Classifications

• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K9/00—Medicinal preparations characterised by special physical form
  • A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate
  • A61K9/50—Microcapsules having a gas, liquid or semi-solid filling; Solid microparticles or pellets surrounded by a distinct coating layer, e.g. coated microspheres, coated drug crystals
  • A61K9/51—Nanocapsules; Nanoparticles
  • A61K9/5107—Excipients; Inactive ingredients
  • A61K9/5123—Organic compounds, e.g. fats, sugars
• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
  • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
  • C12N15/09—Recombinant DNA-technology
  • C12N15/87—Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation
  • C12N15/88—Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation using microencapsulation, e.g. using amphiphile liposome vesicle
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K31/00—Medicinal preparations containing organic active ingredients
  • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
  • A61K31/7088—Compounds having three or more nucleosides or nucleotides
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
  • A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
  • A61K47/08—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing oxygen, e.g. ethers, acetals, ketones, quinones, aldehydes, peroxides
  • A61K47/10—Alcohols; Phenols; Salts thereof, e.g. glycerol; Polyethylene glycols [PEG]; Poloxamers; PEG/POE alkyl ethers
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
  • A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
  • A61K47/08—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing oxygen, e.g. ethers, acetals, ketones, quinones, aldehydes, peroxides
  • A61K47/14—Esters of carboxylic acids, e.g. fatty acid monoglycerides, medium-chain triglycerides, parabens or PEG fatty acid esters
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
  • A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
  • A61K47/20—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing sulfur, e.g. dimethyl sulfoxide [DMSO], docusate, sodium lauryl sulfate or aminosulfonic acids
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
  • A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
  • A61K47/28—Steroids, e.g. cholesterol, bile acids or glycyrrhetinic acid
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K48/00—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K48/00—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
  • A61K48/0008—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy characterised by an aspect of the 'non-active' part of the composition delivered, e.g. wherein such 'non-active' part is not delivered simultaneously with the 'active' part of the composition
  • A61K48/0025—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy characterised by an aspect of the 'non-active' part of the composition delivered, e.g. wherein such 'non-active' part is not delivered simultaneously with the 'active' part of the composition wherein the non-active part clearly interacts with the delivered nucleic acid
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K48/00—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
  • A61K48/0008—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy characterised by an aspect of the 'non-active' part of the composition delivered, e.g. wherein such 'non-active' part is not delivered simultaneously with the 'active' part of the composition
  • A61K48/0025—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy characterised by an aspect of the 'non-active' part of the composition delivered, e.g. wherein such 'non-active' part is not delivered simultaneously with the 'active' part of the composition wherein the non-active part clearly interacts with the delivered nucleic acid
  • A61K48/0033—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy characterised by an aspect of the 'non-active' part of the composition delivered, e.g. wherein such 'non-active' part is not delivered simultaneously with the 'active' part of the composition wherein the non-active part clearly interacts with the delivered nucleic acid the non-active part being non-polymeric
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K48/00—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
  • A61K48/0091—Purification or manufacturing processes for gene therapy compositions
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K9/00—Medicinal preparations characterised by special physical form
  • A61K9/10—Dispersions; Emulsions
  • A61K9/127—Synthetic bilayered vehicles, e.g. liposomes or liposomes with cholesterol as the only non-phosphatidyl surfactant
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K9/00—Medicinal preparations characterised by special physical form
  • A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate
  • A61K9/50—Microcapsules having a gas, liquid or semi-solid filling; Solid microparticles or pellets surrounded by a distinct coating layer, e.g. coated microspheres, coated drug crystals
  • A61K9/51—Nanocapsules; Nanoparticles
  • A61K9/5107—Excipients; Inactive ingredients
  • A61K9/513—Organic macromolecular compounds; Dendrimers
  • A61K9/5138—Organic macromolecular compounds; Dendrimers obtained by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyvinyl pyrrolidone, poly(meth)acrylates
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K9/00—Medicinal preparations characterised by special physical form
  • A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate
  • A61K9/50—Microcapsules having a gas, liquid or semi-solid filling; Solid microparticles or pellets surrounded by a distinct coating layer, e.g. coated microspheres, coated drug crystals
  • A61K9/51—Nanocapsules; Nanoparticles
  • A61K9/5192—Processes
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K48/00—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
  • A61K48/0008—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy characterised by an aspect of the 'non-active' part of the composition delivered, e.g. wherein such 'non-active' part is not delivered simultaneously with the 'active' part of the composition
  • A61K48/0025—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy characterised by an aspect of the 'non-active' part of the composition delivered, e.g. wherein such 'non-active' part is not delivered simultaneously with the 'active' part of the composition wherein the non-active part clearly interacts with the delivered nucleic acid
  • A61K48/0041—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy characterised by an aspect of the 'non-active' part of the composition delivered, e.g. wherein such 'non-active' part is not delivered simultaneously with the 'active' part of the composition wherein the non-active part clearly interacts with the delivered nucleic acid the non-active part being polymeric

Definitions

• Patent No. 4919397 Patent No. 4598908 Patent No. 6093710 International Publication No. 2017/218704 International Publication No. 2021/060440
• Patent Document 5 discloses that lipid nanoparticles that do not contain nucleic acids are prepared in an acidic buffer, further added with a cryoprotectant, lyophilized, and then rehydrated with an aqueous solution containing nucleic acids. It has been shown that lipid nanoparticles can be easily and efficiently encapsulated in lipid nanoparticles. However, since the above technique requires a freeze-drying step, there is room for improvement in terms of simplicity.
• w in X 3 is 1 or 2.
• X 3 is an imidazolidylene group
• X 3 is a piperazylene group.
• R 2a may be the same as or different from R 2b , but preferably R 2a is the same group as R 2b .
• Y a and Y b are each independently an ester bond, an amide bond, a carbamate bond, an ether bond, or a urea bond, preferably each independently an ester bond, an amide bond, or a carbamate bond, and more preferably each Independently, it is an ester bond or an amide bond, most preferably each an ester bond.
• the direction of the bond between Y a and Y b is not limited, but when Y a and Y b are ester bonds, preferably -Z a -CO-O-R 2a - and -Z b -CO-O-R 2b It exhibits a structure of -.
• aromatic rings contained in aromatic compounds having 3 to 16 carbon atoms include benzene rings, naphthalene rings, anthracene rings for aromatic hydrocarbon rings, imidazole rings, pyrazole rings, oxazole rings for aromatic heterocycles, Isoxazole ring, thiazole ring, isothiazole ring, triazine ring, pyrrole ring, furanthiophene ring, pyrimidine ring, pyridazine ring, pyrazine ring, pyridine ring, purine ring, pteridine ring, benzimidazole ring, indole ring, benzofuran ring, quinazoline ring, phthalazine ring, quinoline ring, isoquinoline ring, coumarin ring, chromone ring, benzodiazepine ring, phenoxazine ring, phenothiazine ring, acrid
• each R 4 may be the same or different.
• R 3a may be the same as or different from R 3b , but preferably R 3a is the same group as R 3b .
• R 1a and R 1b are each independently an alkylene group having 1 to 4 carbon atoms (e.g., methylene group, ethylene group);
• X a and X b are each independently an acyclic alkyl tertiary amino group having 1 to 3 carbon atoms and having 1 tertiary amino group (e.g., -N(CH 3 )-); or a cyclic alkylene tertiary amino group having 2 to 5 carbon atoms and 1 tertiary amino group (e.g., piperidylene group);
• R 2a and R 2b are each independently an alkylene group having 6 or less carbon atoms (e.g., methylene group, ethylene group, trimethylene group);
• Y a and Y b are each independently an ester bond or an amide bond;
• Z a and Z b are each independently a divalent group derived from an aromatic compound having 6 to 12 carbon atoms, one aromatic ring, and optionally
• fat-soluble vitamins having a hydroxyl group examples include retinol, ergosterol, 7-dehydrocholesterol, calciferol, colcalciferol, dihydroergocalciferol, dihydrotachysterol, tocopherol, and tocotrienol.
• the fat-soluble vitamin having a hydroxyl group is preferably tocopherol.
• the protection or deprotection reaction of the functional group is carried out using known reagents and methods (for example, the reagents and methods described in GREENE'S PROTECTIVE GROUPS IN ORGANIC SYNTHESIS, 4th edition, WILEY-INTERSCIENCE EM), or by methods known in the art. It is carried out according to the method described in the example.
• PEG Lipid PEG lipid coats the surface of lipid nanoparticles with hydrophilic polyethylene glycol (PEG), and can be used as a stabilizer by suppressing aggregation between particles, or as a stabilizer for interactions between biological components and particles when administered to living organisms.
• PEG polyethylene glycol
• the PEG region can be of any molecular weight. In some embodiments, the PEG region has a molecular weight of 200-10,000 Da and can be linear or branched.
• an acidic buffer containing nucleic acids As the aqueous solution containing nucleic acids, it is preferable to use an acidic buffer containing nucleic acids.
• a buffer having a buffering effect in an acidic region can be used, preferably an acidic buffer having a buffering effect in a pH range of 1 to 6.5, more preferably a pH of 3 to 6.5.
• the acidic buffers used in the preparation of lipid nanoparticles include the same acidic buffers as those exemplified above, such as MES buffer, malic acid buffer, phthalic acid buffer, and maleic acid buffer. , succinate buffer, tartrate buffer, citrate buffer, and the like.
• lipid nanoparticles encapsulating nucleic acids it is possible to introduce nucleic acids into cells not only in vitro but also in vivo. That is, by administering lipid nanoparticles encapsulating nucleic acids to a subject, the lipid nanoparticles reach and contact target cells, and the nucleic acids encapsulated in the lipid nanoparticles are introduced into the cells in vivo. Ru.
• the particle solution (total lipid 20mM, 15 ⁇ L) was mixed with a solution containing luciferase-encoding mRNA (CleanCap (registered trademark) FLuc mRNA - (L-7602)) (nucleic acid concentration 1.5 ⁇ g/135 ⁇ L, buffer conditions MES (pH 6), 135 ⁇ L) and diluted under stirring conditions using a vortex mixer, and incubated at room temperature for 60 minutes, 30 minutes, 15 minutes, 10 minutes, 5 minutes, or 1 minute using an aluminum block incubator. 150 ⁇ L of PBS was added to the solution after incubation to obtain nucleic acid-encapsulated nanoparticles.
• luciferase-encoding mRNA CleanCap (registered trademark) FLuc mRNA - (L-7602)
• Comparative Example 1 Method for Preparing Lyophilized Product
• SS-OP was used as the ionic lipid
• DOPC, Chol, and DMG-PEG2000 were used as other lipids.
• the molar ratio of SS-OP:DOPC:Chol used was 52.5:7.5:40, and 1.5 mol% of DMG-PEG2000 was used with respect to the total of SS-OP, DOPC, and Chol.
• Comparative Example 2 Method for Preparing Lyophilized Product
• SS-OP was used as the ionic lipid
• DOPC, Chol, and DMG-PEG2000 were used as other lipids.
• the molar ratio of SS-OP:DOPC:Chol used was 52.5:7.5:40, and 1.5 mol% of DMG-PEG2000 was used with respect to the total of SS-OP, DOPC, and Chol.
• the obtained nucleic acid-encapsulated nanoparticles or the nucleic acid-encapsulated nanoparticles prepared in Comparative Example 1 using the freeze-drying technique were applied to mice (Balb/c, 6 weeks old, female), and the gene expression activity in the liver was measured using GloMax 20/ 20 Evaluated with Luminometer.
• Each nucleic acid-encapsulated nanoparticle was administered to mice through the tail vein at a dose of 0.05 mg/kg.
• the liver was removed and the expression level of luciferase protein was evaluated.
• higher gene expression activity in the liver was observed in the liquid formulation and the frozen-thawed version compared to the nucleic acid-encapsulated nanoparticles produced by freeze-drying technology (see Figure 18).
• the particle solution (total lipid 20mM, 15 ⁇ L) (liquid) was mixed with a solution containing luciferase-encoding mRNA (CleanCap® FLuc mRNA - (L-7602)) (nucleic acid concentration 3 ⁇ g/135 ⁇ L, buffer condition MES (pH 6)). , 135 ⁇ L) under stirring conditions using a vortex mixer while diluting, and incubated at 37° C. for 5 minutes using an aluminum block incubator. 150 ⁇ L of PBS was added to the solution after incubation to obtain nucleic acid-encapsulated nanoparticles.

Landscapes

• Health & Medical Sciences (AREA)
• Chemical & Material Sciences (AREA)
• Life Sciences & Earth Sciences (AREA)
• Engineering & Computer Science (AREA)
• General Health & Medical Sciences (AREA)
• Pharmacology & Pharmacy (AREA)
• Medicinal Chemistry (AREA)
• Veterinary Medicine (AREA)
• Public Health (AREA)
• Animal Behavior & Ethology (AREA)
• Epidemiology (AREA)
• Bioinformatics & Cheminformatics (AREA)
• Genetics & Genomics (AREA)
• Molecular Biology (AREA)
• Biomedical Technology (AREA)
• General Chemical & Material Sciences (AREA)
• Chemical Kinetics & Catalysis (AREA)
• Physics & Mathematics (AREA)
• Biotechnology (AREA)
• Nanotechnology (AREA)
• Optics & Photonics (AREA)
• Oil, Petroleum & Natural Gas (AREA)
• Organic Chemistry (AREA)
• Biochemistry (AREA)
• Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
• General Engineering & Computer Science (AREA)
• Wood Science & Technology (AREA)
• Zoology (AREA)
• Dispersion Chemistry (AREA)
• Manufacturing & Machinery (AREA)
• Biophysics (AREA)
• Plant Pathology (AREA)
• Microbiology (AREA)
• Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
• Medicinal Preparation (AREA)

Priority Applications (6)

Applications Claiming Priority (2)

Publications (1)

Family

ID=88201455

Family Applications (1)

Country Status (7)

Citations (9)

Family Cites Families (1)

• 2023
  • 2023-03-24 WO PCT/JP2023/011843 patent/WO2023190170A1/ja not_active Ceased
  • 2023-03-24 CA CA3247022A patent/CA3247022A1/en active Pending
  • 2023-03-24 JP JP2024512359A patent/JPWO2023190170A1/ja active Pending
  • 2023-03-24 CN CN202380031621.3A patent/CN119136789A/zh active Pending
  • 2023-03-24 EP EP23780180.8A patent/EP4501317A4/en active Pending
  • 2023-03-24 KR KR1020247035601A patent/KR20240161201A/ko active Pending
  • 2023-03-24 US US18/852,121 patent/US20250205168A1/en active Pending

Patent Citations (11)

Non-Patent Citations (3)

Also Published As

Similar Documents

Legal Events