COMPOSITIONS AND METHODS FOR EFFICIENT DELIVERY OF MOLECULES TO CELLS

摘要

The present invention provides lipid aggregate compositions having at least two cationic lipids in combination with one or optionally more than one non-cationic lipids for the in vitro and/or in vivo delivery of biologically active molecules including nucleic acids, specifically DNA and RNA, and proteins into cells and tissues. Methods are also provided that use the compounds of the present invention to deliver biologically active molecules, into cells and tissues to facilitate the expression of target proteins therein. In some non-limiting embodiments, the subject lipid aggregate compositions can be used to deliver nucleic acid molecules to facilitate the expression of proteins involved in cellular reprogramming and genome editing applications.

主权项

1. A lipid aggregate comprising a first cationic lipid and at least a second cationic lipid, a first neutral lipid and optionally a second neutral lipid, wherein said lipid aggregate is suitable for forming a cationic complex with a nucleic acid under aqueous conditions, wherein said first and second cationic lipids are different from each other, and wherein each of said first and second cationic lipids have the structure:and salts thereof;where:
R1 and R2, independently, are an alkyl, alkenyl or alkynyl groups, having from 8 to 30 carbon atoms; an alkyl, alkenyl or alkynyl groups, having from 8 to 30 carbon atoms and optionally substituted by one or more of an alcohol, an aminoalcohol, an amine, an amide, an ether, a polyether, an ester, a mercaptan, alkylthio, or a carbamoyl group or where R1 is —(CH2)q—N(R6)tR7R8; R3 and R4, independently, are hydrogens, or alkyl, alkenyl or alkynyl groups having from 8 to 30 carbon atoms and optionally substituted by one or more of an alcohol, an aminoalcohol, an amine, an amide, an ether, a polyether, an ester, a mercaptan, alkylthio, or a carbamoyl group; R5-R8, independently, are hydrogens, or alkyl, alkenyl or alkynyl groups; R9 is a hydrogen, or an alkyl, alkenyl or alkynyl group, a carbohydrate or a peptide; r, s and t are 1 or 0 to indicate the presence or absence of the indicated R group, when any of r, s or t are 1 the nitrogen to which the indicated R group is attached is positively charged and wherein at least one of r, s or t is 1; q is an integer ranging from 1 to 6, inclusive; Xv- is an anion, where v is the valency of the anion and A is the number of anions; L is a divalent organic radical capable of covalently linking the two nitrogens selected from: (CH2)n, where n is an integer ranging from 1 to 10, inclusive, which is optionally substituted with one or more ZR10 groups, where Z is O or S, and R10 is hydrogen or an alkyl, alkenyl or alkynyl group; or {—(CH2)k—Y—(CH2)m}p—, where k and m, independently, are integers ranging from 1 to 10, inclusive, and p is an integer ranging from 1 to 6, inclusive, and Y is O, S, CO, COO, CONR11, NR11CO, or NR11COR11N where R11, independent of any other R11, is hydrogen or an alkyl group; wherein one or more CH2 groups of the alkyl, alkenyl or alkynyl groups of R1—R10 can be replaced with an O, S, S—S, CO, COO, NR12CO, NR12COO, or NR12CONR12 where R12, independent of any other R12, is hydrogen or an alkyl, alkenyl or alkynyl group; and wherein the alkyl, alkenyl or alkynyl groups of R1-R12 are optionally substituted with one or more OR13, CN, halogens, N(R13)2, peptide, or carbohydrate groups where R13, independently of other R13, is hydrogen or an alkyl, alkenyl or alkynyl group, and wherein at least one of R3 and R4, when present as alkyl groups, are substituted with both OR13 and N(R13)2 groups.