Environmentally sensitive compositions and methods of use thereof

Patent metadata

Field	Value
Publication number	US-9814781-B2
Application number	US-201615074581-A
Country	US
Kind code	B2
Filing date	Mar 18, 2016
Priority date	Jul 13, 2010
Publication date	Nov 14, 2017
Grant date	Nov 14, 2017

Abstract

An environmentally sensitive membrane binding polypeptide, pH (low)-sensitive membrane peptide (pHLIP) has improved insertion kinetics balanced with solubility to selectively target acidic tissues.

First claim

What is claimed is:

1. An environmentally sensitive composition comprising a pH-triggered membrane peptide comprising
(a) at least 8 contiguous amino acids of SEQ ID NO:40, 285, 287, or 288, 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 peptide 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 cargo attached to one of said flanking domains, wherein said cargo 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 cargo 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. The composition of claim 1 , comprising one or more cargoes 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 cargo 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 cargo 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. The composition of claim 4 , wherein said cargo comprises phalloidin, phallo toxin, amanitin toxin, a DNA intercalator, or a peptide nucleic acid.

13. The composition of claim 1 , wherein said sequence comprises residues 15-22 of SEQ ID NO: 40.

14. The composition of claim 4 , wherein said cargo comprises a dye, a fluorescent protein, a nanoparticle, or a radioactive isotope.

15. The composition of claim 14 , wherein the dye is a fluorescent dye.

16. The composition of claim 14 ,
(a) comprising a dye selected from the group consisting of rhodamine, Alexa750, or Cy5.5;
(b) comprising a fluorescent protein, wherein the fluorescent protein is green fluorescent protein;
(c) comprising a nanoparticle that comprises gold; or
(d) comprising a radioactive isotope selected from the group consisting of Fluorine-18, Copper-64, Thallium-201, Iodine-123, Gallium-67, Strontium-82, Cadmium-113, Tellurium-123, Cobalt-60, or Technetium-99m.

17. The composition of claim 8 , comprising a magnetic resonance, positron emission tomography, single photon emission computed tomography, or fluorescence imaging agent.

18. The composition of claim 8 , comprising a positron emission tomography agent.

19. The composition of claim 18 , wherein said positron emission tomography agent comprises a Copper-64 or Fluorine-18 radioactive isotope.

20. A diagnostic conjugate comprising the composition of claim 1 and a pharmaceutically acceptable detectable marker linked thereto.

21. The conjugate of claim 20 , wherein said detectable marker comprises a dye.

22. The conjugate of claim 21 , wherein the dye comprises a fluorescent dye.

23. The conjugate of claim 20 , wherein said detectable marker comprises a nanoparticle.

24. 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 cargo.

25. The conjugate of claim 24 , wherein said cytotoxic agent is selected from the group consisting of phalloidin, phallo toxin, amanitin toxin, a boron-containing compound, and a DNA intercalator.

26. A method of guiding surgical tumor excision, comprising administering to an anatomical site comprising a tumor the conjugate of claim 4 , removing a primary tumor from said site, detecting residual tumor cells by binding of said conjugate to said residual tumor cells, and excising said residual tumor cells.

27. 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.

28. 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.

Assignees

• Rhode Island Council On Postsecondary Education (Statutory Successor To Rhode Island Board Of Govern
• Univ Yale

Inventors

• Reshetnyak Yana K
• Andreev Oleg A
• Engelman Donald M

Classifications

• A61K38/04

  Peptides having up to 20 amino acids in a fully defined sequence; Derivatives thereof ({enzyme inhibitors A61K38/005;} gastrins {A61K38/2207}, somatostatins A61K38/31, melanotropins A61K38/34 {; protease inhibitors A61K38/55}) · CPC title

• A61K49/0056

  Peptides, proteins, polyamino acids · CPC title

• A61K47/6415

  Toxins or lectins, e.g. clostridial toxins or Pseudomonas exotoxins · CPC title

• A61K47/64 Primary

  Drug-peptide, drug-protein or drug-polyamino acid conjugates, i.e. the modifying agent being a peptide, protein or polyamino acid which is covalently bonded or complexed to a therapeutically active agent (peptidic linkers A61K47/65) · CPC title

• A61K49/085

  conjugated systems · CPC title