WO2012098358A1 - Procédé de lyophilisation - Google Patents

Procédé de lyophilisation Download PDF

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Publication number
WO2012098358A1
WO2012098358A1 PCT/GB2012/000051 GB2012000051W WO2012098358A1 WO 2012098358 A1 WO2012098358 A1 WO 2012098358A1 GB 2012000051 W GB2012000051 W GB 2012000051W WO 2012098358 A1 WO2012098358 A1 WO 2012098358A1
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WIPO (PCT)
Prior art keywords
cells
biological material
freeze
solution
drying
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PCT/GB2012/000051
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English (en)
Inventor
Ward KEVIN
Andrew COWEN
Thomas Peacock
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Biopharma Technology Ltd
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority claimed from GBGB1101002.2A external-priority patent/GB201101002D0/en
Application filed by Biopharma Technology Ltd filed Critical Biopharma Technology Ltd
Priority to GB1314778.0A priority Critical patent/GB2502016B/en
Publication of WO2012098358A1 publication Critical patent/WO2012098358A1/fr

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Classifications

    • F—MECHANICAL ENGINEERING; LIGHTING; HEATING; WEAPONS; BLASTING
    • F26—DRYING
    • F26B—DRYING SOLID MATERIALS OR OBJECTS BY REMOVING LIQUID THEREFROM
    • F26B5/00—Drying solid materials or objects by processes not involving the application of heat
    • F26B5/04—Drying solid materials or objects by processes not involving the application of heat by evaporation or sublimation of moisture under reduced pressure, e.g. in a vacuum
    • F26B5/06—Drying solid materials or objects by processes not involving the application of heat by evaporation or sublimation of moisture under reduced pressure, e.g. in a vacuum the process involving freezing
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N1/00—Preservation of bodies of humans or animals, or parts thereof
    • A01N1/02—Preservation of living parts
    • A01N1/0278—Physical preservation processes
    • A01N1/0284—Temperature processes, i.e. using a designated change in temperature over time

Definitions

  • the present invention relates to a method of freeze drying, in particular to the freeze drying of biological material.
  • the biological material may be cell based and can be from any source such as microbial cells, protozoal cells, animal cells or plant cells.
  • the invention relates to freeze drying mammalian cells, such as blood cells and nucleated cells, as well as to bacterial cells for therapeutic use and use in food products.
  • the present invention further provides a method of reconstitution.
  • freeze drying or lyophilisation is a commonly used technique for a variety of materials allowing preservation and long term storage of such materials. Freeze drying is being used more frequently for the preservation and storage of living materials such as bacteria and cells such as blood cells. However, viability of the freeze dried biological material after re-hydration or reconstitution provides further challenges.
  • pH buffers such as PBS
  • PBS pH buffers
  • buffers employed in freeze-drying should be examined on the basis of their freezing and drying behaviour in addition to their ability to maintain a stable pH range in the liquid state. Tris and citrate buffers have been shown not to undergo pH shifts during freezing with proteins (Carpenter et al, 2002).
  • a further disadvantage is that the cryoprotectant must be removed from the freeze-thawed sample prior to use.
  • Several parameters in a freeze drying process affect the preservation of cells, for example, container type, fill volume, degree of container-shelf contact, sample cooling rate, freezing temperature, primary drying conditions (temperature, pressure, time), extent of desorption of unfrozen water in the 'secondary drying' phase of the lyophilisation cycle, rehydration media and methodology.
  • the formulation medium in which cells are suspended prior to the lyophilisation process is critical in preventing, or at least minimising, damage to cells that may be induced by the freezing or drying processes.
  • Freeze drying biological material such as live bacteria or cells such as red blood cells, so that they are still viable and retain cell integrity after reconstitution, presents some challenges. Maintaining the integrity of the cell membrane after reconstitution is also critical for a cell based therapy to be effective. Many regenerative medicine products rely on the delivery to patients of products that incorporate live cells. This presents a significant challenge in providing viable cells following reconstitution or re-hydration of the freeze dried cells.
  • EP0668013 discloses a freeze drying method involving granulation. This method can result in haemolysis and
  • the present invention relates to an improved method of freeze drying and reconstitution of cell based biological materials which address some of the challenges in this field.
  • a method of freeze drying a cell based biological material comprising the steps of incubating the biological material in a suitable freeze-drying medium and solvent, cooling until frozen, removing the frozen solvent by sublimation, desorption of any unfrozen solvent and where the cooling rate is at or greater than 0.5°C per minute.
  • the cooling rate is between 0.5 °C to 1000 °C per minute. More preferably, the cooling rate is between 10 °C to 100 °C per minute.
  • the cooling can be done by placing the material in a pre-cooled environment such as a freeze-dryer, a freezer or a cold liquid such as liquid nitrogen.
  • the material and medium is cooled down to between -20 °C to -196 °C. More preferably the material and the medium are cooled to between -40 °C and -70 °C.
  • the cell based biological material prior to freeze drying, was incubated in the medium at a temperature of between 15 °C to 45°C for a period of between 1 hour to 24 hours.
  • the incubation temperature was 37 °C. More preferably, the incubation was carried out for 3 to 11 hours.
  • the medium may contain one or more of a buffer solution, bulking agents, pH stabilisers, pH adjusters, thermal stabilisers, cryoprotectants , lyoprotectants, an antioxidant, a biopolymer or an agent that may enable the active 'loading' of the cell or organism with at least one of the aforementioned components.
  • the medium is isotonic (iso-osmotic) with or hypertonic (hyperosmotic) to the biological material (such as the intracellular compartment of a red blood cell).
  • isotonic is defined as the medium having the same concentration or osmotic pressure as the biological material or the medium contained within the biological material
  • hypertonic is defined as the medium having a greater concentration or osmotic pressure compared with the biological material or the medium contained within the biological material.
  • the solvent may be selected from water, tertiary butanol, aqueous or organic solvent.
  • the buffer solution may be selected from phosphate buffered saline, citrate buffer, Tris buffer, amino acid-based buffers (such as histidine buffer, glycine buffer) , sodium dihydrogen orthophosphate, disodium hydrogen orthophosphate, potassium dihydrogen orthophosphate, dipotassium hydrogen orthophosphate, TES, MOPS, PIPES, Cacodylate, SSC, MES and HEPES.
  • the cryoprotectant or lyoprotectant may be selected from disaccharides (such as lactose, trehalose, sucrose, maltose, and mannose), sorbitol, amino acids, peptides, polymers and proteins such as albumins (bovine serum albumin, human serum albumin) or gelatins.
  • disaccharides such as lactose, trehalose, sucrose, maltose, and mannose
  • sorbitol amino acids
  • peptides polymers and proteins
  • albumins bovine serum albumin, human serum albumin
  • the biopolymer may be selected from PLP, PV-50, PL-50, PP-30, PP-50, PP-60 and PP-75.
  • the thermal stabiliser may be select from mannitol, polymers (such as dextran, polyethylene glycol, polyvinyl pyrrolidone) and proteins.
  • the bulking agent may be selected from mannitol, polymers (such as dextran, polyethylene glycol, and polyvinyl pyrrolidone), disaccharides (such as lactose, trehalose, sucrose, maltose, and mannose), sorbitol and proteins such as albumins and gelatins.
  • polymers such as dextran, polyethylene glycol, and polyvinyl pyrrolidone
  • disaccharides such as lactose, trehalose, sucrose, maltose, and mannose
  • sorbitol and proteins such as albumins and gelatins.
  • the cell based biological materials may include mammalian cells and tissue, bacteria, viruses, fungal matter or components thereof.
  • Examples of cells include, but are not limited to stem cells, epithelial cells, endothelial cells, red blood cells.
  • prokaryote microorganisms gram positive bacteria, gram negative bacteria, eukaryotic organisms, fungal spores, Caudovirales, Herpesvirales, Mononegavirales, Nidovirales, Picornavirales, and protozoal cells.
  • the bacterial cells may be selected from Lactobacilli such as Lactobacillus crispatusa and L. plantarum, Bifidobacteria, Bacillus coagulans and Saccharomyces boulardii.
  • the antioxidant may be selected from: Vitamin A (retinol), Vitamin C (ascorbic acid) and Vitamin E (comprising tocotrienol and tocopherol), Vitamin co-factors (such as coenzyme Q10), minerals (such as manganese and iodine), hormones (such as melatonin), carotenoid terpenoids, flavonoid polyphenolics, phenolic acids and esters thereof, and non- fiavonoid phenolics (such as curcumin, silymarin, xanthones and eugenol and biopolymers such as PP-50, PLP, PV-50, PL-50, PP-30, PP-50, PP-60 and PP-75
  • the agent that may enable the active 'loading' of the cell or organism with a cryoprotectant may be a biopolymer selected from PP-50, PLP, PV-50, PL-50, PP-30, PP- 50, PP-60 and PP-75.
  • PP-50, PLP, PV-50, PL-50, PP-30, PP- 50, PP-60 and PP-75 Some recent developments have been made in the cryoprotection of cells by the use of biopolymers that have been shown to render cell membranes permeable on a temporary basis, thus allowing 'loading' of cryoprotectant such as trehalose into the intracellular compartment.
  • Polymer PP-50 has been shown to act as a membrane permeabilising agent to enable intracellular loading of trehalose prior to the freeze drying of red blood cells (Lynch et al (2010); GB 1000999.1).
  • Freezing is defined as the solidification of solvent and the rendering of most or all of the solute phase into a rigidified state by the removal of heat.
  • Sublimation is defined as the direct conversion of solid solvent into the gaseous phase.
  • Desorption is defined as the removal of unfrozen solvent from a material by the application of energy in line with the applicable sorption isotherm.
  • a method for reconstitution of freeze-dried cell based biological material comprising the steps of adding a reconstituting solution to the material that results in the final extracellular concentration being hypertonic to the material and where the reconstituting solution is hypertonic to the cells
  • the hypertonic or hyperosmotic solution and can be added at a temperature between 2°C and 45°C. Preferably, the temperature is between 20 °C and 37 °C.
  • the solution can be given in either a single shot or in several aliquots.
  • the solution may comprise one or more of aqueous solutions of salt (sodium chloride), phosphate buffer or the polymer hydroxy ethyl starch (HES), glucose, or dextran.
  • the solution may be isotonic or iso-osmotic to the cells.
  • Freeze dried cells are usually resuspended in water in order to maintain the same extracellular osmolality as the cell suspension possessed prior to freeze-drying.
  • red blood cells it was noted in studies carried out in the inventors' laboratories on red blood cells that the rehydration in this manner resulted in significant haemolysis, which was attributed to localised regions of hypotonicity during the rehydration step. Therefore, it was acknowledged that water does not represent a suitable rehydrating medium for lyophilised red blood cells but that, instead, the use of solutions that were not significantly hypotonic may lead to a decrease in lysis during the rehydration process.
  • rehydration media comprising aqueous solutions of salt (sodium chloride), phosphate buffer or the polymer hydroxy ethyl starch (HES) demonstrated these solutions to be significantly more effective in maintaining red blood cell counts, which was attributed to the obviation of local regions of hypotonicity during the rehydration process; indeed, the resulting solutions were significantly hypertonic, which may have led to reversible crenulation (temporary shrinking of cells) but not lysis.
  • standard rehydration media used in infusion drips in a medical environment such as glucose or dextran.
  • a method of concentrating a biological material in an aqueous medium by contacting the medium with an osmotic driver solution where the medium and the osmotic driver solution are separated by a semi-permeable membrane and an osmotic gradient is created across the membrane.
  • the semi-premeable membrane is a viral-excluding membrane.
  • the membrane allows water and a range of solutes to pass, but the overall ratio of the buffer components remains isotonic with the biological material. This allows reduction of volume of biological sample thereby concentrating it.
  • the biological material is raw blood, red blood cell suspension or blood washed in phosphate buffered saline.
  • the osmotic drive solution can be selected from saccharides such as sucrose or proteins such as bovine serum albumin or metal salts such as sodium chloride. Concentrations may be greater than 278 mOsm (milliosmoles) and up to the saturation point of the solute in question. Preferably, the concentration may be 70% (w/v).
  • the concentration step may be used prior to the freeze drying method described, thereby reducing freeze drying time and making the process more efficient and cost effective.
  • Concentration can mean the reduction in volume of a suspension to provide for increased cell concentration but with a proportionally lower level of increase in the concentration of solutes.
  • a composition comprising freeze dried biological material and one or more of a freeze dried buffer solution, bulking agents, pH stabilisers, pH adjusters, thermal stabilisers, cryoprotectant, lyoprotectant, an antioxidant, a biopolymer or an agent that may enable the active 'loading' of the cell or organism with at least one of the aforementioned components.
  • the biological material may be cell based biological material and may have been freeze dried in a hypertonic solution.
  • the pH of the hypertonic solution may be acidic and may be pH 7.2 or below.
  • the osmolarity of the hypertonic solution may be greater than 300mOsmol.
  • the biological material is selected from bacterial cells or mammalian cells. More preferably, the mammalian cells are red blood cells or nucleated cells.
  • the bacteria] cells are selected from Lactobacilli such as Lactobacillus crispatusa and L.
  • composition comprising biological material that has been reconstituted according to the method of the present invention.
  • the biological material is cell based and may be selected from bacterial cells or mammalian cells. More preferably, the mammalian cells are red blood cells or nucleated cells.
  • the bacterial cells are selected from Lactobacilli such as Lactobacillus crispatus and L. plantarum, Bifidobacteria, Bacillus coagulans and Saccharomyces boulardii.
  • composition comprising intact, functional or viable biological material that has been freeze dried by the method of the present invention and reconstituted by the method of the present invention.
  • the biological material is cell based and may be selected from bacterial cells or mammalian cells. More preferably, the mammalian cells are red blood cells or nucleated cells.
  • the bacterial cells are selected from Lactobacilli such as Lactobacillus crispatusa. and L. plantarum, Bifidobacteria, Bacillus coagulans and Saccharomyces boulardii
  • the composition may additionally comprise an anti-oxidant.
  • the antioxidant is a biopolymer such as PP-50.
  • the composition may further comprise a cryoprotectant selected from trehalose, lactose, sucrose, maltose or mannose.
  • the antioxidant may be selected from: Vitamin A (retinol), Vitamin C (ascorbic acid) and Vitamin E (comprising tocotrienol and tocopherol), Vitamin co-factors (such as coenzyme Q10), minerals (such as manganese and iodine), hormones (such as melatonin), carotenoid terpenoids, flavonoid polyphenolics, phenolic acids and esters thereof, and non- flavonoid phenolics (such as curcumin, silymarin, xanthones and eugenol), biopolymers such as PLP, PV-50, PL-50, PP-30, PP-50, PP-60 and PP-75.
  • the antioxidant is PP-50.
  • the biological material is cell based and may comprise mammalian cells, such as stem cells, epithelial cells, endothelial cells, or red blood cells.
  • the biological material may comprise bacterial cells such as Lactobacilli such as Lactobacillus crispatus and L. plantarum, Bifidobacteria, Bacillus coagulans and Saccharomyces boulardii.
  • Antioxidants are not widely considered in the development of formulations for freeze-drying.
  • De Paz et al, (1988) used ascorbic acid and monosodium glutamate to reduce oxidation of Streptococcus lactis for the sole purpose reducing the bitterness that can occur due to the use of fast acid-producing strains of bacterial starter cultures in cheese production.
  • the ability to prevent or reduce oxidation of haemoglobin by biopolymers such as PP-50 has not been identified previously. This surprising effect was first noted in the inventors' laboratory following the freeze-drying and reconstitution of red blood cell suspensions. The use of antioxidants to confer cell stability and viability in the freeze-drying process has not previously been contemplated.
  • anti-oxidants to prevent or reduce oxidation within a cell, or to enhance the antioxidant properties of other solutes by enabling intracellular loading, is a surprising new effect in the field of freeze drying and can be utilised to provide better cell viability and the ability of the cells to function normally even after being subjected to freeze drying and reconstitution.
  • the reduced oxidation of haemoglobin allowed the cells to carry oxygen to the same or similar extent as the normal unprocessed blood cells or blood cells in the natural state.
  • a method of providing intact, functional or viable reconstituted freeze dried biological material comprising the steps of adding an antioxidant to the material prior to freeze drying.
  • kits comprising freeze dried cell based biological material prepared by the methods of the invention and a reconstituting medium.
  • the rehydrating or reconstituting medium is hypertonic to the cells in the biological material.
  • the methods of the invention have allowed the development of formulations that will enable long-term storage and transport of a wide range of cell-based therapeutics without the need for ultralow temperature storage facilities or a refrigerated transport "cold chain".
  • the methods of the present invention have also allowed better survivability, functionality or viability of the cells by ensuring they remain intact through the process. Thus, the cells have a greater ability to function normally, or as desired, after freeze drying and reconstitution.
  • Figure 1 is a graph showing "freeze-drying survival" for 30 RBC formulations incubated in the absence of the polymer PP-50, frozen by immersion in liquid nitrogen and subsequent primary drying carried out at a shelf temperature of 0°C.
  • Figure 2 is a graph showing "freeze-drying survival" for 30 RBC formulations incubated in the presence of the polymer PP-50, frozen by immersion in liquid nitrogen and subsequent primary drying carried out at a shelf temperature of 0°C.
  • Figure 3 is a graph showing RBC "survival" and percentage haemoglobin oxidation for PBS- based formulations frozen by immersion in liquid nitrogen and subsequent primary drying at a shelf temperature of -40°C
  • Figure 4 is a graph showing RBC "survival" for incubated and freeze-dried PBS-based formulations following reconstitution with three different rehydration solutions.
  • PBS phosphate buffered saline
  • Tris tris (hydroxymethyl)aminomethane High protectant concentration was 0.7M for trehalose and sucrose, 5% w/v for dextran Medium protectant concentration was 0.36M for trehalose and sucrose, 3% w/v for dextran Low protectant concentration was 0.18M for trehalose and sucrose, 1 % w/v for dextran High cooling rate involved dipping samples into liquid nitrogen at -196°C
  • formulations were tested in the presence and absence of a bio-polymer, PP- 50, at 20( ⁇ g/ml, as reflected in the data shown in Figure 1 (absence of PP-50) and Figure 2 (presence of PP-50).
  • Biopolymer PP-50 was donated by the University of Cambridge, Department of Chemical Engineering and Biotechnology.
  • Biopolymers PLP, PV-50, PL-50, PP-30, PP-50, PP-60 and PP-75 can be made by the process defined in Lynch et al (2010); Eccleston et al (2000); Chen et al (2009).
  • PP-50 was designed to increase RBC membrane permeability to buffer and protectant molecules, while also having the side-effect of reducing the extent of intracellular and extracellular solute concentration differential, which can lead to crenulations or lysis.
  • a graph showing the summary of % RBC survival and level of haemoglobin oxidation for different starting solutions (suspensions) frozen by immersion in liquid nitrogen and with primary drying carried out at a shelf temperature of -40°C, are provided in the Figure 3.
  • the freeze-dried cakes produced were reconstituted using PBS solution and samples were taken from each blood solution for UV analysis, to quantify BC survival and resulting haemoglobin status (haemoglobin, oxy-haemoglobin and met-haemoglobin).
  • the resulting data showed that the best average survival was obtained by using the fast freezing rate, with an average of 32.5% across all formulations, and an average haemoglobin oxidation of 18.5%.
  • the medium freezing rate cycle produced the next best survival with an average of 23.8% and 19.0% haemoglobin oxidation; the slow freezing rate led to an average survival rate of 16.9%, with 3.7% haemoglobin oxidation.
  • the freeze-dried cakes produced were reconstituted using PBS solution and samples were taken from each blood solution for UV analysis.
  • the resulting data showed that the best average survival was obtained by using the low temperature primary drying, with an average of 83.1% across all formulations, and an average haemoglobin oxidation of 23.1%.
  • the medium temperature primary drying cycle produced the next best survival with an average of 81.3% and 29.7% haemoglobin oxidation; the high temperature primary drying led to an average survival rate of 32.5% and 18.5% haemoglobin oxidation.
  • the best survival was obtained by the nonpolymer-containing trehalose solution, which achieved a maximum survival of 96.6% in the low temperature cycle, although haemoglobin oxidation was high (61.1 %).
  • the 0.9% NaCl solution is the same as would readily be available sterile in modern hospitals, so would be of benefit in the final product if proved to be acceptable.
  • Hydroxyethyl Starch (HES) is commercially available as a blood plasma substitute and therefore would be beneficial to the final product if the RBCs could be resuspended in this solution prior to injection to maximise the benefit of the blood to the patient.
  • freeze-dried cakes produced were reconstituted using one of the three solutions above and samples were taken from each blood solution for UY analysis.
  • the resulting data were reconstituted using one of the three solutions above and samples were taken from each blood solution for UY analysis. The resulting data
  • Reconstitution with saline (prepared by dissolution of sodium chloride powder [source: Sigma Chemical Co. Ltd., Poole, UK, Cat. No. S-7653] in analytical grade water) led to an average survival of 633% and 8.8% haemoglobin oxidation; reconstitution with HES (source: Sigma Chemical Co. Ltd., Poole, UK; Cat. No. H6382, dissolved in BDH NormapuR water to give a 1% w/v solution) led to an average survival rate of 71.4%, with 8.3% haemoglobin oxidation.
  • This method employed purpose-built test chambers consisting of two compartments, one containing the substance to be concentrated, and the other containing a highly concentrated solution of an osmotic driver.
  • the two compartments are separated by a novel viral-excluding membrane (obtained from HATS Ltd., Dorset). Water and a range of solutes are able to traverse the membrane, driven by the osmotic gradient generated by the driver, whilst maintaining the overall ratio of the buffer components largely isotonic with the red blood cells, which are themselves excluded by the membrane.
  • Freeze drying of the concentrated RBC suspensions was carried out, in order to determine whether the volume reduction process would have a significant impact on RBC death or haemoglobin oxidation.
  • the concentrated PBS blood samples were included in the freezing rate study cycle so that a direct comparison could be made between concentrated and non- concentrated blood.
  • haemoglobin oxidation may be achieved that are in keeping with the data shown in sections 1- 3 above, while providing the additional benefit of shorter freeze-drying time due to the smaller volumes of water present.

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Abstract

La présente invention concerne un procédé de lyophilisation, en particulier la lyophilisation d'une matière biologique d'origine cellulaire et sa reconstitution ultérieure. La matière biologique peut provenir d'une source quelconque, telle que des cellules microbiennes, des cellules de protozoaires, des cellules animales ou des cellules végétales. En particulier, l'invention concerne la lyophilisation de cellules de mammifère, telles que des cellules sanguines, des cellules nucléées et des cellules bactériennes, pour l'utilisation thérapeutique et l'utilisation dans des produits alimentaires. L'invention concerne en outre un procédé de reconstitution des cellules lyophilisées conduisant à une meilleure viabilité et une meilleure fonctionnalité des cellules après leur reconstitution.
PCT/GB2012/000051 2011-01-20 2012-01-20 Procédé de lyophilisation WO2012098358A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
GB1314778.0A GB2502016B (en) 2011-01-20 2012-01-20 Method of freeze drying and reconstituting biological material

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
US201161461628P 2011-01-20 2011-01-20
US61/461628 2011-01-20
GBGB1101002.2A GB201101002D0 (en) 2011-01-20 2011-01-20 Freeze drying method
GB1101002.2 2011-01-20

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WO2020210553A1 (fr) 2019-04-09 2020-10-15 Dermbiont, Inc. Compositions et méthodes pour améliorer la santé de la peau et pour le traitement et la prévention de maladies, de troubles et d'états associés à des microbes pathogènes
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WO2021050896A1 (fr) * 2019-09-13 2021-03-18 Lonza Ltd Procédé de production de cellules lyophilisées
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EP3583847B1 (fr) * 2017-02-20 2022-04-20 Xiaoyang Xu Système et procédé de lyophilisation de cellules
WO2022081762A1 (fr) 2020-10-14 2022-04-21 Dermbiont, Inc. Compositions et méthodes pour améliorer la santé de la peau et pour le traitement et la prévention de maladies, troubles et états associés à des champignons et autres microbes pathogènes
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WO2023218019A1 (fr) 2022-05-12 2023-11-16 Etherna Immunotherapies Nv Composition d'arn lyophilisée
EP4282489A2 (fr) 2016-06-14 2023-11-29 Vedanta Biosciences, Inc. Traitement d'une infection par clostridium difficile
EP4386090A1 (fr) 2022-12-12 2024-06-19 Assistance Publique - Hôpitaux De Paris Procédé de détermination et d'amélioration de l'efficacité potentielle d'un traitement anticancéreux

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WO2018172483A1 (fr) 2017-03-22 2018-09-27 Assistance Publique - Hopitaux De Paris Procédé de détermination de l'efficacité potentielle d'un traitement anticancéreux
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WO2020210553A1 (fr) 2019-04-09 2020-10-15 Dermbiont, Inc. Compositions et méthodes pour améliorer la santé de la peau et pour le traitement et la prévention de maladies, de troubles et d'états associés à des microbes pathogènes
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WO2022081762A1 (fr) 2020-10-14 2022-04-21 Dermbiont, Inc. Compositions et méthodes pour améliorer la santé de la peau et pour le traitement et la prévention de maladies, troubles et états associés à des champignons et autres microbes pathogènes
WO2022158434A1 (fr) 2021-01-19 2022-07-28 国立研究開発法人理化学研究所 Composition pour la décomposition de la trypsine ou de tmprss2
WO2022178292A1 (fr) 2021-02-18 2022-08-25 Vedanta Biosciences, Inc. Compositions et méthodes se suppression d'organismes pathogènes
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WO2023278477A1 (fr) 2021-06-28 2023-01-05 Vedanta Biosciences, Inc. Procédés de colonisation par un microbiome, de traitement et/ou de prévention d'une maladie intestinale inflammatoire et d'une maladie du greffon contre l'hôte
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WO2023218019A1 (fr) 2022-05-12 2023-11-16 Etherna Immunotherapies Nv Composition d'arn lyophilisée
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