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WO2020203961A1 - Structure de membrane lipidique et son procédé de fabrication - Google Patents

Structure de membrane lipidique et son procédé de fabrication Download PDF

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Publication number
WO2020203961A1
WO2020203961A1 PCT/JP2020/014526 JP2020014526W WO2020203961A1 WO 2020203961 A1 WO2020203961 A1 WO 2020203961A1 JP 2020014526 W JP2020014526 W JP 2020014526W WO 2020203961 A1 WO2020203961 A1 WO 2020203961A1
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Prior art keywords
lipid
dispersion
coq
lipid membrane
membrane structure
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Japanese (ja)
Inventor
勇磨 山田
原島　秀吉
光恵 日比野
悠介 佐藤
学 渡慶次
正寿 真栄城
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Luca Science Inc
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Luca Science Inc
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    • 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
    • A61K31/12—Ketones
    • A61K31/122—Ketones having the oxygen directly attached to a ring, e.g. quinones, vitamin K1, anthralin
    • 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
    • A61K47/24—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing atoms other than carbon, hydrogen, oxygen, halogen, nitrogen or sulfur, e.g. cyclomethicone or phospholipids
    • 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
    • A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
    • A61K47/42—Proteins; Polypeptides; Degradation products thereof; Derivatives thereof, e.g. albumin, gelatin or zein
    • 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
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A61P1/16—Drugs for disorders of the alimentary tract or the digestive system for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
    • A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A61P9/10—Drugs for disorders of the cardiovascular system for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis

Definitions

  • the present invention relates to a lipid membrane structure and a method for producing the same.
  • CoQ 10 is very lipophilic and cannot be applied to injection preparations, the main dosage form is tablets. However, in general, it is difficult to adapt tablets to the treatment of ischemic diseases that require prompt response, and despite the fact that CoQ 10 has functions such as strong antioxidant activity because the dosage form is tablets. The range of indications is narrowing. In response to this situation, the present inventors encapsulated CoQ 10 in lipid bilayer vesicles (liposomes) to obtain dispersibility in a solution, and further provided CoQ as a nanocapsule having a mitochondrial targeting ability. A 10- mounted MITO-Porter was constructed (Non-Patent Document 1, Patent Document 1).
  • CoQ 10 mounted formulations disclosed as MITO-Porter in these references includes a lipid membrane containing a dioleyl phosphatidyl ethanolamine and phosphatidic acid or sphingomyelin, lipid membrane structure containing octaarginine (R8) Become.
  • This system realizes efficient delivery of CoQ 10 to mitochondria, which is a source of active oxygen, and can suppress injury during ischemia.
  • Non-Patent Document 2 by intravenous administration to the liver ischemia model animals (mice), in the tissue of the animal, it was confirmed delivery and accumulation in the mitochondria into the cell CoQ 10 (Non-Patent Document 2 ).
  • Drug delivery system using nanocapsules such as liposomes allows existing drugs and nucleic acid drugs to be encapsulated or carried on the surface of particles, so that the drugs accumulate at the target site, increasing the therapeutic effect and reducing side effects. It brings about biological effects such as. Furthermore, nanocapsules are also applied to solubilization technology and contribute to improving the dispersibility of poorly water-soluble molecules such as CoQ 10 . It also protects the encapsulated drug from oxidation, can improve stability, and has a physicochemical effect.
  • Typical methods for preparing liposomes include a simple hydration method, a reverse phase evaporation method (REV) method, and an alcohol dilution method. It has been shown that when the above-mentioned CoQ 10- loaded MITO-Porter is prepared by an ethanol dilution method, particles having the highest CoQ 10 loading rate (drug content with respect to lipid) can be obtained (Non-Patent Document 3). ).
  • BCS class 4 Biopharmaceutics Classication System, Table 1
  • BCS class 4 Biopharmaceutics Classication System, Table 1
  • drugs belonging to BCS class 4 have poor gastrointestinal absorption due to poor gastrointestinal membrane permeability, and are generally considered to be difficult to formulate.
  • BCS class 4 drugs are poorly water-soluble, it is difficult to make them into injections, and because they are poorly permeable to the gastrointestinal membrane, it is difficult to tablet them as oral preparations to exert a high effect. is there.
  • CoQ 10 mounted MITO-Porter is the therapeutic effect can be expected.
  • the particle size of the CoQ 10- equipped MITO-Porter obtained repeatedly under the same conditions is in the range of 80 to 120 nm, and the particle size is uniform.
  • the CoQ 10- equipped MITO-Porter can be surface-modified with a polyarginine peptide typified by R8 (octaarginine), and the surface charge can be adjusted so as to have a predetermined zeta potential (for example, in the range of 15 to 25 mV). ..
  • a predetermined zeta potential for example, in the range of 15 to 25 mV.
  • the surface is modified with a polyarginine peptide after the preparation of the nanocapsules, but the reproducibility of the surface charge is poor and it is difficult to stably prepare the nanocapsules having a predetermined zeta potential.
  • the amount of CoQ 10- MITO-Porter prepared in one preparation was also small (for example, about 400 ⁇ L).
  • a new method capable of efficiently encapsulating, stable, and preparing nanocapsules having a smaller particle size than that obtained by the conventional method can be provided. Is desired.
  • MITO-Porter By including a poorly water-soluble compound, MITO-Porter can be efficiently solubilized and can be an efficient means of transport to mitochondria. If it is possible to efficiently encapsulate poorly water-soluble molecules other than CoQ 10 and to prepare nanocapsules that are stable and have a smaller particle size than that obtained by the conventional method, many water-resistant molecules belonging to BCS class 4 are poorly water-soluble. The application of sex compounds to pharmaceuticals can be greatly expanded.
  • lipid membrane structure or lipid
  • lipid that contains a poorly water-soluble compound such as that classified as BCS Class 4 containing CoQ 10 and has a smaller particle size and / or a smaller polydispersity index.
  • a method for producing a lipid film structure (or lipid nanoparticles) capable of preparing (nanoparticles) with good reproducibility and mass production is provided.
  • the present invention is as follows.
  • [1] A dispersion containing a poorly water-soluble compound and having a lipid film structure having an average particle size of 60 nm or less measured by a dynamic light scattering (DLS) method as a dispersoid in a dispersion medium, wherein the lipid film is contained.
  • [2] The dispersion according to [1], wherein the phospholipid is diorail phosphatidylethanolamine, phosphatidic acid and / or sphingomyelin.
  • a method for producing a dispersion which comprises a step of diluting the alcohol solution with an aqueous solvent and recovering a dispersion containing a lipid film structure containing a poorly water-soluble compound as a dispersoid from an outlet of a microchannel.
  • the phospholipid is dioleylphosphatidylethanolamine and phosphatidic acid and / or sphingomyelin
  • the membrane-permeable peptide is a polyarginine peptide consisting of 4 to 20 consecutive arginine residues.
  • the lipid membrane structure contains a phospholipid, a membrane-permeable peptide, and a lipid-modified polyethylene glycol.
  • the dilution flow path has a two-dimensionally bent flow path portion at least in a part thereof, and the bent flow path is formed.
  • the axial direction of the dilution flow path upstream from this or the extension direction thereof is the X direction
  • the width direction of the dilution flow path perpendicular to the X direction is the Y direction.
  • the structure that regulates the flow path width of the dilution flow path is formed at regular intervals d1, d2. .. .. It is a flow path structure formed by providing at least two of them, and the alcohol solution is introduced into the first introduction path, and the aqueous solvent is introduced into the second introduction path, [12] to [18]. ]
  • the manufacturing method according to any one of. [20] The dispersion according to any one of the above [1] to [11].
  • a lipid membrane structure comprising one or more phospholipids selected from the group consisting of phosphatidic acid and sphingomyelin, dioleyl phosphatidylethanolamine, and lipid-modified polyethylene glycol.
  • a lipid membrane structure is a dispersion that contains precursors in the biosynthetic pathway between the inner and outer membranes of ubiquinone or its mitochondria. [21] The dispersion according to any one of the above [1] to [11].
  • a dispersion comprising a lipid membrane structure comprising lipid-modified polyethylene glycol, wherein the lipid membrane structure comprises curcumin.
  • the lipid membrane structure is a dispersion having an average particle size of 20 to 150 nm by the DLS method and a polydispersity index of 0.3 or less.
  • a dispersion in which the membrane-permeable peptide is a peptide selected from the group consisting of octaarginine (R8) and S2 peptides.
  • BCS Biopharmaceutics Classification System
  • lipid membrane structure has a polydispersity index (PDI) of 0.3 or less measured by a dynamic light scattering method.
  • PDI polydispersity index
  • the lipid membrane contains one or more phospholipids selected from the group consisting of phosphatidic acid and sphingomyelin, and dioleylphosphatidylethanolamine. Body or composition.
  • the lipid membrane structure further contains a membrane-permeable peptide and expresses the membrane-permeable peptide.
  • the membrane-permeable peptide can also coexist when producing the lipid nanoparticles, and the lipid nanoparticles containing the membrane-permeable peptide can be produced in one step, and as a result, the physical properties of the obtained lipid nanoparticles can be improved. Be controlled. Moreover, by modifying the surface of the lipid nanoparticles with the cationic polymer, the zeta potential of the lipid nanoparticles could be stably controlled.
  • lipid membrane structure containing a poorly water-soluble compound (eg, a BCS class 4 compound) having a more homogeneous particle size and / or a smaller particle size. Nanoparticles) can be prepared in large quantities.
  • the present invention it is possible to provide poorly water-soluble molecule-encapsulating lipid nanoparticles having a particle size of 60 nm or less, which has not been obtained in the past. Furthermore, the lipid nanoparticles have a smaller polydispersity index, and as a result, have the effect of increasing the efficiency of uptake into cells and mitochondria in combination with the small particle size.
  • a schematic explanatory diagram of a method for preparing a lipid membrane structure (or lipid nanoparticles or nanocapsules) using a microchannel device in an example is shown.
  • the experimental result (before dialysis) when the flow velocity ratio of the lipid phase and the aqueous phase was adjusted in Example 1 is shown.
  • the experimental result (after dialysis) when the flow velocity ratio of the lipid phase and the aqueous phase was adjusted in Example 1 is shown.
  • the result of the influence test of the dialysis temperature in Example 1 is shown.
  • the result of the stability test in Example 1 is shown. Comparative Example 1 shows the physical properties of the lipid nanoparticles obtained by the conventional method.
  • Example 3 The particle size, PdI and zeta potential of the lipid membrane structure (or lipid nanoparticles) obtained by the conventional method (Comparative Example 1) and Example 1 (before and after dialysis) in Example 2 are shown.
  • the cell uptake evaluation by flow cytometry (FACS) and the intracellular localization observation result by confocal laser scanning microscope (CLMS) using cervical cancer HeLa cells in Example 3 are shown.
  • the intracellular localization observation (HeLa cell) image in Example 3 is shown.
  • the intracellular localization observation (model disease cell) image in Example 3 is shown.
  • the intracellular localization observation (Human CDC cell) image in Example 3 is shown.
  • An image of intracellular localization observation (Human pulponary artery smooth muscle cells) in Example 3 is shown.
  • FIG. 5 shows a transmission electron microscope image of the CoQ 10- containing lipid membrane structure (or lipid nanoparticles) in Example 1 by negative staining.
  • OCR oxygen consumption rate
  • Example 7-1 Shows the scheme of therapy experiments with CoQ 10 containing lipid membrane structure of the liver injury model mice in Example 7-2 (or lipid nanoparticle). The results of the treatment experiment with the CoQ 10- containing lipid membrane structure (or lipid nanoparticles) of the liver disorder model mouse in Example 7-2 are shown. The average particle size and PdI of the curcumin-containing lipid membrane structure (or lipid nanoparticles) before and after dialysis in Example 8 are shown.
  • the dispersion is a composition containing a dispersoid and a dispersion medium.
  • the dispersoid is a lipid membrane structure containing a poorly water-soluble compound, and the dispersion medium can be an aqueous solvent.
  • the dispersion can preferably be a colored or uncolored clear solution.
  • the dispersoid is a lipid membrane structure (or lipid nanoparticles) containing a phospholipid and a lipid-modified uncharged hydrophilic polymer (eg, lipid-modified polyethylene glycol).
  • the lipid nanoparticles may preferably represent a membrane-permeable molecule (eg, a peptide) that promotes permeability to the cell membrane.
  • the lipid membrane structure has a lipid membrane structure containing a lipid-modified uncharged hydrophilic polymer in addition to phospholipids, whereby the dispersibility of the dispersoid in the dispersion is improved, and the dispersion (for example, a solution) ) Is prevented from becoming turbid.
  • the phospholipid the following phospholipids can be used, and preferably, one or more phospholipids selected from the group consisting of phosphatidic acid and sphingomyelin, and dioleylphosphatidylethanolamine can be contained.
  • the lipid membrane structure may further comprise the charged substances and / or membrane permeable molecules described below.
  • the uncharged hydrophilic polymer that can be used in the present invention can be a polymer that has no charge as a whole molecule and is hydrophilic.
  • the uncharged hydrophilic polymer is not particularly limited as long as it does not significantly inhibit the particle formation of the present invention, and examples thereof include polyoxazolines and polyalkylene glycols.
  • polyalkylene glycol polyethylene glycol can be preferably used.
  • polymer for example, a polymer having a number average molecular weight of 1,000 to 150,000 Da, preferably 1,000 to 5,000 Da (for example, about 2,000 Da) can be used.
  • the uncharged hydrophilic polymer can be linked to the lipid and complexed into a lipid membrane structure.
  • lipid membrane structure From the viewpoint of facilitating detachment from the lipid membrane structure, it can be linked to a lipid that is easily detached from the lipid membrane.
  • diacylglycerol for example, 1,2-dimyristoyl-sn-glycerol
  • Lipid-modified uncharged hydrophilic polymers can be added to prevent turbidity in the dispersion.
  • the lipid membrane structure is obtained as nanoparticles by mixing a lipid phase in which lipids are dissolved and an aqueous phase on a microchannel device containing a baffle mixer. Details of the manufacturing method will be described later.
  • an organic solvent for example, alcohol can be used as the solvent.
  • alcohols for example, t-butanol, 1-propanol, 2-propanol and 2-butoxyethanol can be used as long as biotoxicity is not significantly caused, and ethanol can be preferably used.
  • the poorly water-soluble compound can be made soluble in the alcohol. If heating is required for dissolution, the poorly water-soluble compound may be dissolved in alcohol by heating. Alcohol may be selected from the viewpoint of increasing solubility depending on the type of the poorly water-soluble compound to be included.
  • Lipid membrane structures include phospholipids and lipid-modified uncharged hydrophilic polymers (eg, lipid-modified polyethylene glycol).
  • the dispersoid has a particle size measured by the DLS method of 150 nm or less, 140 nm or less, 130 nm or less, 120 nm or less, 100 nm or less, 90 nm or less, 80 nm or less, 70 nm or less, 60 nm or less, preferably 55 nm or less, or more preferably 50 nm.
  • the lipid membrane structure may have a particle size of 10 nm or more, 20 nm or more, 30 nm or more, 40 nm or more, or 50 nm or more in the particle size measured by the DLS method.
  • the number average particle size of the lipid membrane structure can be, for example, 30 nm or more, 35 nm or more, 40 nm or more, 45 nm or more, or 50 nm or more, 150 nm or less, 140 nm or less, 130 nm or less, 120 nm or less, It can be 110 nm or less, 100 nm or less, 95 nm or less, 90 nm or less, 85 nm or less, 80 nm or less, 75 nm or less, 70 nm or less, 65 nm or less, or 60 nm or less. In some embodiments, the number average particle size can be 30 nm to 150 nm or less, for example 50 nm to 100 nm. As described above, the particle size may be the particle size obtained by the DLS method.
  • the lipid membrane structure (or lipid nanoparticles) has a polydispersity index (PDI) obtained by the DLS method of 0.5 or less, 0.45 or less, 0.4 or less, 0.35 or less, 0.3 or less. , 0.25 or less, or 0.2 or less.
  • PDI means that 1 is the maximum, and the smaller the value, the more uniform the particle size (the particle size distribution becomes sharper and / or monodisperse).
  • the PDI of the lipid membrane structure (or lipid nanoparticles) is 0.3 or less.
  • the lipid membrane structure (or lipid nanoparticles) is The particle size measured by the DLS method is 30 nm to 150 nm. It contains a lipid membrane structure (or lipid nanoparticles) having a PDI determined by the DLS method of 0.3 or less.
  • the particle size is, for example, 30 nm to 40 nm, 40 nm to 150 nm, 40 nm to 140 nm, 40 nm to 130 nm, 40 nm to 120 nm, 40 nm to 110 nm, 40 nm to 100 nm, 40 nm to 90 nm, 40 nm to 80 nm, 40 nm to 70 nm, 40 nm.
  • the lipid membrane structure (or lipid nanoparticles) is The particle size measured by the DLS method is 30 nm to 150 nm. It contains 50% or more, 60% or more, 70% or more, or 80% or more of lipid membrane structures (or lipid nanoparticles) having a PDI determined by the DLS method of 0.3 or less.
  • the particle size is, for example, 30 nm to 40 nm, 40 nm to 150 nm, 40 nm to 140 nm, 40 nm to 130 nm, 40 nm to 120 nm, 40 nm to 110 nm, 40 nm to 100 nm, 40 nm to 90 nm, 40 nm to 80 nm, 40 nm to 70 nm, 40 nm.
  • the lipid membrane structure (or lipid nanoparticles) is The number average (number average particle size) of the particle size measured by the DLS method is 30 nm to 150 nm. It contains a lipid membrane structure (or lipid nanoparticles) having a PDI determined by the DLS method of 0.3 or less.
  • the number average particle size is, for example, 30 nm to 40 nm, 40 nm to 150 nm, 40 nm to 140 nm, 40 nm to 130 nm, 40 nm to 120 nm, 40 nm to 110 nm, 40 nm to 100 nm, 40 nm to 90 nm, 40 nm.
  • the lipid membrane structure (or lipid nanoparticles) comprises phospholipids and lipid-modified polyethylene glycol.
  • the lipid membrane structure is a cationic polymer such as a cell membrane penetrating peptide (eg, a polymer of cationic amino acids, eg, polyarginine).
  • the polymer may be expressed.
  • a conjugate of the cationic polymer and a lipid for example, a myristol group
  • the lipid membrane is formed on the lipid portion of the conjugate.
  • the membrane-permeable peptides include tat peptide (having a peptide sequence corresponding to the 48 to 60th position of the amino acid sequence of AAF35362.1, GenBank registration number of tat protein of human immunodeficiency virus), oligoarginine (R9), oligolysine.
  • tat peptide having a peptide sequence corresponding to the 48 to 60th position of the amino acid sequence of AAF35362.1, GenBank registration number of tat protein of human immunodeficiency virus
  • R9 oligoarginine
  • oligolysine examples thereof include peptides rich in basic amino acids such as (K10), amphipathic peptides having a basic portion and a hydrophobic portion such as penetratin, and peptides such as transportin and TP10.
  • Membrane-permeable peptides having mitochondrial orientation include lipophilic cations such as octaarginine (R8), lipophilic triphenylphosphonium cation (TPP) or rhodamine 123, and mitochondria.
  • Target sequence Mitochondrial Targeting Sequence; MTS
  • MTS Mitochondrial Targeting Sequence
  • S2 peptide Szeto, H. R. 2011, 28, pp. 2669-2679
  • the S2 peptide Dmt-D-Arg-FK-Dmt-D-Arg-FK-NH 2 ⁇ Here, Dmt is 2,6-dimethyltyrosine, and D-Arg is D-form arginine. Yes, F is L-form phenylalanine and K is L-form lysine ⁇ .
  • the zeta potential of the lipid membrane structure can be 5 mV or higher, 10 mV or higher, 15 mV or higher, 16 mV or higher, 17 mV or higher, 18 mV or higher, 19 mV or higher, or 20 mV or higher, for example, about 20 mV. ..
  • the dispersions of the invention contain less than 1%, less than 0.5%, less than 0.1%, or less than 0.05% alcohol that can be used during production in the dispersion medium. Or does not contain alcohol.
  • the dispersion can be dialyzed with an alcohol-free external dialysis solution.
  • the external dialysis solution can be, for example, an aqueous solution such as physiological saline.
  • the dispersion of the present invention comprises a lipid membrane structure containing a poorly water soluble compound (eg, a BCS class 4 compound such as curcumin and / or CoQ 10 ).
  • a poorly water soluble compound eg, a BCS class 4 compound such as curcumin and / or CoQ 10
  • the poorly water-soluble compound can be a compound exhibiting solubility in ethanol.
  • the lipid membrane structure contains 10-40 mol% (preferably 20-40 mol%) of CoQ 10 relative to the total amount of lipid membrane, eg 20. Included in the range of ⁇ 30 mol%).
  • the lipid membrane structure containing CoQ 10 may use ubiquinone (oxidized ubiquinone) instead of CoQ 10 , or may use a precursor of ubiquinone or CoQ 10 in the biosynthetic pathway between the inner and outer membranes of mitochondria. May be good.
  • Ubiquinone is called CoQ n , corresponding to the number n of repeating units of the isoprene.
  • n can be a natural number in the range of 4 to 15 or 6 to 12, for example, a natural number in the range of 6 to 10, for example, in the range of 8 to 12. It can be a natural number, eg, a natural number in the range 8-10, eg, 10.
  • Examples of these precursors include dimethoxyubiquinone (DMQ) and 5-hydroxyubiquinone (5-HQ).
  • DMQ dimethoxyubiquinone
  • 5-HQ 5-hydroxyubiquinone
  • the R8-expressing lipid membrane structure of the present invention was considered to have delivered ubiquinone to the inner mitochondrial membrane, and was able to improve the oxygen consumption rate of cells. Therefore, the R8 expressive lipid membrane structure of the present invention can be used to deliver ubiquinone from extracellular to the inner mitochondrial membrane.
  • the dispersion comprises one or more phospholipids selected from the group consisting of phosphatidic acid and sphingomyelin, dioleylphosphatidylethanolamine, and steallylated polyethylene glycol.
  • the lipid membrane structure comprises precursors (preferably CoQ n , particularly CoQ 10 ) in the biosynthetic pathway between the inner and outer membranes of ubiquinone or its mitochondria.
  • the lipid membrane structure includes a lipid membrane structure having a particle size of 20 to 100 nm according to the DLS method, and has a PDI of less than 0.3.
  • the dispersion (composition) in this preferred embodiment can have a number average particle size of 50 nm to 70 nm.
  • the dispersion medium can be saline, preferably less than 1%, 0.5% or less, 0.4% or less, 0.3% or less, 0.2% or less, 0 alcohol. .Contains less than 1%, or less than 0.01%, or less than the detection limit, or does not contain alcohol.
  • the lipid membrane structure may further express a cell membrane penetrating peptide (eg, R8). The lipid membrane structure is non-hollow.
  • the dispersion comprises one or more phospholipids selected from the group consisting of phosphatidic acid and sphingomyelin, dioleylphosphatidylethanolamine, and steallylated polyethylene glycol.
  • the lipid membrane structure comprises curcumin, the lipid membrane structure comprises a lipid membrane structure having a particle size of 40-300 nm according to the DLS method, and the PDI is less than 0.3. ..
  • the dispersion (composition) in this preferred embodiment can have a number average particle size of 100 nm to 120 nm.
  • the dispersion medium can be saline, preferably less than 1%, 0.5% or less, 0.4% or less, 0.3% or less, 0.2% or less, 0 alcohol. .Contains less than 1%, or less than 0.01%, or less than the detection limit, or does not contain alcohol.
  • the lipid membrane structure may further express the cell membrane penetrating peptide and / or mitochondrial directional peptide (eg, R8 and / or S2 peptide).
  • the method for producing a dispersion of the present invention can be used to produce the above-mentioned dispersion.
  • an alcohol solution in which a phospholipid, a membrane-permeable peptide, a lipid-modified polyethylene glycol, and a poorly water-soluble compound are dissolved and an aqueous solvent are continuously connected to the entrance of the microchannel of the microchannel structure.
  • the alcohol solution is mixed (that is, diluted) with an aqueous solvent in the microchannel, and a dispersion containing a lipid membrane structure containing a poorly water-soluble compound as a dispersoid is provided in the microchannel.
  • the lipid membrane structure (dispersant) thus obtained is lipid nanoparticles.
  • the nanoparticles are non-hollow structures filled with phospholipids or lipid membranes (non-hollow structures; or non-hollow lipid nanoparticles; or non-hollow if they represent mitochondrial directional molecules). It can be a hollow MITO-Porter).
  • the dispersoid in the present invention may be referred to as lipid nanoparticles (or nanocapsules).
  • the phospholipid is not particularly limited, and is, for example, phosphatidylcholine (for example, dioleoil phosphatidylcholine, dilauroylphosphatidylcholine, dipalmitoylphosphatidylcholine, dipalmitoylphosphatidylcholine, distearoylphosphatidylcholine, etc.) Phosphatidylglycerol, dipalmitoylphosphatidylglycerol, dipalmitoylphosphatidylglycerol, dipalmitoylphosphatidylglycerol, distearoylphosphatidylglycerol, etc.
  • phosphatidylcholine for example, dioleoil phosphatidylcholine, dilauroylphosphatidylcholine, dipalmitoylphosphatidylcholine, dipalmitoylphosphatidylcholine, distearoylphosphatid
  • phosphatidylserine phosphatidylinositol, phosphatidylic acid, cardiolipin, sphingomyelin, ceramide phosphorylethanolamine, ceramidephosphorylglycerol, ceramidephosphorylglycerol phosphate, 1,2-dipalmitoyl-1,2- It can be deoxyphosphatidylcholine, plasmalogen, egg yolk lecithin, soybean lecithin, hydrogenated additives thereof and the like.
  • Phospholipids are preferably phosphatidylethanolamine (eg, diolaylphosphatidylethanolamine, dilauroylphosphatidylethanolamine, dimyristylphosphatidylethanolamine, dipalmitoylphosphatidylethanolamine, distearoylphosphatidylethanolamine, etc.) and sphingomyelin. More preferably, diorail phosphatidylethanolamine and sphingomyelin.
  • phosphatidylethanolamine eg, diolaylphosphatidylethanolamine, dilauroylphosphatidylethanolamine, dimyristylphosphatidylethanolamine, dipalmitoylphosphatidylethanolamine, distearoylphosphatidylethanolamine, etc.
  • sphingomyelin More preferably, diorail phosphatidylethanolamine and sphingomyelin.
  • a lipid membrane containing a phospholipid can contain a charged substance in addition to the phospholipid, and the charged substance is a component of the lipid membrane capable of imparting a positive charge or a negative charge to the lipid membrane, and is a lipid.
  • the amount of charged substance contained in the membrane is usually 30% (molar ratio) or less, preferably 25% (molar ratio) or less, and more preferably 20% (molar ratio) or less of the total amount of substances constituting the lipid membrane. ..
  • the lower limit of the content of the charged substance is 0.
  • Examples of the charged substance that imparts a positive charge include saturated or unsaturated aliphatic amines such as stearylamine and oleylamine; saturated or unsaturated cationic synthetic lipids such as dioleoyltrimethylammonium propane, and the like, and negative charges are given.
  • Examples of the charged substance to be imparted include disetyl phosphate, cholesteryl hemisuccinate, phosphatidylserine, phosphatidyl inositol, phosphatidyl acid and the like.
  • the zeta potential of the lipid membrane structure can be 5 mV or higher, 10 mV or higher, 15 mV or higher, 16 mV or higher, 17 mV or higher, 18 mV or higher, 19 mV or higher, or 20 mV or higher, for example, about 20 mV. ..
  • the phospholipids are diorail phosphatidylethanolamine and phosphatidic acid and / or sphingomyelin (ie, one or more phospholipids selected from the group consisting of phosphatidic acid and sphingomyelin, and diorail phosphatidylethanolamine. Included) is preferred for effective delivery of the poorly water-soluble compound, which is the substance of interest, into the mitochondria.
  • the membrane permeable molecule can be, for example, a cationic polymer.
  • the membrane permeable molecule can be, for example, a membrane permeable peptide.
  • Membrane-permeable peptides are membrane-permeable domains that are effective for the effective delivery of poorly water-soluble compounds of interest into mitochondria.
  • the membrane-permeable peptide can be the membrane-permeable peptide described in paragraphs 0052 to 0092 of Patent Document 1, preferably a polyarginine peptide consisting of 4 to 20 consecutive arginine residues.
  • the polyarginine peptide preferably consists of 6-12, more preferably 7-10 contiguous arginine residues, or 8 contiguous arginine residues.
  • Membrane-permeable peptides can be linked to lipids. Thereby, the membrane-permeable peptide can be contained in the lipid membrane structure, and the membrane-permeable peptide can be expressed on the lipid membrane structure.
  • the zeta potential of the lipid membrane structure can be 5 mV or higher, 10 mV or higher, 15 mV or higher, 16 mV or higher, 17 mV or higher, 18 mV or higher, 19 mV or higher, or 20 mV or higher, for example, about 20 mV. ..
  • Lipid-modified polyethylene glycol is a component that imparts hydrophilicity to lipid membranes.
  • Lipid-modified polyethylene glycol (PEG) is a compound obtained by lipid-modifying polyethylene glycol (PEG), and the molecular weight of polyethylene glycol is, for example, about 300 to 10,000, preferably about 500 to 10,000, and more preferably 1. It is about 000 to 5,000.
  • the molecular weight of PEG is represented by a number average molecular weight.
  • Examples of the lipid-modified polyethylene glycol can be dioleoylglycerol-modified PEG, dilauroylglycerol-modified PEG, dimyristoylglycerol-modified PEG, dipalmitoylglycerol-modified PEG, distearoylglycerol-modified PEG and the like. More specifically, as the lipid-modified polyethylene glycol (lipid-modified PEG), stearyllated polyethylene glycol (for example, PEG45 stearate (STR-PEG45)) can be used.
  • stearyllated polyethylene glycol for example, PEG45 stearate (STR-PEG45)
  • a polyethylene glycol derivative such as amine can also be used. However, it is not limited to these.
  • the poorly water-soluble compound can be, for example, a compound belonging to Biopharmaceutics Classification System (BCS) class 4 without limitation.
  • Poorly water-soluble compound is not particularly limited, for example, terfenadine (Terfenadine), furosemide (furosemide), cyclosporine (Cyclosporin), acetazolamide (acetazolamide), colistin (Colistin), mebendazole (Mebendazole), coenzyme Q 10 (CoQ 10) And so on.
  • the alcohol in the alcohol solution is not particularly limited, and examples thereof include ethanol, t-butanol, 1-propanol, 2-propanol, and 2-butoxyethanol.
  • the concentration of each component of the alcohol solution can be appropriately determined according to the desired lipid membrane structure (or lipid nanoparticles), and is not particularly limited.
  • Phospholipids are, for example, in the range of 50-80 mol%
  • Membrane-permeable peptides are, for example, in the range of 5-20 mol%
  • Lipid-modified polyethylene glycols are, for example, in the range of 1-10 mol%.
  • the poorly water-soluble compound can be in the range of 10 to 40 mol%, for example.
  • aqueous solvent examples include an aqueous solution, for example, water or basically water as a main component, for example, a physiological saline solution, a buffer aqueous solution (for example, a phosphate buffer solution, an acetate buffer solution, a citrate buffer solution, etc.). ), Etc., which can be preferably used in the present invention.
  • phosphate buffer may be preferably used.
  • an alcohol solution containing a phospholipid or the like is diluted with an aqueous solvent, and a dispersion containing a lipid membrane structure containing a poorly water-soluble compound as a dispersoid is provided as a microchannel. Collect from the exit of.
  • the microchannel structure used in the method for producing a dispersion of the present invention is prepared by diluting an alcohol solution containing a phospholipid or the like with an aqueous solvent to prepare a dispersion containing a lipid membrane structure as a dispersoid.
  • Any microchannel structure that can be used can be used without particular limitation. Such a microchannel structure is described in, for example, Patent No.
  • Patent Document 2 Japanese Patent No. 6234971, Non-Patent Document 4, Non-Patent Document 5, WO2018 / 190423 A1 (Patent Document 2) and the like. Can be exemplified.
  • a lipid membrane having a uniform particle size with a small degree of dispersion while controlling the particle size of the lipid membrane structure to a desired value. It can be advantageously used to obtain a dispersion containing the structure as a dispersoid.
  • the microchannel structure described in Patent Document 2 has a first introduction path for introducing a first fluid and a second introduction path for introducing a second fluid, which are independent of each other on the upstream side.
  • Each of which has a constant length and merges to form one dilution channel toward the downstream side thereof, and the dilution channel is bent (for example, two-dimensionally bent) at least in a part thereof.
  • the bent flow path portion has a flow path portion, and the axial direction of the dilution flow path upstream from this or the extension direction thereof is the X direction, and the width direction of the dilution flow path perpendicular to the X direction is Y.
  • the direction is approximately Y (approximately + Y) toward the center of the flow path alternately from both side walls of the dilution flow paths facing each other in the Y direction.
  • And has a constant width x1 and x2 in the X direction. .. ..
  • the structure that regulates the flow path width of the dilution flow path is formed at regular intervals d1, d2. .. .. It is a flow path structure formed by providing at least two of them.
  • the flow path width y0 is preferably 20 to 1000 ⁇ m, more preferably 100 to 400 ⁇ m, and further preferably 150 to 300 ⁇ m.
  • the heights of each structure h1, h2. .. .. Is preferably 1 / 2y0 or more and 3/4y0 or less.
  • the structure is preferably provided in the range of 10 to 100, more preferably 10 to 50, and even more preferably 15 to 30.
  • an alcohol solution is introduced into the first introduction path as a lipid phase, and an aqueous solvent is introduced into the second introduction path as an aqueous phase.
  • the distance from the confluence of the first introduction path and the second introduction path to the upstream end of the first structure is appropriately set, but the diluted fluid at the set speed flowing between them is 0.1 seconds or less. It is preferable that it is specified according to the set speed of the diluting fluid so as to pass through.
  • the flow path may be heated.
  • the amount of each supply of the alcohol solution and the aqueous solvent to the microchannel is controlled so that the alcohol concentration of the dispersion recovered from the outlet of the microchannel is 40% or less, which is the desired particle size and dispersion degree. It is preferable because it is possible to obtain a dispersion containing the lipid film structure having the above as a dispersoid.
  • the alcohol concentration of the dispersion recovered from the outlet of the microchannel is preferably controlled to be in the range of 5 to 35%, more preferably in the range of 10 to 30%.
  • the operation in the microchannel structure can be carried out at a temperature in the range of 0 to 70 ° C., for example, while considering the boiling point of the solvent.
  • the alcohol solution is diluted with an aqueous solvent in the microchannel, and the dispersion containing the lipid membrane structure containing the poorly water-soluble compound as a dispersoid is recovered from the outlet of the microchannel.
  • a step of removing alcohol from the dispersion recovered from the outlet of the microchannel can be further included.
  • Alcohol removal from the dispersion can be performed, for example, by dialysis, distillation, or the like. Dialysis can be performed, for example, from 0 ° C. to room temperature.
  • the dispersion or the dispersion from which the alcohol has been removed can be further subjected to a concentration step.
  • the concentration step can be, for example, ultrafiltration, centrifugation, evaporation of solvent (water) or dialysis.
  • Ultrafiltration can be performed using, for example, an ultrafiltration membrane.
  • the ultrafiltration membrane has a predetermined nominal molecular weight cutoff (NMCO), and the predetermined NMCO can be any NMCO in the range of 50 kDa to 200 kDa, and is included in the dispersion to be obtained. It can be appropriately selected according to the particle size of the lipid membrane structure.
  • the present invention is a dispersion containing a poorly water-soluble compound and containing a lipid film structure having an average particle diameter of 60 nm or less measured by a dynamic light scattering (DLS) method as a dispersant in a dispersion medium.
  • the lipid membrane of the lipid membrane structure is the dispersion containing phospholipids and lipid-modified polyethylene glycol.
  • the lipid membrane of the lipid membrane structure can contain a membrane-permeable peptide.
  • the membrane-permeable peptide is not contained as an alcohol solution (lipid phase introduced into the microchannel) containing phospholipids and the like. Use an alcohol solution.
  • Phospholipids are preferably dioleylphosphatidylethanolamine and phosphatidic acid and / or sphingomyelin for effective delivery of the poorly water-soluble compound which is the target substance into the mitochondria.
  • the content of the membrane-permeable peptide is, for example, 5 to 20 mol%, preferably 10 to 15%, based on the total amount of the lipid membrane. It is in the range of mol%.
  • the content of lipid-modified PEG in the lipid membrane structure (or lipid nanoparticles) is, for example, in the range of 1 to 10 mol%, preferably 3 to 5 mol%, based on the total amount of the lipid membrane.
  • the amount of the poorly water-soluble compound in the lipid membrane structure (or lipid nanoparticles) is, for example, in the range of 10 to 40 mol%, preferably 15 to 30 mol%, or 20 to 25 mol%, based on the total amount of the lipid membrane. Is the range of.
  • the polydispersity index (PDI or PdI) of the lipid membrane structure (or lipid nanoparticles) measured by the DLS method is preferably 0.3 or less, preferably 0.25 or less.
  • the zeta potential of the lipid membrane structure is not particularly limited, but is, for example, in the range of 10 mV or more, 11 mV or more, 12 mV or more, 13 mV or more, 14 mV or more, or preferably 15 mV or more. Is.
  • the zeta potential of the lipid membrane structure (or lipid nanoparticles) of the dispersion of the present invention can be measured by the Zetasizer Nano ZS (Malvern, Worcestershire, UK) using laser Doppler electrophoresis.
  • the dispersion medium of the dispersion of the present invention is an aqueous solvent.
  • aqueous solvents are as described above.
  • the dispersion medium does not contain alcohol, or alcohol is removed from the dispersion medium.
  • the dispersion of the present invention is used to transfer a poorly water-soluble compound to intracellular and intracellular mitochondria.
  • the dispersion of the present invention is not particularly limited, but is, for example, a dispersion in which the phospholipids are dioleylphosphatidylethanolamine, phosphatidylic acid and / or sphingomyelin, and CoQ 10 as a poorly water-soluble compound is used as the total amount of the lipid membrane.
  • lipid nanoparticles as a dispersoid.
  • the dispersion of the present invention comprises a lipid membrane structure (or lipid nanoparticles) comprising one or more phospholipids selected from the group consisting of phosphatidic acid and sphingomyelin and dioleylphosphatidylethanolamine.
  • CoQ 10 is contained in the range of 10 to 40 mol% with respect to the total amount of the lipid membrane, and the lipid membrane structure having a particle size measured by the DLS method of 60 nm or less, preferably 55 nm or less, more preferably 50 nm or less ( Or lipid nanoparticles) and may further have a PDI of less than 0.3 when measured by the DLS method.
  • the lipid membrane structure (or lipid nanoparticles) is a dispersoid.
  • the average particle size is preferably small to some extent from the viewpoint of efficiency of uptake into cells, and is not particularly limited, but the lower limit value can be about 10 nm, preferably about 20 nm.
  • the dispersion of the present invention comprises a lipid membrane structure (or lipid nanoparticles) comprising one or more phospholipids selected from the group consisting of phosphatidic acid and sphingomyelin and dioleylphosphatidylethanolamine.
  • the lipid membrane structure (or lipid nanoparticles) contained CoQ 10 in the range of 10 to 40 mol% with respect to the total amount of the lipid membrane, and the lipid membrane structure (or lipid nanoparticles) was measured by the DLS method. It can have an average particle size of 50 nm to 60 nm and a PDI of less than 0.3.
  • the lipid membrane structure (or lipid nanoparticles) is a dispersoid.
  • the poorly water-soluble compound can be a fat-soluble compound.
  • the poorly water soluble compound is soluble in alcohol (eg, ethanol).
  • the poorly water-soluble compound can be a fat-soluble BCS class 4 compound.
  • the poorly water soluble compound can be a BCS class 4 compound that is soluble in ethanol. In order to improve or improve the solubility of the poorly water-soluble compound in alcohol, it may further include dissolving the compound in alcohol under heating conditions.
  • CoQ 10 can be dissolved in ethanol under heating conditions (eg, 50 ° C.) for use in the preparation of lipid membrane structures (or lipid nanoparticles), and in the dissolved state the lipid membrane structures.
  • Heating conditions eg, 50 ° C.
  • it can be incorporated into the membrane in a state close to nature, and therefore, the original functionality is fully exhibited at the place where CoQ 10 delivered intracellularly is delivered. Can be.
  • a pharmaceutical preparation containing a dispersion containing the poorly water-soluble compound of the present invention is provided.
  • a pharmaceutical preparation containing the dispersion of the present invention containing CoQ 10 is provided.
  • the composition in which the poorly water-soluble compound is CoQ 10 can be administered to, for example, a subject having a dysfunction in the respiratory chain complex I.
  • the composition in which the poorly water-soluble compound is CoQ 10 is a subject having a dysfunction in the respiratory chain complex (for example, any one or more of the respiratory chain complexes I, II, III, and IV). It can be used to improve ATP production in cells in subjects suffering from mitochondrial disease.
  • Subjects having dysfunction in respiratory chain complex I are not particularly limited, but are, for example, subjects suffering from mitochondrial encephalomyopathy (MELAS), subjects suffering from Leigh encephalopathy, and subjects suffering from Leber's hereditary optic neuropathy (LHON). Can be mentioned.
  • MELAS mitochondrial encephalomyopathy
  • LHON Leber's hereditary optic neuropathy
  • the composition in which the poorly water-soluble compound is CoQ 10 includes, for example, a subject suffering from CoQ 10 deficiency.
  • the subject can be an animal, particularly a mammal, particularly a primate, particularly preferably a human.
  • a method for administering a poorly water-soluble compound to a subject which comprises administering to the subject a dispersion containing the poorly water-soluble compound of the present invention.
  • a method for delivering a poorly water-soluble compound into cells in a subject's body which comprises administering to the subject a dispersion containing the poorly water-soluble compound of the present invention. ..
  • a method for delivering a poorly water-soluble compound to mitochondria in a cell in a subject body which comprises administering to the subject a dispersion containing the poorly water-soluble compound of the present invention. Will be done.
  • a method for treating mitochondrial disease in a subject suffering from mitochondrial disease which comprises administering to the subject a dispersion containing the poorly water-soluble compound of the present invention.
  • a method for treating mitochondrial disease in a subject suffering from mitochondrial disease which comprises administering the dispersion of the present invention containing CoQ 10 to the subject.
  • the use of a poorly water-soluble compound in the production of a pharmaceutical preparation containing the dispersion of the present invention containing the poorly water-soluble compound is provided.
  • the use of CoQ 10 is provided in the manufacture of a pharmaceutical formulation comprising a dispersion dispersion of the present invention containing CoQ 10.
  • the present invention provides the use of phospholipids and lipid-modified uncharged hydrophilic polymers in the manufacture of pharmaceutical formulations comprising the dispersions of the present invention containing poorly water soluble compounds.
  • the present invention provides the use of phospholipids and lipid-modified uncharged hydrophilic polymers in the manufacture of pharmaceutical formulations comprising the dispersion dispersions of the present invention containing CoQ 10 .
  • INDUSTRIAL APPLICABILITY The present invention provides the use of poorly water-soluble compounds, phospholipids and lipid-modified uncharged hydrophilic polymers in the manufacture of pharmaceutical formulations comprising the dispersions of the present invention containing poorly water-soluble compounds.
  • the present invention in the manufacture of a pharmaceutical formulation comprising a dispersion dispersion of the present invention containing CoQ 10, CoQ 10, the use of phospholipids and lipid-modified uncharged hydrophilic polymer.
  • the dispersion of the present invention can be made into a pharmaceutical formulation.
  • Pharmaceutical formulations containing the dispersions of the invention may further contain pharmaceutically acceptable excipients.
  • Excipients include, but are not limited to, buffering agents, tonicity agents, pharmaceutically acceptable salts, dispersants, antioxidants, preservatives, and soothing agents.
  • the dispersion of the present invention can be prepared as a cosmetic or supplement. Therefore, according to the present invention, cosmetics and supplements containing the dispersion of the present invention may be provided.
  • 2. Make 500 mL of PBS (-) for dialysis and store at 25 ° C. with stirring.
  • 3. Put 300-400 mL of DDW in a beaker, cut the dialysis membrane Spectra / Por 4 dialysis membrane (MWCO 12k-14k) to an appropriate length, stir vigorously so that the membrane does not stick, and hydrate (30 minutes or more). ).
  • 4. Add the following amounts of each solution to the Eppendorf tube to prepare a lipid solution (lipid phase).
  • Example 1 1-1 Preparation method of CoQ 10- MITO-Porter using microchannel
  • the particles were prepared using the microchannel device shown in FIG.
  • As the syringe pump a Standard Infusion Only Pump 11 Elite manufactured by HARVARD APPARATUS was used.
  • As the syringe 1 mL (lipid phase) and 2.5 mL (aqueous phase) of a glass syringe manufactured by HAMILTON were used.
  • (I) Lipid phase An ethanol solution containing 7.5 mM DOPE (EtOH), 7.5 mM SM (EtOH), and 7.5 mM DMG-PEG 2000 (EtOH) was prepared and brought to room temperature. An ethanol suspension containing a lipid material was prepared in an Eppendorf tube so as to have the volume ratio shown in the above figure, and was used as (i) a lipid phase. Further, the lipid phase contained 12.6 ⁇ L of 20 mg / mL STR-R8 so as to be modified by 10 mol% with respect to the lipid concentration.
  • (Iv) Flow Velocity Ratio The flow velocity ratio was adjusted so that the ethanol dilution concentration was 10%, 20%, 30%, or 40% in the lipid phase and the aqueous phase.
  • the flow rates of the lipid phase and the aqueous phase are 50 ⁇ L / min and 450 ⁇ L / min, 100 ⁇ L / min and 400 ⁇ L / min, 150 ⁇ L / min and 350 ⁇ L / min, or 200 ⁇ L / min and 300 ⁇ L / min, respectively. did.
  • Particles were prepared under each of the above flow rate ratio conditions (Fig. 2). As shown in FIG. 2, the particle size decreased as the ethanol dilution concentration decreased. Further, as shown in FIG.
  • the CoQ 10- MITO-Porter solution prepared in the microchannel was dialyzed for 2 hours (Fig. 1).
  • dialysis clips Spectra / Por Closures and dialysis membrane Spectra / Por 4 dialysis membrane (MWCO 12k-14k) manufactured by Spectram Laboratories were used.
  • the lipid nanoparticle solution tended to have a smaller particle size and an increased PdI due to dialysis as compared with before dialysis (FIG. 3).
  • DDS targeting the liver it has been shown that lipid nanoparticles having a particle size of 100 nm or less enable efficient delivery of the target substance, and that the smaller the particle size, the higher the efficiency.
  • the particles were used in the following examples. Neither the preparation of a dispersion having such a small particle size nor the preparation of a dispersion having such a small PdI was possible by the conventional method.
  • Stability Test In order to evaluate the stability of the prepared particles, a stability test was conducted at 4 ° C. and 25 ° C. under shading for 14 days (Fig. 5). During the storage period, the particle size increased over time at 25 ° C. On the other hand, at 4 ° C., the particle size was maintained at around 50 nm and was stable.
  • Comparative Example 1 Preparation method of CoQ 10- MITO-Porter using the conventional method Particles were prepared using the ethanol dilution method (Non-Patent Document 2). Prepare 7.5 mM DOPE (EtOH), 7.5 mM SM (EtOH), and 7.5 mM DMG-PEG 2000 (EtOH) and bring to room temperature. An ethanol suspension containing a lipid material was prepared in an Eppendorf tube so as to have the volume ratio shown on the right. Since CoQ 10 is in a suspended state and precipitation proceeds, ultrasonic treatment was performed immediately before use. PBS (-) buffer was added to the suspension and diluted to an ethanol concentration of 90%.
  • the CoQ 10 concentration and the CoQ 10 recovery rate contained in the prepared CoQ 10- MITO-Porter solution (conventional method) were quantified using HPLC and calculated from the calibration curve.
  • HPLC Agilent 1200 series was used.
  • the CoQ 10- MITO-Porter obtained using the microchannel was negatively stained and observed using a transmission electron microscope. The results were as shown in FIG. As shown in FIG. 16, the obtained CoQ 10- MITO-Porter contained a structure considered to be a lipid membrane inside the particles. As described above, it was clear that the CoQ 10- MITO-Porter obtained by using the microchannel was a non-hollow lipid structure.
  • CoQ 10 is poorly water-soluble, it is considered that CoQ 10 was incorporated into the lipid membrane, and the non-hollow lipid structure whose inside is filled with the lipid membrane is suitable for encapsulation of a large amount of CoQ 10. Was thought to be.
  • Example 2 2-1 Accuracy of particle preparation using microchannel
  • the results of Example 1 and Comparative Example 1 were compared.
  • the particle size tended to be small
  • the obtained dispersion was able to maintain uniform and highly accurate particles.
  • the obtained dispersion was about +20 mV even at the (C) zeta potential, and it is considered that there was no variation between preparations and that the dispersion could be modified with STR-R8 almost constantly.
  • the dispersion was modified with STR-R8 after the dispersion was formed, and it was difficult to reproducibly prepare the zeta potential at about +20 mV. Furthermore, the (D) CV value was used as an accuracy parameter for particle preparation.
  • the particle size CV of the dispersion was 0.082 in Comparative Example 1, whereas the particle size CV of the dispersion before and after dialysis of the method of the present invention was 0.062 and 0.065, respectively. there were.
  • the CV of the PdI of the dispersion was 0.192 in Comparative Example 1, whereas the CV of the PdI of the dispersion before and after dialysis of the method of the present invention was 0.148 and 0.066, respectively. ..
  • the CV of the zeta potential of the obtained dispersion was 0.171 in the conventional method, whereas the CV of the zeta potential of the dispersion before and after dialysis by the method of the present invention was 0.112 and 0.114, respectively. Met.
  • the particle size is 48.3 ⁇ 3.1 nm, which is 1/2 times smaller than that of the prior art.
  • the PdI of the method of the present invention was also smaller than that of Comparative Example 1.
  • the zeta potential was +21.5 ⁇ 1.8 mV, and the modifying effect by STR-R8 was expected, which was considered to be useful for introduction into cells and administration to animals.
  • the microchannel is useful for producing a dispersion and also as a device for stably modifying the dispersion with a functional element.
  • the dispersion obtained by the method of the present invention was smaller than the dispersion obtained in Comparative Example 1 at each CV value of particle size, PdI and zeta potential. This means that according to the method of the present invention, more uniform particles can be prepared, that is, the accuracy of preparation is high.
  • CoQ 10- MITO-Porter obtained in the microchannel was prepared by removing lipid-modified polyethylene glycol from the lipid composition, the obtained liquid became cloudy. Lipid-modified polyethylene glycol is considered to contribute to improving the solubility of CoQ 10- MITO-Porter in the solution.
  • the amount of preparation was also on the order of ⁇ L in Comparative Example 1 (conventional technique), but the method of the present invention allows unlimited preparation, and a large amount of preparation on the order of L can be expected even at the laboratory level.
  • the method of the present invention can be prepared in a shorter time as the preparation time becomes larger. Therefore, the preparation by the method of the present invention using the microchannel could prepare a dispersion of a lipid membrane structure containing a sparingly soluble compound having a small particle size.
  • the dispersion also had a small PdI.
  • the lipid nanoparticle formation time was significantly reduced. Therefore, it is considered that the production time and cost of the preparation are reduced.
  • a continuous dispersion of lipid membrane structures could be obtained.
  • Example 3 3-1 Evaluation of intracellular uptake (1)
  • fluorescent labeling nitrogenbenzoxadiazole (NBD) -DOPE modified with 0.5 mol% of lipid amount
  • Carriers were prepared, uptake into cervical cancer HeLa cells was evaluated using flow cytometry (FACS), and intracellular localization was observed with a confocal laser scanning microscope (CLMS). Evaluation of cell uptake using flow cytometry showed that the dispersion obtained by the method of the present invention had a large amount of uptake into cells as compared with the conventional method (Fig. 8).
  • Example 4 Change in CoQ 10 concentration
  • the CoQ 10 concentration is halved. It was verified by doubling (0.75 mM, hereinafter, 1/2 CoQ10) and doubling (3 mM, hereinafter, 2 CoQ10).
  • the ethanol dilution concentration in the microchannel was 20%.
  • the particle size before dialysis was the smallest in the concentration of Example 1 (1 CoQ 10 ), 70.5 ⁇ 0.3 nm, and the PdI was also the smallest (FIG. 13).
  • the particle size of all CoQ 10 concentrations was about 50 nm, and the PdI was about 0.2, which tended to increase from that before dialysis.
  • the CoQ 10 concentration was 58.1 ⁇ 5.0 ⁇ M before 1/2 CoQ10 dialysis, 36.0 ⁇ 1.5 ⁇ M after 1/2 CoQ10 dialysis, and 113.3 ⁇ 12.7 ⁇ M before CoQ10 dialysis, respectively. It was 76.1 ⁇ 7.5 ⁇ M after CoQ10 dialysis, 261.8 ⁇ 14.8 ⁇ M before CoQ10 dialysis, and 176.4 ⁇ 10.8 ⁇ M after CoQ10 dialysis.
  • the recovery rates were 83.3 ⁇ 7.1% before 1/2 CoQ10 dialysis, 67.2 ⁇ 3.4% after 1/2 CoQ10 dialysis, and 81.2 ⁇ 9.1% before 1 CoQ10 dialysis, respectively. 1, 72.8 ⁇ 6.7% after CoQ10 dialysis, 93.8 ⁇ 5.3% before 2 CoQ10 dialysis, and 80.5 ⁇ 4.7% after 2 CoQ10 dialysis.
  • the Drag / Lipid (w / w) is 0.10 ⁇ 0.01 before 1/2 CoQ10 dialysis, 0.09 ⁇ 0.00 after 1/2 CoQ10 dialysis, and 0.20 before 1 CoQ10 dialysis, respectively.
  • Example 5 Comparison by Preparation Amount It was verified whether the physical properties changed depending on the preparation amount between the conventional method (Comparative Example 1) and the method of the present invention (preparation method using a microchannel).
  • the particle size, PdI, and zeta potential (ZP) fluctuate greatly depending on the amount prepared at one time.
  • both the particle size and PdI were less affected by the amount of preparation as compared with the conventional method, and stable preparation was possible (Table 2).
  • the unit in order to perform stable modification of STR-R8, the unit must be as small as 100 ⁇ L.
  • the zeta potential becomes 9.8 ⁇ 0.9 mV, which is lower than that at the time of preparation of a small volume. This is a major barrier to scaling up lipid nanoparticle drugs.
  • the zeta potential shows a good value of about 20 mV regardless of the prepared amount, and the CV value (index of variation) is 0.072 before dialysis and 0.072 after dialysis. It is 0.031, which is smaller than the conventional method. From the above, it was shown that the preparation method using the microchannel can stably modify functional elements such as STR-R8 and is indispensable for the production of lipid nanoparticle pharmaceuticals.
  • Example 6 6-1 Concentration of CoQ 10- MITO-Porter An attempt was made to concentrate the dispersion obtained in Example 1. In the conventional method, the CoQ 10 concentration can be concentrated to 506.9 ⁇ 134.1 ⁇ M, but the ultrafiltration filter is likely to be clogged, and an efficient concentration operation may not be possible. Therefore, in this example, in order to increase the drug titer per lipid nanoparticles, an attempt was made to prepare a high-concentration CoQ 10- MITO-Porter by concentrating the dispersion obtained in Example 1. .. The post-dialysis solution of the dispersion obtained in Example 1 was applied to Amicon (MWCO: 100 kDa) and ultrafiltered (1000 g, 25 ° C., 25 minutes).
  • Amicon MWCO: 100 kDa
  • Example 7 Example 7-1. Effect of improving mitochondrial respiration by CoQ 10- MITO-Porter of the present invention
  • MELAS fibroblasts with mitochondrial dysfunction Mitochondrial respiration was evaluated using Leight fibroblasts, LHON fibroblasts, and normal fibroblasts.
  • the number of cells was 2.5 ⁇ 10 4 cells / well for MELAS fibroblasts and 2 ⁇ 10 4 cells / well for other cells. The measurement was carried out by a conventional method.
  • the seeds were sown on Agilent Technologies XFp Cell Culture Minilates (Agilent Technologies, Santa Clara, CA, USA) 24 ⁇ 3 hours before the measurement (about 10000 cells / well).
  • a running medium containing a final concentration of 24.75 mM glucose and 4 mM glutamine was prepared by adding 1.0 M Glucose Solution and 200 mM Glutamine Solution (Agilent) in advance to XF DMEM Medium (Agilent). Cells were cultured using this medium (CO 2 free, 37 ° C.) from 1 hour before the measurement, and measured using Agilent Technologies XFp extracellular flux analysers (Agilent Technologies, Santa Clara, CA, USA).
  • FCCP conjugating agent + 3 mM pyruvate after 15 minutes, measure maximal respiration, and after 15 minutes 0.5 ⁇ M rotenone and 0.5 ⁇ M antimycin.
  • A an inhibitor of electron transport chain respiratory chain complexes I and III was added and baseline oxygen consumption was measured again.
  • the maximum respiratory capacity was determined by subtracting the oxygen consumption rate (%) after the addition of rotenone and antimycin A from the maximum value of the oxygen consumption rate (%).
  • the oxygen consumption rate (%) over time is shown in FIG. 17A, and the maximum respiratory capacity is shown in FIG. 17B.
  • FIGS. 17A and 17B The results were as shown in FIGS. 17A and 17B.
  • PBS of CoQ 10 unencapsulated (-) in the addition of the suspension oxygen consumption rate also improves the maximum breathing capacity is not observed in normal cells.
  • the addition of a PBS (-) suspension of CoQ 10 showed a slight improvement in oxygen consumption and maximal respiration.
  • CoQ 10- MITO-Porter produced using the microchannel greatly improved the oxygen consumption rate and maximum respiration capacity of each cell type.
  • Acetaminophen was dissolved in PBS ( ⁇ ) under heating conditions of 60 ° C.
  • PBS (-) containing acetaminophen (Fuji Film Wako Pure Chemical Industries, Ltd.) was intraperitoneally administered at a dose.
  • CoQ 10- MITO-Porter or PBS (-) (negative control) of the present invention obtained in the microchannel was administered at a dose of 8 ⁇ L / g, respectively, and after 24 hours, cardiac blood sampling and liver removal were performed. It was.
  • the collected blood was left at room temperature for 1 to 2 hours and centrifuged (4 ° C., 15 minutes, 3500 rpm). Then, serum was collected and ALT was measured using transaminase CII-Test Wako (Fuji Film Wako Pure Chemical Industries, Ltd., Osaka, Japan) to evaluate the degree of liver damage.
  • the results were as shown in the left panel of FIG. 18B. As shown in the left panel of FIG. 18B, the amount of ALT, which is a marker of liver damage, was less than 1000 IU / L in the negative control, whereas the CoQ 10- MITO of the present invention obtained by the microchannel was obtained. -The ALT value was significantly reduced in the Porter-administered group.
  • the removed liver was subjected to histological evaluation.
  • the liver was drained with 4% paraformaldehyde (Fuji Film Wako Pure Chemical Industries, Ltd.) and allowed to stand at 4 ° C. overnight. Then, every 4 hours, the liver was treated with a solution containing 10% sucrose, 20% sucrose, and 30% sucrose in order at 4 ° C., and then the liver was allowed to stand overnight with a solution containing 30% sucrose.
  • the liver tissue was placed in an implantation dish, and the tissue section implanter O. C. T. It was filled with Compound (Sakura Finetech Japan Co., Ltd.). In addition, liquid nitrogen was used to freeze the tissue.
  • Frozen samples were sliced with LEICA CM3050S (Leica Biosystems) to a thickness of 20 ⁇ m to prepare frozen sections.
  • the excised tissue sections were thoroughly dried and subjected to hematoxylin-eosin staining (HE staining).
  • HE staining hematoxylin-eosin staining
  • the tissue sections were treated with hematoxylin for 8 minutes, stained, and subjected to running water for 10 minutes.
  • the tissue sections were then treated with eosin for 3 minutes, stained and lightly washed with water.
  • the tissue section was treated with 70% ethanol, 90% ethanol, and 100% ethanol solution in this order, and dehydrated. Further, the tissue section was immersed in xylene.
  • tissue sections were encapsulated using a soft mount. Observation of the tissue section was performed using a stereomicroscope TYPE101M (SHIMADZU). The results were as shown in the right panel of FIG. 18B.
  • a non-hollow lipid structure containing a poorly water-soluble compound could be prepared.
  • CoQ 10 was used as the poorly water-soluble compound
  • CoQ 10 was introduced into the cells and could improve the respiratory ability of the cells.
  • Hepatic disorder could be treated therapeutically in liver disorder model mice.
  • the obtained lipid structure could be imparted with cell membrane permeability, mitochondrial directivity could be imparted, and the introduced CoQ 10 was functionally effective. ..
  • Example 8 In this example, curcumin was used as the poorly water-soluble compound.
  • the prepared particles were dialyzed as described in Example 1 to obtain a solution containing curcumin-containing lipid nanoparticles as curcumin-MITO-Porter.
  • Curcumin is yellow and the residue of curcumin in the solution can be visually confirmed. Since curcumin outside the nanoparticles is removed by solution replacement by dialysis, it is considered that curcumin remains only in the nanoparticles after dialysis.
  • yellow color is confirmed after dialysis, the yellow color reflects curcumin incorporated in the nanoparticles, and the amount of curcumin incorporated in the nanoparticles can be estimated from the shade of yellow.
  • the final dilution concentration of ethanol on the microchannel device was set to 10%, 20%, 30%, and 40%, curcumin-containing lipid nanoparticles were prepared, and the color of the solution was confirmed before and after dialysis. As a result, it was observed that the yellow coloring tended to be very light at the final ethanol dilution concentration of 10%, and the coloring tended to be dark at 20% or more.
  • the particle size and PDI of the curcumin-containing lipid nanoparticles obtained by the DLS method were measured.
  • the results were as shown in FIG. As shown in FIG. 19, after dialysis, curcumin-containing lipid nanoparticles having particularly good PDI were obtained at final ethanol dilutions of 30% and 40%, with a PDI of 0.3 at a final ethanol dilution of 40%. It was smaller. Further, nanoparticles having a particle size of 100 nm to 150 nm were obtained at final ethanol dilution concentrations of 30% and 40%.
  • the lipid membrane structure containing a phospholipid and a lipid-modified uncharged hydrophilic polymer can be freely used not only for CoQ 10 but also for inclusion of a poorly water-soluble compound such as curcumin. Became clear.
  • the lipid membrane structure containing phospholipids and lipid-modified uncharged hydrophilic polymers is non-hollow lipid nanoparticles and has a lipid membrane structure inside the particles. Then, it is considered that the poorly water-soluble compound is incorporated into the lipid membrane structure inside the lipid nanoparticles.
  • the present invention is useful in the field related to the production of lipid nanoparticles of poorly water-soluble compounds.

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Abstract

La présente invention concerne un procédé de fabrication de nanoparticules lipidiques qui permet une production de masse et peut, avec une bonne reproductibilité, utiliser un porteur MITO pour fabriquer un composé peu soluble à l'eau comprenant du CoQ10 dans des nanoparticules lipidiques ayant une taille de particule plus petite souhaitée, ayant un indice de polydispersité souhaité et ayant un potentiel zêta souhaité.
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Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2021132735A2 (fr) 2019-12-27 2021-07-01 Luca Science Inc. Mitochondries isolées ayant une taille plus petite et vésicules à base de membrane lipidique encapsulant les mitochondries isolées
WO2022246280A1 (fr) * 2021-05-21 2022-11-24 University Of Iowa Research Foundation Particules contenant un antioxydant et procédés d'utilisation
WO2024010862A1 (fr) 2022-07-07 2024-01-11 Luca Science Inc. Complexes d'organites
WO2024010866A1 (fr) 2022-07-07 2024-01-11 Luca Science Inc. Complexes d'organites de modulation redox
WO2024030441A1 (fr) 2022-08-02 2024-02-08 National University Corporation Hokkaido University Procédés d'amélioration d'une thérapie cellulaire avec des complexes d'organites
US11904006B2 (en) 2019-12-11 2024-02-20 University Of Iowa Research Foundation Poly(diaminosulfide) particle-based vaccine
WO2024206862A1 (fr) 2023-03-31 2024-10-03 Luca Science Inc. Complexes d'organites pégylés

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2018190423A1 (fr) * 2017-04-13 2018-10-18 国立大学法人北海道大学 Structure de canal d'écoulement et procédé de formation de micelles ou particules lipidiques faisant appel à ladite structure

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2018190423A1 (fr) * 2017-04-13 2018-10-18 国立大学法人北海道大学 Structure de canal d'écoulement et procédé de formation de micelles ou particules lipidiques faisant appel à ladite structure

Non-Patent Citations (5)

* Cited by examiner, † Cited by third party
Title
ANONYMOUS: "Stop Aging Now CUR-Q10 Ultra Curcumin CoQ10 Complex Veggie Capsules", AMAZON.COM, 5 June 2020 (2020-06-05), pages 1 - 7, XP055746040, Retrieved from the Internet <URL:https://www.amazon.com/Stop-Aging-Now-Curcumin-Antioxidant/dp/B00ZYIPNTQ> *
ODA YUSUKE ET AL.: "Reviews on useful reagents for liposome research", DRUG DELIVERY SYSTEM, vol. 30, no. 5, 2015, pages 486 - 488 *
TOKESHI MANABU: "Can be manufactured quickly, easily in any size-microfluidic device for manufacturing lipid nanoparticle", JAPAN SCIENCE AND TECHNOLOGY AGENCY, BRIEFING MATERIALS OF LIFE SCIENCE NEW TECHNOLOGY PRESENTATION MEETINGS, 2017, pages 4, 7, - 15 *
YAMADA YUMA, BURGER LAILA, KAWAMURA ERIKO, HARASHIMA HIDEYOSHI: "Packaging of the Coenzyme Q10 into a Liposome for Mitochondrial Delivery and the Intracellular Observation in Patient Derived Mitochondrial Disease Cells", BIOL. PHARM. BULL., vol. 40, no. 12, 2017, pages 2183 - 2190, XP055746034 *
YUMA YAMADA ET AL.: "Mitochondrial delivery of Coenzyme Q10 via systemic administration using a MITO-Porter prevents ischemia/reperfusion injury in the mouse liver", J CONTROL RELEASE, vol. 213, 2015, pages 86 - 95, XP029259356 *

Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US11904006B2 (en) 2019-12-11 2024-02-20 University Of Iowa Research Foundation Poly(diaminosulfide) particle-based vaccine
WO2021132735A2 (fr) 2019-12-27 2021-07-01 Luca Science Inc. Mitochondries isolées ayant une taille plus petite et vésicules à base de membrane lipidique encapsulant les mitochondries isolées
WO2022246280A1 (fr) * 2021-05-21 2022-11-24 University Of Iowa Research Foundation Particules contenant un antioxydant et procédés d'utilisation
WO2024010862A1 (fr) 2022-07-07 2024-01-11 Luca Science Inc. Complexes d'organites
WO2024010866A1 (fr) 2022-07-07 2024-01-11 Luca Science Inc. Complexes d'organites de modulation redox
WO2024030441A1 (fr) 2022-08-02 2024-02-08 National University Corporation Hokkaido University Procédés d'amélioration d'une thérapie cellulaire avec des complexes d'organites
WO2024206862A1 (fr) 2023-03-31 2024-10-03 Luca Science Inc. Complexes d'organites pégylés

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