KR20240066978A - Lipid nanoparticle formulation for transpulmonary delivery of nucleic acid drugs and uses thereof - Google Patents

Abstract

The present invention relates to a composition for transpulmonary delivery of nucleic acid drugs and its use. The composition for transpulmonary delivery of the present invention is specific for lung tissue and its specific cells, has excellent biocompatibility, and can deliver gene therapeutics, etc. with high efficiency. It can be usefully used in related technical fields such as treatment of lipid nanoparticle-mediated lung diseases.

Description

Lipid nanoparticle formulation for transpulmonary delivery of nucleic acid drugs and uses thereof}

The present invention relates to compositions for transpulmonary delivery of nucleic acid drugs and uses thereof.

Lung fibrosis is an irreversible chronic disease based on excessive production of extracellular matrix caused by overexpressed inflammatory activity in environmentally or chemically damaged lung tissue. Abnormal accumulation of fibrous tissue narrows the airways and thickens the epithelium in the alveoli, reducing the oxygen supply to the blood and causing serious breathing problems.

The incidence of pulmonary fibrosis is increasing, but current treatment methods are very limited, and there is an urgent need to uncover its pathophysiological mechanisms and develop new treatments. Recently, in a radiation-induced lung fibrosis model, Gtse1 (G2 and S-phase expressed 1) was found to be associated with lung fibrosis, meaning that Gtse1 and its downstream genes may be potential targets for the treatment of the disease. H Jin et al., J Mol Med., 2019, Jan, 97(1):37-47).

Nucleic acids such as antisense RNA and siRNA are substances that can inhibit the expression of specific proteins in vivo, and are attracting attention as important tools in the treatment of cancer, genetic diseases, infectious diseases, and autoimmune diseases (Novina and Sharp, Nature, 430, 161-164, 2004). However, since nucleic acids such as siRNA are difficult to deliver directly into cells and are easily degraded by enzymes in the blood, many studies are being conducted to overcome this problem.

Drug Delivery System (DDS) is a technology designed to efficiently deliver the required amount of drugs by reducing the side effects of drugs and maximizing their efficacy and effectiveness. In particular, conventional viral carriers have proven to be effective as drug carriers in gene therapy, but the use of viruses as a gene delivery system has been discouraged due to several drawbacks such as immunogenicity, limitations in the size of the injected DNA, and difficulties in mass production. It is being restricted.

Accordingly, as an alternative to the viral system, the method of transporting nucleic acids into cells is currently mixing them with positively charged lipids or polymers (named lipid-DNA conjugates (lipoplex) and polymer-DNA conjugates (polyplex), respectively). ) is mainly used (Hirko et al., Curr, Med, Chem., 10, 1185-1193, 2003; Merdan et al., Adv. Drug. Deliv.Rev., 54, 715-758, 2002; Spagnou et al. al., Biochemistry, 43, 13348-13386, 2004). In particular, lipid-DNA conjugates are widely used at the cellular level because they bind to nucleic acids and deliver nucleic acids well into cells, but when injected locally in vivo, they often cause inflammation in the body (Filonand and Phillips, Biochim. Biophys/Acta , 1329, 345-356, 1997), it has the disadvantage of accumulating in tissues such as the lung, liver, and spleen, which are the first passage organs during intravascular injection (Ren et al., Gene Therapy. 7, 764-768, 2000 ).

Under this background, the present inventors have made diligent efforts to develop a carrier that can efficiently deliver anionic drugs such as nucleic acids to lung organs or cells in order to treat pulmonary fibrosis, etc., and as a result, the lipid nanoparticle formulation containing mannose of the present invention has been developed. The present invention was completed by confirming the excellent drug delivery effect.

One object of the present invention is to provide a composition for transpulmonary delivery of a drug containing lipid nanoparticles containing a mannose-PEG (polyethyleneglycol)-lipid conjugate.

Another object of the present invention is to provide a pharmaceutical composition for preventing or treating lung diseases containing the lipid nanoparticles and anionic drugs.

This is explained in detail as follows. Meanwhile, each description and embodiment disclosed in the present invention may also be applied to each other description and embodiment. That is, all combinations of the various elements disclosed in the present invention fall within the scope of the present invention. Additionally, the scope of the present invention cannot be considered limited by the specific description described below.

As one aspect for achieving the above object, the present invention provides a composition for transpulmonary delivery of a drug containing lipid nanoparticles containing a mannose-PEG-lipid conjugate.

Lipid nanoparticles containing the mannose-PEG-lipid conjugate of the present invention are specifically delivered to lung tissue, specifically fibrotic lungs or their cells (e.g., lung epithelial cells, endothelial cells, etc.), and thus are delivered with high efficiency. Anionic drugs can be delivered specifically to fibrotic lungs. That is, the lipid nanoparticles of the present invention may specifically target fibrotic lung tissue, and may specifically target lung epithelial cells and/or endothelial cells of fibrotic lung tissue.

In the present invention, the term "targeting" may mean internalization within the tissue or cell, and may also mean penetration into the nuclear membrane and internalization within the nucleus.

The composition of the present invention can be delivered topically within the lung, specifically to fibrotic lungs.

In the present invention, "mannose-PEG-lipid conjugate" refers to a form in which mannose, polyethylene glycol (PEG), and lipid are conjugated, with mannose at one end of the lipid and mannose at the other end. may refer to a lipid to which PEG is bound.

The mannose-PEG-lipid conjugate may be taken together with other PEG-lipids to form a mixture of PEG-lipid conjugates, wherein a "PEG-lipid conjugate mixture" It may refer to a mixture containing a lipid conjugate. The PEG-lipid may be, for example, PEG-ceramide, PEG-DMG, PEG-c-DOMG, PEG-DLPE, PEG-DMPE, PEG-DPPC, PEG-DSPE, or a mixture thereof, but is not limited thereto.

The PEG-lipid conjugate mixture contributes to the stability of nanoparticles in serum within lipid nanoparticles, prevents aggregation between nanoparticles, and can play a role in increasing the targeting effect to lung tissue or specific cells. . In addition, the PEG-lipid conjugate mixture protects nucleic acids from degrading enzymes when delivering anionic drugs such as nucleic acids in vivo, enhances the stability of the drug in the body, and can increase the half-life of the drug encapsulated in nanoparticles.

In the PEG-lipid conjugate or mannose-PEG-lipid conjugate, PEG or mannose may be conjugated directly to the lipid or may be connected to the lipid through a linker moiety. Any linker moiety suitable for linking PEG to a lipid can be used, including, for example, ester-free linker moieties and ester-containing linker moieties. The ester-free linker moiety is amido (-C(O)NH-), amino (-NR-), carbonyl (-C(O)-), carbamate (-NHC(O)O- ), urea (-NHC(O)NH-), disulfide (-SS-), ether (-O-), succinyl (-(O)CCH ₂ CH ₂ C(O)-), succinamidyl ( -NHC(O)CH ₂ CH ₂ C(O)NH-), ethers, disulfides, as well as combinations thereof (e.g., linkers containing both carbamate linker moieties and amido linker moieties). However, it is not limited to this. The ester-containing linker moiety includes, for example, carbonate (-OC(O)O-), succinoyl, phosphate ester (-O-(O)POH-O-), sulfonate ester and their Including but not limited to combinations.

The average molecular weight of the PEG-lipid conjugate mixture (e.g., mannose-PEG lipid conjugate and/or PEG-ceramide conjugate) is 100 to 10,000 daltons, 200 to 10,000 daltons, 500 to 10,000 daltons, 1,000 to 10,000 daltons, 1,500 to 10,000 daltons. , 2000 to 10000 daltons, 100 to 7500 daltons, 200 to 7500 daltons, 500 to 7500 daltons, 1000 to 7500 daltons, 1500 to 7500 daltons, 2000 to 7500 daltons, 100 to 5000 daltons, 200 to 5000 daltons , 500 to 5000 daltons , 1000 to 5000 daltons, 1500 to 5000 daltons, 2000 to 5000 daltons, 100 to 3000 daltons, 200 to 3000 daltons, 500 to 3000 daltons, 1000 to 3000 daltons, 1500 to 3000 daltons, 2000 to 3000 daltons ton, 100 to 2600 daltons , 200 to 2600 daltons, 500 to 2600 daltons, 1000 to 2600 daltons, 1500 to 2600 daltons, 2000 to 2600 daltons, 100 to 2500 daltons, 200 to 2500 daltons, 500 to 2500 daltons, 1000 to 2500 daltons, 1500 to 2500 daltons , or 2000 to 2500 daltons.

The lipid in the PEG-lipid conjugate can be used without limitation as long as it is a lipid that can bind to polyethylene glycol, and specifically includes ceramide, dimyristoylglycerol (DMG), succinoyl-diacylglycerol, It may be, but is not limited to, s-DAG), distearoylphosphatidylcholine (DSPC), distearoylphosphatidylethanolamine (DSPE), or cholesterol.

In addition to the mannose-PEG-lipid conjugate, the PEG-lipid conjugate mixture is bound to phospholipids such as PEG (PEG-DAA) bound to dialkyloxypropyl, PEG (PEG-DAG) bound to diacylglycerol, and phosphatidylethanolamine. PEG (PEG-PE), PEG conjugated to ceramide (PEG-CER, or ceramide-PEG conjugate), PEG conjugated to cholesterol or its derivatives, PEG-c-DOMG, PEG-DMG, PEG-DLPE, PEG- It may include DMPE, PEG-DPPC, PEG-DSPE, and mixtures thereof, and specifically may include C16-PEG2000 ceramide, DMG-PEG 2000, and 14:0 PEG2000 PE. According to one embodiment of the present invention, PEG -ceramide conjugates and/or mannose-PEG-lipid conjugates (e.g., mannose-PEG-DSPE conjugates).

The PEG-lipid conjugate mixture is 0.1 to 15 mol%, 0.25 to 15 mol%, 0.5 to 15 mol%, 1 to 15 mol%, 1.5 to 15 mol%, 2 to 15 mol%, 2.5 to 15 mol%, 3 to 15 mol%, 0.1 to 10 mol%, 0.25 to 10 mol%, 0.5 to 10 mol%, 1 to 10 mol%, 1.5 to 10 mol%, 2 to 10 mol%, 2.5 to 10 mol%, 3 to 10 Mol%, 0.1 to 9 mol%, 0.25 to 9 mol%, 0.5 to 9 mol%, 1 to 9 mol%, 1.5 to 9 mol%, 2 to 9 mol%, 2.5 to 9 mol%, 3 to 9 mol% , 0.1 to 7.5 mol%, 0.25 to 7.5 mol%, 0.5 to 7.5 mol%, 1 to 7.5 mol%, 1.5 to 7.5 mol%, 2 to 7.5 mol%, 2.5 to 7.5 mol%, 3 to 7.5 mol%, 0.1 to 5 mol%, 0.25 to 5 mol%, 0.5 to 5 mol%, 1 to 5 mol%, 1.5 to 5 mol%, 2 to 5 mol%, 2.5 to 5 mol%, 3 to 5 mol%, 0.1 to 4.5 Mol%, 0.25 to 4.5 mol%, 0.5 to 4.5 mol%, 1 to 4.5 mol%, 1.5 to 4.5 mol%, 2 to 4.5 mol%, 2.5 to 4.5 mol%, 3 to 4.5 mol%, 0.1 to 4 mol% , 0.25 to 4 mol%, 0.5 to 4 mol%, 1 to 4 mol%, 1.5 to 4 mol%, 2 to 4 mol%, 2.5 to 4 mol%, 3 to 4 mol%, 0.1 to 3.5 mol%, 0.25 to 3.5 mol%, 0.5 to 3.5 mol%, 1 to 3.5 mol%, 1.5 to 3.5 mol%, 2 to 3.5 mol%, 2.5 to 3.5 mol%, 3 to 3.5 mol%, 0.1 to 3 mol%, 0.25 to 3 It may be included in the lipid nanoparticles at mol%, 0.5 to 3 mol%, 1 to 3 mol%, 1.5 to 3 mol%, 2 to 3 mol%, or 2.5 to 3 mol%.

The mannose-PEG-lipid conjugate is 0.1 to 10 mol%, 0.25 to 10 mol%, 0.5 to 10 mol%, 1 to 10 mol%, 1.5 to 10 mol%, 2 to 10 mol%, 0.1 to 7.5 mol%, 0.25 to 7.5 mol%, 0.5 to 7.5 mol%, 1 to 7.5 mol%, 1.5 to 7.5 mol%, 2 to 7.5 mol%, 0.1 to 5 mol%, 0.25 to 5 mol%, 0.5 to 5 mol%, 1 to 5 mol%, 1.5 to 5 mol%, 2 to 5 mol%, 0.1 to 4.5 mol%, 0.25 to 4.5 mol%, 0.5 to 4.5 mol%, 1 to 4.5 mol%, 1.5 to 4.5 mol%, 2 to 4.5 mol% %, 0.1 to 4 mol%, 0.25 to 4 mol%, 0.5 to 4 mol%, 1 to 4 mol%, 1.5 to 4 mol%, 2 to 4 mol%, 0.1 to 3 mol%, 0.25 to 3 mol%, 0.5 to 3 mol%, 1 to 3 mol%, 1.5 to 3 mol%, 2 to 3 mol%, 0.1 to 2.5 mol%, 0.25 to 2.5 mol%, 0.5 to 2.5 mol%, 1 to 2.5 mol%, 1.5 to It may be included in the lipid nanoparticles at 2.5 mol%, 2 to 2.5 mol%, 0.1 to 2 mol%, 0.25 to 2 mol%, 0.5 to 2 mol%, 1 to 2 mol%, or 1.5 to 2 mol%. there is.

Lipid nanoparticles containing the mannose-PEG-lipid conjugate of the present invention may further include any one or more selected from the group consisting of ionizable lipids, phospholipids, and structural lipids.

In the present invention, "ionizable lipid" may refer to an amine-containing lipid that can be easily protonated, for example, a lipid whose charge state changes depending on the surrounding pH. The ionizable lipid may be positively charged at ambient pH below the pKa and may be progressively neutral at ambient pH above the pKa. When positively charged, the ionizable lipid can associate with a negatively charged drug (e.g., anionic drug and/or nucleic acid), and electrostatic interaction with the drug allows the drug to be incorporated into the lipid nanoparticle. It can play a role in ensuring sealing with high efficiency.

The ionizable lipid may be any known lipid that possesses a net positive charge at a selective pH, such as physiological pH, without limitation, such as 9-heptadecanyl 8-{(2-hydroxyethyl)[6-oxo-6- (undecyloxy)hexyl]amino}octanoate (SM-102), [(4-hydroxybutyl)azanediyl]di(hexane-6,1-diyl)bis(2-hexyldecanoate) ( ALC-0315), 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), N-(1-(2,3-dioleoyloxy)propyl)-N,N,N-trimethylammonium chloride (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 prefluoroacetate (DOSPA), dioctadecyl Amidoglycyl spermine (DOGS), 3-dimethylamino-2-(cholest-5-en-3-beta-oxybutane-4-oxy)-1-(cis,cis-9,12-octadecadi Enoxy)propane (CLinDMA), 2-[5'-(cholest-5-en-3β-doxy)-3'-oxapentoxy)-3-dimethyl-1-(cis,cis-9',1 -2'-octadecadienoxy)propane (CpLinDMA), N,N-dimethyl-3,4-dioleyloxybenzylamine (DMOBA), 1,2-N,N'-dioleylcarbamyl-3- Dimethylaminopropane (DOcarbDAP), 2,3-dilinoleoyloxy-N,N-dimethylpropylamine (DLinDAP), 1,2-N,N'-dilinoleylcarbamyl-3-dimethylaminopropane (DLincarbDAP) ), 1,2-dilinoleoylcarbamyl-3-dimethylaminopropane (DLinCDAP), 1,2-dilinoleyloxy-N,N-dimethylaminopropane (DLinDMA), 1,2-dilinoleyloxy -N,N-dimethylaminopropane (DLenDMA), heptatriaconta-6,9,28,31-tetraen-19-yl 4-(dimethylamino)butaoate (DLin-MC3-DMA), 1, 1',1'',1'''-[1,4-piperazinediylbis(3,1-propanediylnitrilo)]tetrakis-2-dodecanol (246-C10), (Z)- non-2-en-1-yl 9,9'-((2-(4-(9-(((Z)-non-2-en-1-yl)oxy)-9-oxononyl)pipe It may be razin-1-yl) ethyl) azanediyl) dinonanoate (244-cis) or a combination thereof.

The phospholipids can be any phospholipid that can promote the fusion of lipid nanoparticles without limitation, such as dioleoylphosphatidylethanolamine (DOPE), distearoylphosphatidylcholine (DSPC), and palmitoyloleoylphosphatidylcholine. (palmitoyloleoylphosphatidylcholine, POPC), egg phosphatidylcholine, EPC, dioleoylphosphatidylcholine, DOPC, dipalmitoylphosphatidylcholine, DPPC, dioleoylphosphatidylglycerol ylglycerol, DOPG), dipalmitoylphosphatidyl Glycerol (dipalmitoylphosphatidylglycerol, DPPG), distearoylphosphatidylethanolamine (DSPE), 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP) , Phosphatidylethanolamine (PE), dipalmitoylphosphatidylethanolamine, 1,2-dioleoyl-sn-glycero-3-phosphate (1,2-dioleoyl-sn-glycero-3-phosphate , 18-PA), 1,2-dilinoleoyl-sn-glycero-3-phosphocholine (1,2-dilinoleoyl-sn-glycero-3-phosphocholine), 1,2-diarachidonoyl- sn-glycero-3-phosphocholine (1,2-diarachidoyl-sn-glycero-3-phosphocholine), 1,2-didocosahexaenoyl-sn-glycero-3-phosphocholine (1,2 -didocosahexaenoyl-snglycero-3-phosphocholine), 1,2-diphytanoyl-sn-glycero-3-phosphoethanolamine (1,2-diphytanoyl-sn-glycero-3-phosphoethanolamine, ME 16:0 PE) , 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine (1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine, POPE), 1,2-dioleoyl- sn-glycero-3-[phospho-L-serine] (1,2-dioleoyl-sn-glycero-3-[phospho-L-serine], DOPS), 1,2-dilinoleoyl-sn- Glycero-3-phosphocholine (1,2-dilinoleoyl-sn-glycero-3-phosphocholine, DLPC), 1,2-dimyristoyl-sn-glycero-phosphocholine (1,2-dimyristoyl- sn-glycero-3-phosphocholine, DMPC), 1,2-diundecanoyl-sn-glycero-phosphocholine (1,2-diundecanoyl-sn-glycero-phosphocholine, DUPC), 1,2-di- O-octadecenyl-sn-glycero-3-phosphocholine (1,2-di-O-octadecenyl-sn-glycero-3-phosphocholine, 18:0 diether PC), 1-oleoyl-2- Cholesterylhemisuccinoyl-sn-glycero-3-phosphocholine (1-oleoyl-2-cholesterylhemisuccinoyl-sn-glycero-3-phosphocholine, OChemsPC), 1-hexadecyl-sn-glycero-3-phospho Forcholine (1-hexadecyl-sn-glycero-3-phosphocholine, C16 Lyso PC), 1,2-distearoyl-sn-glycero-3-phosphoethanolamine (1,2-distearoyl-sn-glycero -3-phosphoethanolamine), 1,2-dilinoleoyl-sn-glycero-3-phosphoethanolamine (1,2-dilinoleoyl-sn-glycero-3-phosphoethanolamine), 1,2-dilinoleoyl -sn-glycero-3-phosphoethanolamine (1,2-dilinoleoyl-sn-glycero-3-phosphoethanolamine), 1,2-diarachidonoyl-sn-glycero-3-phosphoethanolamine (1 ,2-diarachidonoyl-sn-glycero-3-phosphoethanolamine), 1,2-didocosahexaenoyl-sn-glycero-3-phosphoethanolamine (1,2-didocosahexaenoyl-sn-glycero-3-phosphoethanolamine) , sphingomyelin, or a mixture thereof.

The structural lipid maintains the particle shape within the lipid nanoparticle and is dispersed on the core and surface of the nanoparticle to improve the stability of the nanoparticle. The structural lipid is, for example, cholesterol, cholesterol, spinasterol, fecosterol, sitosterol, ergosterol, ergostenol, campesterol, stigmasterol, brassicasterol, tomatidine, ursolic acid, alpha-tocopherol or these. It may be a mixture of, but is not limited thereto.

According to one embodiment, when lipid nanoparticles are prepared by mixing the ionizable lipid of the present invention with phospholipid, structural lipid, and PEG-lipid, the molar ratio of the ionizable lipid: phospholipid: structural lipid: PEG-lipid conjugate mixture is 20. It may be from 60:10 to 30:30 to 60:0.1 to 10. Specifically, the molar ratio may be 30 to 50:10 to 20:35 to 50:0.5 to 8, and more specifically, 40 to 45:10 to 15:38 to 45:1 to 6, but is not limited thereto. No. Among the components included in the lipid nanoparticles, the sum of the moles of the PEG-lipid conjugate mixture and the structural lipid is kept constant, and the number of moles of the structural lipid is reduced as much as the number of moles of the PEG-lipid conjugate mixture is increased, so that the number of moles of the structural lipid is reduced. The molar ratio can be maintained.

Since the lipid nanoparticles of the present invention exhibit a positive charge under acidic pH conditions, they easily form a complex with the drug through electrostatic interaction with therapeutic agents such as nucleic acids and anionic drugs that exhibit a negative charge, thereby encapsulating the anionic drug with high efficiency. It can be used as an intracellular or in vivo drug delivery composition. Therefore, the lipid nanoparticles of the present invention can be useful for the delivery of not only nucleic acids but also all types of drugs with anionic properties. In other words, the lipid nanoparticles of the present invention can be manufactured in a form (encapsulated form) that additionally contains an anionic drug, and the encapsulated anionic drug can be specifically delivered to the lungs, specifically to fibrotic lungs. there is.

In the present invention, the term "encapsulation" refers to encapsulating the delivery material to surround it and efficiently incorporate it into the body, and the drug encapsulation efficiency (encapsulation efficiency) refers to the total drug content used in manufacturing. It refers to the content of drug encapsulated in lipid nanoparticles.

The anionic drug may be a nucleic acid, low molecular weight compound, peptide, protein, protein-nucleic acid structure, or anionic biopolymer-drug conjugate, but can be delivered stably and efficiently by forming lipid nanoparticles with the ionizable lipid of the present invention. It is not limited to this as long as possible.

In the present invention, the nucleic acids include small interfering ribonucleic acid (siRNA), ribosomal ribonucleic acid (rRNA), deoxyribonucleic acid (DNA), complementary deoxyribonucleic acid (cDNA), aptamer, messenger ribonucleic acid (mRNA), and transport ribonucleic acid. (tRNA), sgRNA, antisense oligonucleotide, shRNA, miRNA, ribozyme, PNA, and DNAzyme, or a mixture thereof, and is specifically selected from the group consisting of siRNA, mRNA, antisense oligonucleotide, and miRNA. It may be one or more, but is not limited thereto. According to one embodiment of the present invention, the nucleic acid drug may be siGtse1, an siRNA that inhibits the expression of Gtse1 protein.

The weight ratio of total lipid/nucleic acid in the lipid nanoparticles may be 1 to 20, specifically 5 to 15, and more specifically 7 to 12, but is not limited thereto.

In the present invention, the lipid nanoparticles may have a diameter of, for example, 40 to 150 nm, specifically 50 to 140 nm, and more specifically 60 to 130 nm, but are not limited thereto. .

In another aspect for achieving the above object, the present invention provides a pharmaceutical composition for preventing or treating lung diseases containing the lipid nanoparticles and an anionic drug.

Lipid nanoparticles and anionic drugs are as described above.

The lipid nanoparticles of the present invention form a stable complex with anionic drugs such as nucleic acids and exhibit low cytotoxicity and effective cell absorption, so they are effective in delivering anionic drugs. In addition, the lipid nanoparticles can be specifically delivered to the lungs, specifically fibrotic lungs, or specific cells thereof when administered, and thus can be usefully used in the prevention or treatment of lung diseases.

The lipid nanoparticles of the present invention can target the lungs and deliver drugs specifically to them, and according to one embodiment of the present invention, they are more specific to fibrotic lungs and their cells (e.g., lung epithelial cells and endothelial cells). It can be delivered. Accordingly, the lung disease may specifically be pulmonary fibrosis.

In the present invention, the term "treatment" refers to intervention to alter the natural processes of an individual or cell having a disease, which may be carried out during the progression of the pathology or to prevent it. The desired therapeutic effects include preventing the occurrence or recurrence of the disease, alleviating symptoms, reducing all direct or indirect pathological consequences of the disease, preventing metastasis, reducing the rate of disease progression, and alleviating the disease state. Or it includes temporary relief, remission, or improvement of prognosis. In particular, the present invention includes all actions to improve the course of lung-related diseases by administering a composition containing lipid nanoparticles containing mannose-PEG-lipid conjugates and anionic drugs as active ingredients. Additionally, the term "prevention" refers to all actions that suppress or delay the onset of a disease by administering the lipid nanoparticles. When the lipid nanoparticles of the present invention are used for treatment or prevention purposes, they are administered to an individual in a therapeutically effective amount.

The term "therapeutically effective amount" used in the present invention refers to an effective amount of anionic drug-containing lipid nanoparticles. Specifically, a "therapeutically effective amount" means an amount sufficient to treat a disease with a reasonable benefit/risk ratio applicable to medical treatment, and the effective dose level is determined by the type and severity of the individual, age, gender, and severity of the disease. It can be determined based on factors including the type, activity of the drug, sensitivity to the drug, time of administration, route of administration and excretion rate, duration of treatment, drugs used simultaneously, and other factors well known in the medical field. The pharmaceutical composition of the present invention may be administered as an individual therapeutic agent or in combination with other therapeutic agents, and may be administered sequentially or simultaneously with commercially available therapeutic agents. And it can be administered single or multiple times. Considering all of the above factors, it is important to administer an amount that can achieve maximum effect with the minimum amount without side effects, and can be easily determined by a person skilled in the art. The administered dose of the pharmaceutical composition of the present invention may be determined by an expert depending on various factors such as the patient's condition, age, gender, and complications. Since the active ingredient of the composition of the present invention has excellent safety, it can be used beyond the determined administration dose.

The composition containing the lipid nanoparticles can be administered orally, intramuscularly, intravenously, arterially, subcutaneously, intraperitoneally, pulmonaryly, and intranasally, and specifically can be administered locally directly into the lungs, and more specifically, into fibrotic lungs. Can be administered topically.

The composition of the present invention may further include one or more pharmaceutically acceptable carriers for administration. Pharmaceutically acceptable carriers can be saline solution, sterile water, Ringer's solution, buffered saline solution, dextrose solution, maltodextrin solution, glycerol, ethanol, and a mixture of one or more of these ingredients, and if necessary, antioxidants, buffer solutions, Other common additives such as bacteriostatic agents can be added.

The lipid nanoparticles of the present invention are specific for lung tissue and its specific cells, have excellent biocompatibility, and can deliver gene therapeutic agents with high efficiency, so they can be usefully used in related technical fields such as lipid nanoparticle-mediated gene therapy.

Figure 1 is a schematic diagram showing an exemplary structure of mannose-modified lipid nanoparticles and a mechanism of action against lung diseases using the same.
Figure 2 is a graph measuring the intracellular luminescence intensity of mannose-modified lipid nanoparticles encapsulated with mRNA encoding luciferase using various types of ionizable lipids.
Figure 3 is an image of an ex vivo organ in which bioluminescence was confirmed to confirm specific delivery of mannose-modified lipid nanoparticles in vivo.
Figure 4 is an ex vivo organ image confirming bioluminescence of lipid nanoparticles not modified with mannose.
Figure 5 is a graph comparing the fluorescence expression of mannose-modified lipid nanoparticles in each lung cell in LSL-tdTomato mice (blank: general model, colored: lung fibrosis model).
Figure 6 is a graph confirming the cytotoxicity of mannose-modified lipid nanoparticles using CellTiter-Fluor™ Cell Viability Assay.
Figure 7 is a graph measuring the levels of AST (aspartate aminotransferase), ALT (alanine aminotransferase), BUN (blood urea nitrogen), and creatinine to confirm the presence or absence of liver and kidney toxicity following administration of mannose-modified lipid nanoparticles.
Figure 8 shows GOT (Glutamic Oxaloacetic Transaminase), GPT (Glutamic Pyruvic Transaminase), BUN, and creatinine (CRE) to confirm the presence or absence of liver and kidney damage following administration of mannose-modified lipid nanoparticles using a bleomycin-induced pulmonary fibrosis model. ) This is a graph measuring the values.
Figures 9 and 10 show histopathological changes in the spleen, liver, kidneys, and lungs using hematoxylin-eosin (H&E) staining to confirm histopathological changes in major organs following administration of mannose-modified lipid nanoparticles. It is an image.
Figure 11 shows the results of measuring the inhibitory effect of mannose-modified lipid nanoparticles on the Gtse1 gene by western blot.
Figure 12 shows the results of confirming changes in collagen deposition and Gtse1 protein expression level by administration of mannose-modified lipid nanoparticles in an animal model of bleomycin-induced lung fibrosis.
Figure 13 shows the results of confirming changes in collagen deposition and Gtse1 protein expression level by administration of mannose-modified lipid nanoparticles in an animal model of radiation-induced lung fibrosis.
Figure 14 shows the results of confirming histopathological changes and lung volume caused by administration of mannose-modified lipid nanoparticles in an animal model of radiation-induced lung fibrosis.
Figure 15 is a graph measuring lung capacity indices using Flexivent® in an animal model of radiation-induced pulmonary fibrosis.

Hereinafter, the configuration and effects of the present invention will be described in more detail through examples. These examples are only for illustrating the present invention and the scope of the present invention is not limited by these examples.

Example 1. Preparation of lipid nanoparticles

Example 1-1. Manufacturing of mannose-modified lipid nanoparticles

Ionizable lipid (246-C10), cholesterol (Cholesterol powder, BioReagent, suitable for cell culture, ≥99%, Sigma, Korea), phospholipid (DSPC; Avanti, USA), PEG-lipid conjugate (C16-PEG2000 ceramide or ceramide -PEG conjugate; C16 PEG2000 Ceramide, Avanti, USA) and mannose-PEG-lipid conjugate (mannose-PEG-DSPE conjugate; Biochempeg, USA) were dissolved in ethanol at a molar ratio of 42.5:13:43 to 39.5:0.5:1.0 to 4.5. It was dissolved in a molar ratio of cholesterol and total PEG-lipid conjugate content (= PEG-ceramide conjugate content + mannose-PEG-DSPE conjugate content) adjusted so that the total sum was 100. Ionizable lipids, cholesterol, phospholipids, PEG-ceramide conjugates, and mannose-PEG-lipid conjugates were dissolved in ethanol and acetate buffer at a volume ratio of 1:3 at a flow rate of 12 mL/min using a microfluidic mixing device (Benchtop Nanoassemblr; PNI, Canada) to prepare lipid nanoparticles modified with mannose (hereinafter referred to as 'mannose-LNP').

Example 1-2. Manufacturing of mannose-modified lipid nanoparticles encapsulated with nucleic acids

SM-102, ALC-0315, 246-C10, and 244-cis were prepared as ionizable lipids, and 244-cis was synthesized according to the following reaction scheme.

The organic phase (ethanol) in which the ionizable lipids, cholesterol, phospholipids, PEG-lipid conjugates (C16-PEG2000 ceramide) and mannose-PEG-lipid conjugates (mannose-PEG-DSPE conjugates) are dissolved and nucleic acids (mRNA or siRNA) are dissolved. The aqueous phase (sodium acetate or sodium citrate) was mixed through a microfluidic mixing device (Benchtop Nanoassemblr; PNI, Canada) at a flow rate of 12 ml/min, and specifically, lipid nanoparticles encapsulated with nucleic acid were prepared by the following method. did.

(i) To prepare mannose-modified lipid nanoparticles encapsulated with mRNA, ionizable lipid: phospholipid (DOPE or DSPC): cholesterol: C16-PEG2000 ceramide (PEG-ceramide conjugate), DMG-PEG (1,2-Dimyristoyl -rac-glycero-3-methoxypolyethylene glycol-2000) or ALC-0159 (Methoxypolyethyleneglycoloxy(2000)-N,N-ditetradecylacetamide): Mannose-PEG-lipid conjugate (mannose-PEG-DSPE conjugate) from 26.5 to 50:10 to Ethanol at a molar ratio of 20:38.5 to 52:0.5:1.0 to 4.5 (adjust cholesterol and total PEG-lipid content (= PEG-ceramide conjugate content + mannose-PEG-DSPE conjugate content) so that the total sum of the molar ratios is 100) was dissolved in, and lipid nanoparticles were prepared by mixing the organic phase and the aqueous phase so that the weight ratio of mRNA (luciferase mRNA; SEQ ID NO: 1 or Cre; SEQ ID NO: 2):ionizable lipid was 1:10.

(ⅱ) To prepare siRNA-encapsulated mannose-modified lipid nanoparticles, ionizable lipid (246-C10):phospholipid (DSPC):cholesterol:C16-PEG2000 ceramide:mannose-PEG-DSPE conjugate was mixed at 42.5:13:43 to 42.5:13:43. It was dissolved in ethanol at a molar ratio of 39.5:0.5:1.0 to 4.5 (adjustment of cholesterol and total PEG-lipid content (= PEG-ceramide conjugate content + mannose-PEG-DSPE conjugate content) so that the total molar ratio is 100), siRNA (siGtse1; SEQ ID NO: 3 and 4): Lipid nanoparticles were prepared by mixing the organic phase and the aqueous phase so that the ionizable lipid had a weight ratio of 1:7.5.

The prepared lipid nanoparticles were dialyzed against PBS for 16 hours using a 3500 MWCO dialysis cassette to remove ethanol and adjust the pH of the lipid nanoparticles to that of the body.

Example 2. Confirmation of nucleic acid delivery effect of mannose-modified lipid nanoparticles depending on the type of ionizable lipid

One day before transfecting cells with mannose-modified lipid nanoparticles, HeLa cells (Korean Cell Line Bank) were distributed at ^0.01x106 cells/well in a white plate (96 well) and grown in DMEM media (SH30022, Hyclone, USA). Cultured at 37°C and 0.5-3% CO ₂ conditions. The mannose-LNP prepared in Example 1-2 containing luciferase mRNA was stirred by pipetting, incubated at room temperature for 10 minutes, and then treated with HeLa cells (20 ng/well based on the mRNA contained in the lipid nanoparticles). .

After 24 hours of treatment, 100 ㎕/well of Bright-Glo™ Luciferase Assay solution (promega, USA) was treated and left at room temperature for 10 minutes. The dissolved cells were measured for luminescence intensity (Tecan, USA) using an Infinite M200 luminescence measuring device (Tecan, USA). luminescence intensity) was measured, and the results are shown in Figure 2.

As confirmed in Figure 2, it was confirmed that mannose-modified lipid nanoparticles showed high luminescence value within cells regardless of the type of ionizable lipid.

Example 3. Confirmation of lung-specific delivery of mannose-modified lipid nanoparticles

Example 3-1. Confirmation of lung-specific delivery using fLuc mRNA

Mannose-LNP encapsulated with luciferase mRNA prepared in Example 1-2 was intratracheally injected into the kidney, spleen, liver, lung, and heart of 7-week-old C57BL/6 mice, respectively, at a dose of 0.1 mg/kg based on mRNA, 3 After some time, 0.25 mg/kg of luciferin was administered intraperitoneally, and bioluminescence was confirmed by ex vivo organ imaging using IVIS (PerkinElmer, USA) equipment. The results are shown in Figure 3 and Table 1. .

[Table 1]

In addition, as LNPs that are not modified with mannose, lipid nanoparticles (lipid-PEG at 1.5 mol% including) were manufactured. The non-mannose-modified LNP was injected intravenously at a dose of 0.1 mg/kg based on mRNA into C57BL/6 female 7-week-old mice (Orient Bio), and 3 hours later, 0.25 mg/kg of luciferin was administered intraperitoneally, and the mouse was sacrificed. The organs were removed and bioluminescence was confirmed in each organ through disembodied organ images using IVIS equipment, and the results are shown in Figure 4.

As confirmed in Figures 3 and 4, mannose-LNPs showed high luminescence intensity in the lungs, while LNPs not modified with mannose showed high luminescence intensities in the liver, through which the formulation of lipid nanoparticles of the present invention It was confirmed that it was delivered specifically to the lungs as it was modified with mannose.

Example 3-2. Confirmation of lung cell-specific delivery using Cre mRNA

Mannose-LNP encapsulated with Cre mRNA (mCre) prepared in Example 1-2 and LNP (Native LNP) not modified with mannose were administered to LSL-tdTomato mice and lungs in which lung fibrosis occurred at a dose of 0.3 mg/kg based on mRNA. Each LSL-tdTomato mouse without fibrosis was administered intratracheally, and the expression of tomato fluorescence in each lung cell (epithelial cells, endothelial cells, immune cells) was measured using flow cytometry (LSRFortessa, BD) in the lungs extracted from sacrificed mice. This was confirmed, and the results are shown in Figure 5 (blank: general model, colored: pulmonary fibrosis model).

As seen in Figure 5, it was confirmed that the mannose-LNP of the present invention was specifically delivered to fibrotic lungs compared to LNPs that were not modified with mannose. In addition, the mannose-LNP of the present invention had a different fluorescence expression pattern for each lung cell compared to the non-mannose-modified LNP, and in particular, it was confirmed that it was most specifically delivered to the epithelial cells of the fibrotic lung.

Example 4. Confirmation of toxicity of mannose-modified lipid nanoparticles

Example 4-1. Confirmation of cytotoxicity

Whether the mannose-LNP of the present invention exhibits cytotoxicity was confirmed through CellTiter-Fluor™ Cell Viability Assay, which shows that the substrate passing through the cell membrane causes cleavage by protease inside the cell, forming fluorescent AFC, which kills the cell through fluorescence analysis. This is an analysis method that can check viability.

Specifically, L132 and MEF cells were seeded in ^a transparent 96 well plate (SPL, 96 Well Cell Culture Plates, 30096) at 0.1 Lipid nanoparticles corresponding to 0.05 μg or 0.1 μg of sugar mRNA were treated in the medium so that the total amount was 100 μl. 24 hours after lipid nanoparticle treatment, 100 μl of CellTiter-Fluor™ Cell Viability Assay (Promega, Cat.# G6080) was added per well, incubated for 30 minutes, and then measured at 450 nm using Infinite® 200 PRO NanoQ (Tecan). The absorbance was measured, and the results are shown in Figure 6.

As confirmed in Figure 6, it was confirmed that the mannose-modified lipid nanoparticles of the present invention did not show cytotoxicity.

Example 4-2. Check for liver and kidney toxicity

In order to confirm the presence or absence of liver and kidney toxicity following administration of the mannose-LNP of the present invention, the levels of aspartate aminotransferase (AST) and alanine aminotransferase (ALT), blood urea nitrogen (BUN), and creatinine were measured, respectively.

Specifically, the lipid nanoparticles prepared in Example 1-2 were intratracheally administered once to 7-week-old C57BL/6 mice at doses of 0.5 and 0.75 mg/kg based on mRNA, and blood was collected 24 hours after administration to determine AST and ALT levels. , BUN and creatinine levels were confirmed, and the results are shown in Figure 7.

As confirmed in Figure 7, it was confirmed that the mannose-modified lipid nanoparticles of the present invention did not show significant liver and kidney toxicity.

Example 4-3. Confirmation of liver and kidney damage indicators in lung fibrosis animal model

Using the bleomycin-induced pulmonary fibrosis model, the presence or absence of liver and kidney damage during administration of the mannose-LNP of the present invention was additionally confirmed.

Specifically, an animal model of pulmonary fibrosis was constructed by intratracheally administering bleomycin at a dose of 1.8 mg/kg to 7-week-old C57BL/6 mice, and the pulmonary fibrosis animal model prepared in Example 1-2 was administered 1 day before and 4 days after bleomycin administration. Mannose-modified lipid nanoparticles encapsulated with siGtse1 were administered intratracheally twice at a dose of 0.5 mg/kg based on mRNA. Then, on the 14th day, the mouse was sacrificed and plasma was separated from the collected blood, and this was used to analyze GOT (Glutamic Oxaloacetic Transaminase) and GPT (Glutamic Pyruvic Transaminase) as liver damage indicators and BUN and creatinine (CRE) as kidney damage indicators. The values were measured, and the results are shown in Figure 8 and Table 2.

[Table 2]

As confirmed in Figure 8 and Table 2, it was confirmed that the mannose-modified lipid nanoparticles of the present invention did not show significant liver and kidney toxicity.

Example 5. Confirmation of pathological phenomena in major organs

In order to confirm the histopathological changes in major organs following administration of the mannose-LNP of the present invention, histopathological changes in the spleen, liver, kidneys, and lungs were observed using hematoxylin-eosin (H&E) staining.

Specifically, 1 day before and 4 days after bleomycin administration to 7-week-old C57BL/6 mice at a dose of 1.8 mg/kg, siGtse1-encapsulated mannose-modified lipid nanoparticles prepared in Example 1-2 were administered at a dose of 0.5 mg/kg based on mRNA. It was administered intratracheally twice at a dose of kg. Then, on the 14th day, the mouse was sacrificed, the spleen, liver, kidney, and lung were stained with H&E, and pathological phenomena were observed through an optical microscope. The results are shown in Figures 9 and 10.

As shown in Figures 9 and 10, damage to major organs was confirmed in the bleomycin administered group (BLM), but no significant histopathological changes were observed in the mannose-LNP administered group.

Example 6. Confirmation of effect on lung fibrosis

Example 6-1. Silencing effect on lung fibrosis-related genes

The inhibitory effect of mannose-LNP on the Gtse1 gene, which is known to be highly associated with lung fibrosis, was confirmed.

Specifically, the siGtse1-encapsulated mannose-LNP prepared in Example 1-2 was intratracheally injected into 7-week-old C57BL/6 mice at a dose of 0.5 mg/kg based on siRNA, and the mice were sacrificed 24 and 48 hours later to infuse the lungs. Gtse1 protein in cells was measured by western blot, and the results are shown in Figure 11.

As shown in Figure 11, it was confirmed that mannose-LNP encapsulated with siGtse1 can significantly inhibit the expression of Gtse1 protein in lung cells.

Example 6-2. Confirmation of effect in bleomycin-induced lung fibrosis animal model

The effect of mannose-LNP was confirmed using a bleomycin-induced lung fibrosis animal model.

Specifically, an animal model of pulmonary fibrosis was constructed by intratracheally administering bleomycin at a dose of 1.8 mg/kg to 7-week-old C57BL/6 mice, and the pulmonary fibrosis animal model prepared in Example 1-2 was administered 1 day before and 4 days after bleomycin administration. Mannose-modified lipid nanoparticles encapsulated with siGtse1 were administered intratracheally twice at a dose of 0.5 mg/kg based on mRNA. Then, on the 14th day, the mouse was sacrificed and collagen deposition was confirmed through Masson Trichrome staining, and the results are shown in Figure 12.

As seen in Figure 12, collagen deposition increased in the bleomycin-administered group (Saline) and the mannose-LNP-administered group (siCon) encapsulated with siControl nucleic acid (Bioneer, Cat.# SN-1013), whereas the siGtse1 nucleic acid drug encapsulated group In the mannose-LNP administered group (siGtse1), collagen was significantly decreased, and tissue immunology staining also confirmed that the expression of Gtse1 protein was decreased.

Example 6-3. Confirmation of effectiveness in radiation-induced lung fibrosis animal model

The effect of mannose-LNP was confirmed using a radiation-induced lung fibrosis animal model.

Specifically, 75 Gy of high-dose focal radiation was irradiated to the left lung of 3 mm volume in 10-week-old C57BL/6 mice to construct an animal model of pulmonary fibrosis, and the mannose-modified lipid containing siGtse1 prepared in Example 1-2 was used. After irradiation, nanoparticles were administered intratracheally twice at a dose of 0.5 mg/kg based on mRNA, 1 day and 3 weeks later in the prevention group, and 4 weeks and 5 weeks later in the treatment group. Then, 6 weeks later, the mouse was sacrificed and collagen deposition was confirmed through Masson Trichrome staining, and the results are shown in Figure 13.

As seen in Figure 13, collagen was significantly decreased in both the prevention and treatment groups administered mannose-LNP encapsulated with siGtse1 nucleic acid drug, and tissue immunology staining also confirmed that the expression of Gtse1 protein was decreased.

In addition, formalin liquid was inserted into the trachea of the sacrificed mouse to fix the entire lung, and histopathological changes in the irradiated area were observed with the naked eye, and the degree of lung damage was evaluated through small animal Mico-CT imaging. And the results are shown in Figure 14.

As shown in Figure 14, it was confirmed that lung volume was recovered in both the prevention and treatment groups administered with mannose-LNP encapsulated with the siGtse1 nucleic acid drug compared to the control group.

Example 6-4. Confirmation of various lung capacity indicators

It is known that as pulmonary fibrosis progresses after radiation, lung volume decreases, and at the same time, it is accompanied by changes in lung tissue elasticity and various lung capacity indicators. Accordingly, the lung capacity indices shown in Table 3 below were measured in the animal model of Example 6-2 using Flexivent®, a test method that allows the results of various lung function tests measured in humans to be obtained in small animals, and the results are as follows. It is shown in Figure 15.

[Table 3]

As can be seen in Figure 15, the irradiation group showed a restirictive tendency in the lung capacity index compared to the control group, but in both the prevention and treatment groups administered mannose-LNP encapsulated with the siGtse1 nucleic acid drug, the pulmonary capacity index was lower than that of the irradiation group. It was confirmed that their restirictive tendency was recovered.

Sequence list electronic file attached

Claims (13)

A composition for transpulmonary delivery of a drug containing lipid nanoparticles containing a mannose-PEG (polyethyleneglycol)-lipid conjugate.
The composition for transpulmonary delivery of a drug according to claim 1, wherein the composition is delivered locally within the lung.
The composition for transpulmonary delivery of a drug according to claim 1, wherein the composition is delivered locally to fibrotic lungs.
The composition for transpulmonary delivery of a drug according to claim 1, wherein the lipid nanoparticles further include at least one selected from the group consisting of ionizable lipids, phospholipids, and structural lipids.
5. The method of claim 4, wherein the ionizable lipid is SM-102, ALC-0315, DODAC, DODMA, DSDMA, DOTMA, DDAB, DOTAP, DC-Chol, DMRIE, DOSPA, DOGS, CLinDMA, CpLinDMA, DMOBA, DOcarbDAP, DLinDAP , DLincarbDAP, DLinCDAP, DLinDMA, DLenDMA, DLin-MC3-DMA, 246-C10, and 244-cis. A composition for transpulmonary delivery of a drug.
The method of claim 4, wherein the phospholipids are DOPE, DSPC, POPC, EPC, DOPC, DPPC, DOPG, DPPG, DSPE, DOTAP, phosphatidylethanolamine, dipalmitoylphosphatidylethanolamine, 1,2-dioleoyl-sn- Glycero-3-phosphate, 1,2-dilinoleoyl-sn-glycero-3-phosphocholine, 1,2-diarachidonoyl-sn-glycero-3-phosphocholine, 1,2 -didocosahexaenoyl-sn-glycero-3-phosphocholine, 1,2-diphytanoyl-sn-glycero-3-phosphoethanolamine, POPE, DOPS, DLPC, DMPC, DUPC, 1, 2-di-O-octadecenyl-sn-glycero-3-phosphocholine, 1-oleoyl-2-cholesterylhemisuccinoyl-sn-glycero-3-phosphocholine, 1-hexadecyl -sn-glycero-3-phosphocholine, 1,2-distearoyl-sn-glycero-3-phosphoethanolamine, 1,2-dilinoleoyl-sn-glycero-3-phos Foethanolamine, 1,2-dilinoleoyl-sn-glycero-3-phosphoethanolamine, 1,2-diarachidonoyl-sn-glycero-3-phosphoethanolamine, 1,2- A composition for transpulmonary delivery of a drug, which is at least one selected from the group consisting of didocosahexaenoyl-sn-glycero-3-phosphoethanolamine, and sphingomyelin.
5. The method of claim 4, wherein the structural lipids are cholesterol, cholesterol, spinasterol, fecosterol, sitosterol, ergosterol, ergostenol, campesterol, stigmasterol, brassicasterol, tomatidine, ursolic acid and alpha - A composition for transpulmonary delivery of a drug, which is at least one selected from the group consisting of tocopherol.
The method of claim 4, wherein the lipid nanoparticles contain a mixture of ionizable lipid: phospholipid: structural lipid: PEG-lipid conjugate in a molar ratio of 20 to 60: 10 to 30: 30 to 60: 0.1 to 10. Composition for delivery.
i) Lipid nanoparticles containing mannose-PEG (polyethyleneglycol)-lipid conjugate; and ii) an anionic drug; a pharmaceutical composition for preventing or treating lung disease.
The pharmaceutical composition of claim 9, wherein the anionic drug is a nucleic acid drug.
The pharmaceutical composition of claim 10, wherein the nucleic acid drug is at least one selected from the group consisting of siRNA, mRNA, antisense oligonucleotide, and miRNA.
The pharmaceutical composition according to claim 9, wherein the lung disease is pulmonary fibrosis.
The pharmaceutical composition according to claim 9, wherein the composition is administered locally in the lung.