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WO2008144365A2 - Procédé de fabrication de compositions de poudre sèche contenant des acides nucléiques - Google Patents

Procédé de fabrication de compositions de poudre sèche contenant des acides nucléiques Download PDF

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Publication number
WO2008144365A2
WO2008144365A2 PCT/US2008/063655 US2008063655W WO2008144365A2 WO 2008144365 A2 WO2008144365 A2 WO 2008144365A2 US 2008063655 W US2008063655 W US 2008063655W WO 2008144365 A2 WO2008144365 A2 WO 2008144365A2
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WO
WIPO (PCT)
Prior art keywords
sirna
solution
supercritical fluid
antisolvent
powder
Prior art date
Application number
PCT/US2008/063655
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English (en)
Other versions
WO2008144365A3 (fr
Inventor
Andrew Geall
Sushma Kommareddy
Gerhard Muhrer
Vishal Saxena
Ranjit Thakur
Original Assignee
Novartis Ag
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Publication date
Application filed by Novartis Ag filed Critical Novartis Ag
Publication of WO2008144365A2 publication Critical patent/WO2008144365A2/fr
Publication of WO2008144365A3 publication Critical patent/WO2008144365A3/fr

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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
    • A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
    • A61K9/16—Agglomerates; Granulates; Microbeadlets ; Microspheres; Pellets; Solid products obtained by spray drying, spray freeze drying, spray congealing,(multiple) emulsion solvent evaporation or extraction
    • A61K9/1682—Processes
    • A61K9/1694—Processes resulting in granules or microspheres of the matrix type containing more than 5% of excipient
    • 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
    • C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
    • C12N15/111—General methods applicable to biologically active non-coding nucleic acids
    • 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
    • A61K9/0012—Galenical forms characterised by the site of application
    • A61K9/0019—Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner
    • 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
    • A61K9/0012—Galenical forms characterised by the site of application
    • A61K9/007—Pulmonary tract; Aromatherapy
    • A61K9/0073—Sprays or powders for inhalation; Aerolised or nebulised preparations generated by other means than thermal energy
    • 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
    • C12N2310/00—Structure or type of the nucleic acid
    • C12N2310/10—Type of nucleic acid
    • C12N2310/14—Type of nucleic acid interfering nucleic acids [NA]
    • 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
    • C12N2320/00—Applications; Uses
    • C12N2320/30—Special therapeutic applications
    • C12N2320/32—Special delivery means, e.g. tissue-specific

Definitions

  • the present invention relates to a method for making pharmaceutical products containing small double stranded nucleic acid molecules. More specifically, the present invention also relates to powder formulations containing short interfering RNA, or "siRNA.”
  • RNA interference is a natural cellular process for selectively turning off the activity of genes by targeting RNA, the key intermediate between DNA and proteins through the use of double stranded nucleic acid.
  • the double stranded nucleic acid molecules that mediate RNA interference include, but are not limited to, double-stranded RNA ("dsRNA”), micro-RNA (“miRNA”), short hairpin RNA (“shRNA”), short interfering nucleic acid (“siNA”) and short interfering ribonucleic acid (“siRNA”).
  • siRNA the mediators of RNAi-induced specific mRNA degradation
  • a major issue in gene therapy is the delivery of therapeutically active siRNA into target tissues and/or cells in vivo.
  • the desirable approach is direct application of catalytically active siRNA.
  • These formulations can be of a solid pure form or with some other delivery system like lipoplexes and polypexes.
  • siRNA is hydrophilic
  • Many existing recrystallization techniques result in non-uniformity of particle size.
  • Milling techniques are frequently used in industrial practice to reduce the particle size of solids. Wet milling processes require the presence of a liquid carrier that could cause product contamination. Additionally, such a liquid carrier would have to be subsequently removed. Dry milling processes often suffer from significant product loss or from operational problems such as product caking or equipment clogging. Spray and freeze drying techniques have been used as alternative processes to produce micronized dry powders. However, these technologies may produce inconsistent average particle sizes. Additionally, common processing temperature exceed what is acceptable for siRNA which has a melting point of approximately 50 0 C.
  • siRNA particle manufacture there is need for processes that overcome these above described limitations for siRNA particle manufacture.
  • the present invention feature processes that overcome some of the aforementioned disadvantages.
  • siRNA is formulated into a dry particulate form from an aqueous solution using a supercritical fluid process.
  • Supercritical fluids provide a means for controlling particle size and shape.
  • the present invention relates to a process for preparing a powder, for example a particulate composition, that includes the steps of contacting a solution containing a therapeutic agent with an antisolvent to precipitate a powder containing dsRNA.
  • the antisolvent for example, may be supercritical fluid.
  • incorporated in the powder may be a pharmaceutically acceptable excipient, such as a nonionic surfactant.
  • the present invention has particular use in creating powder compositions that may be administered parenterally or via inhalation.
  • dsRNA particularly useful forms include siRNA which may comprise from ten to fifty nucleotides.
  • the resulting particulates may have a particle size of less than five microns.
  • the contacting of the solution containing dsRNA is contacted with an antisolvent in a high pressure vessel that is set at the conditions of a temperature from about 0 0 C to about 80 0 C and a pressure from about 70 to about 250 bar.
  • the present invention features a method for making a dry powder, or particulate, formulation of siRNA that can be subsequently formed into a solution, suspension, gel or cream for parenteral, inhalable, topical administration, or any other suitable route of administration.
  • the invention uses supercritical fluid technology to form the dry powder formulation.
  • parenteral administration means an injection administered by routes such as intravenous, subcutaneous, intradermal, intramuscular, intraarticular, intraocular, intracranial, intrathecal or to any other body part or tissue.
  • the term "powder composition” means finely dispersed solid particles that contain a therapeutic agent that can be administered to a mammal, e.g., a human in order to prevent, treat or control a particular disease or condition affecting the mammal.
  • the powder composition can further include a pharmaceutically excipient.
  • the powder composition can be subsequently mixed with a pharmaceutically acceptable vehicle to form a dispersion or solution appropriate for parenteral administration.
  • the powder composition itself or with a pharmaceutically acceptable vehicle can be used for inhalable administration.
  • pharmaceutically acceptable refers to those compounds, materials, compositions and/or dosage forms, which are, within the scope of sound medical judgment, suitable for contact with the tissues of mammals, especially humans, without excessive toxicity, irritation, allergic response and other problem complications commensurate with a reasonable benefit/risk ratio.
  • RNA interference means nucleic acid molecules, single or double stranded, that mediate RNA interference which include, but are not limited to, double stranded nucleic acid (“dsNA”), double-stranded RNA (“dsRNA”), micro-RNA (“miRNA”), short hairpin RNA (“shRNA”), short interfering nucleic acid (“siNA”) and short interfering ribonucleic acid (“siRNA”).
  • dsNA double stranded nucleic acid
  • dsRNA double-stranded RNA
  • miRNA micro-RNA
  • shRNA short hairpin RNA
  • siNA short interfering nucleic acid
  • siRNA short interfering ribonucleic acid
  • dsRNA refers to an oligoribonucleotide or polyribonucleotide, modified or unmodified, and fragments or portions thereof, of genomic or synthetic origin or derived from the expression of a vector, which may be partly or fully double-stranded and which may be blunt-ended or contain a 5'- and/or 3'- overhang, and also may be of a hairpin form comprising a single oligoribonucleotide which folds back up on itself to give a double-stranded region.
  • the dsRNA used in the present invention may also contain modified nucleotide residues.
  • siRNA denotes short interfering RNAs and refers to short double stranded ribonucleic acids useful for RNA interference.
  • siRNAs for example, have lengths between 10 to 50 nucleotides, e.g., 15 to 25 nucleotides.
  • the term "inhibition' of gene expression means the reduction of the expression of said gene by at least 10%, 33%, 50%, 90%, 95% or 99% when compared to a cell not treated with RNA interference.
  • Lower doses of the therapeutic agent, lower concentrations of the therapeutic agent in the cell and/or longer times after administration of the therapeutic agent may result in inhibition at a lower level and/or in a smaller fraction of cells (e.g., at least 10%, 20%, 50%, 75%, 90% or 95% of targeted cells).
  • the term "form" of or “format” of in relation to oligonucleotides refers to different chemical nature of the oligoribonucleotide, in particular to modifications as compared to naturally occurring ribonucleotides, such as for instance chemically modified 2'OH groups of the ribose moiety or the modified intermucleosidic linkages such as phosphothioate linkages, or the modified nucleobases such as for example 5-methyl-C.
  • nucleotide means ribonucleotide or deoxyribonucleotide
  • oligonucleotide and oligoribonucleotide are interchangeable and refer, depending on the context, to modified or unmodified oligonucleotides comprising ribonucleotides and/or deoxyribonucleotides.
  • the therapeutic agent is present in the powder composition of the present invention in a therapeutically effective amount or concentration.
  • a therapeutically effective amount or concentration is known to one of ordinary skill in the art as the amount or concentration varies with the therapeutic agent being used and the indication which is being addressed.
  • the therapeutic agent may be present in an amount by weight from about 1 % to about 100% by weight of the pharmaceutical composition.
  • 100% may be a desired outcome.
  • the weight fraction is established on a case by case basis as is known within the skill in the art.
  • the efficiency i.e., the degree of inhibition of the target gene, is dependent on a number of different factors including the specificity of the dsRNA for its target sequence.
  • specificity means homology, i.e. sequence identity between the dsRNA in the duplex region and the target sequence. It is understood by a person skilled in the art that 100% sequence identity is not required in order to achieve significant inhibition. Normally, at least 75% sequence identity between the dsRNA and the target sequence is sufficient in order to inhibit expression of the target nucleic acid.
  • Useful is a sequence identity of at least 80%, more useful is a sequence identity of at least 90%. Most useful is a sequence identity of at least 95% between the dsRNA and the target sequence.
  • the siRNA reagent should have 100% homology to the target mRNA and at least two mismatched nucleotides to all other genes present in the cell or organism.
  • Methods to analyze and identify dsRNAs with sufficient sequence identity i to effectively inhibit expression of a specific target sequence are known in the art. Sequence identity may be optimized by sequence comparison and alignment algorithms known in the art (see Gribskov and Devereux, Sequence Analysis Primer, Stockton Press, 1991 , and references cited therein) and calculating the percent difference between the nucleotide sequences by, for example, the Smith-Waterman algorithm as implemented in the BESTFIT software program using default parameters (e.g., University of Wisconsin Genetic Computing Group).
  • RNAi reagent Another factor affecting the efficiency of the RNAi reagent is the target region of the target mRNA.
  • the region of a target mRNA effective for inhibition by the RNAi reagent may be determined by experimentation. Most preferred mRNA target region would be the coding region. Also preferred are untranslated regions, particularly the 3'-UTR, splice junctions. For instance, transfection assays as described in Elbashir et al. (2001) may be performed for this purpose.
  • the powder compositions may also further include pharmaceutically acceptable excipients such as carriers, adjuvants, stabilizers, preservatives, dispersing agents, surfactants and other agents conventional in the art having regard to the type of formulation in question.
  • pharmaceutically acceptable excipients such as carriers, adjuvants, stabilizers, preservatives, dispersing agents, surfactants and other agents conventional in the art having regard to the type of formulation in question.
  • Such materials are non-toxic to recipients at the dosages and concentrations employed; buffers such as phosphate, citrate, succinate, acetic acid, and other organic acids or their salts; antioxidants such as ascorbic acid; hydrophilic polymers such as polyvinylpyrrolidone; monosaccharides, disaccharides, and other carbohydrates including cellulose or its derivatives, glucose, manose, or dextrins; chelating agents such as EDTA; sugar alcohols such as mannitol or sorbitol; counterions such as sodium; and/or non
  • lipids e.g., cationic lipids, zwitterionic lipids (such as helper lipids) and anionic lipids.
  • cationic lipids include, but are not limited to 1 ,2-dioleoyl-3-trimethylammonium propane (DOTAP); N-[1- 2(2,3-dioleyloxy)propyl]-N,N,N-trimethylammonium chloride (DOTMA); 2,3-dioleoyloxy-N-[2- (sperminecarboxamido)ethyl]-N,N-deimethyl-1-propanaminium (DOSPA); dioctadecyl amido glycil spermine (DOGS); and 3,[N-N 1 ,N-dimethylethylenediamine)-carbamoyl]cholesterol (D- chol).
  • zwitterionic lipids include, but are not limited to
  • compositions according to the present invention include, but are not limited to, polyethylenimine, chitin, chitosan, poly (L- lysine) and dendrimers.
  • the term “particles” may encompass both microparticles and nanoparticles.
  • microparticles refers to particles having an average particle diameter in the range of about one to five hundred microns, e.g., about one to about ten microns, especially less than six microns.
  • nanoparticles' refers to particles having an average particle diameter in the range from about 0.001 to one micron, e.g., about 0.05 to about 0.5 microns.
  • the particles can comprise solely of the therapeutic agent or mixtures of the therapeutic agent with a pharmaceutically acceptable excipient.
  • the term “supercritical fluid” refers to a fluid at or above its critical pressure (P c ) and critical temperature (T c ) simultaneously.
  • a fluid above its critical pressure and at its critical temperature is in a supercritical state.
  • a fluid at its critical pressure and above its critical temperature is also supercritical.
  • supercritical fluids also encompass both near supercritical fluids and subcritical fluids.
  • a “near supercritical fluid” is above but close to its critical pressure (P c ) and critical temperature (T c ) simultaneously.
  • a “subcritical fluid” is above its critical pressure (P c ) and close to its critical temperature (T c ).
  • an "antisolvent” refers to a supercritical fluid.
  • any material can be used as the supercritical fluid provided that material is suitable for processing under the specific operation conditions contemplated herein.
  • materials that can be compressed into a supercritical fluid include, but are not limited to, carbon dioxide, methane, benzene, methanol, ethane, ethylene, xenon, nitrous oxide, fluroform, dimethyl ether, propane, n-butane, isobutane, n-pentane, isopropanol, methanol, toluene, propylene, chlorotrifluro-methane, sulfur hexafluoride, bromotrifluromethane, chlorodifluoromethane, hexafluoroethane, carbon tetrafluoride, decalin, cyclohexane, xylene, tetralin, aniline, acetylene, monofluoromethane, 1 ,1-difluoroethylene, ammonia, water and mixture
  • supercritical fluid process Various processes for particle formation exist based on supercritical fluid technology (such processes are hereinafter termed “supercritical fluid process.”)
  • supercritical fluid processes include, but are not limited to, rapid expansion of supercritical solutions (“RESS”), gas antisolvent precipitation (“GAS”), supercritical anti-solvent (“SAS”) and solution enhanced dispersion by supercritical fluids (“SEDS”).
  • RES supercritical solutions
  • GAS gas antisolvent precipitation
  • SAS supercritical anti-solvent
  • SEDS solution enhanced dispersion by supercritical fluids
  • a particularly useful supercritical fluid process in the present invention is the SAS process.
  • solvent refers to a material that is able to dissolve, disperse and/or solubilize the therapeutic agent of interest (e.g., solubility of the therapeutic agent in the solvent from about 0.01% to 50% w/v, e.g., 0.1 % to 10%, e.g., 1 % to 5%).
  • a solvent can include a single material or a mixture of materials. Modifiers or co-solvents to enhance the dissolution, dispersion and/or solubilization of the therapeutic agent can also be added to the solvent.
  • solvents and/or modifiers include, but are not limited to, alcohols, ethers, ketones, esters, alkanes, halides or mixtures thereof.
  • solvents include, but are not limited to water, ammonia, dimethylsulfoxide, dimethyl ether, methanol, ethanol, isopropanol, n-propanol, methylene chloride, acetone, ethylacetate, tetrohydrofuran, ethyl ether or mixtures thereof.
  • Particularly useful solvents are those that are polar, i.e., polar solvents.
  • the RESS process involves the dissolution, suspension and/or solubilization of the therapeutic agent and any optional pharmaceutically acceptable excipients in a supercritical fluid to form a homogenous solution.
  • solution may also refer to a mixture if the therapeutic agent and/or the pharmaceutically acceptable excipients are suspended in the solvent.
  • the solution is then depressurized by passing through a heated orifice or nozzle into a low pressure, e.g. atmospheric chamber. When the solution depressurizes, the supercritical fluid vaporizes leaving the substrates (i.e., the powder composition) in the form of particles.
  • the SAS process also known as PCA (precipitation with compressed anti-solvents) uses a solvent and an antisolvent.
  • the therapeutic agent and any pharmaceutically acceptable excipients are, for example, dissolved, suspended and/or solubilized in a solvent to form a homogeneous solution.
  • the solvent is miscible with the supercritical fluid.
  • the solution is then mixed with, for example sprayed into, the supercritical fluid.
  • the supercritical fluid causes the solvent to expand resulting in a lower solvent strength than the pure solvent. There is a rapid mass transfer of solvent from the solution phase to the antisolvent phase which leads to higher supersaturation resulting in homogenous nucleation.
  • the mixing is accomplished through the use of particular nozzle designs, which can be varied, and the particle size and morphology of the therapeutic agent can be controlled by varying the pressure and temperature prior to spraying the solution into the collection chamber as well as by varying the flow rate ratio between the two streams entering the collection vessel through the nozzle, i.e., the solution and antisolvent flow rates.
  • the GAS process is similar to the SAS process; however, the supercritical fluid is added to a solution of the therapeutic agent dissolved, suspended and/or solubilized in an organic cosolvent.
  • the supercritical fluid and solvent are miscible whereas the therapeutic agent has limited solubility in the supercritical fluid.
  • the supercritical fluid functions as an antisolvent to precipitate particles of the therapeutic agent.
  • the SEDS process features the therapeutic agent and any optionally pharmaceutically acceptable excipients dissolved, suspended and/or solubilized in a solvent to form a solution.
  • the solution and the supercritical fluid are each passed through an orifice or nozzle and sprayed into a pressurized collection chamber.
  • the two orifices or nozzles can be arranged separately or co-axially.
  • the high velocity of the supercritical fluid disrupts the solution into very small droplets. Additionally, the conditions are such that the supercritical fluid extracts the solvent from the solution as the supercritical fluid and solution contact each other. Like the preceding processes, as the solvent is extracted, the powder composition remains.
  • the resulting particles from the inventive process of the present invention may be formulated into pharmaceutical compositions for administration.
  • the dry powder particles are formed into suspensions, solutions or emulsions in sterile pyrogen-free water, oily or aqueous vehicles as known in the art.
  • the formulation may then be administered as a bolus injection or continuous infusion.
  • the SAS process is an exemplary process useful for the present invention.
  • a high pressure vessel is pressurized with supercritical carbon dioxide (i.e., the antisolvent) at a desired pressure, for example from about 5 to 350 bar, in particular from about 70 to 250 bar at a desired temperature, for example from about 0 0 C to about 80 0 C in particular from about 25°C to about 45°C.
  • the therapeutic agent and any optional pharmaceutically acceptable excipients are first dissolved in a solvent to form a solution.
  • This solution along with a modifier, for example ethanol, are then injected into the high pressure vessel at specific flow rates as a first stream.
  • a modifier for example ethanol
  • Exemplary flow rates of the solution can be in the range of 0.01 - 10 ml/min.
  • antisolvent can be introduced.
  • the antisolvent can be introduced in a co- current, cross-current or counter-current mode by itself or with a modifier.
  • the antisolvent extracts the water from the solution and causes high supersatu ration leading to particle precipitation of the solute.
  • the resulting particles are then dried first with mixture of modifier and antisolvent and then with fresh supercritical carbon dioxide. After drying, the system is depressurized and the solid dried powder is recovered.
  • a nozzle is used to achieve better mixing and atomization.
  • An exemplary nozzle is a co-axial nozzle.
  • the diameters of the co-axially arranged inner and outer capillaries are 1/16" and 1/8", respectively.
  • the annulus of the nozzle is 0.05 mm.
  • the inner passage carries the solution and the outer passage carries the antisolvent and/or antisolvent with a modifier.
  • siRNA is produced in a dry form using SAS method with D- mannitol as an excipient. Ethanol is injected directly to a carbon dioxide line before expanding in a reactor. Aqueous solution and a carbon dioxide-ethanol mixture is then sprayed using a Schlick atomizing nozzle. At the start of the experiment, a reactor is pressurized using carbon dioxide and the temperature of the reactor is maintained by flowing heated water through the reactor jacket.
  • the system After injecting the solution, the system is first purged with the carbon dioxide-ethanol mixture for fifteen minutes and then with fresh carbon dioxide to remove any residual solvent (ethanol or water) from the system. For purging, 4.1 kg of carbon dioxide is used. After purging, the system is depressurized to obtain a dry powder composition. In this experiment, the yield is high at 98%.
  • Example 1 From Example 1 , spherical particles of siRNA ranging from one to ten microns are obtained. However, the D-mannitol appears to have a high aspect ratio in the form of columns and/or needles which are up to about a hundred microns in size.
  • Example 1 this experiment is conducted to compare particle morphology of D-mannitol and siRNA, only difference being absence of siRNA.
  • 252 mg of D-mannitol is dissolved in 5 ml of water.
  • An aqueous solution is sprayed in the vessel maintained at 150 bar and 35 0 C at flow rate of 0.25 ml/min.
  • the flow rate of carbon dioxide is maintained at ⁇ 47 g/min and ethanol flow was 5.0 ml/min.
  • particles are purged with ethanol-carbon dioxide and then with fresh carbon dioxide. After purging, the system is depressurized to obtain a dry powder composition.
  • the D-mannitol When viewed under a scanning electron microscope, the D-mannitol appears to have a high aspect ratio in the form of columns and/or needles which are up to about a hundred microns in size as seen with Example 1.
  • this experiment studies the effect of different excipient with siRNA.
  • 215 mg of siRNA and 110 mg of poloxamer 188 (a nonionic surfactant) is dissolved in 5 ml of water.
  • the solution obtained is clear.
  • the reactor is pressurized up to 150 bar and temperature is maintained at 35 0 C as before. After stabilizing this condition, ethanol solution is injected first and then aqueous solution is fed using nozzle. Also, the flow conditions remained same as of example 1 and 2. Dry, free flowing powder was obtained with this method.

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Abstract

La présente invention concerne un procédé de fabrication de compositions de poudre sèche comportant des molécules double brin d'acide nucléique. Le procédé selon l'invention peut être utilisé pour former des particules à partir d'un fluide supercritique basé sur un traitement à base d'anti-solvant pouvant être utilisées ultérieurement pour administration pharmaceutique, notamment pour une administration parentérale.
PCT/US2008/063655 2007-05-17 2008-05-15 Procédé de fabrication de compositions de poudre sèche contenant des acides nucléiques WO2008144365A2 (fr)

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US9572897B2 (en) 2012-04-02 2017-02-21 Modernatx, Inc. Modified polynucleotides for the production of cytoplasmic and cytoskeletal proteins
US9597380B2 (en) 2012-11-26 2017-03-21 Modernatx, Inc. Terminally modified RNA
US10323076B2 (en) 2013-10-03 2019-06-18 Modernatx, Inc. Polynucleotides encoding low density lipoprotein receptor
US10815291B2 (en) 2013-09-30 2020-10-27 Modernatx, Inc. Polynucleotides encoding immune modulating polypeptides

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