Patent ID: 6127170
Filing Date: 2000-10-03
Classification: A61K,C07K,C12N

Abstract:
A multifunctional molecular complex for the transfer of a nucleic acid composition to a target cell comprising: A) said nucleic acid composition non-covalently bound to B) a transfer moiety, wherein said transfer moiety comprises one or more cationic polyamine components bound to said nucleic acid composition, each independently comprising a cationic polyamine of the formula (1):NR(R.sup.3)--[--(CR.sup.1 R.sup.2).sub.m --N(R.sup.3)--].sub.n --(CR.sup.1 R.sup.2).sub.m --NR(R.sup.3) (1)wherein:R, R.sup.1 and R.sup.2 are each independently selected from the group consisting of hydrogen and C.sub.1-6 alkyl;m in each occurrence is independently selected from the integers 2 through 5 inclusive;n is selected from the integers 1 through 10 inclusive;R.sup.3 is independently selected from the group consisting of hydrogen; C.sub.1-6 alkyl; or NR(R.sup.3) is guanidino; and at least one endosome membrane disruption promoting component independently selected from the group consisting of:(a) --B--(CR.sup.1 R.sup.2).sub.j --C(R).sub.3, where R is independently selected from the group consisting of hydrogen, C.sub.1-6 alkyl, or C(R).sub.3 is C.sub.6 H.sub.5 aromatic or absent; R.sup.1 and R.sup.2 are each independently selected from the group consisting of hydrogen and C.sub.1-6 alkyl; j is an integer from 0 to 24 inclusive; and B is optionally absent, or is a bridging group of the formula:(i) --(CR.sup.1 R.sup.2).sub.k --C(.dbd.O)--Z--;(ii) --(CR.sup.1 R.sup.2).sub.k --N(R)--C(.dbd.O)--Z--;(iii) --(CR.sup.1 R.sup.2).sub.k --N(R)--{--C(.dbd.O)--CH.sub.2 --O[--(CH.sub.2).sub.2 --O--].sub.1 --(CH.sub.2).sub.k --N(R)}.sub.p --C(.dbd.O)--Z--; or(iv) --(CR.sup.1 R.sup.2).sub.k --C(.dbd.O)--{--N(R)--[--(CH.sub.2).sub.2 --O--].sub.1 --CH.sub.2 --C(.dbd.O)}.sub.p --Z--;where k is, independently, an integer from 1 to 11 inclusive, 1 is an integer from 0 to 30 inclusive, and p is an integer from 1 to 3 inclusive; R is independently defined as above or is absent, R.sup.1 and R.sup.2 are each independently selected from the group consisting of hydrogen and C.sub.1-6 alkyl; and Z is O, OH, S, N(R), or is absent;(b) --B--(R.sup.4)R, where R, R.sup.1 and R.sup.2 are each independently defined as above; B cannot be absent and is a bridging group independently selected from groups (i) through (iv) above, and additionally from the group of the formula:(v) --(CR.sup.1 R.sup.2).sub.j' --X--, where j' is an integer from 1 to 8 inclusive; R.sup.1 and R.sup.2 are each independently defined as above;X is O, S, N(R), or absent; andR.sup.4 is independently selected from the group consisting of:(i) fusogenic peptides comprising spike glycoproteins of enveloped animal viruses;(ii) cholic acid derivatives of the formula (2): ##STR5## where: represents a bond of unspecified stereochemistry;--- represents a single or double bond, forming a saturated or unsaturated portion of the ring system, provided that they cannot both be unsaturated at the same time, whereby the ring system must be either .DELTA.4 or .DELTA.5;R.sup.6 is --H, --OH, --CO.sub.2 H, --C(.dbd.O)NH.sub.2, --OC(.dbd.O)NH.sub.2, --NH.sub.2, or --O(CH.sub.2 CH.sub.2 O).sub.n' H, where n' is an integer from 1 to 6 inclusive;R.sup.7 is a radical that forms the point of attachment of the cholic acid derivative, comprising --C.sub.1-6 alkyl- or --C.sub.1-6 alkylcarbonyl-; andR.sup.8 is C.sub.1-6 alkyl; and(iii) cholesteryl derivatives of the formula (3): ##STR6## where: represents a bond of unspecified stereochemistry;--- represents a single or double bond, forming a saturated or unsaturated portion of the ring system, provided that they cannot both be unsaturated at the same time, whereby the ring system must be either .DELTA.4 or .DELTA.5;R.sup.6a is a radical that forms the point of attachment of the cholesteryl derivative, comprising --C.sub.1-6 alkyl-, --OC(.dbd.O)--, or --OCH.sub.2 C(.dbd.O)--;R.sup.7a is C.sub.1-6 alkyl; andR.sup.8a is C.sub.1-6 alkyl;PROVIDED THAT R.sup.3 is one or more endosome membrane disruption promoting components attached to at least one nitrogen atom of at least one of said cationic polyamine components; andOPTIONALLY, R.sup.3 may be one or more groups defined below, attached either to a further nitrogen atom of at least one of said cationic polyamine components to which said one or more endosome membrane disruption promoting components is attached, or to a nitrogen atom of at least one further polyamine component which does not have attached thereto any endosome membrane disruption promoting component:(c) --B--(R.sup.5)R, where B cannot be absent, and is a bridging group independently selected from groups (i) through (v) inclusive; R is independently defined as above or absent; andR.sup.5 is a receptor specific binding component independently selected from the group consisting of:(i) D-biotin;(ii) .beta.-3'-propionyl galactosyl-.beta.1-4-thioglucoside;(iii) N.sup.2,N.sup.6 -bis(.beta.-3'-propionyl galactosyl-.beta.1-4-thioglucoside)lysine;(iv) N.sup.2,N.sup.6 -bis(.beta.-3'-propionyl galactosyl-.beta.1-4-thioglucoside)lysyl-N.sup.6 -(.beta.-3'-propionyl galactosyl-.beta.1-4-thioglucoside)lysine;(v) 5-methyltetrahydrofolate;(vi) folic acid;(vii) folinic acid;(viii) .alpha.-3'-propionyl thiomannoside; and(ix) .alpha.-3'-propionyl thiomannoside-6-phosphate.