Environmentally sensitive compositions and methods of use in the treatment and diagnosis of tumors
An environmentally sensitive membrane binding polypeptide, pH (low)-sensitive membrane peptide (pHLIP) has improved insertion kinetics balanced with solubility to selectively target acidic tissues.
1. An environmentally sensitive composition comprising
a pH-triggered membrane peptide comprising (a) at least 8 contiguous amino acids of SEQ ID NO:51, 286, 289, 290, 298, or 299, wherein,
(b) at least 6 of the contiguous 8 amino acids of said membrane peptide are non-polar,
(c) at least one of the at least 8 amino acids of said membrane sequence is protonatable, and
(d) the peptide has a higher affinity to a membrane lipid bilayer at pH 5.0 compared to at pH 8.0.
2. The composition of claim 1 , further comprising a single flanking domain at an N-terminus or at a C-terminus of said membrane peptide.
3. The composition of claim 2 , further comprising a first flanking domain at said C-terminus and a second flanking domain at said N-terminus.
4. The composition of claim 3 , further comprising a moiety attached to one of said flanking domains, wherein said moiety is selected from a therapeutic, diagnostic, radiation-enhancing, radiation-sensitizing, imaging, gene regulation, cytotoxic, apoptotic, or research reagent.
5. The composition of claim 4 , wherein said moiety is attached to said flanking region via a thiol linkage.
6. The composition of claim 1 , wherein one or more atoms are replaced by a radioactive isotope or a stable isotope.
7. The composition of claim 1 , wherein one or more of the amino acid side chains are chemically modified to render them radioactive or detectable by probing radiation.
8. A composition of claim 1 , comprising one or more cargo molecules attached to said peptide used as a therapeutic, diagnostic, imaging, immune activation, gene regulation or cell function regulation agent, radiation-enhancing agent, radiation-sensitizing agent, or as a research tool.
9. The composition of claim 1 , for use as an agent to deliver a functional moiety across cell membranes to cells in a diseased tissue with a naturally acidic extracellular environment or in a tissue with an artificially induced acidic extracellular environment relative to normal physiological pH.
10. The composition of claim 9 , wherein said diseased tissue is selected from the group consisting of inflamed tissue, ischemic tissue, arthritic tissue, tissue infected with a microorganism, and atherosclerotic tissue.
11. The composition of claim 1 , for use as an agent to deliver a functional moiety to cell surfaces in a diseased tissue with a naturally acidic extracellular environment or in a tissue with an artificially induced acidic extracellular environment relative to normal physiological pH.
12. A diagnostic conjugate comprising the composition of claim 1 and a pharmaceutically acceptable detectable marker linked thereto.
13. The conjugate of claim 12 , wherein said detectable marker comprises a fluorescent dye or a nanoparticle.
14. A therapeutic conjugate comprising the composition of claim 1 , further comprising a first cargo comprising a cytotoxic agent and a second cargo comprising a hydrophobicity-balancing moiety.
15. The conjugate of claim 14 , wherein said cytotoxic agent is selected from the group consisting of phalloidin, phallo toxin, amanitin toxin, a boron-containing compound, and a DNA intercalator.
16. The composition of claim 4 , wherein said moiety comprises phalloidin, phallo toxin, amanitin toxin, a DNA intercalator, or a peptide nucleic acid.
17. The composition of claim 1 , wherein said sequence comprises residues 15-23 of SEQ ID NO:51.
18. A method of guiding surgical tumor excision, comprising administering to an anatomical site comprising a tumor the conjugate of claim 12 , removing a primary tumor from said site, and detecting residual tumor cells by binding of said conjugate to said residual tumor cells.
19. A method of determining the aggressiveness of a primary tumor, comprising contacting said tumor with the composition of claim 1 , wherein an increased level of binding of said composition compared to a control level of binding indicates an increased risk of metastasis from said primary tumor.
20. A method of preferentially inhibiting proliferation of tumor cells, comprising administering to a subject suffering from a tumor the composition of claim 1 , wherein tumor cells are preferentially inhibited compared to normal non-tumor cells.