# US 9,750,693 B2 · Liposome structures and methods of use thereof > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 9,750,693 B2 - Document ID: US9750693B2 - Status: Granted - Kind: B2 - Application number: 14/499,600 - Filed: 2014-09-29 - Published: 2015-03-26 - Granted: 2017-09-05 - Earliest claimed: 2010-08-13 - Assignees: Rhode Island Council on Postsecondary Education (Statutory Successor to Rhode Island Board of Governors for Higher Education); Yale University - Applicants: Rhode Island Council on Postsecondary Education (statutory successor to Board of Governors for Higher Education, State of Rhode Island and Providence Plantations); Yale University - Inventors: Yana K. Reshetnyak; Oleg A. Andreev; Donald M. Engelman - Web page: https://patentgrep.com/patent/US9750693B2 ## CPC - A61K 9/1271: Non-conventional liposomes, e.g. PEGylated liposomes or liposomes coated or grafted with polymers (liposomes as conjugates {A61K47/6911}) - A61K 9/127: Synthetic bilayered vehicles, e.g. liposomes or liposomes with cholesterol as the only non-phosphatidyl surfactant - A61K 31/20: having a carboxyl group bound to a chain of seven or more carbon atoms, e.g. stearic, palmitic, arachidic acids - A61K 31/337: having four-membered rings, e.g. taxol - A61K 31/704: attached to a condensed carbocyclic ring system, e.g. sennosides, thiocolchicosides, escin, daunorubicin {(digitoxin A61K31/7048)} - A61K 31/7105: Natural ribonucleic acids, i.e. containing only riboses attached to adenine, guanine, cytosine or uracil and having 3'-5' phosphodiester links - A61K 31/711: Natural deoxyribonucleic acids, i.e. containing only 2'-deoxyriboses attached to adenine, guanine, cytosine or thymine and having 3'-5' phosphodiester links - A61K 31/713: Double-stranded nucleic acids or oligonucleotides - A61K 47/6911: {the form being a liposome} - A61P 35/00: Antineoplastic agents - A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D) - A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS ## Abstract The present application relates to compositions comprising and methods of using a liposome comprising a pHLIP polypeptide, wherein a lipid bilayer of the liposome is substantially free of the pHLIP polypeptide. ## Claims ### Claim 1 A liposome comprising non-pore forming pHLIP polypeptide monomers covalently attached to polar headgroups of phospholipids in the lipid bilayer of said liposome, wherein the hydrophobic phospholipid tail region of the lipid bilayer of said liposome is substantially free of said pHLIP polypeptides. ### Claim 2 (depends on 1) The liposome of claim 1, wherein said liposome further comprises a cargo inside of said liposome or inside of said lipid bilayer. ### Claim 3 (depends on 1) The liposome of claim 1, wherein said cargo comprises a therapeutic compound. ### Claim 4 (depends on 1) The liposome of claim 1, wherein said cargo comprises a polar composition. ### Claim 5 (depends on 1) The liposome of claim 1, wherein said liposome further comprises a hydrophobic cargo incorporated into said lipid bilayer. ### Claim 6 (depends on 1) The liposome of claim 1, wherein said liposome further comprises a lipid bilayer-tethered cargo. ### Claim 7 (depends on 6) The liposome of claim 6, wherein said tethered cargo is attached to a lipid by a cleavable or non-cleavable bond. ### Claim 8 (depends on 6) The liposome of claim 6, wherein said tethered cargo is attached to a lipid by a S-S bond. ### Claim 9 (depends on 1) A method of delivering a cargo into a target cell comprising contacting said target cell with cargo-loaded pHLIP+ liposomes according to claim 1, wherein at least 10% more of said cargo is delivered to the cytoplasm of said target cell compared to the amount delivered using pHLIP− liposomes. ### Claim 10 (depends on 9) The method of claim 9, wherein said target cell is characterized by a microenvironment comprising a low pH. ### Claim 11 (depends on 9) The method of claim 9, wherein said pHLIP+liposomes fuse with a cell membrane of said target cell. ### Claim 12 (depends on 9) The method of claim 9, wherein said pHLIP+liposomes both fuse with a cell membrane of said target cell and are taken up by said cell by endocytosis. ### Claim 13 (depends on 9) The method of claim 9, wherein said pHLIP+liposomes preferentially fuse with a membrane of an endosomal compartment and a lysosomal compartment of said target cell after uptake by endocytosis. ### Claim 14 (depends on 9) The method of claim 9, wherein said target cell is a tumor cell, ischemic cell, inflamed cell, bacterially-infected cell, fungus-infected cells, or virally-infected cell. ### Claim 15 (depends on 9) The method of claim 9, wherein said cargo comprises ceramide, a deoxyribonucleotide (DNA) binding agent, a small interfering ribonucleic acid (RNA), a DNA, a polar toxin, an inhibitor, paclitaxel, or doxorubicin. ### Claim 16 (depends on 1) A pharmaceutical composition comprising the liposome of claim 1. ### Claim 17 (depends on 1) The liposome of claim 1, wherein said pHLIP polypeptides are located outside of the lipid bilayer of said liposome. ### Claim 18 (depends on 1) The liposome of claim 1, wherein the amino-terminal end of said pHLIP polypeptide is covalently attached to said phospholipid. ### Claim 19 (depends on 1) The liposome of claim 1, wherein the carboxy-terminal end of said pHLIP polypeptide is covalently attached to said phospholipid.