject, the method comprising: detecting the administered aromatic-cationic peptide in a biological sample from the subject, wherein the aromatic-cationic peptide is selected from the group consisting of: TABLE-US-00011 6-Butyric acid CoQ0-Phe-D-Arg-Phe-Lys-NH.sub.2 6-Decanoic acid CoQ0-Phe-D-Arg-Phe-Lys-NH.sub.2 Arg-Arg-Dmt-Phe Arg-Cha-Lys Arg-Dmt Arg-Dmt-Arg Arg-Dmt-Lys Arg-Dmt-Lys-Phe Arg-Dmt-Lys-Phe-Cys Arg-Dmt-Phe Arg-Dmt-Phe-Lys Arg-Lys-Dmt-Phe Arg-Lys-Phe-Dmt Arg-Phe-Dmt-Lys Arg-Phe-Lys Arg-Trp-Lys Arg-Tyr-Lys Arg-Tyr-Lys-Phe D-Arg-D-Dmt-D-Lys-D-Phe-NH.sub.2 D-Arg-D-Dmt-D-Lys-L-Phe-NH.sub.2 D-Arg-D-Dmt-L-Lys-D-Phe-NH.sub.2 D-Arg-D-Dmt-L-Lys-L-Phe-NH.sub.2 D-Arg-Dmt-D-Lys-NH.sub.2 D-Arg-Dmt-D-Lys-Phe-NH.sub.2 D-Arg-Dmt-Lys-D-Phe-NH.sub.2 D-Arg-Dmt-Lys-NH.sub.2 D-Arg-Dmt-Lys-Phe-Cys D-Arg-Dmt-NH.sub.2 D-Arg-L-Dmt-D-Lys-D-Phe-NH.sub.2 D-Arg-L-Dmt-D-Lys-L-Phe-NH.sub.2 D-Arg-L-Dmt-L-Lys-D-Phe-NH.sub.2 D-Arg-Phe-Lys-NH.sub.2 D-Arg-Trp-Lys-NH.sub.2 D-Arg-Tyr-Lys-NH.sub.2 Dmt-Arg Dmt-Lys Dmt-Lys-D-Phe-NH.sub.2 Dmt-Lys-NH.sub.2 Dmt-Lys-Phe Dmt-Lys-Phe Dmt-Lys-Phe-NH.sub.2 Dmt-Phe-Arg-Lys H-Arg-D-Dmt-Arg-NH.sub.2 H-Arg-D-Dmt-Lys-NH.sub.2 H-Arg-D-Dmt-Lys-Phe-NH.sub.2 H-Arg-D-Dmt-NH.sub.2 H-Arg-Dmt-Lys-Phe-NH.sub.2 H-D-Arg-2,6-dichloro-L-tyrosine-L-Lys-L-Phe-NH.sub.2 H-D-Arg-2,6-dichlorotyrosine-Lys-Phe-NH.sub.2 H-D-Arg-2,6-difluoro-L-tyrosine-L-Lys-L-Phe-NH.sub.2 H-D-Arg-2,6-difluorotyrosine-Lys-Phe-NH.sub.2 H-D-Arg-2,6-dimethyl-L-phenylalanine-L-Lys-L-Phe-NH.sub.2 H-D-Arg-2,6-dimethylphenylalanine-Lys-Phe-NH.sub.2 H-D-Arg-4-methoxy-2,6-dimethyl-L-tyrosine-L-Lys-L-Phe-NH.sub.2 H-D-Arg-4-methoxy-2,6-dimethyltyrosine-Lys-Phe-NH.sub.2 H-D-Arg-Arg-Dmt-Phe-NH.sub.2 H-D-Arg-Cha-Lys-NH.sub.2 H-D-Arg-D-Dmt-D-Lys-D-Phe-NH.sub.2 H-D-Arg-D-Dmt-Lys-Phe-NH.sub.2 H-D-Arg-D-Dmt-NH.sub.2 H-D-Arg-Dmt-D-Lys-D-Phe-NH.sub.2 H-D-Arg-Dmt-Lys-2,6-dimethylphenylalanine-NH.sub.2 H-D-Arg-Dmt-Lys-3-hydroxyphenylalanine-NH.sub.2 H-D-Arg-Dmt-Lys-NH.sub.2 H-D-Arg-Dmt-Lys-OH H-D-Arg-Dmt-Lys-Phe-OH H-D-Arg-Dmt-N6-acetyllysine-Phe-NH.sub.2 H-D-Arg-Dmt-OH H-D-Arg-Dmt-Phe-Lys-NH.sub.2 H-D-Arg-Dmt-Phe-NH.sub.2 H-D-Arg-D-Phe-L-Lys-L-Phe-NH.sub.2 H-D-Arg-D-Trp-L-Lys-L-Phe-NH.sub.2 H-D-Arg-D-Tyr-L-Lys-L-Phe-NH.sub.2 H-D-Arg-L-Dmt-L-Lys-2,6-dimethyl-L-phenylalanine-NH.sub.2 H-D-Arg-L-Dmt-L-Lys-3-hydroxy-L-phenylalanine-NH.sub.2 H-D-Arg-L-Dmt-L-Lys-D-Dmt-NH.sub.2 H-D-Arg-L-Dmt-L-Lys-D-Trp-NH.sub.2 H-D-Arg-L-Dmt-L-Lys-D-Tyr-NH.sub.2 H-D-Arg-L-Dmt-L-Lys-L-Dmt-NH.sub.2 H-D-Arg-L-Dmt-L-Lys-L-Dmt-NH.sub.2 H-D-Arg-L-Dmt-L-Lys-L-Trp-NH.sub.2 H-D-Arg-L-Dmt-L-Lys-L-Tyr-NH.sub.2 H-D-Arg-L-Dmt-L-Phe-L-Lys-NH.sub.2 H-D-Arg-L-Dmt-N6-acetyl-L-lysine-L-Phe-NH.sub.2 H-D-Arg-L-Lys-L-Dmt-L-Phe-NH.sub.2 H-D-Arg-L-Lys-L-Phe-L-Dmt-NH.sub.2 H-D-Arg-L-Phe-L-Dmt-L-Lys-NH.sub.2 H-D-Arg-L-Phe-L-Lys-L-Dmt-NH.sub.2 H-D-Arg-L-Phe-L-Lys-L-Phe-NH.sub.2 H-D-Arg-L-Trp-L-Lys-L-Phe-NH.sub.2 H-D-Arg-L-Tyr-L-Lys-L-Phe-NH.sub.2 H-D-Arg-Lys-Dmt-Phe-NH.sub.2 H-D-Arg-Lys-Phe-Dmt-NH.sub.2 H-D-Arg-Phe-Dmt-Lys-NH.sub.2 H-D-Arg-Phe-Lys-Dmt-NH.sub.2 H-D-Arg-Tyr-Lys-Phe-NH.sub.2 H-D-Dmt-Arg-NH.sub.2 H-D-His-L-Dmt-L-Lys-L-Phe-NH.sub.2 H-D-Lys-L-Dmt-L-Lys-L-Phe-NH.sub.2 H-Dmt-D-Arg-Lys-Phe-NH.sub.2 H-Dmt-D-Arg-NH.sub.2 H-Dmt-D-Arg-Phe-Lys-NH.sub.2 H-Dmt-D-Phe-Arg-Lys-NH.sub.2 H-Dmt-Lys-D-Arg-Phe-NH.sub.2 H-Dmt-Lys-Phe-D-Arg-NH.sub.2 H-Dmt-Phe-D-Arg-Lys-NH.sub.2 H-Dmt-Phe-Lys-D-Arg-NH.sub.2 H-D-N2-acetylarginine-Dmt-Lys-Phe-NH.sub.2 H-D-N8-acetylarginine-Dmt-Lys-Phe-NH.sub.2 H-D-Phe-D-Arg-D-Phe-D-Lys-NH.sub.2 H-L-Dmt-D-Arg-L-Lys-L-Phe-NH.sub.2 H-L-Dmt-D-Arg-L-Phe-L-Lys-NH.sub.2 H-L-Dmt-L-Lys-D-Arg-L-Phe-NH.sub.2 H-L-Dmt-L-Lys-L-Phe-D-Arg-NH.sub.2 H-L-Dmt-L-Phe-D-Arg-L-Lys-NH.sub.2 H-L-Dmt-L-Phe-L-Lys-D-Arg-NH.sub.2 H-L-His-L-Dmt-L-Lys-L-Phe-NH.sub.2 H-L-Lys-D-Arg-L-Dmt-L-Phe-NH.sub.2 H-L-Lys-D-Arg-L-Phe-L-Dmt-NH.sub.2 H-L-Lys-L-Dmt-D-Arg-L-Phe-NH.sub.2 H-L-Lys-L-Dmt-L-Lys-L-Phe-NH.sub.2 H-L-Lys-L-Dmt-L-Phe-D-Arg-NH.sub.2 H-L-Lys-L-Phe-D-Arg-L-Dmt-NH.sub.2 H-L-Lys-L-Phe-L-Dmt-D-Arg-NH.sub.2 H-L-Phe-D-Arg-L-Dmt-L-Lys-NH.sub.2 H-L-Phe-D-Arg-L-Lys-L-Dmt-NH.sub.2 H-L-Phe-L-Dmt-D-Arg-L-Lys-NH.sub.2 H-L-Phe-L-Dmt-L-Lys-D-Arg-NH.sub.2 H-L-Phe-L-Lys-D-Arg-L-Dmt-NH.sub.2 H-L-Phe-L-Lys-L-Dmt-D-Arg-NH.sub.2 H-Lys-D-Arg-Dmt-Phe-NH.sub.2 H-Lys-D-Arg-Phe-Dmt-NH.sub.2 H-Lys-Dmt-D-Arg-Phe-NH.sub.2 H-Lys-Dmt-Phe-D-Arg-NH.sub.2 H-Lys-D-Phe-Arg-Dmt-NH.sub.2 H-Lys-Phe-D-Arg-Dmt-NH.sub.2 H-Lys-Phe-Dmt-D-Arg-NH.sub.2 H-N2-acetyl-D-arginine-L-Dmt-L-Lys-L-Phe-NH.sub.2 H-N7-acetyl-D-arginine-Dmt-Lys-Phe-NH.sub.2 H-Phe(d5)-D-Arg-Phe(d5)-Lys-NH.sub.2 H-Phe-Arg-Phe-Lys-NH.sub.2 H-Phe-D-Arg-Dmt-Lys-NH.sub.2 H-Phe-D-Arg-Dmt-Lys-NH.sub.2 H-Phe-D-Arg-D-Phe-Lys-NH.sub.2 H-Phe-D-Arg-Lys-Dmt-NH.sub.2 H-Phe-D-Arg-Phe-D-Lys-NH.sub.2 H-Phe-D-Arg-Phe-Lys-Glu-Cys-Gly-NH.sub.2 H-Phe-D-Dmt-Arg-Lys-NH.sub.2 H-Phe-Dmt-D-Arg-Lys-NH.sub.2 H-Phe-Dmt-Lys-D-Arg-NH.sub.2 H-Phe-Lys-D-Arg-Dmt-NH.sub.2 H-Phe-Lys-Dmt-D-Arg-NH.sub.2 L-Arg-D-Dmt-D-Lys-D-Phe-NH.sub.2 L-Arg-D-Dmt-D-Lys-D-Phe-NH.sub.2 L-Arg-D-Dmt-D-Lys-L-Phe-NH.sub.2 L-Arg-D-Dmt-D-Lys-L-Phe-NH.sub.2 L-Arg-D-Dmt-L-Lys-D-Phe-NH.sub.2 L-Arg-D-Dmt-L-Lys-D-Phe-NH.sub.2 L-Arg-D-Dmt-L-Lys-L-Phe-NH.sub.2 L-Arg-D-Dmt-L-Lys-L-Phe-NH.sub.2 L-Arg-L-Dmt-D-Lys-D-Phe-NH.sub.2 L-Arg-L-Dmt-D-Lys-D-Phe-NH.sub.2 L-Arg-L-Dmt-D-Lys-L-Phe-NH.sub.2 L-Arg-L-Dmt-D-Lys-L-Phe-NH.sub.2 L-Arg-L-Dmt-L-Lys-D-Phe-NH.sub.2 L-Arg-L-Dmt-L-Lys-D-Phe-NH.sub.2 L-Arg-L-Dmt-L-Lys-L-Phe-NH.sub.2 L-Arg-L-Dmt-L-Lys-L-Phe-NH.sub.2 Lys-Dmt-Arf Lys-Dmt-D-Arg-NH.sub.2 Lys-Phe Lys-Phe-Arg-Dmt Lys-Phe-NH.sub.2 Lys-Trp-Arg Lys-Trp-D-Arg-NH.sub.2 Phe-Arg-Dmt-Lys Phe-Arg-Phe-Lys Phe-Arg-Phe-Lys Phe-Arg-Phe-Lys Phe-Arg-Phe-Lys Phe-Arg-Phe-Lys Phe-Arg-Phe-Lys-Glu-Cys-Gly Phe-Dmt-Arg-Lys Phe-Lys-Dmt Phe-Lys-Dmt-NH.sub.2 Succinic monoester CoQ0-Phe-D-Arg-Phe-Lys-NH.sub.2

8. The method of claim 7, wherein detecting is performed during administration of the peptide.

9. The method of claim 7, wherein detecting is performed after administration of the peptide.

10. The method of any one of claim 7, wherein detecting comprises HPLC.

11. The method of claim 10, wherein the HPLC comprises reverse phase HPLC.

12. The method of claim 10, wherein the HPLC comprises ion exchange HPLC.

13. The method of claim 7, wherein detecting comprises mass spectrometry.

14. The method of claim 7, wherein the biological sample comprises a fluid.

15. The method of claim 7, wherein the biological sample comprises a cell.

16. The method of claim 7, wherein the biological sample comprises a tissue.

17. The method of any one of claims 7, wherein the biological sample comprises a biopsy.

18. An aromatic-cationic peptide selected from the group consisting of: a) an aromatic-cationic peptide comprising formula VII or a stereoisomer thereof ##STR00022## wherein the chiral centers of formula III are defined as H-(R)-Arg-(S)-DMT-(S)-Lys-(S)-Phe-NH.sub.2, and wherein stereoisomers are described by the formulas R-S-S-S, S-R-R-R, S-S-S-S, R-R-R-R, R-R-S-S, S-S-R-R, S-R-S-S, R-S-R-R, R-S-R-S, S-R-S-R, R-R-S-R, S-S-R-S, R-R-R-S, S-S-S-R, R-S-S-R, and S-R-R-S; b) an aromatic-cationic peptide comprising formula VII or a constitutional thereof ##STR00023## selected from the group consisting of Arg-Dmt-Lys-Phe-NH.sub.2, Phe-Dmt-Arg-Lys-NH.sub.2, Phe-Lys-Dmt-Arg-NH.sub.2, Dmt-Arg-Lys-Phe-NH.sub.2, Lys-Dmt-Arg-Phe-NH.sub.2, Phe-Dmt-Lys-Arg-NH, Arg-Lys-Dmt-Phe-NH, or Arg-Dmt-Phe-Lys-N.sub.2; c) an aromatic-cationic peptide comprising formula VIII ##STR00024## wherein R is selected from (i) OMe, and (ii) d) an aromatic-cationic peptide comprising formula IX ##STR00025## wherein R is selected from (i) F, (ii) Cl, and (iii) H; e) an aromatic-cationic peptide comprising formula X ##STR00026## wherein R1-R4 are selected from (i) Ac, (ii) H, (iii) H, (iv) H, (i) H, (ii) Ac, (iii) H, (iv) H, (i) H, (ii) H, (iii) Ac, (iv) H, and (i) H, (ii) H, (iii) H, (iv) OH; f) an aromatic-cationic peptide comprising formula XI ##STR00027##

19.-23. (canceled)

Description

EXAMPLES

[0235] The present invention is further illustrated by the following examples, which should not be construed as limiting in any way.

Example 1

Aromatic-Cationic Peptides of the Present Technology Inhibit Inhibits H.SUB.2.O.SUB.2 .Generation by Isolated Mitochondria

[0236] In this Example, the effects of the aromatic-cationic peptides of the invention on H.sub.2O.sub.2 generation by isolated mitochondria are investigated. H.sub.2O.sub.2 is measured using luminol chemiluminescence as described in Y. Li, H. Zhu, M. A. Trush, Biochim. Biophys. Acta 1428, 1-12 (1999)). Briefly, 0.1 mg mitochondrial protein is added to 0.5 ml potassium phosphate buffer (100 mM, pH 8.0) in the absence or presence of an aromatic-cationic peptide. Luminal (25 mM) and 0.7 IU horseradish peroxidase are added, and chemilumunescence is monitored with a Chronolog Model 560 aggregometer (Havertown, Pa.) for 20 min at 37 C. The amount of H.sub.2O.sub.2 produced is quantified as the area under the curve (AUC) over 20 min, and all data are normalized to AUC produced by mitochondria alone.

[0237] It is predicted that the aromatic-cationic peptide will reduce the spontaneous production of H.sub.2O.sub.2 by isolated mitochondria. As such, the aromatic-cationic peptides are useful for reducing oxidative damage and are useful in the treatment or prevention of diseases or conditions that relate to oxidative damage.

Example 2

Aromatic-Cationic Peptides of the Present Technology Reduce Intracellular ROS and Increase Cell Survival

[0238] To show that the claimed peptides are effective antioxidants when applied to whole cells, neuronal N.sub.2A cells are plated in 96-well plates at a density of 110.sup.4/well and allowed to grow for 2 days before treatment with tBHP (0.5 or 1 mM) for 40 min. Cells are washed twice and replaced with medium alone or medium containing varying concentrations of aromatic-cationic peptides (10.sup.12 M to 10.sup.9 M) for 4 h. Intracellular ROS is measured by carboxy-H.sub.2DCFDA (Molecular Probes, Portland, Oreg.). Cell death is assessed by a cell proliferation assay (MTS assay, Promega, Madison, Wis.).

[0239] Incubation with tBHP will result in an increase in intracellular ROS and decrease in cell viability. However, it is predicted that incubation of these cells with an aromatic-cationic peptide will reduce intracellular ROS and increase cell survival. As such, the aromatic-cationic peptides are useful for reducing oxidative damage and are useful in the treatment or prevention of diseases or conditions that relate to oxidative damage.

Example 3

Aromatic-cationic Peptides of the Present Technology Protect Against MPT Induced by Ca.SUP.2+ .and 3-Nitropropionic Acid

[0240] To isolate mitochondria from mouse liver, mice are sacrificed by decapitation. The liver is removed and rapidly placed into chilled liver homogenization medium. The liver is finely minced using scissors and then homogenized by hand using a glass homogenizer. The homogenate is centrifuged for 10 min at 1000 g at 4 C. The supernatant is aspirated and transferred to polycarbonate tubes and centrifuged again for 10 min at 3000 g, 4 C. The resulting supernatant is removed, and the fatty lipids on the side-wall of the tube are carefully wiped off. The pellet is resuspended in liver homogenate medium and the homogenization repeated twice. The final purified mitochondrial pellet is resuspended in medium. Protein concentration in the mitochondrial preparation is determined by the Bradford procedure.

[0241] To investigate the localization of the aromatic-cationic peptides of the invention, approximately 1.5 mg mitochondria in 400 l buffer is incubated with labeled aromatic-cationic peptide for 5-30 min at 37 C. The mitochondria are then centrifuged down and the amount of label is measured in the mitochondrial fraction and buffer fraction. Assuming a mitochondrial matrix volume of 0.7 l/mg protein (Lim et al., J Physiol 545:961-974, 2002), the concentration of peptide in mitochondria can be determined. It is predicted that the claimed aromatic-cationic peptides will be more concentrated in mitochondria compared to the buffer fraction.

[0242] To investigate the effects of the aromatic-cationic peptides of the invention on mitochondrial membrane potential, isolated mouse liver mitochondria are incubated with 100-200 M aromatic-cationic peptide. Mitochondrial membrane potential is measured using tetramethyl rhodamine methyl ester (TMRM). Addition of mitochondria results in immediate quenching of the TMRM signal which is readily reversed by the addition of FCCP, indicating mitochondrial depolarization. The addition of Ca.sup.2+ (150 M) results in immediate depolarization followed by progressive loss of quenching, indicative of MPT. Addition of aromatic-cationic peptide alone, even at 200 M, is not predicted to cause mitochondrial depolarization or MPT. It is also predicted that the aromatic-cationic peptides will not alter mitochondrial function, including oxygen consumption during state 3 or state 4, or the respiratory ratio (state 3/state 4).

[0243] To show that the claimed peptides are effective at protecting against MPT induced by Ca.sup.2+ overload, isolated mitochondria are pre-treated with aromatic-cationic peptide (10 M) for 2 min prior to addition of Ca.sup.2+. It is predicted that the aromatic-cationic peptides of the invention will increase the tolerance of mitochondria to cumulative Ca.sup.2+ challenges.

[0244] 3-Nitropropionic acid (3NP) is an irreversible inhibitor of succinate dehydrogenase in complex II of the electron transport chain. Addition of 3NP (1 mM) to isolated mitochondria causes dissipation of mitochondrial potential and onset of MPT. Pretreatment of mitochondria with the aromatic-cationic peptides of the invention is predicted to delay the onset of MPT induced by 3NP.

[0245] To demonstrate that the aromatic-cationic peptides of the invention can penetrate cell membranes and protect against mitochondrial depolarization elicited by 3NP, Caco-2 cells are treated with 3NP (10 mM) in the absence or presence of the aromatic-cationic peptides for 4 h, and then incubated with TMRM and examined under LSCM. In control cells, the mitochondria are clearly visualized as fine streaks throughout the cytoplasm. In cells treated with 3NP, the TMRM fluorescence is much reduced, indicating generalized depolarization. In contrast, it is predicted that concurrent treatment with the aromatic-cationic peptides of the invention will protect against mitochondrial depolarization caused by 3NP.

[0246] As such, the aromatic-cationic peptides are useful for preventing MPT and are useful in the treatment or prevention of diseases or conditions that relate to MPT.

Example 4

Aromatic-cationic Peptides of the Present Technology Protect Against Mitochondrial Swelling and Cytochrome c Release

[0247] MPT pore opening results in mitochondrial swelling. This Example examines the effects of the aromatic-cationic peptides of the invention on mitochondrial swelling by measuring reduction in absorbance at 540 nm (A.sub.540). Once the absorbance is measured, the mitochondrial suspension is then centrifuged and cytochrome c in the mitochondrial pellet and supernatant is determined by a commercially-available ELISA kit. It is predicted that pretreatment of isolated mitochondria with the aromatic-cationic peptides of the invention will inhibit swelling and cytochrome c release induced by Ca.sup.2 overload. Besides preventing MPT induced by Ca.sup.2+ overload, it is predicted that the aromatic-cationic peptides of the invention will also prevent mitochondrial swelling induced by MPP.sup.+(1-methyl-4-phenylpyridium ion), an inhibitor of complex I of the mitochondrial electron transport chain.

[0248] As such, the aromatic-cationic peptides are useful for preventing MPT and are useful in the treatment or prevention of diseases or conditions that relate to MPT.

Example 5

The Peptides of the Present Technology Increase the Rate ATP Synthesis in Isolated Mitochondria

[0249] This example will demonstrate the impact of peptides of the present technology on the rate of mitochondrial ATP synthesis.

[0250] The rate of mitochondrial ATP synthesis will be determined by measuring ATP in respiration buffer collected from isolated mitochondria 1 min after addition of 400 mM ADP. ATP will be assayed by HPLC. All experiments will be carried out in triplicate, with n=3. It is predicted that addition of peptides of the present technology to isolated mitochondria will increase the rate of ATP synthesis in a dose-dependent manner.

[0251] This result will demonstrate the peptides of the present technology are useful in methods and compositions for increasing the rate of mitochondrial ATP synthesis.

Example 6

Characterization of Aromatic-Cationic Peptides

[0252] Aromatic-cationic peptides of the present technology can be synthesized using solid phase synthesis and characterized using HPLC and MS. Exemplary HPLC and MS methods are provided in Examples 7 and 8 below.

Example 7

Detection of Aromatic-Cationic Peptides in a Biological Sample

[0253] This example demonstrates the detection of aromatic-cationic peptides in a biological sample by HPLC. Biological samples are collected from subjects in a suitable manner depending on the nature of the sample. Biological samples include any material derived from or contacted by living cells. Examples of biological samples include but are not limited to whole blood, fractionated blood, semen, saliva, tears, urine, fecal material, sweat, buccal, skin, cerebrospinal fluid, and hair. Biological samples also include biopsies of internal organs or cancers. Once obtained, the biological samples are stored in a manner compatible with the methods of detection until the methods are performed to ensure the preservation of aromatic-cationic peptides present in the sample.

[0254] Samples are loaded onto a 2504.6 (i.d.) mm C18 5 m column and subjected to a gradient of 0.1% trifluoroacetic acid in acetonitrile (Solution A) and 0.1% trifluoroacetic acid in HPLC-grade water (Solution B) according to the following scheme:

TABLE-US-00008 TABLE 6 HLPC Methods A B 0.01 min 7% 93% 25 min 32% 68% 25.1 min 100% 0% Flow rate 1.0 ml/min Wave Length 220 nm Load Volume 10 l

[0255] The presence of aromatic-cationic peptides in the biological sample is established by comparison to data obtained for reference samples such as those provided in Example 6.

[0256] The foregoing method is illustrative only, and should not be construed as limiting in any way. One of skill in the art will understand that the aromatic-cationic peptides described herein may be analyzed by a number of HPLC methods, such as those describe in Aguilar, HPLC of Peptides and Proteins: Methods and Protocols, Humana Press, New Jersey (2004).

Example 8

Detection of Aromatic-Cationic Peptides in a Biological Sample by MS

[0257] This example demonstrates the detection of aromatic-cationic peptides in a biological sample by MS. Biological samples are collected from subjects in a suitable manner depending on the nature of the sample. Biological samples include any material derived from or contacted by living cells. Examples of biological samples include but are not limited to whole blood, fractionated blood, semen, saliva, tears, urine, fecal material, sweat, buccal, skin, cerebrospinal fluid, and hair. Biological samples also include biopsies of internal organs or cancers. Once obtained, the biological samples are stored in a manner compatible with the methods of detection until the methods are performed to ensure the preservation of aromatic-cationic peptides present in the sample.

[0258] Samples are loaded in a 20 l volume and analyzed under the following exemplary conditions.

TABLE-US-00009 TABLE 7 MS Methods Probe ESI Nebulizer Gas Flow 1.5 L/min Curved Desolvation Line (CDL) 20.0 v CDL Temp 250 C. Block Temp 200 C. Probe Bias +4.5 kv Detector 1.5 kv T. Flow 0.2 ml/min Buffer 50% H.sub.2O-50% Acetonitrile

[0259] One of skill in the art will understand, that the aromatic-cationic peptides described herein may be analyzed by a number of MS methods, such as those describe in Sparkman, Mass Spectrometry Desk Reference, Pittsburgh: Global View Pub (2000).

[0260] The present invention is not to be limited in terms of the particular embodiments described in this application, which are intended as single illustrations of individual aspects of the invention. Many modifications and variations of this invention can be made without departing from its spirit and scope, as will be apparent to those skilled in the art. Functionally equivalent methods and apparatuses within the scope of the invention, in addition to those enumerated herein, will be apparent to those skilled in the art from the foregoing descriptions. Such modifications and variations are intended to fall within the scope of the appended claims. The present invention is to be limited only by the terms of the appended claims, along with the full scope of equivalents to which such claims are entitled. It is to be understood that this invention is not limited to particular methods, reagents, compounds compositions or biological systems, which can, of course, vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting.

[0261] In addition, where features or aspects of the disclosure are described in terms of Markush groups, those skilled in the art will recognize that the disclosure is also thereby described in terms of any individual member or subgroup of members of the Markush group.

[0262] As will be understood by one skilled in the art, for any and all purposes, particularly in terms of providing a written description, all ranges disclosed herein also encompass any and all possible subranges and combinations of subranges thereof. Any listed range can be easily recognized as sufficiently describing and enabling the same range being broken down into at least equal halves, thirds, quarters, fifths, tenths, etc. As a non-limiting example, each range discussed herein can be readily broken down into a lower third, middle third and upper third, etc. As will also be understood by one skilled in the art all language such as up to, at least, greater than, less than, and the like, include the number recited and refer to ranges which can be subsequently broken down into subranges as discussed above. Finally, as will be understood by one skilled in the art, a range includes each individual member. Thus, for example, a group having 1-3 cells refers to groups having 1, 2, or 3 cells. Similarly, a group having 1-5 cells refers to groups having 1, 2, 3, 4, or 5 cells, and so forth.

[0263] All patents, patent applications, provisional applications, and publications referred to or cited herein are incorporated by reference in their entirety, including all figures and tables, to the extent they are not inconsistent with the explicit teachings of this specification.

[0264] Other embodiments are set forth within the following claims.