WO2014152484A1 - Microbiote fécal lyophilisé à utiliser dans la transplantation microbienne fécale - Google Patents

Microbiote fécal lyophilisé à utiliser dans la transplantation microbienne fécale Download PDF

Info

Publication number
WO2014152484A1
WO2014152484A1 PCT/US2014/027391 US2014027391W WO2014152484A1 WO 2014152484 A1 WO2014152484 A1 WO 2014152484A1 US 2014027391 W US2014027391 W US 2014027391W WO 2014152484 A1 WO2014152484 A1 WO 2014152484A1
Authority
WO
WIPO (PCT)
Prior art keywords
composition
colitis
freeze
optionally
biological material
Prior art date
Application number
PCT/US2014/027391
Other languages
English (en)
Inventor
Michael J. Sadowsky
Alexander KHORUTS
Matthew Hamilton
Aleh BOBR
Alexa Rachel WEINGARDEN
Original Assignee
Regents Of The University Of Minnesota
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
Application filed by Regents Of The University Of Minnesota filed Critical Regents Of The University Of Minnesota
Publication of WO2014152484A1 publication Critical patent/WO2014152484A1/fr
Priority to US14/850,318 priority Critical patent/US20150374761A1/en
Priority to US15/837,834 priority patent/US20180110810A1/en

Links

Classifications

    • 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
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
    • A61K35/37—Digestive system
    • A61K35/38—Stomach; Intestine; Goblet cells; Oral mucosa; Saliva
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/66—Microorganisms or materials therefrom
    • A61K35/74—Bacteria
    • A61K35/741—Probiotics
    • 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
    • A61K9/0031—Rectum, anus
    • 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
    • A61K9/19—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles lyophilised, i.e. freeze-dried, solutions or dispersions
    • 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
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
    • Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Definitions

  • Fecal microbiota transplantation also commonly known as 'fecal
  • bacteriotherapy represents the one therapeutic protocol that allows the fastest reconstitution of a normal composition and functional gut microbial community.
  • FMT has been offered by select centers across the world, typically as an option of last resort for patients with recurrent Clostridium difficle infection (CDI).
  • CDI Clostridium difficle infection
  • the mostly commonly earliest cited report for FMT was by Eiseman and colleagues who in 1958 described the use of fecal enemas for patients who likely had severe or fulminant form of pseudomembranous colitis (Eiseman et al. Surgery 1958;44:854-9). Since this time, well over 500 cases have been reported as individual case reports, small case series, or clinical trials with a -90% cumulative success rate in clearing recurrent CDI, without any noted adverse events.
  • compositions presented herein provide a significant advantage by making useful compositions of intestinal microflora, for instance, colon microflora.
  • Such compositions may be readily available for use by a physician to treat a patient, readily available for use by a patient at home, and/or readily available as an over-the-counter
  • compositions for sale directly to a consumer.
  • the compositions are not limited to:
  • delivery is by oral
  • the material can be delivered via nasogastric tube, enema, or colonoscopy.
  • the composition includes biological material and a cryoprotectant, wherein the freeze-dried composition is friable, wherein optionally the composition includes a pharmaceutically acceptable carrier, and optionally the composition is a formulation for oral administration, and wherein the freeze-dried composition, upon reconstitution with water, includes no greater than about 0.05%, 0.1% , 0.2%> 0.3% , 0.4% , 0.5% , 0.6% , 0.7% , 0.8% , 0.9%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9% or 10% weight non-living material/weight biological material, wherein the biological material includes human gut, colon or intestinal fecal microbes, and optionally the biological material includes human gut, colon or intestinal bacteria.
  • the composition includes an extract or preparation of human feces including human fecal material and a cryoprotectant, wherein the freeze-dried composition is friable, wherein the human fecal material, upon reconstitution with water, consists of, or consists essentially of, particles of non-living material and/or particles of biological material that will pass through a sieve having a sieve size of 2.0 mm, 1.0 mm, 0.5 mm, 0.25 mm, 0.212 mm, 0.180 mm, 0.150 mm, 0.125 mm, 0.106 mm, 0.090 mm, 0.075 mm, 0.063 mm, 0.053 mm, 0.045 mm, 0.038 mm, 0.032 mm, 0.025 mm, 0.020 mm, or 0.01 mm, and wherein optionally the composition includes a pharmaceutically acceptable carrier, and optionally the composition is a formulation for oral administration.
  • the composition includes at least 4 different phyla of gut, colon or intestinal bacteria extracted or prepared from the gut, colon or intestine, and a
  • cryoprotectant wherein the phyla include a Bacteroidetes, a Firmicutes, a Proteobacteria a Tenericutes phylum, or a combination thereof, wherein optionally the phyla are chosen from Bacteroidetes, Firmicutes, Proteobacteria, Tenericutes, or a combination thereof, wherein the composition, upon reconstitution with water, includes no greater than about 0.05%, 0.1% , 0.2% , 0.3% , 0.4% , 0.5% , 0.6% , 0.7% , 0.8% , 0.9% 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%), 9%> or 10%> weight non-living material/weight biological material, wherein the biological material includes human gut, colon or intestinal fecal microbes, and optionally the biological material includes human gut, colon or intestinal bacteria, and wherein optionally the composition includes a pharmaceutically acceptable carrier, and optionally the composition is a formulation for oral administration.
  • the composition includes an extract of human feces and a cryoprotectant, wherein the composition, upon reconstitution with water, is substantially odorless, wherein the composition includes biological material, and optionally wherein the biological material includes microbes, and wherein optionally the composition includes a pharmaceutically acceptable carrier, and optionally the composition is a formulation for oral administration.
  • the fecal material consists of, or consists essentially of, particles that will pass through a 0.25 mm sieve, or equivalent.
  • the composition includes an extract of human feces and a cryoprotectant, wherein the composition, upon reconstitution with water, is substantially odorless, wherein the composition includes biological material, and optionally wherein the biological material includes microbes, and wherein optionally the composition includes a pharmaceutically acceptable carrier, and optionally the composition is a formulation for oral administration.
  • the fecal material consists of, or consists essentially of, particles that will pass through a 0.25 mm sieve, or equivalent.
  • composition includes at least about 1 x 10 , 1.5 x 10 , 2 x 10 , or 2.5 x 10 bacteria.
  • the biological material includes: a plurality of prokaryotic cells, eukaryotic cells, or viruses; or a population of prokaryotic cells, eukaryotic cells, and viruses, that is substantially identical to or representative of or equivalent to a population of prokaryotic cells, eukaryotic cells, and viruses present in gut, intestine, colon, or feces of a normal healthy human.
  • the biological material present includes a population of prokaryotic cells and viruses that is substantially identical to or
  • the biological material includes a population of prokaryotic cells, eukaryotic cells, or viruses that is substantially identical to or representative of or equivalent to a population of prokaryotic cells, eukaryotic cells, and viruses present in the feces of a normal healthy human.
  • the compositions are prepared by a process.
  • the process includes subjecting a fecal sample to a condition or conditions that removes at least about 91%, 92%, 93%, 94% 95%, 96%, 97%, 98%, 99% or more of the non-living material present in the fecal sample prior to the subjecting to result in an extract, adding a cryoprotectant to the extract to result in a mixture, and freeze-drying the mixture to result in the composition.
  • the composition includes a biological material, and optionally the biological material includes bacteria, wherein optionally the composition includes a pharmaceutically acceptable carrier, and optionally the composition is a formulation for oral administration.
  • the subjecting occurs at a temperature of no greater than about 26°C, 27°C, 28°C, 29°C, 30°C, 31°C, 32°C, 33°C, or 34°C.
  • the process includes filtering a fecal sample with a filter medium, wherein the filter medium includes at least one sieve size of no greater than about 2.0 mm, 1.0 mm, 0.5 mm, 0.25 mm, 0.212 mm, 0.180 mm, 0.150 mm, 0.125 mm, 0.106 mm, 0.090 mm, 0.075 mm, 0.063 mm, 0.053 mm, 0.045 mm, 0.038 mm, 0.032 mm, 0.025 mm, 0.020 mm, or 0.01 mm to result in or to generate a filtrate, adding a
  • the composition includes a biological material, and optionally the biological material includes bacteria, wherein optionally the composition includes a pharmaceutically acceptable
  • composition is a formulation for oral administration.
  • the process further includes reconstituting the composition with an aqueous solution.
  • the process further includes filtering the fecal sample with a filter medium, wherein the filter medium includes a sieve size of no greater than 0.25 mm.
  • the filtering occurs at a temperature of no greater than about 26°C, 27°C, 28°C, 29°C, 30°C, 31°C, 32°C, 33°C, or 34°C.
  • the composition includes at least 4 different phyla of bacteria, wherein the phyla include a Bacteroidetes, a Firmicutes, a Proteobacteria, a Tenericutes phyla, or a combination thereof, wherein optionally the phyla are chosen from Bacteroidetes, Firmicutes, Proteobacteria, Tenericutes, or a combination thereof.
  • the composition further includes at least 5, 6, 7, 8, 9, or 10 different classes of bacteria chosen from
  • Betaproteobacteria Gammaproteobacteria, Mollicutes, and Verrucomicrobiae.
  • cryoprotectant present in a composition described herein includes skim milk, gelatin, mannitol, or a combination thereof. In one embodiment, a composition described herein includes at least two cryoprotectants, wherein the first
  • cryoprotectant is sucrose.
  • a second cryoprotectant is selected from skim milk, gelatin, and mannitol.
  • the cryoprotectant is present at a concentration of at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, or 10% (vol/vol).
  • the cryoprotectant of a composition described herein does not include
  • composition described herein does not include surcose as the only cryoprotectant.
  • a composition described herein is encapsulated in a capsule.
  • the capsule includes an acid-resistant enteric coating.
  • composition described herein is for use as a therapeutically active agent.
  • a composition described herein is for use in the treatment of a disease or a pathological or iatrogenic condition of the colon.
  • the disease is a disease or condition characterized by a dysfunctional or pathological composition of colon microbiota.
  • the disease is a Clostridium difficile colitis, such as acute Clostridium difficile colitis, relapsing Clostridium difficile colitis, or severe Clostridium difficile colitis.
  • the method is for replacing or supplementing or modifying a subject's colon microbiota.
  • such a method includes administering to the subject the composition described herein.
  • the method further includes reconstituting the composition with an aqueous solution.
  • the method is for treating a subject.
  • such a method includes administering to a subject in need thereof an effective amount of a composition described herein.
  • the subject has or is at risk for having a colitis, such as an autoimmune colitis.
  • the autoimmune colitis is selected from an inflammatory bowel disease, an ulcerative colitis, a Crohn's disease and an irritable bowel syndrome.
  • the colitis is an infectious colitis.
  • the infectious colitis is selected from a Clostridium difficile colitis and an enterohemorrhagic colitis.
  • the Clostridium difficile colitis is selected from an acute Clostridium difficile colitis, a relapsing Clostridium difficile colitis, and a severe Clostridium difficile colitis.
  • the enterohemorrhagic colitis is caused by a microbe selected from a Shigella spp. and an E. coli.
  • the subject has or is at risk for chronic diarrhea or chronic constipation.
  • a method further includes removal of some, most, or substantially all of the subject's colon, gut or intestinal microbiota prior to the
  • a composition described herein for the manufacture of a medicament is for the manufacture of a medicament for treating or ameliorating or preventing a disease or a pathological or iatrogenic condition of the colon, wherein optionally the disease is a disease or condition characterized by a dysfunctional or pathological composition of colon microbiota, or the disease is a Clostridium difficile colitis, or the disease or condition is a colitis, an autoimmune colitis, an infectious colitis or an enterohemorrhagic colitis.
  • the term "and/or” means one or all of the listed elements or a combination of any two or more of the listed elements.
  • the steps may be conducted in any feasible order. And, as appropriate, any combination of two or more steps may be conducted simultaneously.
  • freeze-dried compositions that include fecal microbes.
  • fecal microbes refers to microorganisms that are present in the gut, intestine, or colon, preferably colon, or feces of a normal healthy adult human.
  • Such a freeze-dried composition may be prepared by processing fecal material.
  • fecal material refers to human stool. Unprocessed fecal material contains nonliving material and biological material.
  • non-living material refers to the non-living material in fecal material, and may include, but is not limited to, dead bacteria, shed host cells, proteins, carbohydrates, fats, minerals, mucus, bile, undigested fiber and other foods, and other compounds resulting from food and metabolic ingestion and waste products and partial or complete digestion of food materials.
  • Non-living material does not include an excipient, e.g., a pharmaceutically inactive substance, such as a cryoprotectant, added to a processed fecal material.
  • Bio material refers to the living material in fecal material, and includes microbes including prokaryotic cells, such as bacteria and archea (e.g., living prokaryotic cells and spores that can sporulate to become living prokaryotic cells), eukaryotic cells such as protozoa and fungi, and viruses.
  • prokaryotic cells such as bacteria and archea
  • eukaryotic cells such as protozoa and fungi
  • viruses such as protozoa and fungi
  • biological material refers to the living material, e.g., the microbes, eukaryotic cells, and viruses, which are present in the colon of a normal healthy human.
  • freeze-dried refers to a composition having the characteristics described herein and further having substantially no water present, and in one
  • no detectable water in any embodiment, no detectable water.
  • Methods for freeze-drying a composition are known and routinely used. The word freeze-drying is used synonymously with lyophilization.
  • a method for freeze-drying a composition may include one or more pretreatments (e.g., concentrating, addition of a cryoprotectant, increasing the surface area of a composition), freezing the composition, and drying (e.g., exposing the composition to a reduced atmospheric pressure to result in sublimation of the water present in the composition).
  • prokaryotic cells that may be present in a freeze-dried composition described herein include cells that are members of the class Actinobacteria, such as the subclass Actinobacteridae and subclass Coriobacteridae.
  • Examples of the subclass Actinobacteridae include members of the order Actinomycetales, and members of the order Bifidobacteriales.
  • Members of the order Bifidobacteriales include members of the family Bifidobacteriaceae.
  • Examples of the subclass Coriobacteridae include members of the order Coriobacteriales.
  • Members of the order Coriobacteriales include members of the family Coriobacteriaceae.
  • prokaryotic cells include members of the phylum Bacteroidetes, such as class Bacteroidia.
  • Members of class Bacteroidia include order Bacteroidales.
  • Members of order Bacteroidales include members of the family Bacteroidaceae, members of the family Porphyromonadaceae, members of the family Prevotellaceae, and members of the family Rikenellaceae.
  • prokaryotic cells include members of the phylum Firmicutes, such as class Bacilli, Clostridia, Erysipelotrichi, and Negativicutes.
  • class Bacilli include members of the order Bacillales (including members of the family
  • Lactobacillales including members of the family Aerococcaceae, Enterococcaceae, Lactobacillaceae, and Streptococcaceae.
  • Clostridia include members of the order Clostridiales
  • examples of the order Colstridiales include the family Catabacteriaceae, Peptococcaceae, Peptostreptococcaceae, Ruminococcaceae, Clostridiaceae, Eubacteriaceae, and Lachnospiraceae.
  • Examples of the class 1 include members of the family Aerococcaceae, Enterococcaceae, Lactobacillaceae, and Streptococcaceae.
  • examples of the order Colstridiales include the family Catabacteriaceae, Peptococcaceae, Peptostreptococcaceae, Ruminococcaceae, Clostridiaceae, Eubacteriaceae, and Lachnospiraceae.
  • Erysipelotrichi include members of the family Erysipelotrichaceae.
  • Examples of the class Negativicutes include members of the family Veillonellaceae.
  • Other examples of the order Bacillales include Bacillales Family Xl. Incertae Sedis, and Bacillaceae 1.
  • Other examples of the order Clostridiales include Clostridiales Family XI. Incertae Sedis, Clostridiales Family XIII. Incertae Sedis, and Clostridiaceae 1
  • prokaryotic cells include members of the phylum Proteobacteria, such as class Alphaproteobacteria, Betaproteobacteria, Deltaproteobacteria,
  • Epsilonproteobacteria Epsilonproteobacteria, and Gammaproteobacteria. Examples of the class
  • Alphaproteobacteria include members of the order Rhizobiales, and examples of members of the order Rhizobiales includes members of the family Rhodobiaceae, members of the family Brucellaceae, and members of the family Hyphomicrobiaceae.
  • Examples of the class Betaproteobacteria include members of the order Burkholderiales, and examples of members of the order Burkholderiales include members of the family Alcaligenaceae, members of the family Burkholderiaceae, and members of the family Sutterellaceae.
  • Examples of the class Deltaproteobacteria include members of the order
  • Desulfovibrionales and examples of members of this order include members of the family Desulfovibrionaceae and Desulfomicrobiaceae. Examples of the class
  • Epsilonproteobacteria include members of the order Desulfobacterales, and examples of members of this order include members of the family Desulfobacteraceae.
  • Examples of the class Gammaproteobacteria includes members of the order Alteromonadales and Enterobacteriales.
  • Examples of members of the order Alteromonadales include members of the family Shewanellaceae, and examples of members of the order Enterobacteriales include members of the family Enterobacteriaceae.
  • prokaryotic cells include members of the phylum
  • Tenericutes include members of the class Mollicutes.
  • Examples of the class Mollicutes include members of the order Entomoplasmatales, and members of the order Entomoplasmatales include members of the family Spiroplasmataceae.
  • prokaryotic cells include members of the class Verrucomicrobiae include members of the order Verrucomicrobiales, and examples of members of the order Verrucomicrobiales includes members of the family Verrucomicrobiaceae.
  • prokaryotic cells include members of the family Fusobacteriaceae.
  • a freeze-dried composition may include prokaryotic bacteria that are members of at least 1 phylum, at least 2 phyla, at least 3 phyla, at least 4 phyla, at least 5 phyla, at least 6 phyla, at least 7 phyla, at least 8 phyla, at least 9 phyla, or at least 10 phyla.
  • a composition of the present invention may include prokaryotic bacteria that are members of at least 1 class, at least 2 classes, at least 3 classes, at least 4 classes, at least 5 classes, at least 6 classes, or at least 7 classes.
  • a composition of the present invention may include prokaryotic bacteria that are members of at least 1 order, at least 2 orders, at least 3 orders, at least 4 orders, at least 5 orders, at least 6 orders, or at least 7 orders.
  • a composition of the present invention may include prokaryotic bacteria that are members of at least 1 family, at least 2 families, at least 3 families, at least 4 families, at least 5 families, at least 6 families, at least 7 families.
  • a composition of the present invention may include at least 5, at least 10, at least 20, or at least 30 different genera of prokaryotic bacteria.
  • a composition of the present invention may include at least 10, at least 50, at least 100, at least 200, at least 300, at least 400, at least 500, at least 600, or at least 700 different species of prokaryotic bacteria.
  • a freeze-dried composition described herein includes, when reconstituted with water, no greater than 10% weight of non-living material/weight biological material (wt/wt), no greater than 5 % (wt/wt), no greater than 2.5 % (wt/wt), no greater than 1% (wt/wt), no greater than 0.1% (wt/wt), no greater than 0.01% (wt/wt), or no greater than 0.001% (wt/wt) non-living material.
  • the amount of non-living material in a composition of the present invention is undetectable using currently available techniques. For instance, living material can be stained for biological activity, electron transport, DNA and RNA for specific genes.
  • the fecal material present in a freeze-dried composition described herein does not include, when reconstituted with water, particles (e.g., particles of non-living material and/or particles of biological material) having a size of greater than 2.0 millimeters (mm), greater than 1.0 mm, greater than 0.5 mm, greater than 0.4 mm, greater than 0.3 mm, greater than 0.25 mm, greater than 0.212 mm, greater than 0.180 mm, greater than 0.150 mm, greater than 0.125 mm, greater than 0.106 mm, greater than 0.090 mm, greater than 0.075 mm, greater than 0.063 mm, greater than 0.053 mm, greater than 0.045 mm, greater than 0.038 mm, greater than 0.032 mm, greater than 0.025 mm, greater than 0.020 mm, or greater than 0.01 mm.
  • particles e.g., particles of non-living material and/or particles of biological material having a size of greater than 2.0 millimeters (mm), greater than
  • Non- fecal material present in a composition may include particles having a size of greater than 2.0 mm, greater than 1.0 mm, greater than 0.5 mm, greater than 0.4 mm, greater than 0.3 mm, greater than 0.25 mm, greater than 0.212 mm, greater than 0.180 mm, greater than 0.150 mm, greater than 0.125 mm, greater than 0.106 mm, greater than 0.090 mm, greater than 0.075 mm, greater than 0.063 mm, greater than 0.053 mm, greater than 0.045 mm, greater than 0.038 mm, greater than 0.032 mm, greater than 0.025 mm, greater than 0.020 mm, or greater than 0.01 mm.
  • the fecal material present in a composition of the present invention consists of, or consists essentially of, particles of non-living material and/or biological material having a size that will pass through a sieve having a sieve size of 2.0 mm, 1.0 mm, 0.5 mm, 0.4 mm, 0.3 mm, 0.25 mm, 0.212 mm, 0.180 mm, 0.150 mm, 0.125 mm, 0.106 mm, 0.090 mm, 0.075 mm, 0.063 mm, 0.053 mm, 0.045 mm, 0.038 mm, 0.032 mm, 0.025 mm, 0.020 mm, or 0.01 mm.
  • the fecal material present in a composition has a size that is less than or equal to 2.0 mm, less than or equal to 1.0 mm, less than or equal to 0.5 mm, less than or equal to 0.4 mm, less than or equal to 0.3 mm, less than or equal to 0.25 mm, less than or equal to 0.212 mm, less than or equal to 0.180 mm, less than or equal to 0.150 mm, less than or equal to 0.125 mm, less than or equal to 0.106 mm, less than or equal to 0.090 mm, less than or equal to 0.075 mm, less than or equal to 0.063 mm, less than or equal to 0.053 mm, less than or equal to 0.045 mm, less than or equal to 0.038 mm, less than or equal to 0.032 mm, less than or equal to 0.025 mm, less than or equal to 0.020 mm, or less than or equal to 0.01 mm.
  • the sieve size may be based on the US Standard
  • a freeze-dried composition described herein includes a cryoprotectant.
  • Cryoprotectants useful in freeze-drying microbes include, for instance, D- Mannitol, D-Sorbitol, D-Glucose, casein hydrolysate, sucrose, gelatin, non-fat skim milk, starch hydolysate, fetal calf serum, bovine serum albumin, or combinations of 1, 2, 3, or 4 of the above cryoprotectants.
  • Other cryoprotectants are also known.
  • a cryoprotectant useful herein maintains the viability of fecal microbes when subjected to freeze-drying conditions, milling or grinding, and/or when stored as a freeze-dried composition.
  • Milling also referred to as grinding, is a process that physically changes a material into smaller particles.
  • Methods for milling freeze-dried compositions are known to the skilled person, and can occur at various temperatures, e.g., at or below 0°C, or above 0°C.
  • a cryoprotectant useful herein results in a freeze-dried composition that is friable.
  • a "friable" composition refers to a composition that can be easily milled to result in a fine powder.
  • a freeze-dried composition described herein that is friable is one that results in a powder that can be subsequently used to produce a tablet.
  • a useful powder may have size, density, flow, and compression characteristics suitable for production of tablets or encapsulation.
  • Useful cryoprotectants include those that result in a composition that is friable, does not crystallize during freeze-drying, and maximizes survival of microbes.
  • examples of useful cryoprotectants include, but are not limited to, skim milk, gelatin, and mannitol, and sucrose.
  • more than one cryoprotectant may be used, such as the combination of sucrose (5%) and skim milk (5%), or the combination of sucrose (10%) and gelatin (0.1%).
  • a useful cryopotectant is not sucrose alone, which unexpectedly crystallizes and hardens during freeze-drying, or glycerol, which unexpectedly results in an oily and viscous composition upon freeze-drying with a fecal material as described herein.
  • the total cryoprotectant used to produce a freeze-dried composition may be 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, or 30% (vol/vol) of the final concentration of a mixture of fecal microbes and the cryoprotectant before freeze- drying the composition.
  • a composition having a cryoprotectant at a final concentration of 10% equal volumes of a 20% solution of the cryoprotectant and a mixture of microbes derived from fecal material can be combined and mixed, and then freeze-dried.
  • a freeze-dried composition of the present invention does not include pathogenic biological material.
  • fecal material is from a person that has undergone a medical history, a physical examination, and laboratory testing.
  • the evaluation of medical history may include, but is not limited to, risk of infectious agents, presence of gastrointestinal co-morbidities, factors that can or do affect the composition of the intestinal microbiota, and systemic medical conditions (see Sadowsky et al, WO 12/122478).
  • Exclusion criteria regarding risk of infectious agents may include, but are not limited to, known viral infection with Hepatitis B, C or HIV; known exposure to HIV or viral hepatitis at any time; high risk behaviors including sex for drugs or money, men who have sex with men, more than one sexual partner in the preceding 12 months, any past use of intravenous drugs or intranasal ***e, history of incarceration; tattoo or body piercing within 12 months; travel to areas of the world where risk of traveler's diarrhea is higher than the US; and current communicable disease, e.g., upper respiratory viral infection.
  • Exclusion criteria regarding gastrointestinal co-morbidities include, but are not limited to, history of irritable bowel syndrome, wherein specific symptoms may include frequent abdominal cramps, excessive gas, bloating, abdominal distension, fecal urgency, diarrhea, constipation; history of inflammatory bowel disease such as Crohn's disease, ulcerative colitits, microscopic colitis; chronic diarrhea; chronic constipation or use of laxatives; history of gastrointestinal malignancy or known colon polyposis; history of any abdominal surgery, e.g., gastric bypass, intestinal resection, appendectomy,
  • cholecystectomy and the like; use of probiotics or any other over the counter aids used by the potential donor for purpose of regulating digestion, but yogurt and kefir products may be allowed if taken merely as food rather than nutritional supplements.
  • Exclusion criteria regarding factors that can or do affect the composition of the intestinal microbiota include, but are not limited to, antibiotics for any indication within the preceding 6 months; any prescribed immunosuppressive or anti-neoplastic
  • Exclusion criteria regarding systemic medical conditions include, but are not limited to, established or emerging metabolic syndrome, where criteria used for definition here are stricter than established criteria, including history of increased blood pressure, history of diabetes or glucose intolerance; known systemic autoimmunity, e.g., connective tissue disease, multiple sclerosis; known atopic diseases including asthma or eczema; chronic pain syndromes including fibromyalgia, chronic fatigue syndrome; ongoing (even if intermittent) use of any prescribed medications, including inhalers or topical creams and ointments; neurologic, neurodevelopmental, and neurodegenerative disorders including autism, Parkinson's disease.
  • criteria used for definition here are stricter than established criteria, including history of increased blood pressure, history of diabetes or glucose intolerance
  • known systemic autoimmunity e.g., connective tissue disease, multiple sclerosis
  • known atopic diseases including asthma or eczema
  • chronic pain syndromes including fibromyalgia, chronic fatigue syndrome
  • Exclusion criteria on physical examination may include, but are not limited to, general, such as body mass index ⁇ 30 kg/m 2 , central obesity defined by waste:hip ratio > 0.90 (male) and > 0.85 (female); blood pressure > 135 mmHg systolic and > 85 mmHg diastolic; skin - presence of a rash, tattoos or body piercing placed within a year, jaundice; enlarged lymph nodes; wheezing on auscultation; hepatomegaly or stigmata of liver disease; swollen or tender joints; muscle weakness; abnormal neurologic examination.
  • Exclusion criteria on laboratory testing may include, but is not limited to, positive stool Clostridium difficile toxin B tested by PCR; positive stool cultures for any of the routine pathogens including Salmonella, Shigella, Yersinia, Campylobacter, E.
  • coli 0157:H7 abnormal ova and parasites examination; positive Giardia, Cryptosporidium, or Helicobacter pylori antigens; positive screening for any viral illnesses, including HIV 1 and 2, Viral Hepatitis A IgM, Hepatitis surface antigen and core Ab; abnormal RPR (screen for syphilis); any abnormal liver function tests including alkaline phosphatase, aspartate aminotransaminase, alanine aminotransferase; raised serum triglycerides > 150 mg/dL; HDL cholesterol ⁇ 40 mg/dL (males) and ⁇ 50 mg/dL (females); high sensitivity CRP > 2.4 mg/L; raised fasting plasma glucose (> 100 mg/dL).
  • compositions of the present invention may be included in a diversity of pharmaceutically acceptable formulations.
  • a formulation may be a solid composition.
  • Solid compositions include, but are not limited to, powder, granule, compressed tablet, pill, capsule, microsphere, wafer, and the like.
  • Those formulations may include a pharmaceutically acceptable carrier to render the composition appropriate for administration to a subject.
  • pharmaceutically acceptable carrier includes pharmacologically inactive compounds compatible with pharmaceutical administration.
  • the compositions of the present invention may be formulated to be compatible with its intended route of administration.
  • a composition of the present invention may be administered by any method suitable for depositing in the
  • administration include rectal administration (e.g., by suppository, enema, upper endoscopy, upper push enteroscopy, or colonoscopy), intubation through the nose or the mouth (e.g., by nasogastric tube, nasoenteric tube, or nasal jejunal tube), or oral administration (e.g., by a solid such as a pill, tablet, or capsule, or by liquid).
  • a liquid form of the composition is delivered to a subject
  • the freeze- dried composition is reconstituted with an aqueous solution, such as by adding water or saline, or exposing the freeze-dried composition to a body fluid.
  • composition described herein may be conveniently administered in a form containing one or more
  • Suitable carriers are well known in the art and vary with the desired form and mode of administration of the composition. For example, they may include diluents or excipients such as fillers, binders, wetting agents, disintegrators, surface-active agents, glidants, lubricants, and the like.
  • the carrier may be a solid (including powder), liquid, or combinations thereof.
  • Each carrier is preferably "acceptable” in the sense of being compatible with the other ingredients in the composition and not injurious to the subject.
  • the carrier is preferably biologically acceptable and inert, i.e., it permits the composition to maintain viability of the biological material until delivered to the appropriate site.
  • Oral compositions may include an inert diluent or an edible carrier.
  • the freeze-dried composition can be any suitable carrier.
  • compositions can also be prepared by combining a composition of the present invention with a food.
  • a food used for administration is chilled, for instance, ice cream or milk.
  • Pharmaceutically compatible binding agents, and/or adjuvant materials can be included as part of the composition.
  • the tablets, pills, capsules, and the like can contain any of the following ingredients, or compounds of a similar nature: a binder such as microcrystalline cellulose, gum tragacanth or gelatin; an excipient such as starch or lactose, a disintegrating agent such as alginic acid, Primogel, or corn starch; a lubricant such as magnesium stearate or Sterotes; a glidant such as colloidal silicon dioxide; a sweetening agent such as sucrose or saccharin; or a flavoring agent such as peppermint, methyl salicylate, or orange flavoring.
  • a binder such as microcrystalline cellulose, gum tragacanth or gelatin
  • an excipient such as starch or lactose, a disintegrating agent such as alginic acid, Primogel, or corn starch
  • a lubricant such as magnesium stearate or Sterotes
  • a glidant such as colloidal silicon dioxide
  • a sweetening agent such
  • the freeze-dried composition can also be prepared in the form of suppositories (e.g., with conventional suppository bases such as cocoa butter and other glycerides) or retention enemas for rectal delivery.
  • suppositories e.g., with conventional suppository bases such as cocoa butter and other glycerides
  • retention enemas for rectal delivery.
  • the freeze-dried composition may be prepared with carriers that will protect the microbes against rapid elimination from the body, such as a controlled release
  • Biodegradable, biocompatible polymers can be used, such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen,
  • polyorthoesters and polylactic acid.
  • Such formulations can be prepared using standard techniques.
  • the materials can also be obtained commercially from, for instance, Alza Corporation and Nova Pharmaceuticals, Inc.
  • Liposomal suspensions can also be used as pharmaceutically acceptable carriers. These can be prepared according to methods known to those skilled in the art.
  • the freeze-dried composition may be present in a formulation that permits passage to the small intestine or colon.
  • the dosage form may be formulated so the composition is not exposed to conditions prevalent in the gastrointestinal tract before the small intestine or colon, e.g., high acidity and digestive enzymes present in the stomach and/or intestine.
  • the dosage form may be formulated so the composition passes through the stomach and is released in conditions that include a pH of greater than 5.5, greater than 6, greater than 6.5, or greater than 7.
  • a freeze-dried composition may be prepared with an enteric coating.
  • an enteric coating is acid- resistant to protect the composition from the low pH of the stomach and break down when exposed to a pH greater than present in the stomach.
  • Materials used for enteric coatings include fatty acids, waxes, shellac, plastics, and plant fibers. Examples include, but are not limited to, methyl acrylate-methacrylic acid copolymers, cellulose acetate succinate, hydroxy propyl methyl cellulose phthalate, enteric coatings (hydroxypropyl methylcellulose (HPMC), hydroxy propyl methyl cellulose acetate succinate
  • Encapsulation may include hard-shelled capsules, which may be used for dry, powdered ingredients, or soft-shelled capsules.
  • Capsules may be made from aqueous solutions of gelling agents such as animal protein (e.g., gelatin), plant polysaccharides or derivatives like carrageenans and modified forms of starch and cellulose.
  • Other ingredients may be added to a gelling agent solution such as plasticizers (e.g., glycerin and or sorbitol), coloring agents, preservatives, disintegrants, lubricants, and surface treatment.
  • Enteric coated capsules can be co-combined to provide for release of the freeze dried composition within the large bowel or colon.
  • a composition may be prepared by obtaining a fecal sample from an appropriate donor and blending with a diluent as described in Sadowsky et al.
  • a composition may be prepared by obtaining a fecal sample from an appropriate donor and using ballistic disruption with a horizontal or vertical shaker, a suitable diluent and stainless steel beads of 3.2 mm in diameter. The mixture is shaken to break up the sample and the beads are removed from the suspension by filtration a stainless steel strainer. The suspension is centrifuged at a suitable speed to pellet the microbes, for instance, 500-1,000 rpm, the supernatant poured off and the resulting microbial fraction obtained by selective filtration.
  • conditions for preparing a freeze-dried composition include the use of temperatures that decrease the replication of the microbes and eukaryotic cells.
  • the conditions used for preparation are maintained below 37°C.
  • the conditions used for preparation are maintained at a temperature of no greater than 30°C, no greater than 20°C, no greater than 10°C, or no greater than 5°C.
  • conditions are used such that replication of the microbes and eukaryotic cells is undetectable, and preferably does not occur.
  • the biological material present in a composition includes a population of microbes, eukaryotic cells, and viruses that is essentially identical to a population of microbes, eukaryotic cells, and viruses present in the colon or feces of a normal healthy human, e.g., the donor from whom the fecal sample was obtained.
  • the conditions used for preparation decrease exposure of the microbes and eukaryotic cells to oxygen, both before and after purification of microbiota.
  • Removal of non-living material may be achieved by passing the blended sample through a sieve with a sieve size of no greater than 2.0 mm, no greater than 1.0 mm, no greater than 0.5 mm, no greater than 0.25 mm, no greater than 0.212 mm, no greater than 0.180 mm, no greater than 0.150 mm, no greater than 0.125 mm, no greater than 0.106 mm, no greater than 0.090 mm, no greater than 0.075 mm, no greater than 0.063 mm, no greater than 0.053 mm, no greater than 0.045 mm, no greater than 0.038 mm, no greater than 0.032 mm, no greater than 0.025 mm, no greater than 0.020 mm, no greater than 0.01 mm, or no greater than 0.2 mm.
  • the blended sample is prepared by passing it through a sieve with a sieve size of 0.25 mm and collecting the filtrate.
  • the blended sample is passed through sieves with progressively smaller sieve sizes until final passage through a sieve size of 0.25 mm. For instance, if a total of four sieves are used the sieve size of the first sieve may be 2 mm, followed by 1 mm, followed by 0.5 mm, and then followed by 0.25 mm.
  • the final filtrate may be collected in a centrifuge tube, washed, and the cells resuspened in an aqueous solution (e.g., diluent, cryoprotectant, and the like, or a combination thereof).
  • the volume of the blended mixture is decreased through the steps of sieving and washing to result in at least 1 x 10 12 cells, for instance, at least 1.5 x 10 12 cells, at least 2 x 10 12 cells, or at least 2.5 x 10 12 cells in a volume that is subsequently freeze-dried. Since most biological material is difficult or impossible to culture, a hemocytometer may be used to determine the number of cells. This process results in an extract of feces that is highly enriched for all colon microbiota that are able to pass through a sieve as described above, and can be centrifuged at ⁇ , ⁇ g for 10 minutes.
  • enriched refers to increasing the abundance of biological material relative to non-living material, such that biological material constitutes a significantly higher proportion compared to the fecal material before the enrichment.
  • enriched refers to those situations in which a person has intervened to elevate the proportion of biological material.
  • odorless means there is a decreased amount of volatile organic molecules present, and the decreased amount of volatile organic molecules present can be easily detected by a person comparing the material before processing with the material after processing.
  • a composition described herein of fecal microbes is freeze- dried to form solid dried powder.
  • a composition of fecal microbes is mixed with a cryoprotectant and subjected to conditions that result in freeze-drying. Such conditions typically include freezing the sample, and reducing the pressure surrounding the frozen sample to remove water from the sample.
  • the composition may be further processed by subjecting the dried material to force sufficient to break up the material into a powder that can be easily stored until used.
  • the powder may be used to form granules, compressed tablets, pills, capsules, wafers, and the like.
  • the freeze dried material can be formulated such that it is released from a capsule into the small or large intestine or the colon, and not the stomach.
  • the present invention is further directed to methods of using the freeze-dried compositions described herein.
  • One method includes administering to a subject in need thereof an effective amount of a composition described herein.
  • the administering is under conditions suitable for deposition of the composition in a region of the large or small intestine such that the biological material in the composition colonizes the colon.
  • administration may be into upper gastrointestinal tract, as well as lower gastrointestinal tract, e.g., the terminal ileum, cecum, colonic areas containing
  • the administering may be oral, such as by tablet.
  • the administering may be by intubation, such as by nasogastric tube, of a freeze-dried composition that has been reconstituted.
  • the administering may be rectal, for instance by a colonoscope, enema, or suppository.
  • an "effective amount” relates to a sufficient amount of a composition described herein, to provide the desired effect.
  • an "effective amount” is an amount effective to alleviate one or more symptoms and/or signs of the disease as described herein.
  • an effective amount is an amount that is sufficient to effect a reduction in a symptom and/or sign associated with a disease, such as diarrhea or C. difficile.
  • a reduction in a symptom and/or a sign is, for instance, at least 10%, at least 20%, at least 30%>, at least
  • an effective amount is an amount sufficient to result in at least 1 x 10 12 , at least 1.5 x 10 12 , at least 2 x 10 12 , or at least 2.5 x 10 12 cells administered to the subject. In one embodiment, an effective amount is an amount sufficient to result in at least 1 x 10 12 , at least 1.5 x 10 12 , at least 2 x 10 12 , or at least 2.5 x 10 12 cells delivered to the colon.
  • an effective amount is an amount sufficient to result in 1 x 10 12 to 3 x 10 12 cells delivered to the colon, or 1.5 x 10 12 to 2.5 x 10 12 cells delivered to the colon. It will be understood, however, that the total dosage of the compositions as disclosed herein will be decided by the attending physician within the scope of sound medical judgment. The exact amount required will vary depending on factors such as the type and extent of disease being treated.
  • a method of the present invention includes treating certain diseases in a subject in need of treatment.
  • the subject may be a mammal, such as a human.
  • animal models may be used, such as a mammal, including a rat, a mouse, a hamster, a gerbil, or a primate.
  • the term "disease” refers to any deviation from or interruption of the normal structure or function of a part, organ, or system, or combination thereof, of a subject that is manifested by a characteristic symptom or clinical sign. Diseases include those characterized by dysfunctional composition of colon microbiota.
  • Such diseases include, but are not limited to, colitis, including autoimmune colitis (e.g., inflammatory bowel disease, ulcerative colitis, Crohn's disease, irritable bowel syndrome) and infectious colitis.
  • infectious colitis include, but are not limited to Clostridium difficile colitis (e.g., acute C. difficile colitis, relapsing C. difficile colitis, or severe C. difficile colitis) and enterohemorrhagic colitis (e.g., a colitis caused by Shigella spp. or E. coli).
  • diseases include, but are not limited to, chronic diarrhea; chronic constipation, metabolic syndrome and obesity, atopic diseases including asthma, eczema, eosinophilic disorders of the GI tract, systemic autoimmunity including rheumatoid arthritis, systemic lupus erythematosis, multiple sclerosis, etc., chronic pain disorders such fibromyalgia, chronic fatigue syndrome, neurodegenerative disorders, eating disorders, and malnutrition.
  • symptom refers to subjective evidence of disease or condition experienced by the patient and caused by disease.
  • clinical sign or simply “sign,” refers to objective evidence of a disease present in a subject. Symptoms and/or signs associated with diseases referred to herein and the evaluation of such signs are routine and known in the art. Typically, whether a subject has a disease, and whether a subject is responding to treatment, may be determined by evaluation of signs associated with the disease.
  • Treatment of a disease can be prophylactic or, alternatively, can be initiated after the development of a disease.
  • Treatment that is prophylactic, for instance, initiated before a subject manifests signs of a disease is referred to herein as treatment of a subject that is "at risk" of developing a disease.
  • An example of a subject that is at risk of developing a disease is a person having a risk factor.
  • An example of a risk factor for Clostridium difficile colitis is antibiotic therapy of the gastrointestinal tract. Treatment can be performed before, during, or after the occurrence of the diseases described herein.
  • a method of the present invention includes transplanting a microbiota from a donor to a recipient.
  • a method of the present invention includes increasing the relative abundance of members of the phylum Firmicutes, such as a non-pathogenic member of the class Clostridia, and/or members of the phylum Bateriodetes, in a recipient's colon.
  • the phrase "relative abundance" refers to number of members of a phylum or class compared to the number of members of all other taxa in a recipient's colon. Such a comparison can be expressed as a percent.
  • the relative abundance of non-pathogenic members of the class Clostridia in a recipient's colon after the administration may be increased by at least 5%, at least 10%, at least 20%, or at least 50%, compared to the recipient's colon before the administration.
  • the relative abundance of members of the phylum Firmicutes in a recipient's colon after the administration may be increased by at least 5%, at least 10%, at least 20%, or at least 50% compared to the recipient's colon before the administration.
  • the change in the abundance may be determined at, for instance, 3 days, 10 days, 15 days, or 25 days after the administration of fecal microbiota.
  • a method of the present invention includes decreasing the relative abundance of members of the phylum Proteobacteria in a recipient's colon.
  • the relative abundance of members of the phylum Proteobacteria in a recipient's colon after the administration may be decreased by at least 10%, at least 20%, at least 30%, or at least 40% compared to the recipient's colon before the administration.
  • the change in the abundance of members of the phylum Proteobacteria may be determined at, for instance, 3 days, 10 days, 15 days, or 25 days after the administration.
  • the existing microbiota does not need to be cleared prior to administration of a freeze-dried composition of the present invention.
  • clearance of the microbiota may be necessary. Methods for clearance of existing microbiota are known and routine. In one example, clearance can be
  • a cocktail of antibiotics for one week until a day prior to transplant.
  • An example of a useful cocktail is Metronidazole (1000 mg twice daily), Rifaximin (550 mg twice daily), Vancomycin (500 mg twice daily), and Neomycin (1000 mg twice daily).
  • Metronidazole 1000 mg twice daily
  • Rifaximin 550 mg twice daily
  • Vancomycin 500 mg twice daily
  • Neomycin 1000 mg twice daily
  • the processing equipment container (that was eventually used to place blenders and sieves that come into contact with feces) was filled with a 10% bleach solution.
  • the donor fecal material was removed from a transport container.
  • Donor fecal material was transferred to a sterile weighing container using a sterile tongue depressor. Up to three multiples of 50 grams of fecal material were weighed for a final weight of 150 grams and transferred to a sterile blending chamber.
  • 250 ml of sterile phosphate buffered saline (PBS) was added for every 50 grams of fecal material, up to a total of 750 ml.
  • PBS sterile phosphate buffered saline
  • a nitrogen gas hose was attached to the tubing connector on the blender lid.
  • An outflow tube was attached to the other connection on the blender lid, and the outflow hose was attached to a vacuum flask.
  • the gas valve on the nitrogen tank was opened and set to a flow rate of 1.5 liters/min and the blending chamber purged for 3 minutes.
  • the nitrogen gas valve was left open while blending.
  • the sample was blended three times, each for 20 seconds, with a 10 second pause between blending.
  • a sterilized sieve stack containing 2.0, 1.0, 0.5, and 0.25 mm sieves was assembled from top to bottom, respectively.
  • the collection pan was at the bottom.
  • the blended fecal slurry was poured on the 2.0 mm sieve at the top of stack. As much of the sieve surface area as possible was covered to prevent clogging.
  • the material was allowed to pass through the 2.0 mm sieve, and the filter stack was tilted as necessary to allow material to run through the sieve.
  • the 2.0 mm sieve was removed from the filter stack and held over the stack at an angle to allow material on the bottom of the sieve to run onto the sieve stack.
  • the material was allowed to pass through the 1.0 mm sieve, and the filter stack was tilted as necessary to allow material to run through the sieve.
  • the 1.0 mm sieve was removed from the filter stack and held over the stack at an angle to allow material on the bottom of the sieve to run onto the sieve stack. The same procedure was followed for the 0.5 mm sieve and the 0.25 mm sieve, and the final material, the intermediate fecal slurry, collected in the collection pan.
  • the intermediate fecal slurry from the collection pan was transferred to a sterile 250 ml centrifuge bottle using sterile conditions. The approximate volume in each bottle was recorded, and centrifuged for 15 minutes at 4500 rpm in GSA rotor at 4°C.
  • sterile PBS Approximately one third of the original volume sterile PBS was added to each centrifuge bottle and the pellet resuspended. The washed material was combined into a single centrifuge bottle and mixed to ensure the slurry was well mixed. The slurry was transferred to a sterile 250 ml graduated cylinder, and transfer of large unsuspended particles from the centrifuge bottle to the cylinder was avoided. The volume of the slurry was recorded and transferred into a new 250ml centrifuge bottle.
  • fecal bacteria slurry was removed from the centrifuge bottle and transferred to a microfuge tube for microscopy and total protein assay, and stored in a refrigerator. Total protein measured by boiling preparations followed by the BCA assay. Two sets of serial dilutions of the fecal bacterial slurry were prepared in sterile saline (0.85% NaCl). The sample was diluted to 1 : 10, 1 : 100, 1 : 1000, and 1 : 10000, and a microbial cell count performed using a hemocytometer.
  • the volume of the intermediate fecal slurry necessary to provide 2.5xl0 12 total microbes was calculated and the appropriate volume of the intermediate fecal slurry was transferred into a 100 ml bottle. An equal volume of 20% mannitol was added, to give a final mannitol concentration of 10%. After mixing, the suspension was transferred to a freeze drying jar which was then covered. A dry ice / ethanol bath was used to rapidly freeze the sample while holding the freeze drying jar at an angle and rotating it by hand in the dry ice / ethanol bath so that the fecal bacteria suspension froze in a thin layer on the sides of the jar.
  • the frozen suspension in the jar was stored at -80°C until starting the freeze drying process.
  • a freeze dryer was used for 24-48 hours to completely dry the material. After the material was completely dry, the freeze drying jar was removed from the freeze drying machine and transferred to a BSL-2 safety hood to protect from aerosolization of the freeze dried material.
  • a sterile scoop was used to scrape the freeze dried material off the walls of the jar and break up large chunks of the material.
  • the freeze dried material was transferred to a disinfected bone grinder cup, and ground with three 10 second pulses. The resulting material was a fine powder.
  • the powdered material was transferred to a 100ml storage container and stored at -80°C until use.
  • Example 2 Clinical case histories of two patients treated with fecal microbiota prepared on the same day in liquid frozen form and freeze-dried form.
  • Fecal Microbiota Tranpslantation The patient continued on vancomycin until 2 days before the procedure. One day before the procedure she took Moviprep to cleanse her colon and wash out any residual antibiotics.
  • One unit of donor fecal microbiota (2.5 x 10 12 bacteria), frozen in 10% glycerin, was thawed in an ice bath for two hours and resuspended in room temperature non-bacteristatic, sterile normal saline to 240 mL. This material was injected via the biopsy port of a colonoscope in the cecum. Colonoscopy was otherwise unremarkable.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Medicinal Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Epidemiology (AREA)
  • Animal Behavior & Ethology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Microbiology (AREA)
  • Biotechnology (AREA)
  • Zoology (AREA)
  • Virology (AREA)
  • Mycology (AREA)
  • Cell Biology (AREA)
  • Organic Chemistry (AREA)
  • Wood Science & Technology (AREA)
  • Biomedical Technology (AREA)
  • Genetics & Genomics (AREA)
  • Developmental Biology & Embryology (AREA)
  • Immunology (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Molecular Biology (AREA)
  • Physiology (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • Nutrition Science (AREA)
  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)

Abstract

La présente invention concerne des compositions lyophilisées qui comprennent un extrait de matières fécales humaines et un cryoprotecteur, et des méthodes de fabrication et d'utilisation desdites compositions, notamment des méthodes qui permettent de remplacer, compléter ou modifier le microbiote du côlon d'un patient, ainsi que des méthodes pour traiter une maladie, un état pathologique et/ou un état iatrogène du côlon.
PCT/US2014/027391 2011-03-09 2014-03-14 Microbiote fécal lyophilisé à utiliser dans la transplantation microbienne fécale WO2014152484A1 (fr)

Priority Applications (2)

Application Number Priority Date Filing Date Title
US14/850,318 US20150374761A1 (en) 2011-03-09 2015-09-10 Freeze dried fecal microbiota for use in fecal microbial transplantation
US15/837,834 US20180110810A1 (en) 2011-03-09 2017-12-11 Freeze dried fecal microbiota for use in fecal microbial transplantation

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US201361782222P 2013-03-14 2013-03-14
US61/782,222 2013-03-14

Related Parent Applications (2)

Application Number Title Priority Date Filing Date
PCT/US2012/028484 Continuation-In-Part WO2012122478A1 (fr) 2011-03-09 2012-03-09 Compositions et procédés de transplantation du microbiote du côlon
US14/003,411 Continuation-In-Part US9968638B2 (en) 2011-03-09 2012-03-09 Compositions and methods for transplantation of colon microbiota

Related Child Applications (1)

Application Number Title Priority Date Filing Date
US14/850,318 Continuation-In-Part US20150374761A1 (en) 2011-03-09 2015-09-10 Freeze dried fecal microbiota for use in fecal microbial transplantation

Publications (1)

Publication Number Publication Date
WO2014152484A1 true WO2014152484A1 (fr) 2014-09-25

Family

ID=51581222

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2014/027391 WO2014152484A1 (fr) 2011-03-09 2014-03-14 Microbiote fécal lyophilisé à utiliser dans la transplantation microbienne fécale

Country Status (1)

Country Link
WO (1) WO2014152484A1 (fr)

Cited By (39)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20150216917A1 (en) * 2013-06-05 2015-08-06 Rebiotix, Inc. Microbiota restoration therapy (mrt), compositions and methods of manufacture
CN105662921A (zh) * 2015-12-25 2016-06-15 吉林农业大学 一种冷冻干燥油体的制备方法
WO2016191356A1 (fr) * 2015-05-22 2016-12-01 Arizona Board Of Regents On Behalf Of Arizona State University Procédés pour le traitement d'un trouble du spectre de l'autisme et de symptômes associés
WO2016201114A1 (fr) * 2015-06-09 2016-12-15 Rebiotix, Inc. Compositions pour thérapie de restauration du microbiote (mrt) et procédés de fabrication
WO2017075098A1 (fr) 2015-10-26 2017-05-04 Crestovo Llc Compositions et procédés de thérapie associée au microbiote fécal
WO2017103550A1 (fr) * 2015-12-18 2017-06-22 Maat Pharma Procede de lyophilisation d'un echantillon de microbiote fecal
WO2017103225A1 (fr) * 2015-12-18 2017-06-22 Institut National De La Recherche Agronomique Composition lyophilisee pour la conservation de microbiote dans son ecosysteme
CN107530280A (zh) * 2015-04-24 2018-01-02 玛阿特制药公司 粪便微生物群样品的制备方法
WO2018006088A1 (fr) * 2016-07-01 2018-01-04 Regents Of The University Of Minnesota Compositions et procédés de traitement de c. difficile
WO2018026913A1 (fr) * 2016-08-03 2018-02-08 Crestovo Llc Méthodes de traitement de la rectocolite hémorragique (rch)
WO2018046485A1 (fr) 2016-09-09 2018-03-15 Swecure Ab Utilisation de butyricimonas et/ou d'oxalobacter pour traiter le syndrome de l'intestin irritable
WO2018071536A1 (fr) 2016-10-11 2018-04-19 Crestovo Holdings Llc Compositions et procédés pour traiter la cholangite sclérosante primitive et des troubles associés
WO2018071532A1 (fr) 2016-10-11 2018-04-19 Crestovo Holdings Llc Compositions et méthodes permettant de traiter le syndrome de fatigue chronique et les troubles associés
WO2018071534A1 (fr) 2016-10-11 2018-04-19 Crestovo Holdings Llc Compositions et méthodes permettant de traiter le syndrome du côlon irritable et les troubles associés
WO2018071537A1 (fr) 2016-10-11 2018-04-19 Crestovo Holdings Llc Compositions et méthodes pour traiter la sclérose en plaques et des troubles associés
US9962413B2 (en) 2010-08-04 2018-05-08 Crestovo Holdings Llc Compositions for fecal floral transplantation and methods for making and using them and devices for delivering them
RU2661624C1 (ru) * 2017-09-28 2018-07-17 Федеральное государственное бюджетное учреждение науки Институт химической биологии и фундаментальной медицины Сибирского отделения Российской академии наук (ИХБФМ СО РАН) Способ лечения синдрома раздражённого кишечника
WO2018187464A1 (fr) 2017-04-05 2018-10-11 Crestovo Holdings Llc Compositions et méthodes pour traiter la diverticulite et des troubles associés
WO2018187467A1 (fr) 2017-04-05 2018-10-11 Crestovo Holdings Llc Compositions et méthodes pour traiter la maladie de parkinson (pd) et des troubles associés
EP3294307A4 (fr) * 2015-05-14 2019-01-23 Crestovo Holdings Llc Compositions permettant la transplantation de flore fécale et méthodes de préparation et d'utilisation de ces dernières, et dispositifs pour leur administration
WO2019051380A1 (fr) * 2017-09-08 2019-03-14 Evelo Biosciences, Inc. Vésicules extracellulaires (ev) bactériennes
US10251914B2 (en) 2011-03-09 2019-04-09 Regents Of The University Of Minnesota Compositions and methods for transplantation of colon microbiota
US10369175B2 (en) 2000-07-25 2019-08-06 Crestovo Holdings Llc Probiotic recolonisation therapy
WO2020069280A1 (fr) 2018-09-27 2020-04-02 Crestovo Holdings Llc Compositions et méthodes de traitement de l'épilepsie et de troubles apparentés
CN111534466A (zh) * 2020-05-14 2020-08-14 深圳市融思生物科技有限公司 一种人类粪便肠道菌群样本的处理方法及处理试剂
US10799539B2 (en) 2015-06-09 2020-10-13 Rebiotix, Inc. Microbiota restoration therapy (MRT) compositions and methods of manufacture
US10828340B2 (en) 2015-06-09 2020-11-10 Rebiotix, Inc. Microbiota restoration therapy (MRT) compositions and methods of manufacture
US10849936B2 (en) 2016-07-01 2020-12-01 Regents Of The University Of Minnesota Compositions and methods for C. difficile treatment
US10905726B2 (en) 2015-06-09 2021-02-02 Rebiotix, Inc. Microbiota restoration therapy (MRT) compositions and methods of manufacture
WO2021142358A1 (fr) 2020-01-10 2021-07-15 Finch Therapeutics Holdings Llc Compositions et méthodes pour le traitement de l'encéphalopathie hépatique (eh)
WO2021142347A1 (fr) 2020-01-10 2021-07-15 Finch Therapeutics Holdings Llc Compositions et procédés relatifs à la stéatohépatite non alcoolique (nash)
WO2021142353A1 (fr) 2020-01-10 2021-07-15 Finch Therapeutics Holdings Llc Compositions et méthodes pour le traitement de l'hépatite b (vhb) et de l'hépatite d (vhd)
US11166990B2 (en) 2018-07-13 2021-11-09 Finch Therapeutics Holdings Llc Methods and compositions for treating ulcerative colitis
US11242544B2 (en) * 2018-07-11 2022-02-08 Wisconsin Alumni Research Foundation Microbiomes and methods for producing medium-chain fatty acids from organic substrates
US11357801B2 (en) 2016-06-15 2022-06-14 Arizona Board Of Regents On Behalf Of Arizona State University Methods for treating autism spectrum disorder and associated symptoms
US11446337B2 (en) 2010-02-01 2022-09-20 Rebiotix, Inc. Bacteriotherapy for clostridium difficile colitis
US11542560B2 (en) 2012-05-25 2023-01-03 Board of Regents on Behalf of Arizona State University Microbiome markers and therapies for autism spectrum disorders
US11865145B2 (en) 2017-08-07 2024-01-09 Finch Therapeutics Holdings Llc Compositions and methods for maintaining and restoring a healthy gut barrier
US11890306B2 (en) 2017-05-26 2024-02-06 Finch Therapeutics Holdings Llc Lyophilized compositions comprising fecal microbe-based therapeutic agents and methods for making and using same

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20070059296A1 (en) * 2005-09-13 2007-03-15 Liang-Jung Chen Probiotic composition having acid-resistant enteric coating
WO2011033310A1 (fr) * 2009-09-17 2011-03-24 William John Martin Flore intestinale encapsulée extraite de fèces destinée à être utilisée dans le traitement de troubles gastro-intestinaux
US20120039853A1 (en) * 2009-04-30 2012-02-16 Actogenix Nv Cryoprotectants for freeze drying of lactic acid bacteria
WO2012122478A1 (fr) * 2011-03-09 2012-09-13 Regents Of The University Of Minnesota Compositions et procédés de transplantation du microbiote du côlon

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20070059296A1 (en) * 2005-09-13 2007-03-15 Liang-Jung Chen Probiotic composition having acid-resistant enteric coating
US20120039853A1 (en) * 2009-04-30 2012-02-16 Actogenix Nv Cryoprotectants for freeze drying of lactic acid bacteria
WO2011033310A1 (fr) * 2009-09-17 2011-03-24 William John Martin Flore intestinale encapsulée extraite de fèces destinée à être utilisée dans le traitement de troubles gastro-intestinaux
WO2012122478A1 (fr) * 2011-03-09 2012-09-13 Regents Of The University Of Minnesota Compositions et procédés de transplantation du microbiote du côlon

Cited By (116)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US10369175B2 (en) 2000-07-25 2019-08-06 Crestovo Holdings Llc Probiotic recolonisation therapy
US10772919B2 (en) 2000-07-25 2020-09-15 Crestovo Holdings Llc Probiotic recolonisation therapy
US11446337B2 (en) 2010-02-01 2022-09-20 Rebiotix, Inc. Bacteriotherapy for clostridium difficile colitis
US11986500B2 (en) 2010-02-01 2024-05-21 Rebiotix Inc Bacteriotherapy for clostridium difficile colitis
US11944654B2 (en) 2010-02-01 2024-04-02 Rebiotix, Inc. Bacteriotherapy for clostridium difficile colitis
US11065284B2 (en) 2010-08-04 2021-07-20 Finch Therapeutics Holdings Llc Compositions for fecal floral transplantation and methods for making and using them and devices for delivering them
US10987385B2 (en) 2010-08-04 2021-04-27 Finch Therapeutics Holdings Llc Compositions for fecal floral transplantation and methods for making and using them and devices for delivering them
US10610551B2 (en) 2010-08-04 2020-04-07 Crestovo Holdings, Inc. Compositions for fecal floral transplantation and methods for making and using them and devices for delivering them
US10617724B2 (en) 2010-08-04 2020-04-14 Crestovo Holdings Llc Compositions for fecal floral transplantation and methods for making and using them and devices for delivering them
US10463702B2 (en) 2010-08-04 2019-11-05 Crestovo Holdings Llc Compositions for fecal floral transplantation and methods for making and using them and devices for delivering them
US10675309B2 (en) 2010-08-04 2020-06-09 Crestovo Holdings Llc Compositions for fecal floral transplantation and methods for making and using them and devices for delivering them
US11541080B2 (en) 2010-08-04 2023-01-03 Finch Therapeutics Holdings Llc Compositions for fecal floral transplantation and methods for making and using them and devices for delivering them
US11504403B2 (en) 2010-08-04 2022-11-22 Finch Therapeutics Holdings Llc Compositions for fecal floral transplantation and methods for making and using them and devices for delivering them
US11491193B2 (en) 2010-08-04 2022-11-08 Finch Therapeutics Holdings Llc Compositions for fecal floral transplantation and methods for making and using them and devices for delivering them
US11890307B2 (en) 2010-08-04 2024-02-06 Finch Therapeutics Holdings Llc Compositions for fecal floral transplantation and methods for making and using them and devices for delivering them
US11207356B2 (en) 2010-08-04 2021-12-28 Finch Therapeutics Holdings Llc Compositions for fecal floral transplantation and methods for making and using them and devices for delivering them
US10328107B2 (en) 2010-08-04 2019-06-25 Crestovo Holdings Llc Compositions for fecal floral transplantation and methods for making and using them and devices for delivering them
US11173183B2 (en) 2010-08-04 2021-11-16 Finch Therapeutics Holdings Llc Compositions for fecal floral transplantation and methods for making and using them and devices for delivering them
US11129859B2 (en) 2010-08-04 2021-09-28 Finch Therapeutics Holdings Llc Compositions for fecal floral transplantation and methods for making and using them and devices for delivering them
US11890308B2 (en) 2010-08-04 2024-02-06 Finch Therapeutics Holdings Llc Compositions for fecal floral transplantation and methods for making and using them and devices for delivering them
US9962413B2 (en) 2010-08-04 2018-05-08 Crestovo Holdings Llc Compositions for fecal floral transplantation and methods for making and using them and devices for delivering them
US11103541B2 (en) 2010-08-04 2021-08-31 Finch Therapeutics Holdings Llc Compositions for fecal floral transplantation and methods for making and using them and devices for delivering them
US10278997B2 (en) 2010-08-04 2019-05-07 Crestovo Holdings Llc Compositions for fecal floral transplantation and methods for making and using them and devices for delivering them
US10849937B2 (en) 2010-08-04 2020-12-01 Crestovo Holdings Llc Compositions for fecal floral transplantation and methods for making and using them and devices for delivering them
US11850269B2 (en) 2010-08-04 2023-12-26 Finch Therapeutics Holdings Llc Compositions for fecal floral transplantation and methods for making and using them and devices for delivering them
US10064899B1 (en) 2010-08-04 2018-09-04 Crestovo Holdings Llc Compositions for fecal floral transplantation and methods for making and using them and devices for delivering them
US10857188B2 (en) 2010-08-04 2020-12-08 Crestovo Holdings Llc Compositions for fecal floral transplantation and methods for making and using them and devices for delivering them
US10251914B2 (en) 2011-03-09 2019-04-09 Regents Of The University Of Minnesota Compositions and methods for transplantation of colon microbiota
US11801269B2 (en) 2011-03-09 2023-10-31 Regents Of The University Of Minnesota Compositions and methods for transplantation of colon microbiota
US10286011B2 (en) 2011-03-09 2019-05-14 Regents Of The University Of Minnesota Compositions and methods for transplantation of colon microbiota
US10286012B2 (en) 2011-03-09 2019-05-14 Regents Of The University Of Minnesota Compositions and methods for transplantation of colon microbiota
US11542560B2 (en) 2012-05-25 2023-01-03 Board of Regents on Behalf of Arizona State University Microbiome markers and therapies for autism spectrum disorders
US20150216917A1 (en) * 2013-06-05 2015-08-06 Rebiotix, Inc. Microbiota restoration therapy (mrt), compositions and methods of manufacture
US10603341B2 (en) * 2013-06-05 2020-03-31 Rebiotix, Inc. Microbiota restoration therapy (MRT), compositions and methods of manufacture
US11554143B2 (en) 2013-06-05 2023-01-17 Rebiotix, Inc. Microbiota restoration therapy (MRT), compositions and methods of manufacture
CN107530280A (zh) * 2015-04-24 2018-01-02 玛阿特制药公司 粪便微生物群样品的制备方法
US10980839B2 (en) * 2015-04-24 2021-04-20 Maat Pharma Method of preparing a faecal microbiota sample
US10821138B2 (en) 2015-05-14 2020-11-03 Crestovo Holdings Llc Compositions for fecal floral transplantation and methods for making and using them and device for delivering them
AU2016262615B2 (en) * 2015-05-14 2021-01-07 Finch Therapeutics Holdings Llc Compositions for fecal floral transplantation and methods for making and using them and devices for delivering them
AU2016262615C1 (en) * 2015-05-14 2021-06-10 Finch Therapeutics Holdings Llc Compositions for fecal floral transplantation and methods for making and using them and devices for delivering them
US11123377B2 (en) 2015-05-14 2021-09-21 Finch Therapeutics Holdings Llc Compositions for fecal floral transplantation and methods for making and using them and device for delivering them
EP3294307A4 (fr) * 2015-05-14 2019-01-23 Crestovo Holdings Llc Compositions permettant la transplantation de flore fécale et méthodes de préparation et d'utilisation de ces dernières, et dispositifs pour leur administration
US11202808B2 (en) 2015-05-22 2021-12-21 Arizona Board Of Regents On Behalf Of Arizona State University Methods for treating autism spectrum disorder and associated symptoms
WO2016191356A1 (fr) * 2015-05-22 2016-12-01 Arizona Board Of Regents On Behalf Of Arizona State University Procédés pour le traitement d'un trouble du spectre de l'autisme et de symptômes associés
US10828340B2 (en) 2015-06-09 2020-11-10 Rebiotix, Inc. Microbiota restoration therapy (MRT) compositions and methods of manufacture
AU2016274757B2 (en) * 2015-06-09 2019-02-07 Rebiotix, Inc. Microbiota restoration therapy (MRT) compositions and methods of manufacture
AU2020220084B2 (en) * 2015-06-09 2022-07-21 Rebiotix, Inc. Microbiota restoration therapy (MRT) compositions and methods of manufacture
EP3939601A1 (fr) * 2015-06-09 2022-01-19 Rebiotix, Inc. Compositions de thérapie de restauration du microbiote (mrt) et procédés de fabrication
KR20200006181A (ko) * 2015-06-09 2020-01-17 리바이오틱스, 인코퍼레이티드 미생물상 복원 치료(ｍｒｔ) 조성물 및 제조 방법
AU2019203164B2 (en) * 2015-06-09 2020-05-21 Rebiotix, Inc. Microbiota restoration therapy (MRT) compositions and methods of manufacture
US10391064B2 (en) 2015-06-09 2019-08-27 Rebiotix, Inc. Microbiota restoration therapy (MRT) compositions and methods of manufacture
CN107921072A (zh) * 2015-06-09 2018-04-17 雷柏奥提斯有限公司 微生物群恢复治疗（mrt）组合物和制造方法
WO2016201114A1 (fr) * 2015-06-09 2016-12-15 Rebiotix, Inc. Compositions pour thérapie de restauration du microbiote (mrt) et procédés de fabrication
KR102342925B1 (ko) * 2015-06-09 2021-12-23 리바이오틱스, 인코퍼레이티드 미생물상 복원 치료(ｍｒｔ) 조성물 및 제조 방법
US10799539B2 (en) 2015-06-09 2020-10-13 Rebiotix, Inc. Microbiota restoration therapy (MRT) compositions and methods of manufacture
US11654164B2 (en) 2015-06-09 2023-05-23 Rebiotix, Inc. Microbiota restoration therapy (MRT) compositions and methods of manufacture
KR102175829B1 (ko) * 2015-06-09 2020-11-06 리바이오틱스, 인코퍼레이티드 미생물상 복원 치료(ｍｒｔ) 조성물 및 제조 방법
KR20200127060A (ko) * 2015-06-09 2020-11-09 리바이오틱스, 인코퍼레이티드 미생물상 복원 치료(ｍｒｔ) 조성물 및 제조 방법
US10226431B2 (en) 2015-06-09 2019-03-12 Rebiotix, Inc. Microbiota restoration therapy (MRT) compositions and methods of manufacture
KR20180016539A (ko) * 2015-06-09 2018-02-14 리바이오틱스, 인코퍼레이티드 미생물상 복원 치료(mrt) 조성물 및 제조 방법
IL281424B1 (en) * 2015-06-09 2023-06-01 Rebiotix Inc Microbiota restoration treatment preparations and production methods
IL256033A (en) * 2015-06-09 2018-01-31 Rebiotix Inc Microbiotic reconstruction treatment preparations and methods of manufacture
JP2020037573A (ja) * 2015-06-09 2020-03-12 レビオティクス インコーポレイテッドＲｅｂｉｏｔｉｘ，Ｉｎｃ． マイクロバイオータ回復治療（ｍｒｔ）組成物
US10905726B2 (en) 2015-06-09 2021-02-02 Rebiotix, Inc. Microbiota restoration therapy (MRT) compositions and methods of manufacture
JP2018516960A (ja) * 2015-06-09 2018-06-28 レビオティクス インコーポレイテッドＲｅｂｉｏｔｉｘ，Ｉｎｃ． マイクロバイオータ回復治療（ｍｒｔ）組成物及び製造方法
US11642381B2 (en) 2015-06-09 2023-05-09 Rebiotix, Inc. Microbiota restoration therapy (MRT) compositions and methods of manufacture
WO2017075098A1 (fr) 2015-10-26 2017-05-04 Crestovo Llc Compositions et procédés de thérapie associée au microbiote fécal
FR3045383A1 (fr) * 2015-12-18 2017-06-23 Maat Pharma Procede de lyophilisation d'un echantillon de microbiote fecal
FR3045384A1 (fr) * 2015-12-18 2017-06-23 Agronomique Inst Nat Rech Composition lyophilisee pour la conservation de microbiote dans son ecosysteme
US10736849B2 (en) 2015-12-18 2020-08-11 Maat Pharma Method of lyophilization of a sample of faecal microbiota
AU2016370600B2 (en) * 2015-12-18 2021-07-15 Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement Method of lyophilization of a sample of faecal microbiota
IL260017B2 (en) * 2015-12-18 2023-06-01 Institut National De Rech Pour Lagriculture Lalimentation Et Lenvironnement The lyophilized composition for preserving microbiota in the microbiota ecosystem
JP2018537504A (ja) * 2015-12-18 2018-12-20 アンスティトゥー ナショナル ドゥ ラ ルシェルシュ アグロノミーク 生態系のままの微生物叢を保存するために凍結乾燥された組成物
CN108430481A (zh) * 2015-12-18 2018-08-21 玛阿特制药公司 冻干粪便微生物群样品的方法
KR102341203B1 (ko) 2015-12-18 2021-12-20 매트 파마 분변 미생물총의 샘플의 동결건조 방법
CN108430481B (zh) * 2015-12-18 2021-12-28 玛阿特制药公司 冻干粪便微生物群样品的方法
WO2017103225A1 (fr) * 2015-12-18 2017-06-22 Institut National De La Recherche Agronomique Composition lyophilisee pour la conservation de microbiote dans son ecosysteme
US11110133B2 (en) 2015-12-18 2021-09-07 Institut National De Recherche Pour L'agriculture, L'alimentation Et L'environnement Lyophilized composition for preserving microbiota in its ecosystem
KR20180086506A (ko) * 2015-12-18 2018-07-31 매트 파마 분변 미생물총의 샘플의 동결건조 방법
WO2017103550A1 (fr) * 2015-12-18 2017-06-22 Maat Pharma Procede de lyophilisation d'un echantillon de microbiote fecal
CN105662921A (zh) * 2015-12-25 2016-06-15 吉林农业大学 一种冷冻干燥油体的制备方法
US11357801B2 (en) 2016-06-15 2022-06-14 Arizona Board Of Regents On Behalf Of Arizona State University Methods for treating autism spectrum disorder and associated symptoms
WO2018006088A1 (fr) * 2016-07-01 2018-01-04 Regents Of The University Of Minnesota Compositions et procédés de traitement de c. difficile
US10849936B2 (en) 2016-07-01 2020-12-01 Regents Of The University Of Minnesota Compositions and methods for C. difficile treatment
CN109789172A (zh) * 2016-07-01 2019-05-21 明尼苏达大学董事会 用于难辨梭菌治疗的组合物和方法
US11819523B2 (en) 2016-07-01 2023-11-21 Regents Of The University Of Minnesota Compositions and methods for C. difficile treatment
WO2018026913A1 (fr) * 2016-08-03 2018-02-08 Crestovo Llc Méthodes de traitement de la rectocolite hémorragique (rch)
US11071759B2 (en) 2016-08-03 2021-07-27 Finch Therapeutics Holdings Llc Methods for treating ulcerative colitis
US10561690B2 (en) 2016-08-03 2020-02-18 Crestovo Holdings Llc Methods for treating ulcerative colitis
WO2018046485A1 (fr) 2016-09-09 2018-03-15 Swecure Ab Utilisation de butyricimonas et/ou d'oxalobacter pour traiter le syndrome de l'intestin irritable
WO2018071534A1 (fr) 2016-10-11 2018-04-19 Crestovo Holdings Llc Compositions et méthodes permettant de traiter le syndrome du côlon irritable et les troubles associés
US11213549B2 (en) 2016-10-11 2022-01-04 Finch Therapeutics Holdings Llc Compositions and method for treating primary sclerosing cholangitis and related disorders
WO2018071536A1 (fr) 2016-10-11 2018-04-19 Crestovo Holdings Llc Compositions et procédés pour traiter la cholangite sclérosante primitive et des troubles associés
WO2018071532A1 (fr) 2016-10-11 2018-04-19 Crestovo Holdings Llc Compositions et méthodes permettant de traiter le syndrome de fatigue chronique et les troubles associés
WO2018071537A1 (fr) 2016-10-11 2018-04-19 Crestovo Holdings Llc Compositions et méthodes pour traiter la sclérose en plaques et des troubles associés
US11026978B2 (en) 2016-10-11 2021-06-08 Finch Therapeutics Holdings Llc Compositions and methods for treating multiple sclerosis and related disorders
US11040073B2 (en) 2017-04-05 2021-06-22 Finch Therapeutics Holdings Llc Compositions and methods for treating diverticulitis and related disorders
US11433102B2 (en) 2017-04-05 2022-09-06 Finch Therapeutics Holdings Llc Compositions and methods for treating Parkinson's disease (PD) and related disorders
US11529375B2 (en) 2017-04-05 2022-12-20 Finch Therapeutics Holdings Llc Compositions and methods for treating diverticulitis and related disorders
WO2018187467A1 (fr) 2017-04-05 2018-10-11 Crestovo Holdings Llc Compositions et méthodes pour traiter la maladie de parkinson (pd) et des troubles associés
WO2018187464A1 (fr) 2017-04-05 2018-10-11 Crestovo Holdings Llc Compositions et méthodes pour traiter la diverticulite et des troubles associés
US11890306B2 (en) 2017-05-26 2024-02-06 Finch Therapeutics Holdings Llc Lyophilized compositions comprising fecal microbe-based therapeutic agents and methods for making and using same
US11865145B2 (en) 2017-08-07 2024-01-09 Finch Therapeutics Holdings Llc Compositions and methods for maintaining and restoring a healthy gut barrier
US11524037B2 (en) 2017-09-08 2022-12-13 Evelo Biosciences, Inc. Bacterial extracellular vesicles
TWI814739B (zh) * 2017-09-08 2023-09-11 美商艾弗洛生物科技股份有限公司 細菌胞外囊泡
WO2019051380A1 (fr) * 2017-09-08 2019-03-14 Evelo Biosciences, Inc. Vésicules extracellulaires (ev) bactériennes
RU2661624C1 (ru) * 2017-09-28 2018-07-17 Федеральное государственное бюджетное учреждение науки Институт химической биологии и фундаментальной медицины Сибирского отделения Российской академии наук (ИХБФМ СО РАН) Способ лечения синдрома раздражённого кишечника
US11242544B2 (en) * 2018-07-11 2022-02-08 Wisconsin Alumni Research Foundation Microbiomes and methods for producing medium-chain fatty acids from organic substrates
US11166990B2 (en) 2018-07-13 2021-11-09 Finch Therapeutics Holdings Llc Methods and compositions for treating ulcerative colitis
US11911419B2 (en) 2018-09-27 2024-02-27 Finch Therapeutics Holdings Llc Compositions and methods for treating epilepsy and related disorders
WO2020069280A1 (fr) 2018-09-27 2020-04-02 Crestovo Holdings Llc Compositions et méthodes de traitement de l'épilepsie et de troubles apparentés
WO2021142358A1 (fr) 2020-01-10 2021-07-15 Finch Therapeutics Holdings Llc Compositions et méthodes pour le traitement de l'encéphalopathie hépatique (eh)
WO2021142353A1 (fr) 2020-01-10 2021-07-15 Finch Therapeutics Holdings Llc Compositions et méthodes pour le traitement de l'hépatite b (vhb) et de l'hépatite d (vhd)
WO2021142347A1 (fr) 2020-01-10 2021-07-15 Finch Therapeutics Holdings Llc Compositions et procédés relatifs à la stéatohépatite non alcoolique (nash)
CN111534466B (zh) * 2020-05-14 2021-06-04 深圳市融思生物科技有限公司 一种人类粪便肠道菌群样本的处理方法及处理试剂
CN111534466A (zh) * 2020-05-14 2020-08-14 深圳市融思生物科技有限公司 一种人类粪便肠道菌群样本的处理方法及处理试剂

Similar Documents

Publication Publication Date Title
WO2014152484A1 (fr) Microbiote fécal lyophilisé à utiliser dans la transplantation microbienne fécale
AU2019219826B2 (en) Compositions and methods for transplantation of colon microbiota
US20180110810A1 (en) Freeze dried fecal microbiota for use in fecal microbial transplantation
JP6692897B2 (ja) 便フローラを移植するための組成物、並びにそれを調製及び使用する方法、並びにそれを送達するためのデバイス
CN114099553A (zh) 菌群恢复疗法（mrt）、组合物和制造方法
WO2018006088A1 (fr) Compositions et procédés de traitement de c. difficile

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 14771123

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 14771123

Country of ref document: EP

Kind code of ref document: A1