Cationic lipids

Details Cationic lipids Cationic lipids

2000-04-25

2002-08-20

Powers, Fiona T. (Department: 1626)

Organic compounds -- part of the class 532-570 series

Organic compounds

Chalcogen in the nitrogen containing substituent

C544S139000, C544S168000, C548S312700, C548S338100, C564S153000, C564S157000, C564S197000

Reexamination Certificate

active

06437121

ABSTRACT:

BACKGROUND OF THE INVENTION
The present invention relates to new cationic lipids and synthetic intermediates therefor as well as their use for delivering therapeutic compounds, particularly anionic and polyanionic polymers such as nucleic acids and peptide compounds into eukaryotic and prokaryotic cells.
Workers have described lipids that are useful for delivering or transfecting into cells nucleic acids, peptides, proteins and other compounds such as lipophilic and anionic therapeutic agents (WO 96/10390; WO 96/01841; WO 96/01840; WO 95/35094; WO 95/12386; WO 94/05624; WO 94/00569; WO 93/24640; WO 91/16024; WO 90/14074; WO 90/11092; U.S. Pat. Nos. 5,459,127, 5,283,185, 5,171,687, 5,286,634, 4,880,635, 4,857,319 and 4,229,360; Boussif et al.
Proc. Natl. Acad. Sci
. (
U.S.A
.) 92:7297-7301, 1995; Budker et al.
Nature Biotech
. 14:760-764; Felgner et al.
J. Biol. Chem
. 2,6:2550-2561, 1994; Koff et al.
Science
224:1007-1009 1980; Jaaskelainen et al.
Biochim, Biophys. Acta
1195:115-123, 1994; Leserman et al.
J. Liposome Res
. 4:107-119, 1994; Lewis et al.
Proc. Natl. Acad. Sci
. (
U.S.A
.) 93:3176-3181, 1996; Nabel et al.
Proc. Natl. Acad. Sci
. (
U.S.A
.) 90:11307, 1993; Nabel et al.
Hum. Gene Ther
. 3:649, 1992; Nabel et al.
Science
249:1285-1288, 1990; Philip et al.
J. Biol. Chem
. 268:16087-16090, 1993; Puyal et al.
Eur. J. Biochem
. 228:697-703, 1995; Remy et al.
Bioconjugate Chem
. 5:647-654, 1994; Ropert et al.
Biochem. Biophys. Res. Commun
. 183:879-885, 1992; Stribling et al.
Proc. Natl. Acad. Sci
. (
U.S.A
.) 89:11277-11281, 1992; Tong et al.
Acta Pharm. Sinica
27:15-21, 1991; van Borssum Waalkes et al.
Biochim, Biophys. Acta
1148:161-172, 1993; Walker et al.
Proc. Natl. Acad. Sci
. (
U.S.A
.) 89:7915-7919, 1992; Zalipsky et al.
FEBS Letters
353:71-74, 1994; Zhu et al.
Science
261:209-211, 1993; Xu et al.
Biochem
. 35:5616-5623, 1996; D. D. Lasic
Liposomes: From Physics to Applications
, Elsevier, Amsterdam, 1993.
OBJECTS OF THE INVENTION
The invention lipids or methods include one or more compounds or methods that accomplish one or more of the following objects.
It is an object of the invention to provide cationic lipids and intermediates for making such lipids.
Another object of the invention is to provide cationic lipids that are suitable for delivering or transfecting compounds such as nudeic acids, peptides, and anionic therapeutic agents into cell cytoplasm or cell nuclei in vitro or in vivo in the presence or absence of serum or blood.
Another object of the invention is to provide cationic lipids that are suitable for efficiently delivering polyanionic polymers such as nucleic acids into cells using cells in tissue culture at a cell confluency of about 50% to 100%.
Another object of the invention is to provide cationic lipids that are suitable for efficiently delivering a large amount of polyanionic polymers such as nucleic acids, proteins, peptides or anionic therapeutic agents into cells.
Another object of the invention is to provide cationic lipids having improved pharmacological or other properties such as, improved storage stability, reduced toxicity or increased efficacy in the presence of serum or in the presence of components found in tissue culture medium.
Another object of the invention is to obviate the need to use a colipid such as DOPE in the intracellular delivery of nucleic acids, oligonucleotides or other anionic compounds into cells in vitro or in vivo.
Another object of the invention is to provide cationic lipids that are suitable for efficiently delivering polyanionic polymers such as nucleic acids or peptides sytemically to the lung, spleen or other organs of a mammal such as rodents, non-human primates or humans.
Another object of the invention is to provide methods to deliver anionic compounds or hydrophobic compounds into cells in vitro or in vivo.
Another object of the invention is to provide compositions comprising cationic lipids and anionic compounds or therapeutic agents such as nucleic acids, peptides, proteins, oligonucleotides, or antiviral agents. Such compositions optionally contain colipid(s).
Another object of the invention is to provide compositions comprising cationic lipids and hydrophobic compounds or therapeutic agents such as antitumor or antifungal agents. Such compositions optionally contain colipid(s).
SUMMARY OF THE INVENTION
Invention embodiments include cationic lipids and intermediates therefor having the structure A
wherein
each R is independently hydrogen or a lipophilic moiety, the lipophilic moieties typically consisting of 1 or 2 groups, usually 2, containing at least about 10 linked carbon atoms, typically about 10-50 linked carbon atoms, usually about 10-22 linked carbon atoms and R is optionally selected from alkyl (C
10-22
), a mono-, di- or tri-unsaturated alkenyl (C
10-22
) group, or one R is a cholesteryl moiety and the other R is hydrogen, provided that both R are not hydrogen;
R
1
and R
2
are independently hydrogen, —(CH
2
)
z
—N(R
4
)
2
, —(CH
2
)
z
NR
4
—C(═NH)—N(R
5
)
2
, or W
1
, provided that at least one of R
1
and R
2
is W
1
;
each R
3
is independently hydrogen, alkyl (C
1-10
), —CH
2
—(CF
2
)
p
—CF
3
, aryl (e.g., phenyl or naphthyl), a protecting group, or both R
3
together are a protecting group, or one R
3
is hydrogen and the other R
3
is —C(O)CH
2
NH
2
or —C(O)CH(CH
3
)NH
2
, provided that both R
3
are not aryl;
each R
4
is independently hydrogen, alkyl (C
1-6
) (e.g., methyl, ethyl, propyl, isopropyl), a protecting group, —CH
2
—(CF
2
)
p
—CF
3
, or both R
4
together are a protecting group;
each R
5
is independently hydrogen, alkyl (C
1-6
) (e.g., methyl, ethyl, propyl, isopropyl), a protecting group, or both R
5
together are a protecting group;
each W
1
is independently a cationic group, at least one of which has a pKa of about 6.0-7.5, W
1
is optionally selected from
m is an integer having the value 0, 1, 2, 3 or 4, usually 0 or 1;
n is an integer having the value 0, 1, 2, 3 or 4, usually 0 or 1;
p is an integer having the value 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10;
s is an integer having the value 0, 1 or 2, usually 0 or 1;
z is an integer having the value 1, 2, 3, or 4, usually 0 or 1;
positions designated * are carbon atoms with linked substituents in the R, S or RS configuration; and
the salts, tautomers, solvates, resolved, partially resolved and unresolved enantiomers, purified, partially purified and unpurified positional isomers or diastereomers thereof. Structure A1 cationic lipids are compounds where each W
1
independently has structures shown for A, with the remaining portions of the structure being the same as A.
Invention embodiments include cationic lipids and intermediates therefor having the structure E
wherein q and r are each independently 0, 1, 2 or 3, usually q is 0 and r is 1 or q is 1 and r is 0;
R
11
is a moiety with a pKa of about 6.0-10, usually about 7.0-8.5; and
R
12
is a W
1
moiety with a pKa of about 6.0-7.5 such as B, C or D
In structure E compounds, when q is 0, R
11
is —N(R
4
)
2
or —N(R
14
)(R
4
) where R
14
is hydrogen or alkyl (C
1-6
) and when q is 1, 2, or 3, R
11
is an amine substituted with an electron withdrawing substituent, e.g., R
11
is —NH—CH
2
—CN, —NH—CH
2
—NO
2
, —NH—CH
2
—SO
2
R
15
, —NH—CH
2
—C(O)(CH
2
)
m
CH
3
, —NH—CH
2
(CF
2
)
m
CF
3
, or —NH—CH
2
O(CH
2
)
m
CH
3
, where R
15
is hydrogen or alkyl (C
1-6
). R
11
has a greater positive charge at a pH about 7 than R
12
. At a pH of about 7, R
12
has a charge that is significantly less than +1.0 (about 0.1-0.6) and a charge of about +0.8-1.0 at lower pH of about 5-6. The presence of an electron withdrawing substituent at R
11
reduces the pKa of the charged moiety when q is 1, 2, or 3 and the charged moiety is located farther from the carbonyl group.
In some embodiments, the structure A or E lipids have 1, 2, or 3 moieties, usually 1 or 2, with a pKa of about 6.0-7.5 wherein the pKa is determined by the process of: (a) preparing a water solution containing a suspension of the HCl salt of the cationic lipid at its CMC

Affiliated with

Also associated with

Rating

Cationic lipids does not yet have a rating. At this time, there are no reviews or comments for this patent.

If you have personal experience with Cationic lipids, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Cationic lipids will most certainly appreciate the feedback.

Rate now

Comments { 0 }

Profile ID: LFUS-PAI-O-2919493