Lipid nanoparticles and methods of use
Provided herein are, inter alia, lipid nanoparticles, lipid nanoparticles comprising nucleic acids encapsulated therein, pharmaceutical compositions comprising lipid nanoparticles which comprise nucleic acids encapsulated therein, and methods of treating diseases, such as coronavirus infections (e.g., SARS-CoV-2, SARS-CoV-2, MERS-CoV), COVID, and MERS.
1 . A lipid nanoparticle comprising:
(i) about 36 mole % to about 44 mole % of a cationic lipid; wherein the cationic lipid is N,N-dioleyl-N,N-dimethylammonium chloride (DODAC), 1,2-dioleyloxy-N,N-dimethylaminopropane (DODMA), 1,2-distearyloxy-N,N-dimethylaminopropane (DSDMA), N-(1-(2,3-dioleyloxy)propyl)-N,N,N-trimethylammonium chloride (DOTMA), N,N-distearyl-N,N-dimethylammonium bromide (DDAB), 1,2-dioleoyloxy-3-(trimethylammonium)propane (DOTAP), 3-(N—(N′,N′-dimethylaminoethane)-carbamoyl)cholesterol (DC-Chol), N-(1,2-dimyristyloxyprop-3-yl)-N,N-dimethyl-N-hydroxyethyl ammonium bromide (DMRIE), 2,3-dioleyloxy-N-[2 (spermine-carboxamido)ethyl]-N,N-dimethyl-1-propanaminium-trifluoroacetate (DOSPA), dioctadecylamidoglycyl spermine (DOGS), N,N-dimethyl-3,4-dioleyloxybenzylamine (DMOBA), 1,2-N,N′-dioleylcarbamyl-3-dimethylaminopropane (DOcarbDAP), 3-(N,N-dioleylamino)-1,2-propanedio (DOAP), or a mixture of two or more thereof;
(ii) about 21 mole % to about 29 mole % of a dilinoleic cationic lipid; wherein the dilinoleic cationic lipid is dilinoleyl-methyl-4-dimethylaminobutyrate (MC3), 1,2-dilinoleyloxy-N,N-dimethylaminopropane (DLinDMA), 2,2-dilinoleyl-4-(2-dimethylaminoethyl)-[1,3]-dioxolane (DLin-KC2-DMA), 2,2-dilinoleyl-4-(3-dimethylaminopropyl)-[1,3]-dioxolane (DLin-K-3-DMA), 2,2-dilinoleyl-4-(4-dimethylaminobutyl)-[1,3]-dioxolane (DLin-KC4-DMA), 2,2-dilinoleyl-5-dimethylaminomethyl-[1,3]-dioxane (DLin-K6-DMA), 2,2-dilinoleyl-4-N-methylpepiazino-[1,3]-dioxolane (DLin-K-MPZ), 2,2-dilinoleyl-4-dimethylaminomethyl-[1,3]-dioxolane (DLin-K-DMA), 1,2-dilinoleylcarbamoyloxy-3-dimethylaminopropane (DLin-C-DAP), 1,2-dilinoleyoxy-3-(dimethylamino)acetoxypropane (DLin-DAC), 1,2-dilinoleyoxy-3-morpholinopropane (DLin-MA), 1,2-dilinoleoyl-3-dimethylaminopropane (DLinDAP), 1,2-dilinoleylthio-3-dimethylaminopropane (DLin-S-DMA), 1-linoleoyl-2-linoleyloxy-3-dimethylaminopropane (DLin-2-DMAP), 1,2-dilinoleyloxy-3-trimethylaminopropane chloride salt (DLin-TMA.Cl), 1,2-dilinoleoyl-3-trimethylaminopropane chloride salt (DLin-TAP.Cl), 1,2-dilinoleyloxy-3-(N-methylpiperazino)propane (DLin-MPZ), 3-(N,N-dilinoleylamino)-1,2-propanediol (DLinAP), 3-(N,N-dioleylamino)-1,2-propanedio (DOAP), 1,2-dilinoleyloxo-3-(2-N,N-dimethylamino)ethoxypropane (DLin-EG-DMA), 1,2-N,N′-dilinoleylcarbamyl-3-dimethylaminopropane (DLincarbDAP), or a combination of two or more thereof;
(iii) about 6 mole % to about 14 mole % of a phospholipid; wherein the phospholipid is dipalmitoylphosphatidylcholine (DPPC), distearoylphosphatidylcholine (DSPC), dioleoylphosphatidylethanolamine (DOPE), palmitoyloleoyl-phosphatidylcholine (POPC), palmitoyloleoyl-phosphatidylethanolamine (POPE), palmitoyloleyol-phosphatidylglycerol (POPG), dipalmitoyl-phosphatidylethanolamine (DPPE), dimyristoyl-phosphatidylethanolamine (DMPE), distearoyl-phosphatidylethanolamine (DSPE), monomethyl-phosphatidylethanolamine (MMPE), dimethyl-phosphatidylethanolamine (DMPE), dielaidoyl-phosphatidylethanolamine (DEPE), stearoyloleoyl-phosphatidylethanolamine (SOPE), egg phosphatidylcholine (EPC), hydrogenated soy phosphatidylcholine (HSPC), or a mixture of two or more thereof;
(iv) about 18 mole % to 26 mole % of a sterol; wherein the sterol is cholesterol, cholestanol, cholestanone, cholestenone, coprostanol, cholesteryl-2′-hydroxyethyl ether, cholesteryl-4′-hydroxybutyl ether, or a mixture of two or more thereof; and
(v) about 1 mole % to about 4 mole % of a polyethylene glycol-lipid conjugate; wherein polyethylene glycol-lipid conjugate is N-palmitoyl-sphingosine-1-{succinyl[methoxy(polyethylene glycol)]} (C16 PEG ceramide), 1,2-dimyristoyl-rac-glycero-3-methoxypolyethylene glycol (DMG-PEG), 1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine-N-[azido(polyethylene glycol) (DPPE-PEG), 1,2-dipalmitoyl-rac-glycero-3-methylpolyoxyethylene (DPG-PEG), distearoyl-rac-glycerol (polyethylene glycol) (DSG-PEG), 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[amino(polyethylene glycol) (DSPE-PEG), or a mixture of two or more thereof; wherein the polyethylene glycol has an average molecular weight from about 1,000 Daltons to about 6,000 Daltons.
2 . The lipid nanoparticle of claim 1 , comprising:
(i) about 38 mole % to about 42 mole % of the cationic lipid;
(ii) about 23 mole % to about 27 mole % of the dilinoleic cationic lipid;
(iii) about 8 mole % to about 12 mole % of the phospholipid;
(iv) about 20 mole % to 24 mole % of the sterol; and
(v) about 2 mole % to about 4 mole % of the polyethylene glycol-lipid conjugate.
3 . The lipid nanoparticle of claim 1 , comprising:
(i) about 40 mole % of the cationic lipid;
(ii) about 25 mole % of the dilinoleic cationic lipid;
(iii) about 10 mole % of the phospholipid;
(iv) about 22 mole % of the sterol; and
(v) about 3 mole % of the polyethylene glycol-lipid conjugate.
4 . The lipid nanoparticle of claim 1 , wherein:
(i) the cationic lipid is DOTAP;
(ii) the dilinoleic cationic lipid is MC3;
(iii) the phospholipid is DSPC, DPPC, DOPE, POPC, POPE, POPG, DPPE, DMPE, DSPE, MMPE, DMPE, DEPE, SOPE, EPC, or HSPC;
(iv) the sterol is cholesterol, cholestanol, cholestanone, cholestenone, coprostanol, cholesteryl-2′-hydroxyethyl ether, or cholesteryl-4′-hydroxybutyl ether;
(v) the polyethylene glycol-lipid conjugate is C16 PEG ceramide, DMG-PEG, DPPE-PEG, DPG-PEG, DSG-PEG, or DSPE-PEG, wherein the polyethylene glycol has an average molecular weight from about 1,000 Daltons to about 6,000 Daltons.
5 . The lipid nanoparticle of claim 4 , wherein the polyethylene glycol-lipid conjugate is C16 PEG ceramide.
6 . The lipid nanoparticle of claim 1 , wherein
(i) the cationic lipid is DOTAP;
(ii) the dilinoleic cationic lipid is MC3;
(iii) the phospholipid is DSPC;
(iv) the sterol is cholesterol; and
(v) the polyethylene glycol-conjugated lipid is C16 PEG2000 ceramide.
7 . The lipid nanoparticle of claim 1 , further comprising a nucleic acid encapsulated within the lipid nanoparticle.
8 . The lipid nanoparticle of claim 7 , wherein the nucleic acid is RNA or DNA.
9 . The lipid nanoparticle of claim 8 , wherein the RNA is siRNA or mRNA, and wherein the DNA is a plasmid or a minigene.
10 . The lipid nanoparticle of claim 7 , wherein the nucleic acid comprises 15 nucleotides to 30 nucleotides and is capable of hybridizing to SEQ ID NO:57, SEQ ID NO: 51, SEQ ID NO:63, SEQ ID NO:55, SEQ ID NO:45, SEQ ID NO:50, or SEQ ID NO:62.
11 . The lipid nanoparticle of claim 7 , wherein the nucleic acid comprises SEQ ID NO: 1 hybridized to SEQ ID NO:3; SEQ ID NO:2 hybridized to SEQ ID NO: 4; SEQ ID NO:9 hybridized to SEQ ID NO: 10; SEQ ID NO:11 hybridized to SEQ ID NO: 12; SEQ ID NO:5 hybridized to SEQ ID NO:7; SEQ ID NO:6 hybridized to SEQ ID NO:8; SEQ ID NO:13 hybridized to SEQ ID NO:14; SEQ ID NO:15 hybridized to SEQ ID NO:16; SEQ ID NO:17 hybridized to SEQ ID NO:18; SEQ ID NO: 19 hybridized to SEQ ID NO:20; SEQ ID NO:21 hybridized to SEQ ID NO:22; SEQ ID NO:23 hybridized to SEQ ID NO:24; SEQ ID NO:25 hybridized to SEQ ID NO:26; SEQ ID NO:27 hybridized to SEQ ID NO:28; SEQ ID NO:29 hybridized to SEQ ID NO:30; SEQ ID NO:31 hybridized to SEQ ID NO:32; SEQ ID NO:33 hybridized to SEQ ID NO:34; SEQ ID NO:35 hybridized to SEQ ID NO:36; SEQ ID NO:37 hybridized to SEQ ID NO:38; SEQ ID NO:39 hybridized to SEQ ID NO:40; SEQ ID NO:41 hybridized to SEQ ID NO:42; SEQ ID NO:43 hybridized to SEQ ID NO:44; or a combination of two or more of the foregoing.
12 . A plurality of the lipid nanoparticles of claim 1 .
13 . The lipid nanoparticle of claim 12 , wherein a plurality of the lipid nanoparticles have an average size from about 50 nm to about 110 nm.
14 . The lipid nanoparticle of claim 12 , wherein a plurality of the lipid nanoparticles have a zeta potential from about 10 mV to about 30 mV.
15 . The lipid nanoparticle of claim 12 , wherein a plurality of the lipid nanoparticles have a polydispersity from about 0.12 to 0.2.
16 . A pharmaceutical composition comprising the lipid nanoparticle of claim 1 and a pharmaceutically acceptable excipient.
17 . A method for administering a lipid nanoparticle to a lung of a patient in need thereof, the method comprising administering to the patient the lipid nanoparticle of claim 1 .
18 . A method for treating a SARS coronavirus infection in a patient in need thereof, the method comprising administering to the patient the lipid nanoparticle of claim 1 .
19 . A lipid nanoparticle comprising:
(i) about 38 mole % to about 42 mole % of 1,2-dioleoyloxy-3-(trimethylammonium)propane (DOTAP);
(ii) about 23 mole % to about 27 mole % of dilinoleyl-methyl-4-dimethylaminobutyrate (MC3);
(iii) about 6 mole % to about 14 mole % of distearoylphosphatidylcholine (DSPC);
(iv) about 18 mole % to about 26 mole % of cholesterol; and
(v) about 1 mole % to about 4 mole % of N-palmitoyl-sphingosine-1-{succinyl[methoxy(polyethylene glycol)]}, wherein the polyethylene glycol has an average molecular weight of about 2,000 Daltons.
20 . A plurality of the lipid nanoparticle of claim 19 ; wherein the plurality of the lipid nanoparticles have an average size from about 50 nm to about 110 nm, a zeta potential from about 10 mV to about 30 mV, and a polydispersity from about 0.12 to 0.2.