METHODS, COMPOSITIONS, AND SYSTEMS FOR DELIVERING THERAPEUTIC AND DIAGNOSTIC AGENTS INTO CELLS
Disclosed are methods for delivering a therapeutic or diagnostic agent to the cytosol of a cell in a subject. The disclosed methods generally include administering to the subject an effective amount of a lipid nanoparticle comprising the therapeutic or diagnostic agent and an effective amount of a membrane-destabilizing polymer. Also disclosed are related compositions and delivery systems.
1 - 272 . (canceled)
273 . A pH-sensitive, membrane-destabilizing polymer of formula V:
T1-L-[PEGMA m -M2 n ] v -[DMAEMA 1 -PAA r -BMA s ] w V
wherein
PEGMA is polyethyleneglycol methacrylate residue with 2-20 ethylene glycol units;
M2 is a methacrylate residue selected from the group consisting of
a (C4-C18)alkyl-methacrylate residue;
a (C4-C18)branched alkyl-methacrylate residue;
a cholesteryl methacrylate residue;
a (C4-C18)alkyl-methacrylate residue substituted with one or more fluorine atoms; and
a (C4-C18)branched alkyl-methacrylate residue substituted with one or more fluorine atoms;
BMA is butyl methacrylate residue;
PAA is propyl acrylic acid residue;
DMAEMA is dimethylaminoethyl methacrylate residue;
m and n are each a mole fraction greater than 0, wherein m is greater than n and m+n=1;
q is a mole fraction of 0.2 to 0.75;
r is a mole fraction of 0.05 to 0.6;
s is a mole fraction of 0.2 to 0.75;
q+r+S=1;
v is 1 to 25 kDa;
w is 1 to 25 kDa;
T1 is a first targeting ligand comprising an N-acetylgalactosamine (NAF) residue; and
L is a linking moiety comprising polyethylene glycol (PEG) moiety.
274 . The polymer of claim 273 , wherein M2 is a methacrylate residue selected from the group consisting of
a (C4-C18)alkyl-methacrylate residue;
a (C4-C18)branched alkyl-methacrylate residue; and
a (C4-C18)alkyl-methacrylate residue substituted with one or more fluorine atoms.
275 . The polymer of claim 274 , wherein M2 is a (C4-C18)alkyl-methacrylate residue.
276 . The polymer of claim 273 , wherein M2 is a methacrylate residue selected from the group consisting of 2,2,3,3,4,4,4-heptafluorobutyl methacrylate residue; 3,3,4,4,5,6,6,6-octafluoro-5(trifluoromethyl)hexyl methacrylate residue; 2,2,3,3,4,4,5,5,6,6,7,7,8,8,8-pentadecafluorooctyl 2-methylacrylate residue; 3,3,4,4,5,5,6,6,6-nonafluorohexyl methacrylate residue; 3,3,4,4,5,5,6,6,7,7,8,8,8-tridecafluorooctyl methacrylate residue; 2-ethylhexyl methacrylate residue; octyl methacrylate residue; n-decyl methacrylate residue; lauryl methacrylate residue; myristyl methacrylate residue; stearyl methacrylate residue; cholesteryl methacrylate residue; neopentyl methacrylate residue; 5-nonyl methacrylate residue; 2-butyl-1-octyl methacrylate residue; and 2-hexyl-1-decyl methacrylate residue.
277 . The polymer of claim 276 , wherein M2 is a methacrylate residue selected from the group consisting of 2-ethylhexyl methacrylate residue; 5-nonyl methacrylate residue; octyl methacrylate residue; n-decyl methacrylate residue; lauryl methacrylate residue; myristyl methacrylate residue; and stearyl methacrylate residue.
278 . The polymer of claim 277 , wherein M2 is 2-ethylhexyl methacrylate residue or 5-nonyl methacrylate residue.
279 . The polymer of claim 273 , wherein PEGMA is polyethyleneglycol methacrylate residue having 4-5 ethylene glycol units or 7-8 ethylene glycol units.
280 . The polymer of claim 273 , wherein L includes a polyethylene glycol (PEG) moiety having 2-20 ethylene glycol units.
281 . The polymer of claim 273 , wherein T1-L- in formula V comprises the structure
282 . The polymer of claim 273 , wherein the polymer of formula V is selected from the group consisting of
NAG-PEG 12 -[PEGMA300 m -(F1-BMA) n ] v -[D q -P r -B s ] w Vb;
NAG-PEG 12 -[PEGMA300 m -(OF1-5TFM-HMA) n ] v -[D q -P r -B s ] w Vc;
NAG-PEG 12 -[PEGMA300 m -(F115-OMA) n ] v -[D q -P r -B s ] w Vd;
NAG-PEG 12 -[PEGMA300 m -(B-F1-HMA) n ] v -[D q -P r -B s ] w Ve;
NAG-PEG 12 -[PEGMA300 m -(B-F1-OMA) n ] v -[D q -P r -B s ] w Vf;
NAG-PEG 12 -[PEGMA300 m -EHMA n ] v -[D q -P r -B s ] w Vg;
NAG-PEG 12 -[PEGMA300 m -B n ] v -[D q -P r -B s ] w Vh;
NAG-PEG 12 -[PEGMA300 m -C8MA n ] v -[D q -P r -B s ] w Vj;
NAG-PEG 12 -[PEGMA300 m -C12MA n ] v -[D q -P r -B s ] w Vk;
NAG-PEG 12 -[PEGMA300 m -Bu1-OMA n ] v -[D q -P r -B s ] w Vl; and
NAG-PEG 12 -[PEGMA300 m -NMA n ] v -[D q -P r -B s ] w Vm;
wherein
D is DMAEMA;
P is PAA;
B is BMA;
NAG is an N-acetylgalactosamine residue;
PEG 12 is a linking moiety comprising polyethylene glycol having 12 ethylene glycol units;
F1-BMA is 2,2,3,3,4,4,4-heptafluorobutyl methacrylate residue;
OF1-5TFM-HMA is 3,3,4,4,5,6,6,6-octafluoro-5(trifluoromethyl)hexyl methacrylate residue;
F115-OMA is 2,2,3,3,4,4,5,5,6,6,7,7,8,8,8-pentadecafluorooctyl 2-methylacrylate residue;
B-F1-HMA is 3,3,4,4,5,5,6,6,6-nonafluorohexyl methacrylate residue;
B-F1-OMA is 3,3,4,4,5,5,6,6,7,7,8,8,8-tridecafluorooctyl methacrylate residue;
EHMA is 2-ethylhexyl methacrylate residue;
CSMA is octyl methacrylate residue;
C12MA is lauryl methacrylate residue;
2-Bu1-OMA is 2-butyl-1-octyl methacrylate residue;
5-NMA is 5-nonyl methacrylate residue;
m and n are each a mole fraction greater than 0, wherein m is greater than n and m+n=1;
q is a mole fraction of 0.2 to 0.75;
r is a mole fraction of 0.05 to 0.6;
s is a mole fraction of 0.2 to 0.75;
q+r+s=1;
v is 1 to 25 kDa; and
w is 1 to 25 kDa.
283 . The polymer of claim 282 , wherein m is 0.7 to 0.85.
284 . The polymer of claim 282 , wherein n is 0.15 to 0.3.
285 . The polymer of claim 282 , wherein s is 0.5 to 0.65.
286 . The polymer of claim 282 , wherein v is 2.5 to 7 kDa.
287 . The polymer of claim 282 , wherein w is 4 to 7 kDa.
288 . The polymer of claim 282 , wherein q is 0.25 to 0.4.
289 . The polymer of claim 282 , wherein r is 0.07 to 0.15.
290 . The polymer of claim 273 , wherein the polymer of Formula V is selected from the group consisting of
P79: NAG-C5N-PEG12-[PEGMA (300, 69.9%)-HMA(30.1%)] 2.93 KDa-b-[DMAEMA (34.4%)-BMA (53.6%)-PAA (12%)]4.43 Kda;
P90: NAG-C5N-PEG12-[PEGMA (300, 72.6%)-HMA (27.4%)]3.58 KDa-b-[DMAEMA (30.6%)-BMA(56.2%)-PAA (13.3%)]5.6 KDa;
P106: NAG-C5N-PEG12-[PEGMA(300, 74%)-HMA(26%)]4.1 KDa-b-[DMAEMA(31%)-BMA(56%)-PAA(13%)]5 KDa;
P107: NAG-C5N-PEG12-[PEGMA(300, 74%)-HMA(26%)]4.1 KDa-b-[DMAEMA(31%)-BMA(56%)-PAA(13%)]4.2 KDa;
P108: NAG-C5N-PEG12-[PEGMA(300, 80%)-HMA(20%)]4.96 KDa-b-[DMAEMA(31%)-BMA(56%)-PAA(13%)]5.5 KDa;
P109: NAG-C5N-PEG12-[PEGMA(300, 80%)-HMA(20%)]4.96 KDa-b-[DMAEMA(31%)-BMA(56%)-PAA(13%)]6.5 KDa;
P114: NAG-C5N-PEG12-[PEGMA(300, 67%)-HMA(33%)]5.7 KDa-b-[DMAEMA(31%)-BMA(56%)-PAA(13%)]6.15 KDa;
P115: NAG-C5N-PEG12-[PEGMA(300, 67%)-HMA(33%)]5.7 KDa-b-[DMAEMA(31%)-BMA(56%)-PAA(13%)]6 KDa;
P118: NAG-C5N-PEG12-[PEGMA(300, 70%)-HMA(30%)]5.2 KDa-b-[DMAEMA(31%)-BMA(56%)-PAA(13%)]5.7 KDa;
P119: NAG-C5N-PEG12-[PEGMA(300, 70%)-HMA(30%)]5.2 KDa-b-[DMAEMA(31%)-BMA(56%)-PAA(13%)]5 KDa;
P124: NAG-C5N-PEG12-[PEGMA(300, 74%)-HMA(26%)]4.15 KDa-b-[DMAEMA(31%)-BMA(56%)-PAA(13%)]5 KDa; and
P125: NAG-C5N-PEG12-[PEGMA(300, 74%)-HMA(26%)]4.15 KDa-b-[DMAEMA(31%)-BMA(56%)-PAA(13%)]5 KDa.
291 . The polymer of claim 273 , wherein the polymer of Formula V is selected from the group consisting of
P74: NAG-C5N-PEG 12-[PEGMA(300, 80.3%)-BMA(23.3%)]3.8 KDa-b-[DMAEMA(38.2%)-BMA(51.5%)-PAA(10.3%)]3.5 KDa;
P89: NAG-C5N-PEG12-[PEGMA (300, 73.8%)-BMA (26.2%)]3.5 KDa-b-[DMAEMA (30.7%)-BMA(58.9%)-PAA (10.4%)]4.9 KDa;
P80: NAG-C5N-PEG12-[PEGMA (300, 85.4%)-EHMA(14.6%)] 3.36 KDa-b-[DMAEMA (36.5%)-BMA (53.7%)-PAA (9.7%)]4.18 KDa;
P87: NAG-C5N-PEG12-[PEGMA (300, 69.7%)-EHMA(30.3%)] 3.9 KDa-b-[DMAEMA (31.1%)-BMA(56.7%)-PAA (12.1%)]5.1 KDa;
P110: NAG-C5N-PEG12-[PEGMA(300, 77.7%)-EHMA(22.3%)]4.37 KDa-b-[DMAEMA (31%)-BMA(56%)-PAA(13%)]6 KDa;
P88: NAG-C5N-PEG12-[PEGMA (300, 76%)-5-NMA (24%)] 3.0 KDa-b-[DMAEMA (34.4%)-BMA (54%)-PAA (11.6%)]5.6 KDa;
P99: NAG-C5N-PEG12-[PEGMA(300, 73.3%)-5-NMA(26.7%)]4.05 KDa-b-[DMAEMA(31.5%)-BMA(55.2%)-PAA(13.3%)]5.20 KDa;
P76: NAG-C5N-PEG12-[PEGMA(300, 74.9%)-isoA-MA(25.1%)]2.9 KDa-b-[DMAEMA(38%)-BMA(53%)-PAA(9.1%)]5.2 KDa;
P86: NAG-C5N-PEG12-[PEGMA (300, 78.1%)-C12MA(21.9%)]3.67 KDa-b-[DMAEMA (32.1%)-BMA (53.7%)-PAA (14.2%)]4.7 KDa;
P98: NAG-C5N-PEG12-[PEGMA(300, 75%)-2-Bul-OMA(25%]4.26 KDa-b-[DMAEMA(32.1%)-BMA(55.7%)-PAA(12.2%)]5.69 KDa;
P123: NAG-C5N-PEG12-[PEGMA(300, 79%)-Bul-O-MA(21%)]4.88 KDa-b-[DMAEMA(31%)-BMA(56%)-PAA(13%)]4.6 KDa;
P94: NAG-C5N-PEG12-[PEGMA(300, 72.2%)-B-F1-HMA (27.8%)]4.2 KDa-b-[DMAEMA(35.7%)-BMA(54.4%)-PAA (9.9%)]4.7 KDa;
P103: NAG-C5N-PEG12-[PEGMA(300, 70.3%)-F1-BMA(29.7%)]3.6 KDa-b-[DMAEMA(32.2%)-BMA(57.6%)-PAA(10.2%)]5 KDa; and
P104: NAG-C5N-PEG12-[PEGMA(300, 68%)-F1-BMA(32%)]3.7 KDa-b-[DMAEMA(31%)-BMA(56%)-PAA(13%)]5.3 KDa.