BIODEGRADABLE LIPIDS FOR THE DELIVERY OF ACTIVE AGENTS
The present invention relates to a cationic lipid having one or more biodegradable groups located in a lipidic moiety (e.g., a hydrophobic chain) of the cationic lipid. These cationic lipids may be incorporated into a lipid particle for delivering an active agent, such as a nucleic acid. The invention also relates to lipid particles comprising a neutral lipid, a lipid capable of reducing aggregation, a cationic lipid of the present invention, and optionally, a sterol. The lipid particle may further include a therapeutic agent such as a nucleic acid.
1 . A compound of formula (A):
or a salt thereof, wherein
R′ is absent, hydrogen, or alkyl;
with respect to R 1 and R 2 ,
(i) R 1 and R 2 are each, independently, optionally substituted alkyl, alkenyl, alkynyl, cycloalkylalkyl, heterocycle, or R 10 ;
(ii) R 1 and R 2 , together with the nitrogen atom to which they are attached, form an optionally substituted heterocylic ring; or
(iii) one of R 1 and R 2 is an optionally substituted alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkylalkyl, or heterocycle, and the other forms a 4-10 membered heterocyclic ring or heteroaryl with (a) the adjacent nitrogen atom and (b) the (R) a group adjacent to the nitrogen atom;
each occurrence of R is, independently, —(CR 3 R 4 )—;
each occurrence of R 3 and R 4 are, independently H, halogen, OH, alkyl, alkoxy, —NH 2 , R 10 , alkylamino, or dialkylamino;
each occurrence of R 10 is independently selected from PEG and polymers based on poly(oxazoline), poly(ethylene oxide), poly(vinyl alcohol), poly(glycerol), poly(N-vinylpyrrolidone), poly[N-(2-hydroxypropyl)methacrylamide] and poly(amino acid)s, wherein (i) the PEG or polymer is linear or branched, (ii) the PEG or polymer is polymerized by n subunits, (iii) n is a number-averaged degree of polymerization between 10 and 200 units, and (iv) the compound has at most two R 10 groups;
the dashed line to Q is absent or a bond;
when the dashed line to Q is absent then Q is absent or is —O—, —NH—, —N(R 5 )—, —S—, —C(O)—, —C(O)O—, —OC(O)—, —C(O)N(R 4 )—, —N(R 5 )C(O)—, —S—S—, —OC(O)O—, —O—N═C(R 5 )—, —C(R 5 )═N—O—, —OC(O)N(R 5 )—, —N(R 5 )C(O)N(R 5 )—, —N(R 5 )C(O)O—, —C(O)S—, —C(S)O— or —C(R 5 )═N—O—C(O)—; or
when the dashed line to Q is a bond then (i) b is 0 and (ii) Q and the tertiary carbon adjacent to it (C*) form a substituted or unsubstituted, mono- or bi-cyclic heterocyclic group having from 5 to 10 ring atoms;
each occurrence of R 5 is, independently, H or alkyl;
X is alkylene or alkenylene;
M 1 is a biodegradable group;
a is 1, 2, 3, 4, 5 or 6;
b is 0, 1, 2, or 3;
Z 1 is a C 6 -C 14 branched alkyl group; and
Z 2 is C 4 -C 20 alkenyl, wherein the alkenyl group may optionally be substituted with one or two fluorine atoms at the alpha position to a double bond which is between the double bond and the terminus of Z 2 .
2 . The compound of claim 1 , wherein R′R 1 R 2 N—(R) a -Q-(R) b — is (CH 3 ) 2 N—(CH 2 ) 2 —, (CH 3 ) 2 N—(CH 2 ) 3 —C(O)O—, (CH 3 ) 2 N—(CH 2 ) 2 —NH—C(O)O—, (CH 3 ) 2 N—(CH 2 ) 2 —OC(O)—NH—, or (CH 3 ) 2 N—(CH 2 ) 3 —C(CH 3 )═N—O—.
3 . The compound of claim 1 , wherein R 1 and R 2 are both alkyl.
4 . The compound of claim 1 or 2 , wherein M 1 is —OC(O)— or —C(O)O—.
5 . The compound of any one of the preceding claims, wherein Z 1 is a C 6 -C 10 branched alkyl group
6 . The compound of claim 5 , wherein Z 1 is —CH(CH 2 CH 3 )(CH 2 CH 2 CH 2 CH 3 ), —CH 2 CH( i Pr)(CH 2 CH 2 i Pr) or —CH 2 CH(n-Bu) 2 .
7 . The compound of any one of the preceding claims, wherein Z 2 is a C 19 alkenyl containing one or two double bonds.
8 . The compound of claim 7 , wherein Z 2 is —(CH 2 ) 9 CH═CHCH 2 CH═CH(CH 2 ) 4 CH 3 .
9 . A compound selected from selected from:
and salts thereof.
10 . A compound of formula (A-I):
or a salt thereof, wherein
s, t, u, v and q are each, independently, 0, 1, 2, 3, 4, 5, 6 or 7; and
W is a head group.
11 . The compound of claim 10 , wherein the head group is a protonatable amine group having a pKa of between about 4 and about 11.
12 . The compound of claim 10 or 11 , wherein the head group is (CH 3 ) 2 N—(CH 2 ) 2 —, (CH 3 ) 2 N—(CH 2 ) 3 —C(O)O—, (CH 3 ) 2 N—(CH 2 ) 2 —NH—C(O)O—, (CH 3 ) 2 N—(CH 2 ) 2 —OC(O)—NH—, or (CH 3 ) 2 N—(CH 2 ) 3 —C(CH 3 )═N—O—.
13 . The compound of any one of claims 10 - 12 , wherein s is 3 to 5.
14 . The compound of claim 13 , wherein s is 4.
15 . The compound of any one of claims 10 - 14 , wherein t is 4 to 6.
16 . The compound of claim 15 , wherein t is 5.
17 . The compound of any one of claims 10 - 16 , wherein q is 2 to 4.
18 . The compound of claim 17 , wherein q is 3.
19 . The compound of any one of claims 10 - 18 , wherein u is 0 to 2.
20 . The compound of claim 19 , wherein u is 1.
21 . The compound of any one of claims 10 - 20 , wherein v is 0 to 2.
22 . The compound of claim 21 , wherein v is 1.
23 . A compound of formula (A-II):
or a salt thereof, wherein
b is 0, 1, 2 or 3;
s is 0, 1, 2, 3, 4 or 5;
R 200 is C 12 -C 22 alkyl, C 12 -C 22 alkyenyl, or C 12 -C 22 alkynyl; and
R 100 is C 5 -C 15 alkyl, C 5 -C 15 alkyenyl, or C 5 -C 15 alkynyl.
24 . The compound of claim 23 , wherein b is 1.
25 . The compound of claim 23 or 24 , wherein s is 2, 3, or 4.
26 . The compound of claim 24 , wherein s is 3.
27 . The compound of any one of claims 23 - 26 , wherein R 100 is a C 8 -C 12 alkyl.
28 . The compound of any one of claims 23 - 27 , wherein R 200 is C 12 -C 14 alkyl, C 12 -C 14 alkyenyl, or C 12 -C 14 alkynyl.
29 . The compound of any one of claims 23 - 27 , wherein R 200 is C 18 -C 20 alkyl, C 18 -C 20 alkyenyl, or C 18 -C 20 alkynyl.
30 . The compound of any of the preceding claims, wherein the compound is in the form of a pharmaceutically acceptable salt.
31 . The compound of any of the preceding claims, wherein the compound is in the form of a cationic lipid.
32 . A lipid particle comprising a neutral lipid, a lipid capable of reducing aggregation, and a cationic lipid of claim 31 .
33 . The lipid particle of claim 32 , wherein the neutral lipid is selected from DSPC, DPPC, POPC, DOPE, or SM; the lipid capable of reducing aggregation is a PEG lipid; and the lipid particle further comprises a sterol.
34 . The lipid particle of any one of claims 32 and 33 , wherein the cationic lipid is present in a mole percentage of about 20% and about 60%; the neutral lipid is present in a mole percentage of about 5% to about 25%; the sterol is present in a mole percentage of about 25% to about 55%; and the PEG lipid is PEG-DMA, PEG-DMG, or a combination thereof, and is present in a mole percentage of about 0.5% to about 15%.
35 . The lipid particle of claim 32 , wherein the lipid capable of reducing aggregation is PEG-DMG.
36 . The lipid particle of claim 35 , wherein the lipid particle comprises about 50 mole % of the cationic lipid, about 10% DSPC, about 38.5% cholesterol, and about 1.5% PEG-DMG (based on 100% of the lipid components in the lipid particle).
37 . The lipid particle of claim 35 , wherein the lipid particle comprises about 58 mole % of the cationic lipid, about 10% DSPC, about 30% cholesterol, and about 2% PEG-DMG (based on 100% of the lipid components in the lipid particle)
38 . The lipid particle of any of claims 33 - 37 , further comprising an active agent.
39 . The lipid particle of claim 38 , wherein the active agent is a nucleic acid selected from a plasmid, an immunostimulatory oligonucleotide, an siRNA, an antisense oligonucleotide, a microRNA, an antagomir, an aptamer, and a ribozyme.
40 . A pharmaceutical composition comprising a lipid particle of any one of claims 33 - 39 and a pharmaceutically acceptable carrier.
41 . A method of modulating the expression of a target gene in a cell, comprising providing to the cell a lipid particle of any one of claims 38 - 39 .
42 . The method of claim 41 , wherein the active agent is a nucleic acid is an siRNA.
43 . A method of treating a disease or disorder characterized by the overexpression of a polypeptide in a subject, comprising providing to the subject the pharmaceutical composition of claim 40 , wherein the active agent is a nucleic acid selected from the group consisting of an siRNA, a microRNA, and an antisense oligonucleotide, and wherein the siRNA, microRNA, or antisense oligonucleotide includes a polynucleotide that specifically binds to a polynucleotide that encodes the polypeptide, or a complement thereof.
44 . A method of treating a disease or disorder characterized by underexpression of a polypeptide in a subject, comprising providing to the subject the pharmaceutical composition of claim 40 , wherein the active agent is a plasmid that encodes the polypeptide or a functional variant or fragment thereof.
45 . A method of inducing an immune response in a subject, comprising providing to the subject the pharmaceutical composition of claim 40 , wherein the active agent is an immunostimulatory oligonucleotide.
46 . A lipid particle comprising a biodegradable cationic lipid, a neutral lipid, a sterol, and a lipid capable of reducing aggregation, wherein (i) the molar ratio of the biodegradable cationic lipid to the sterol ranges from about 1.6:1 to about 2.0:1 and/or the molar ratio of the biodegradable cationic lipid to the neutral lipid ranges from about 5.5:1 to about 5.9:1, and (ii) the biodegradable cationic lipid comprises a lipid moiety, where the lipid moiety has one or more biodegradable groups.
47 . The lipid particle of claim 46 , wherein the lipid capable of reducing aggregation is 1,2-dimyristoyl-sn-glycerol-methoxy polyethylene glycol.
48 . The lipid particle of claim 47 , wherein the lipid capable of reducing aggregation is 1,2-dimyristoyl-sn-glycerol-methoxy polyethylene glycol with an average polyethylene glycol molecular weight of 2000.
49 . The lipid particle of any one of claims 46 - 48 , wherein the molar ratio of the biodegradable cationic lipid to the sterol is from about 1.7 to about 1.9:1.
50 . The lipid particle of claim 49 , wherein the molar ratio of the biodegradable cationic lipid to the sterol is about 1.9:1.
51 . The lipid particle of any one of claims 46 - 50 , wherein the molar ratio of the biodegradable cationic lipid to the neutral lipid ranges from about 5.5:1 to about 5.8:1.
52 . The lipid particle of claim 51 , wherein the molar ratio of the biodegradable cationic lipid to the neutral lipid is about 5.8:1.
53 . The lipid particle of any one of claims 46 - 52 , wherein the lipid particle comprises from about 55 to about 60 mol % of the biodegradable cationic lipid (based on 100 mol % of the lipid components in the lipid particle).
54 . The lipid particle of claim 53 , wherein the lipid particle comprises about 58 mol % of the biodegradable cationic lipid (based on 100 mol % of the lipid components in the lipid particle).
55 . The lipid particle of any one of claims 46 - 54 , wherein the lipid particle comprises from about 28 to about 33 mol % of the sterol (based on 100 mol % of the lipid components in the lipid particle).
56 . The lipid particle of any one of claims 46 - 55 , wherein the lipid particle comprises about 10 mol % of the neutral lipid (based on 100 mol % of the lipid components in the lipid particle).
57 . The lipid particle of any one of claims 46 - 56 , wherein the lipid particle comprises about 2 mol % of the lipid capable of reducing aggregation (based on 100 mol % of the lipid components in the lipid particle).
58 . The lipid particle of any one of claims 46 - 57 , wherein the lipid particle comprises from about 55 to about 60 mol % of the cationic lipid, from about 3 to about 12% of the neutral lipid, from about 28 to about 33 mol % of the sterol, and from about 0.5 to about 10 mol % of the lipid capable of reducing aggregation (based on 100 mol % of the lipid components in the lipid particle).
59 . The lipid particle of claim 58 , wherein the lipid particle comprises about 58% of the cationic lipid, about 10% of the neutral lipid, about 30% of the sterol, and about 2% of the lipid capable of reducing aggregation (based on 100 mol % of the lipid components in the lipid particle).
60 . The lipid particle of claim 58 , wherein the lipid particle comprises about 55% of the cationic lipid, about 10% of the neutral lipid, about 33% of the sterol, and about 2% of the PEG-modified lipid (based on 100 mol % of the lipid components in the lipid particle).
61 . The lipid particle of any of claims 46 - 60 , further comprising an active agent.
62 . The lipid particle of claim 61 , wherein the active agent is a nucleic acid selected from a plasmid, an immunostimulatory oligonucleotide, an siRNA, an antisense oligonucleotide, a microRNA, an antagomir, an aptamer, and a ribozyme.
63 . A pharmaceutical composition comprising a lipid particle of any one of claims 61 - 62 and a pharmaceutically acceptable carrier.
64 . A method of modulating the expression of a target gene in a cell, comprising providing to the cell a lipid particle of any one of claims 46 - 62 .
65 . The method of claim 64 , wherein the active agent is a nucleic acid is an siRNA.
66 . A method of treating a disease or disorder characterized by the overexpression of a polypeptide in a subject, comprising providing to the subject the pharmaceutical composition of claim 63 , wherein the active agent is a nucleic acid selected from the group consisting of an siRNA, a microRNA, and an antisense oligonucleotide, and wherein the siRNA, microRNA, or antisense oligonucleotide includes a polynucleotide that specifically binds to a polynucleotide that encodes the polypeptide, or a complement thereof.
67 . A method of treating a disease or disorder characterized by underexpression of a polypeptide in a subject, comprising providing to the subject the pharmaceutical composition of claim 63 , wherein the active agent is a plasmid that encodes the polypeptide or a functional variant or fragment thereof.
68 . A method of inducing an immune response in a subject, comprising providing to the subject the pharmaceutical composition of claim 63 , wherein the active agent is an immunostimulatory oligonucleotide.