# US 11,441,118 B2 · Nucleic acid and other compositions and methods for the modulation of cell membranes > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 11,441,118 B2 - Document ID: US11441118B2 - Status: Granted - Kind: B2 - Application number: 16/179,214 - Filed: 2018-11-02 - Published: 2019-05-02 - Granted: 2022-09-13 - Earliest claimed: 2017-11-02 - Assignees: The Board of Trustees of the University of Illinois; Cambridge Enterprise Limited - Applicants: The Board of Trustees of the University of Illinois; Cambridge Enterprise Limited - Inventors: Aleksei Aksimentiev; Ulrich F. Keyser - Web page: https://patentgrep.com/patent/US11441118B2 ## CPC - C12N 5/0006: {Modification of the membrane of cells, e.g. cell decoration} - A61K 9/5146: {obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyethylene glycol, polyamines, polyanhydrides} - 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 - C12N 15/11: DNA or RNA fragments; Modified forms thereof (DNA or RNA not used in recombinant technology, C07H21/00); {Non-coding nucleic acids having a biological activity} - C12N 15/111: {General methods applicable to biologically active non-coding nucleic acids} - C12N 2310/3515: Lipophilic moiety, e.g. cholesterol - C12N 2500/40: Nucleotides, nucleosides or bases - C12N 2500/50: Soluble polymers, e.g. polyethyleneglycol [PEG] - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) - 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) ## Abstract The present invention provides compositions and methods for transferring phospholipids and other molecules between the leaflets of a cell membrane. The compositions comprise at least one nucleic acid or compound having a hydrophilic region, where the composition is able to form a nanostructure that forms a toroidal pore in a lipid membrane. The nucleic acid or hydrophilic region-containing compound further contains an attached molecule capable of inserting the nanostructure into the lipid membrane. The invention also provides methods for scrambling lipids and other molecules in a cell membrane, which can be used to alter the function of a selected cell or to facilitate the death of the cell. The scrambling activity of synthetic scramblases described herein outperforms previously known enzymatically active DNA nanostructures and naturally occurring scramblases, in some cases by several orders of magnitude. ## Claims ### Claim 1 A method of altering a biological state of a cell having a lipid membrane with a first leaflet and a second leaflet, the method comprising: (a) contacting the lipid membrane of the cell with a composition comprising one or more hydrophilic regions forming a nanostructure and one or more hydrophobic or amphiphilic molecules attached to the one or more hydrophilic regions, wherein the composition comprises one or more nucleic acids which form the nanostructure comprising at least one interconnected nucleic acid duplex, and wherein said one or more hydrophobic or amphiphilic molecules are able to insert the nanostructure into the lipid membrane; and (b) inserting the nanostructure into the lipid membrane and forming a toroidal pore in the lipid membrane surrounding the nanostructure, where the hydrophilic surfaces of the first and second leaflets are fused together to form a continuous structure, wherein selected lipids, other molecules, or both, in the first leaflet are transported to the second leaflet at a rate greater than a transport rate of a control lipid membrane which was not contacted with the composition, and wherein the increased rate of transportation alters the amount of the selected lipids, other molecules, or both, in the first and second leaflets, thereby altering the biological state of the cell. ### Claim 2 (depends on 1) The method of claim 1, wherein the increased rate of transportation of the selected lipids homogenizes the lipid composition between the first and second leaflet. ### Claim 3 (depends on 2) The method of claim 2, wherein the homogenization of the lipid composition induces or marks cells for apoptosis. ### Claim 4 (depends on 1) The method of claim 1, wherein the increased rate of transportation increases the amount of antigens, receptors, or signaling peptides in the outer leaflet of the membrane, thereby increasing a cellular activity. ### Claim 5 (depends on 1) The method of claim 1, wherein the increased rate of transportation decreases the amount of antigens, receptors, or signaling peptides in the outer leaflet of the membrane, thereby decreasing a cellular activity. ### Claim 6 (depends on 1) The method of claim 1, wherein the composition further comprises an activation mechanism able to cause a conformal change in the composition structure when exposed to stimuli, wherein said conformal change activates the nanostructure to be inserted into the lipid membrane and to form the toroidal pore. ### Claim 7 (depends on 6) The method of claim 6 further comprising administering the composition to a plurality of cells in a subject and exposing a selected portion of the plurality of cells to the stimuli thereby activating nanostructures within the selected portion of the plurality of cells. ### Claim 8 (depends on 6) The method of claim 6 wherein the activating mechanism comprises an azobenzene molecule attached to the composition and the stimuli comprises ultraviolet light. ### Claim 9 (depends on 6) The method of claim 6 wherein the activating mechanism comprises one or more molecules attached to the composition, wherein the one or more molecules are able to undergo conformational changes in response to a change in pH, and the stimuli comprises a local decrease in pH. ### Claim 10 (depends on 1) The method of claim 1, wherein the selected lipids or other molecules comprise a therapeutic drug or chemical agent. ### Claim 11 A method of scrambling lipids or other molecules in a first and second leaflet of a biological membrane, the method comprising: (a) contacting the biological membrane with a composition comprising one or more hydrophilic regions forming a nanostructure and one or more hydrophobic or amphiphilic molecules attached to the one or more hydrophilic regions, wherein the composition comprises one or more nucleic acids which form the nanostructure comprising at least one interconnected nucleic acid duplex, and wherein said one or more hydrophobic or amphiphilic molecules are able to insert the nanostructure into the biological membrane; and (b) inserting the nanostructure into the biological membrane and forming a toroidal pore in the biological membrane surrounding the nanostructure, where the hydrophilic surfaces of the first and second leaflets are fused together to form a continuous structure, wherein lipids, other molecules, or both, in the first leaflet are transported to the second leaflet at a rate greater than a transport rate of a control biological membrane which was not contacted with the composition. ### Claim 12 (depends on 11) The method of claim 11, wherein lipids, other molecules, or both, in the second leaflet are transported to the first leaflet at a rate greater than a transport rate of a control biological membrane which was not contacted with the composition. ### Claim 13 (depends on 11) The method of claim 11, wherein the composition is modified to selectively transport specific lipids or other molecules from one of the leaflets to the other. ### Claim 14 A method of altering a biological state of a cell having a lipid membrane with a first leaflet and a second leaflet, the method comprising: (a) contacting the lipid membrane of the cell with a composition comprising one or more hydrophilic regions forming a nanostructure and one or more hydrophobic or amphiphilic molecules attached to the one or more hydrophilic regions, wherein said one or more hydrophobic or amphiphilic molecules are able to insert the nanostructure into the lipid membrane; and (b) inserting the nanostructure into the lipid membrane and forming a toroidal pore in the lipid membrane surrounding the nanostructure, where the hydrophilic surfaces of the first and second leaflets are fused together to form a continuous structure, wherein the composition further comprises an activation mechanism able to cause a conformal change in the composition structure when exposed to stimuli, wherein said conformal change activates the nanostructure to be inserted into the lipid membrane and to form the toroidal pore, and the activating mechanism comprises an azobenzene molecule attached to the composition and the stimuli comprises ultraviolet light, wherein selected lipids, other molecules, or both, in the first leaflet are transported to the second leaflet at a rate greater than a transport rate of a control lipid membrane which was not contacted with the composition, and wherein the increased rate of transportation alters the amount of the selected lipids, other molecules, or both, in the first and second leaflets, thereby altering the biological state of the cell. ### Claim 15 (depends on 14) The method of claim 14, wherein the increased rate of transportation of the selected lipids homogenizes the lipid composition between the first and second leaflet. ### Claim 16 (depends on 15) The method of claim 15, wherein the homogenization of the lipid composition induces or marks cells for apoptosis. ### Claim 17 (depends on 14) The method of claim 14, wherein the increased rate of transportation increases the amount of antigens, receptors, or signaling peptides in the outer leaflet of the membrane, thereby increasing a cellular activity. ### Claim 18 (depends on 14) The method of claim 14, wherein the increased rate of transportation decreases the amount of antigens, receptors, or signaling peptides in the outer leaflet of the membrane, thereby decreasing a cellular activity.