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CN-122005494-A - Lipid nanoparticle modified based on taurochenodeoxycholic acid and preparation method and application thereof

CN122005494ACN 122005494 ACN122005494 ACN 122005494ACN-122005494-A

Abstract

The application provides lipid nano particles based on taurochenodeoxycholic acid modification, a preparation method and application thereof, wherein the lipid nano particles comprise nucleic acid and a lipid carrier for encapsulating the nucleic acid, the lipid carrier comprises, by mole percent, 40% -65% of ionized lipid, 5% -20% of structural phospholipid, 20% -50% of sterol component and 0.5% -5% of PEG-lipid, and the sterol component comprises cholesterol and taurochenodeoxycholic acid. The membrane order degree and fluidity of the lipid carrier are effectively regulated by adopting taurochenodeoxycholic acid to replace cholesterol, the targeting delivery and endosome release efficiency of LNP to liver cells is remarkably improved, the LNP delivery system can obtain more remarkable and durable Lp (a) reducing effect under the same or lower dosage, and the LNP preparation is ensured to have higher controllability and reliability in the production and storage process by optimizing the LNP formula.

Inventors

  • ZHENG LEMIN
  • NIU CHENGUANG
  • JIAO HAIQING
  • ZHANG QI
  • Su Shenge

Assignees

  • 福建普立辰生物技术有限公司
  • 郑乐民

Dates

Publication Date
20260512
Application Date
20260306

Claims (10)

  1. 1. A lipid nanoparticle modified by taurochenodeoxycholic acid, comprising a nucleic acid and a lipid carrier encapsulating the nucleic acid; the lipid carrier comprises, by mole percent, 40% -65% of ionized lipid, 5% -20% of structural phospholipids, 20% -50% of sterol components and 0.5% -5% of PEG-lipids, wherein the sterol components comprise cholesterol and taurochenodeoxycholic acid.
  2. 2. The lipid nanoparticle according to claim 1, wherein the taurochenodeoxycholic acid comprises 0.1% -100% of the total moles of the sterol component.
  3. 3. The lipid nanoparticle of claim 1, wherein the nitrogen to phosphorus molar ratio of the ionized lipid to the nucleic acid is from 3:1 to 10:1.
  4. 4. The lipid nanoparticle of claim 1, wherein the nucleic acid is an siRNA.
  5. 5. The lipid nanoparticle of claim 1, wherein the ionized lipids comprise one or more of DLin-MC3-DMA, 5A2-SC8, SM-102, C12-200, ALC-0315, dotap, and dotma.
  6. 6. The lipid nanoparticle of claim 1, wherein the structural phospholipid comprises a neutral helper phospholipid including, but not limited to, 1, 2-distearoyl-sn-glycero-3-phosphorylcholine, 1, 2-dipalmitoyl-sn-glycero-3-phosphorylcholine, 1, 2-dioleoyl-sn-glycero-3-phosphoethanolamine, 1, 2-palmitoyl-sn-glycero-3-phosphoethanolamine, 1, 2-dimyristoyl-sn-glycero-3-phosphoethanolamine, 2-dioleoyl-sn-glycero-3-phosphoethanolamine, 1' -rac-glycero, oleoyl phosphatidylcholine, 1-palmitoyl-2-oleoyl phosphatidylethanolamine.
  7. 7. The lipid nanoparticle of claim 1, wherein the PEG-lipid comprises one or more of ALC-0159, DSPE-PEG2000, DMG-PEG 2000.
  8. 8. The preparation method of the lipid nanoparticle modified by taurochenodeoxycholic acid is characterized by comprising the following steps of: dissolving the ionizable lipid, the auxiliary phospholipid, the sterol component and the pegylated lipid in an organic solvent according to a first preset proportion to obtain a lipid organic phase; dissolving the nucleic acid in a buffer solution according to a second preset proportion to obtain a nucleic acid water phase; mixing the lipid organic phase with the nucleic acid water to self-assemble lipid to form a lipid nanoparticle coarse dispersion encapsulating the nucleic acid; And dialyzing the lipid nanoparticle coarse dispersion to displace the solvent and neutralize the pH to obtain the target lipid nanoparticles.
  9. 9. Use of the lipid nanoparticle according to any one of claims 1-7 in the manufacture of a medicament for reducing serum lipoprotein (a) [ Lp (a) ] levels; the medicament is suitable for preventing or treating diseases associated with elevated Lp (a), selected from the group consisting of atherosclerotic cardiovascular disease, ischemic heart disease, ischemic stroke, peripheral arterial disease and/or calcified aortic stenosis; preferably, the drug is administered by a method including, but not limited to, oral, enteral, subcutaneous, intramuscular, intravenous, nasal, transdermal, subconjunctival, intra-ocular, orbital, retrobulbar, retinal, choroidal, intrathecal; Preferably, the dosage form of the medicament includes, but is not limited to, tablets, capsules, pills, injections, inhalants, lozenges, suppositories, emulsions, microemulsions, sub-microemulsions, nanoparticles, gels, powders, suspoemulsions, creams, jellies, sprays.
  10. 10. Use of the lipid nanoparticle of any one of claims 1-7, the lipid nanoparticle prepared by the preparation method of claim 8, and the drug of claim 9 in any one of the following: 1) The application in preparing the medicine for treating and/or preventing the LPA gene expression related diseases; 2) Use in inhibiting LPA gene expression; Preferably, the medicament is used in a method of inhibiting LPA target activity or reducing LPA expression in a subject by using the lipid nanoparticle according to claims 1-7, the lipid nanoparticle prepared by the preparation method according to claim 8, or the medicament according to claim 9; Preferably, the disease associated with LPA gene expression comprises an atherosclerotic cardiovascular disease, an ischemic heart disease, an ischemic stroke, a peripheral arterial disease and/or a calcified aortic valve stenosis.

Description

Lipid nanoparticle modified based on taurochenodeoxycholic acid and preparation method and application thereof Technical Field The invention relates to the technical field of medicines, in particular to lipid nano particles based on modification of taurochenodeoxycholic acid, and a preparation method and application thereof. Background Apolipoprotein (a) [ Lp (a) ] is an independent risk factor for atherosclerotic cardiovascular disease, the level of which is primarily determined by LPA gene polymorphism, and existing lipid lowering drugs reduce cholesterol in statin drugs by inhibiting HMG-CoA reductase, but are almost ineffective for Lp (a). RNA interference technology provides a new strategy for targeting LPA genes. Small interfering RNAs (sirnas) can silence LPA gene expression from post-transcriptional levels. However, the core bottleneck in successful development of LPA sirnas as drugs has been shifted from sequence discovery to delivery systems. Lipid Nanoparticles (LNPs) are the most mature nucleic acid delivery platform currently in clinical validation. Typical LNPs consist of ionizable lipids, helper phospholipids, sterols, and pegylated lipids, where cholesterol is an essential sterol component to maintain LNP stability. The traditional LNP using cholesterol as the sole sterol has the problems of insufficient liver uptake efficiency, large dosage burden and limited stability and batch-to-batch consistency when LPA siRNA is delivered. Therefore, under the premise of not remarkably increasing the process complexity and the toxicity, the lipid nanoparticle system capable of efficiently, stably and directionally delivering the siRNA to the target gene Lp (a) in a large scale and the preparation method thereof are developed, and the technical problem to be solved in the field is urgent. Disclosure of Invention In view of the problems, the present application has been made in order to provide lipid nanoparticles based on modification of tauchenodeoxycholic acid, which overcome the problems or at least partially solve the problems, and a preparation method and application thereof, including: a lipid nanoparticle based on modification of taurochenodeoxycholic acid comprising a nucleic acid and a lipid carrier encapsulating the nucleic acid; the lipid carrier comprises, by mole percent, 40% -65% of ionized lipid, 5% -20% of structural phospholipids, 20% -50% of sterol components and 0.5% -5% of PEG-lipids, wherein the sterol components comprise cholesterol and taurochenodeoxycholic acid. Preferably, the taurochenodeoxycholic acid accounts for 0.1% -100% of the total mole number of the sterol component. Preferably, the molar ratio of the ionized lipid to nitrogen to phosphorus of the nucleic acid is from 3:1 to 10:1. Preferably, the nucleic acid is an siRNA. Preferably, the ionized lipids include one or more of DLin-MC3-DMA, 5A2-SC8, SM-102, C12-200, ALC-0315, DODAP and DODMA. Preferably, the structural phospholipids include neutral helper phospholipids including, but not limited to, 1, 2-distearoyl-sn-glycero-3-phosphorylcholine, 1, 2-dipalmitoyl-sn-glycero-3-phosphorylcholine, 1, 2-dioleoyl-sn-glycero-3-phosphoethanolamine, 1, 2-palmitoyl-sn-glycero-3-phosphoethanolamine, 1, 2-dimyristoyl-sn-glycero-3-phosphoethanolamine, 2-dioleoyl-sn-glycero-3-phospho- (1' -rac-glycero), oleoyl phosphatidylcholine, 1-palmitoyl-2-oleoyl phosphatidylethanolamine. Preferably, the PEG-lipid comprises one or more of ALC-0159, DSPE-PEG2000, and DMG-PEG 2000. The application also provides a preparation method of the lipid nanoparticle modified by taurochenodeoxycholic acid, which comprises the following steps: dissolving the ionizable lipid, the auxiliary phospholipid, the sterol component and the pegylated lipid in an organic solvent according to a first preset proportion to obtain a lipid organic phase; dissolving the nucleic acid in a buffer solution according to a second preset proportion to obtain a nucleic acid water phase; mixing the lipid organic phase with the nucleic acid water to self-assemble lipid to form a lipid nanoparticle coarse dispersion encapsulating the nucleic acid; And dialyzing the lipid nanoparticle coarse dispersion to displace the solvent and neutralize the pH to obtain the target lipid nanoparticles. The application also provides the use of the above lipid nanoparticle for the manufacture of a medicament for reducing serum lipoprotein (a) [ Lp (a) ] levels; the medicament is suitable for preventing or treating diseases associated with elevated Lp (a), selected from the group consisting of atherosclerotic cardiovascular disease, ischemic heart disease, ischemic stroke, peripheral arterial disease and/or calcified aortic stenosis; preferably, the drug is administered by a method including, but not limited to, oral, enteral, subcutaneous, intramuscular, intravenous, nasal, transdermal, subconjunctival, intra-ocular, orbital, retrobulbar, retinal, choroidal, intrathecal; Preferably, the dosage form of