# US 8,785,200 B2 · Transfection reagents > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 8,785,200 B2 - Document ID: US8785200B2 - Status: Granted - Kind: B2 - Application number: 13/420,091 - Filed: 2012-03-14 - Published: 2012-09-20 - Granted: 2014-07-22 - Earliest claimed: 1998-11-12 - Assignees: Life Technologies Corporation - Applicants: Yongliang Chu; Malek Masoud; Gulilat Gebeyehu - Inventors: Yongliang Chu; Malek Masoud; Gulilat Gebeyehu - Web page: https://patentgrep.com/patent/US8785200B2 ## CPC - A61K 9/16: Agglomerates; Granulates; Microbeadlets {; Microspheres; Pellets; Solid products obtained by spray drying, spray freeze drying, spray congealing,(multiple) emulsion solvent evaporation or extraction (A61K9/20 takes precedence if the final form is a tablet; microspheres with drug-free outer coating, microcapsules A61K9/50; mixture of different granules, microcapsules, (coated) microparticles A61K9/5084; nanoparticles A61K9/51)} - A61K 31/14: Quaternary ammonium compounds, e.g. edrophonium, choline (betaines A61K31/205) - A61K 38/28: Insulins - A61K 38/40: Transferrins, e.g. lactoferrins, ovotransferrins - A61K 47/543: {Lipids, e.g. triglycerides; Polyamines, e.g. spermine or spermidine} - C07C 211/21: Monoamines - C07C 211/63: having quaternised nitrogen atoms bound to acyclic carbon atoms - C07C 211/64: having quaternised nitrogen atoms bound to carbon atoms of six-membered aromatic rings - C07C 229/26: having more than one amino group bound to the carbon skeleton, e.g. lysine - C07D 209/48: with oxygen atoms in positions 1 and 3, e.g. phthalimide - C07D 241/12: with only hydrogen atoms, hydrocarbon or substituted hydrocarbon radicals, directly attached to ring carbon atoms - C12N 15/88: using microencapsulation, e.g. using {amphiphile} liposome vesicle - Y10S 435/81: Packaged device or kit - 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) - C07C: ACYCLIC OR CARBOCYCLIC COMPOUNDS - C07D: HETEROCYCLIC COMPOUNDS (macromolecular compounds C08) - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) - Y10S: TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS ## Abstract Disclosed are cationic lipid compounds and compositions of lipid aggregates for delivery of macromolecules and other compounds into cells. The compounds can be used alone or in combination with other compounds to prepare liposomes and other lipid aggregates suitable for transfection or delivery of compounds to target cells, either in vitro or in vivo. The compounds are preferably polycationic and preferably form highly stable complexes with various anionic macromolecules, particularly polyanions such as nucleic acids. These compounds have the property, when dispersed in water, of forming lipid aggregates which associate strongly, via their cationic portion, with polyanions. Also disclosed are intermediates for preparing the compound and compositions of the invention and methods of using the compounds to introduce other compounds into cells. ## Claims ### Claim 1 A method for transfecting a target cell with an expressible nucleic acid, said method comprising: a) obtaining a target cell; b) forming a liposome complex comprising an expressible nucleic acid, at least one transfection enhancer, and a compound having the structure: or a polycation thereof; wherein R4 is a straight-chain, branched or cyclic, alkyl or alkenyl group having 8 to 24 carbon atoms, R5 is an alkyl group substituted with one or more amines and one or more alcohols, and R7 is H or a carbohydrate; and c) contacting the liposome complex with the target cell. ### Claim 2 (depends on 1) The method according to claim 1, wherein R5 is an alkyl group substituted with one amine and one or more alcohols. ### Claim 3 (depends on 2) The method according to claim 2, wherein the amine substituent of the R5 alkyl group comprises an aminoethanol, an aminopropanol or an aminobutanol group. ### Claim 4 (depends on 3) The method according to claim 3, wherein the amine substituent of the R5 alkyl group has the structure: wherein n is an integer from 7-23. ### Claim 5 (depends on 1) The method according to claim 1, wherein R5 is an alkyl group having 3-30 carbon atoms and is substituted by one amine and one or more alcohols. ### Claim 6 (depends on 5) The method according to claim 5, wherein the amine substituent of the R5 alkyl group comprises an aminoethanol, an aminopropanol or an aminobutanol group. ### Claim 7 (depends on 6) The method according to claim 6, wherein the amine substituent of the R5 alkyl group has the structure: wherein n is an integer from 7-23. ### Claim 8 (depends on 5) The method according to claim 5, wherein R5 is a propyl or a butyl group which is substituted by one amine and one or more alcohols. ### Claim 9 (depends on 8) The method according to claim 8, wherein the amine substituent of the R5 alkyl group comprises an aminoethanol, an aminopropanol or an aminobutanol group. ### Claim 10 (depends on 9) The method according to claim 9, wherein the amine substituent of the R5 alkyl group has the structure: wherein n is an integer from 7-23. ### Claim 11 (depends on 1) The method according to claim 1, wherein R7 is H. ### Claim 12 (depends on 11) The method according to claim 11, wherein R4 is a straight-chain alkyl group having 8 to 24 carbon atoms. ### Claim 13 (depends on 12) The method according to claim 12, wherein R5 is an alkyl group having 3-30 carbon atoms and is substituted with one amine and one or more alcohols. ### Claim 14 (depends on 13) The method according to claim 13, wherein the amine substituent of the R5 alkyl group comprises an aminoethanol, an aminopropanol or an aminobutanol group. ### Claim 15 (depends on 14) The method according to claim 14, wherein the amine substituent of the R5 alkyl group has the structure: wherein n is an integer from 7-23. ### Claim 16 (depends on 13) The method according to claim 13, wherein R5 is a propyl or a butyl group which is substituted by one amine and one or more alcohols. ### Claim 17 (depends on 16) The method according to claim 16, wherein the amine substituent of the R5 alkyl group comprises an aminoethanol, an aminopropanol or an aminobutanol group. ### Claim 18 (depends on 17) The method according to claim 17, wherein the amine substituent of the R5 alkyl group has the structure: wherein n is an integer from 7-23. ### Claim 19 (depends on 1) The method according to claim 1, wherein R4 is a straight-chain alkyl group having 8 to 24 carbon atoms, R5 is a butyl group substituted with one amine and one or more alcohols, and R7 is H. ### Claim 20 (depends on 19) The method of claim 19, wherein the amine substituent of the R5 alkyl group comprises an aminoethanol, an aminopropanol or an aminobutanol group. ### Claim 21 (depends on 20) The method according to claim 20, wherein the amine substituent of the R5 alkyl group has the structure: wherein n is an integer from 7-23. ### Claim 22 (depends on 1) The method according to claim 1, wherein R4 is a straight-chain alkyl group having 8 to 24 carbon atoms, R5 is a propyl group substituted with one amine and one alcohol, and R1 is H. ### Claim 23 (depends on 22) The method according to claim 22, wherein the amine substituent of the R5 alkyl group comprises an aminoethanol, an aminopropanol or an aminobutanol group. ### Claim 24 (depends on 23) The method according to claim 23, wherein the amine substituent of the R5 alkyl group has the structure: wherein n is an integer from 7-23. ### Claim 25 (depends on 1) The method according to claim 1, wherein the liposome further comprises one or more neutral lipids. ### Claim 26 (depends on 25) The method according to claim 25, wherein the lipid is DOPE, DOPC or cholesterol. ### Claim 27 (depends on 1) The method according to claim 1, wherein the liposome complex is formed in an aqueous medium. ### Claim 28 (depends on 1) The method according to claim 1, wherein the transfection enhancer is a protein, a peptide or a growth factor that enhances cell-targeting, uptake, internalization, nuclear targeting or expression.