WO2021130181A1 - Novel faecal composition - Google Patents

Novel faecal composition Download PDF

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Publication number
WO2021130181A1
WO2021130181A1 PCT/EP2020/087491 EP2020087491W WO2021130181A1 WO 2021130181 A1 WO2021130181 A1 WO 2021130181A1 EP 2020087491 W EP2020087491 W EP 2020087491W WO 2021130181 A1 WO2021130181 A1 WO 2021130181A1
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Prior art keywords
faecal
cryoprotectant
faecal composition
less
stool
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PCT/EP2020/087491
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French (fr)
Inventor
Hengameh Chloe LAURIDSEN
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Lauridsen Hengameh Chloe
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Publication of WO2021130181A1 publication Critical patent/WO2021130181A1/en

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate
    • A61K9/4891—Coated capsules; Multilayered drug free capsule shells
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K10/00—Animal feeding-stuffs
    • A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
    • A23K10/16—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions
    • A23K10/18—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions of live microorganisms
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K10/00—Animal feeding-stuffs
    • A23K10/20—Animal feeding-stuffs from material of animal origin
    • A23K10/26—Animal feeding-stuffs from material of animal origin from waste material, e.g. feathers, bones or skin
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K20/00—Accessory food factors for animal feeding-stuffs
    • A23K20/10—Organic substances
    • A23K20/105—Aliphatic or alicyclic compounds
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K20/00—Accessory food factors for animal feeding-stuffs
    • A23K20/10—Organic substances
    • A23K20/163—Sugars; Polysaccharides
    • 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
    • A61K35/24—Mucus; Mucous glands; Bursa; Synovial fluid; Arthral fluid; Excreta; Spinal fluid
    • 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
    • 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
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
    • A61K47/08—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing oxygen, e.g. ethers, acetals, ketones, quinones, aldehydes, peroxides
    • A61K47/10—Alcohols; Phenols; Salts thereof, e.g. glycerol; Polyethylene glycols [PEG]; Poloxamers; PEG/POE alkyl ethers
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
    • A61K47/26—Carbohydrates, e.g. sugar alcohols, amino sugars, nucleic acids, mono-, di- or oligo-saccharides; Derivatives thereof, e.g. polysorbates, sorbitan fatty acid esters or glycyrrhizin
    • 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
    • A61K9/0012—Galenical forms characterised by the site of application
    • A61K9/0053—Mouth and digestive tract, i.e. intraoral and peroral administration
    • 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
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system

Definitions

  • the present invention relates to a faecal composition for gut microbiota transplantation.
  • the present invention relates to a faecal composition for gut microbiota transplantation wherein the side effects has been reduced.
  • the human body contains trillions of bacteria cells, protozoa and viruses, known as the microbiome.
  • the microbiome has been linked to many aspects of human health, from gastrointestinal related diseases to obesity.
  • GMT refers to the process of implanting of faecal suspension from a healthy donor with the right gut microbiota composition into the recipient's intestinal tract to restore the community and function of intestinal microbiota and/or intestinal functionality.
  • GMT Gut Microbiota Transplantation
  • FMT faecal microbiota transplantation
  • a faecal preparation from a carefully screened healthy stool-donor is transplanted into the colon of the patient.
  • routes of administration e.g., via colonoscopy, naso-enteric tube, each of which has unique risks and benefits.
  • Gut microbiome plays an important role in human and animal health by helping digestion and benefiting the immune system and many other aspects of health.
  • An imbalance of gut microbiota/dysbiosis might be restored by gut microbiota transplantation from a healthy gut , which may contribute to the treatment of various diseases, such as ulcerative colitis and other gastrointestinal conditions.
  • Jun Hu et al. describes the preparation of faecal compositions, where fresh feces are obtained from a healthy donor and diluted with sterile saline and homogenized in a standard blender. The slurry is then filtered three times through gauze, strainer, or 0.25 mm stainless steel sieves to eliminate the undigested and small particulate matter in the faecal suspension.
  • the faecal suspension may be centrifuged at 6,000 x g for 15 min.
  • the precipitate, without the supernatant, is re-suspended in fresh sterile saline.
  • the resulting suspension should be transferred to the recipients immediately if a fresh faecal composition is to be used to ensuring faecal microbial viability.
  • fresh stool samples should be diluted with sterile saline homogenized and filtered using the protocol used in the preparation of the fresh faecal material above. Subsequently, the resulting suspension should be added to glycerol (a cryoprotectant) to get a final concentration of 10%. Finally, the faecal suspensions are labeled accurately and then stored, as soon as possible, at -80°C to ensure the faecal microbial survival.
  • glycerol a cryoprotectant
  • the frozen faecal suspension should be thawed at and mixed with saline solution to obtain the required concentration and the infusion of faecal suspension should be implemented as soon as possible at room temperature.
  • the disadvantage of the prior art exemplified by Jun Hu et al. as described above, is that high amount of glycerol (cryoprotectant) is added to the faecal composition and that other beneficial products in the product, such as fungi, protozoa, viruses, vitamins and minerals produced by the healthy gut microbiota and the nutrition suitable for survival of the healthy gut microbiota in the diseased gut, are discharged together with the supernatant.
  • the process is performed under aerobic condition and using standard blender adds more air to the product, which means there are no-anaerobic bacteria in the final product.
  • cryoprotectant such as glycerol
  • glycerol has several disadvantages side effects like being harmful to the mucous membranes by increasing tissue osmolarity; or causing headaches, dizziness, bloating, nausea, vomiting, thirst, or diarrhoea, in particular the negative effects on the mucous membranes has been found to occur frequently and is undesirable.
  • an improved faecal composition would be advantageous, and in particular an efficient, a durable, uniform and viable faecal composition for GMT with no or reduced number of side effects would be advantageous.
  • an object of the present invention relates to a faecal composition for gut microbiota transplantation.
  • one aspect of the invention relates to a faecal composition
  • a faecal composition comprising a stool fraction and a diluent, wherein the stool fraction comprises one or more non-pathogenic microorganisms and the diluent comprises a cryoprotectant resulting in a final concentration of cryoprotectant in the faecal composition below 8% (v/v) cryoprotectant.
  • Another aspect of the present invention relates to a method for providing a faecal composition, the method comprises the steps of: (i) obtaining at least one sample of stool from a donor,
  • step (ii) placing said sample obtained in step (i) in a sterile, preferably an anaerobic sterile, collection device
  • step (iii) mixing (preferably gently mixing) the sample obtained in step (ii) with at least one diluent comprises a cryoprotectant resulting in a final concentration of cryoprotectant in the faecal composition below 8% (v/v) cryoprotectant
  • step (iv) filtering the mixture obtained in step (iii), providing the faecal composition comprising a stool fraction and a diluent, wherein the diluent comprises a cryoprotectant resulting in a final concentration of cryoprotectant in the faecal composition below 8% (v/v) cryoprotectant.
  • a further aspect of the present invention relates to a faecal composition according to the present invention for use as a medicament.
  • An even further aspect of the present invention relates to a faecal composition according to the present invention for use in the treatment and/or prophylaxis of intestinal imbalance, such as chronic Inflammatory Bowel Disease; irritable bowel syndrome; dysbiosis; Clostridium difficile infections; diarrhea; idiopathic constipation; celiac disease; chronic urinary tract infection; infertility; type II diabetes; food allergies; cancer; refractory GvHD; obesity and morbid obesity; autism, sclerosis; constipation; chronic urinary tract infection or vaginal infection (including cystitis, mycoses); bone and joint infections; Parkinson's disease; Alzheimer's disease; schizophrenia and bipolar disorders; and gut dysbiosis associated with anti-cancer chemotherapy or immunotherapy.
  • intestinal imbalance such as chronic Inflammatory Bowel Disease; irritable bowel syndrome; dysbiosis; Clostridium difficile infections; diarrhea; idiopathic constipation; celiac disease; chronic urinary tract infection; infertility; type II diabetes;
  • a faecal composition may be provided which has a significant reduction in the side effects and without compromising the efficiency, durability, uniformity and viability of the faecal composition.
  • a preferred embodiment of the present invention relates to a faecal composition
  • a faecal composition comprising a stool fraction and a diluent, wherein the stool fraction comprises one or more non-pathogenic microorganisms and the diluent comprises a cryoprotectant resulting in a final concentration of cryoprotectant in the faecal composition below 8% (v/v) cryoprotectant.
  • the faecal composition according to the present invention relates to a composition obtained from a human or animal donor, preferably, from a healthy human donor.
  • the term "donor” relates to a healthy human or animal-with the suitable gut microbiota delivering a faecal material.
  • the term "recipient” relates to a sick human or animal receiving the healthy gut microbiota composition, vitamins and mineral produced by healthy gut microbiota, antimicrobial peptides, bacteria, protozoa, viruses, nutrition necessary for the healthy gut bacteria to survive in the disease prepared from faecal material delivered.
  • the faecal material may be a healthy human or animal faeces or stool, preferably healthy human faeces or stool with the suitable gut microbiota.
  • tools and “faeces” may be used interchangeably.
  • the stool fraction may comprise one or more non-pathogenic microorganisms.
  • the one or more non-pathogenic microorganisms relates to one or more healthy gut bacteria, fungi, protozoa, viruses, present in the faecal material.
  • the faecal composition comprises vitamins and mineral produced by healthy gut microbiota, antimicrobial peptides, and nutrition necessary for the healthy gut bacteria to survive in a gut.
  • non-pathogenic microorganism relates to a microorganism that do not cause disease, harm or death to another organism, such as a human or an animal.
  • at least 50% of the non-pathogenic microorganisms in the faecal composition may be viable non-pathogenic microorganisms relative to the total number of non-pathogenic microorganisms in the faecal composition, such as at least 75%, e.g. at least 90%, such as at least 95%.
  • the faecal composition may comprise in the range of 10 4 -10 12 viable non-pathogenic facultative anaerobe, anaerobe and aerobe microorganisms, such as in the range of 10 4 -10 12 , e.g. in the range of 10 6 -10 10 , such as in the range of 10 8 -10 9 .
  • the viable non-pathogenic microorganisms may be viable non-pathogenic facultative anaerobe, anaerobe and aerobe microorganisms.
  • the faecal composition according to the present invention may be subjected to removal of undigested food. Furthermore, the faecal composition according to the present invention may preferably comprise vitamins and mineral produced by healthy gut microbiota, antimicrobial peptides, and nutrition necessary for the healthy gut bacteria to survive in a gut.
  • a 50-gram stool fraction results in 12-28 gram faecal composition after filtration from undigested food, depending on the diet during the donation time.
  • the faecal composition does not comprises antibiotics.
  • the faecal composition according to the present invention comprises no worms; no pathogenic parasites; and/or no pathogenic microorganisms.
  • the faecal composition may not comprise one or more of Clostridium difficile, ⁇ Clostridium Perfringens, Vibrio, Pleisiomonas, Aeromonas, enteric pathogens, including Salmonella, Shigella, ⁇ Campylobacter, Escherichia coli (0157 H7, Intimin-producing Escherichia coli, STEC, EPEC, ETEC, EIEC); Carbapenemase-producing enterobacteriae (CPE), Yersinia, ⁇ vancomycin-resistant enterococci (VRE); methicillin-resistant Staphylococcus aureus (MRSA); Helicobacter, Gram/negative multidrug-resistant bacteria; Enterobacteriaceae, such as Spectrum beta-lactamase (ESBL) producing Enterobacteriaceae, or carbapenem-resistant Enterobacteriaceae (CRE); Norovirus; Antigens and/or acid-fast staining for Giardia lamblia
  • the faecal composition may not comprises above mentioned Clostridium species, ⁇ enteric pathogens, including Salmonella, Shigella, ⁇ Campylobacter, above mentioned pathogenic Escherichia co//spp.; Yersinia, ⁇ vancomycin-resistant enterococci (VRE); methicillin-resistant Staphylococcus aureus (MRSA); Gram/negative multidrug- resistant bacteria; Enterobacteriaceae, such as Spectrum beta-lactamase (ESBL) producing Enterobacteriaceae, or carbapenem-resistant Enterobacteriaceae (CRE); Norovirus; Antigens and/or acid-fast staining for Giardia lamblia and Criptosporidium parvum;
  • VRE vancomycin-resistant enterococci
  • MRSA methicillin-resistant Staphylococcus aureus
  • MRSA methicillin-resistant Staphylococcus aureus
  • Gram/negative multidrug- resistant bacteria Enterobacteriace
  • Protozoa including Blastocystis hominis and helminths; and faecal occult blood, Entamoeba histolytica, Sapovirus, Rotavirus, Astrovirus, Adenovirus, Coronavirus, worms, worm eggs and cysts, Bacillus cereus.
  • the faecal composition may be a fresh faecal composition.
  • the faecal may be a fresh or a frozen faecal composition.
  • the faecal composition may comprise a diluent which is used to make the stool fraction more viscus.
  • the diluent may comprise an aqueous solution and/or a cryoprotectant.
  • the aqueous solution may comprise a salt, a mineral, and acid or an alkaline.
  • the aqueous solution comprises a salt, such as sodium chloride providing a saline solution.
  • the saline solution comprises sodium chloride, preferably, the concentration of sodium chloride in the saline solution may be 2-30 g/l; such as 4-20 g/l; e.g. 7-19 g/l; such as about 9 g/l sodium chloride (0.9%).
  • the aqueous solution may preferably be a sterile aqueous solution.
  • the diluent may be a saline containing diluent, thus resulting in a diluent that in addition to the cryoprotectant, preferably glycerol, comprises a saline solution.
  • cryoprotectant preferably glycerol
  • the faecal composition may be provided in dosage forms comprising 50 gram stool product and in the range of 50-150 ml saline comprising at most 10% (w/w) glycerol, such as at most 8% (w/w) glycerol, e.g. at most 5% (w/w) glycerol; such as at most 4% (w/w) glycerol, e.g. at most 3% (w/w) glycerol; such as at most 2% (w/w) glycerol, e.g.
  • glycerol at most 1% (w/w) glycerol, relative to the saline solution; such as in the range of 60-125 ml, e.g. in the range of 65-100 ml, such as in the range of 70-80 ml, e.g. about 75 ml.
  • the faecal composition according to the present invention may be provided in dosage forms consisting essentially of:
  • 50 gram stool product and in the range of 50-150 ml saline comprising at most 10% (w/w) glycerol, such as at most 8% (w/w) glycerol, e.g. at most 5% (w/w) glycerol; such as at most 4% (w/w) glycerol, e.g. at most 3% (w/w) glycerol; such as at most 2% (w/w) glycerol, e.g. at most 1% (w/w) glycerol, relative to the saline solution; such as in the range of 60-125 ml, e.g. in the range of 65-100 ml, such as in the range of 70- 80 ml, e.g. about 75 ml.
  • tool product relates to stool present in the faecal composition where undigested and small particulate material have been reduced or removed.
  • the diluent may be added to the stool in several sequences, such as addition in 2 sequences, e.g. addition in 3 sequences, such as addition in 4 sequences.
  • addition of diluent to the stool fraction may be done by an addition of a saline solution, preferably a sterile saline solution, to increase viscosity followed by addition of a cryoprotectant before freezing.
  • a saline solution preferably a sterile saline solution
  • the addition of saline solution may involve several additions of saline solutions.
  • the sterile cryoprotectant may be added to the stool fraction in combination with an sterile aqueous solution, such as a silane solution.
  • the diluent comprises a cryoprotectant resulting in a final concentration of cryoprotectant in the faecal composition below 8%
  • cryoprotectant such as 7% (v/v) cryoprotectant or below; e.g. 6% (v/v) cryoprotectant or below; such as 5% (v/v) cryoprotectant or below; e.g. 4% (v/v) cryoprotectant or below; such as 3% (v/v) cryoprotectant or below; e.g. 2% (v/v) cryoprotectant or below; such as 1% (v/v) cryoprotectant or below; e.g. 0.5% (v/v) cryoprotectant or below; such as 0.1% (v/v) cryoprotectant or below; e.g.
  • cryoprotectant or below such as 0.001% (v/v) cryoprotectant or below; e.g. 0.0001% (v/v) cryoprotectant or below; such as in the range of 0-8% (v/v) cryoprotectant; e.g. in the range of 0.0001-7% (v/v) cryoprotectant; such as in the range of 0.001-6% (v/v) cryoprotectant; e.g. in the range of 0.01-5% (v/v) cryoprotectant; such as in the range of 0.1-4% (v/v) cryoprotectant; e.g. in the range of 1-3% (v/v) cryoprotectant; such as in the range of 1.5-2.5% (v/v) cryoprotectant; e.g. in the range of 0.1-2% (v/v) cryoprotectant.
  • the content of the diluent may constitute less than 5 times the content of the stool fraction on a (w/w) basis; such as less than 4 times the content of the stool fraction on a (w/w) basis; e.g. less than 3 times content of the content of the stool fraction on a (w/w) basis; such as less than 2.5 times the content of the stool fraction on a (w/w) basis; e.g. less than 2 times content of the content of the stool fraction on a (w/w) basis; such as less than 1.8 times the content of the stool fraction on a (w/w) basis; e.g.
  • cryoprotectant may be selected from the group consisting of glycerol; mannitol; sorbitol; propylene glycol; ethylene glycol; trehalose and its analogues; saccharose; galactose-lactose and mixtures hereof.
  • the cryoprotectant may be glycerol.
  • the stool fraction and/or the faecal composition may comprise a particulate material as beneficial bacteria such as: Faecalbacterium, Prevotella bacteria, Lactobacilli species, Weissella spp., Bifidobacterium spp., Streptococcus thermophilus.
  • the stool fraction also contains antimicrobial peptides, healthy gut microbiota composition, short chain fatty acid, which serves as nutrition for host cell functionality (only produced by healthy gut microbiota), Vitamins and mineral produced by healthy gut microbiota, bacteria, protozoa, viruses, nutrition necessary for the healthy gut bacteria to survive in the disease.
  • the particulate material may comprise a particle size of the stool fraction and/or the faecal composition is below 0.4 mm; such as 0.3 mm or below; e.g. 0.25 mm or below; such as 0.20 mm or below.
  • the faecal composition may be a liquid faecal composition.
  • the faecal composition may be formulated into an enema formulation, a probe formulation or a colonoscopy formulation.
  • the stool fraction may be provided from a stool obtained from a mammal (donor).
  • the mammal may be a human; a dog; a cat; a horse; cattle; sheep; goat; a chicken; a duck; and pig.
  • the selection of a suitable and healthy donor may follow a very strict scheme for collecting and inspecting donor faecal sample to ensure quality of the stool.
  • the process of donor selection may be described below and may be demonstrated in the figure 1. Initially the donor should pass the preliminary donor health questionnaire regarding various health questions.
  • the donor stool may be obtained and tested. If approved (e.g. not comprising pathogenic microorganisms, antibiotics, presence of medicine, etc.) the donor may be accepted.
  • the donor stool may be placed in quarantine for 3 months at 5-8 degrees to investigate if the donor intestinal bacteria produce any toxins in the stool. If the donor is approved, the donor may be supplied with a stool collection kit, e.g. comprising a zip-bag for collecting the stool until processed.
  • a stool collection kit e.g. comprising a zip-bag for collecting the stool until processed.
  • the donor may deliver stool 3 times a week. Once the donor is accepted, they will be supplied with the collection kit and they must regularly donate up to 3 times a week.
  • the quality of the stool may be checked frequently, such as 4 times a year. E.g. once a year, such as each second month, e.g. each month, such as each 14 days, e.g. each week, e.g. every time a stool donation is made.
  • a preferred embodiment of the present invention relates to a method for providing a faecal composition, the method comprises the steps of:
  • step (ii) placing said sample obtained in step (i) in a sterile collection device
  • step (iii) mixing the sample obtained in step (ii) with at least one diluent comprises a cryoprotectant resulting in a final concentration of cryoprotectant in the faecal composition below 8% (v/v) cryoprotectant (iv) filtering the mixture obtained in step (iii), providing the faecal composition comprising a stool fraction and a diluent, wherein the diluent comprises a cryoprotectant resulting in a final concentration of cryoprotectant in the faecal composition below 8% (v/v) cryoprotectant.
  • the donor, healthy donor may be analyzed and determined to be an acceptable healthy donor as described herein.
  • the liquid phase obtained from separating the solid and liquid phase obtained from the filtering performed in step (iv) may be included as part of the faecal composition.
  • the faecal composition is not added a water immiscible compound.
  • the water immiscible compound is an oil compound.
  • Polyols like glycerol, may be a water miscible compound.
  • the faecal composition is not added a carbohydrate, an oligosaccharide or a polysaccharide compound.
  • the faecal composition according to the present invention may be used immediately or it may be frozen before use (in particular if not used immediately.
  • the faecal composition may be stored for at least 6 hours at a temperature of -80°C or less, such as for at least 12 hours, e.g. for at least 24 hours, such as for at least 3 days, e.g. for at least 5 days, such as for at least 7 days, e.g. for at least 10 days, such as for at least 15 days, e.g. for at least 25 days, such as for at least 1 month, e.g. for at least 3 months, such as for at least 6 months, e.g. for at least 9 months, such as for at least 12 months or 2 till 3 years.
  • the faecal composition may be immediately stored at a temperature of -80°C or less.
  • the faecal composition may be stored (preferably immediately) at a temperature of -80°C or less, such as a temperature of -100°C or less, e.g. a temperature of -120°C or less, such as a temperature of -150°C or less.
  • the process of freezing the faecal composition to a temperature of -100°C or less may be performed in a stepwise procedure comprising a first freezing step to about -80C followed by a second freezing step to a temperature of - 120°C or less, such as a temperature of -150°C or less.
  • the faecal composition may be stored at the final storage temperature for a period of at least 40 minutes, e.g. at least 1 hour, e.g. at least 24 hours, e.g. at least 1 week, e.g. at least 2 weeks, e.g. at least 1 month, e.g. at least 3 months, e.g. at least 5 months, e.g. at least 1 year, e.g. at least 2 years, e.g. at least 3 years, preferably about 1 year.
  • at least 40 minutes e.g. at least 1 hour, e.g. at least 24 hours, e.g. at least 1 week, e.g. at least 2 weeks, e.g. at least 1 month, e.g. at least 3 months, e.g. at least 5 months, e.g. at least 1 year, e.g. at least 2 years, e.g. at least 3 years, preferably about 1 year.
  • the stool desired for the purpose of the present invention may be characterised as a type 2-5 on the Bristol Stool Chart; such as a type 2- 5; e.g. a type 2-4; such as a type 3-4; e.g. a type 4.
  • a donor suitable for delivering gut microbiota transplantation may be tested (preferably from blood samples) for one or more, preferably all, of: ⁇ Cytomegalovirus, IgM, IgG
  • Creatinine and electrolytes (Calcium, K, Na, Cl, AST, ALT) ⁇ Aminotransferases, bilirubin, gamma-glutamyltransferase, alkaline phosphatase
  • the stool from a donor suitable for delivering the stool sample may be tested for one or more, preferably all, of:
  • enteric pathogens including Salmonella, Shigella ⁇ Campylobacter, Escherichia coli (0157 H7, Intimin-producing Escherichia coli,
  • Protozoa including Blastocystis hominis
  • helminths Faecal occult blood testing
  • stool from the donor should be tested on a regularly basis, such as each 2 nd months, preferably, for a period of at least 6 months after stool donation, e.g. at least 8 months, such as at least 10 months, e.g. at least 12 months, such as at least 18 months.
  • the filtering performed in step (iv) may be performed using a filter having pores of diameter less than or equal to 0.4 mm; such as less than or equal to 0.3 mm; e.g. less than or equal to 0.25 mm; such as less than or equal to 0.20 mm.
  • the filtering performed in step (iv) may be performed in a two-step filtering procedure.
  • the two-step filtering procedure may involve a first filtering process using a filter with a pore size in the range of 0.5-1 mm and/or a second filtering process using a filter with a pore size in the range of 0.01-0.25 mm.
  • the method after the sample obtained in (i) has been places in the sterile collection device (step (iii), may be performed anaerobically.
  • the sterile collection device may be a zip bag or a container.
  • the zip bag or the container is a plastic zip bag or a plastic container.
  • the mixing of the sample obtained in (ii) with at least one diluent (step (iii)) may be a gentle mixing.
  • mixing may be performed manually or mechanically.
  • Mechanically mixing may be performed using a grinder or a blender, preferably by grinding.
  • the faecal composition according to the present invention may be formulated into different products to make is easily used.
  • a preferred embodiment of the present invention relates to a device comprising a flexible container and a syringe wherein the flexible container comprises a faecal composition according to the present invention.
  • This device may be provided for medical use, either by the doctor, or by a recipient himself.
  • the device may be a rectal bulb syringe, rectal enema syringe or the like.
  • the device may be a disposable device.
  • Another preferred embodiment of the present invention relates to an animal feed product comprising the faecal composition according to the present invention. By using the faecal composition according to the present invention, the use of antibiotics in a heard of pigs, cattle, sheep; and/or poultry may be reduced or even avoided.
  • An even further preferred embodiment of the present invention relates to a faecal composition according to the present invention for use as a medicament.
  • a preferred embodiment of the present invention relates to a faecal composition according to the present invention for use in the treatment and/or prophylaxis of intestinal imbalance, such as chronic Inflammatory Bowel Disease; irritable bowel syndrome; dysbiosis; Clostridium difficile infections; diarrhea; idiopathic constipation; celiac disease; chronic urinary tract infection; infertility; type II diabetes; food allergies; cancer; refractory GvHD; obesity and morbid obesity; autism, sclerosis; constipation; chronic urinary tract infection or vaginal infection (including cystitis, mycoses); bone and joint infections; Parkinson's disease; Alzheimer's disease; schizophrenia and bipolar disorders; and gut dysbiosis associated with anti-cancer chemotherapy or immunotherapy.
  • intestinal imbalance such as chronic Inflammatory Bowel Disease; irritable bowel syndrome; dysbiosis; Clostridium difficile infections; diarrhea; idiopathic constipation; celiac disease; chronic urinary tract infection; infertility; type II diabetes;
  • intestinal induced disease relates to an imbalance in the gastrointestinal track, such as in the intestine caused by an inflammation in the gastrointestinal track, such as in the intestine, or in one or more parts of the intestine.
  • the disease caused by an imbalance in the intestine may be selected from the group consisting of chronic Inflammatory Bowel Disease; irritable bowel syndrome; dysbiosis; Clostridium difficile infections; diarrhea; idiopathic constipation; celiac disease; chronic urinary tract infection; infertility; type II diabetes; food allergies; cancer; refractory GvHD; obesity and morbid obesity; autism, sclerosis; constipation; chronic urinary tract infection or vaginal infection (including cystitis, mycoses); bone and joint infections; Parkinson's disease; Alzheimer's disease; schizophrenia and bipolar disorders; and gut dysbiosis associated with anti-cancer chemotherapy or immunotherapy.
  • chronic Inflammatory Bowel Disease irritable bowel syndrome
  • dysbiosis Clostridium difficile infections
  • diarrhea idiopathic constipation
  • celiac disease chronic urinary tract infection
  • infertility type II diabetes
  • food allergies cancer
  • cancer refractory GvHD
  • autism, sclerosis constipation
  • This imbalance may be caused by an inflammation in the intestine or in one or more parts of the intestine.
  • the faecal composition is an enema and is suitable for the treatment of chronic urinary tract infection or vaginal infection (including cystitis, mycoses)
  • an inflammation in the descending colon of a mammal such as a human
  • the faecal composition for the oral administration may be provided in the form the rectal administration may be provided in the form of enema.
  • an inflammation in the sigmoid colon of a mammal may be treated and/or suppressed by administering an effective amount of the faecal composition by rectal administration of the faecal composition to the mammal.
  • the faecal composition for the rectal administration may be provided in the form of enema.
  • an inflammation in the rectum of a mammal such as a human
  • a mammal such as a human
  • an effective amount of the faecal composition mainly by rectal administration.
  • the faecal composition may be provided in the form of enema.
  • an inflammation in the anal canal of a mammal may be treated and/or suppressed by administering to the mammal an effective amount of the faecal composition mainly by rectal administration.
  • the faecal composition may be provided in the form of enema.
  • the faecal composition according to the present invention may be suitable for the treatment of infertility. Without being bound by theory the inventor of the present invention believe that the treatment of infertility may be provided by stimulating female egg production.
  • the faecal composition may be particular suitable for the treatment of infertility when:
  • the stool donor may preferably be a woman of childbearing potential (between 20- 25 years old), who is not using any birth control medication; and/or
  • the stool donor may be taking a daily dosage of 100-500, preferably about 400 pg folic acid.
  • the recipient should preferably also receive folic acid for a period before the treatment, preferably, for a couple week before the treatment; and/or the donor may preferably donate the stool, at her ovulation days;
  • a rectal administration preferably using enema, may be used.
  • the enema is used every night before bedtime, preferably for a period of at least 1 day before the ovulation period, even more preferably 2 days before the ovulation period; and during the ovulation days.
  • a fresh faecal composition was prepared following the below procedure:
  • a healthy donor with suitable intestinal microbiota was selected, and tested according to the present invention, and a fresh stool sample was obtained from an approved donor and subjected to a first microbial analysis,
  • the fresh stool sample (comprising the saline solution) was mixed manually for 5 minutes, but a BagMixer could also have been used 4.
  • the mixture was gradually filtered down to ⁇ 0.2 millimeter resulting in a fresh faecal composition.
  • the fresh faecal composition may be used immediately or stored in a plastic bottle at a temperature about 6-8°C if used within 6 hours after preparation. If stored for longer periods the faecal composition may be frozen to a temperature of -80°C or below.
  • the fresh faecal composition was subjected to a second microbial analysis and compared with the first microbial analysis performed and more than 95% viability was observed.
  • a frozen faecal composition was prepared following the below procedure: 1.
  • a suitable healthy donor was selected, and tested and approved according to the present invention, and a fresh stool sample was obtained and subjected to a first microbial analysis,
  • the fresh stool sample (comprising the saline solution) was mixed manually for 5 minutes, but a BagMixer could also have been used
  • the faecal composition comprising 4% (v/v) glycerol was immediately stored at - 80°C and stored for 1 year.
  • the frozen faecal composition was thawed in a warm water bath (0 °C). 8. The thawed faecal composition was subjected to a second microbial analysis and compared with the first microbial analysis performed and about 90% viability was observed.

Abstract

The present invention relates to a novel faecal composition comprising a stool fraction and a diluent, wherein the stool fraction comprises one or more non-pathogenic microorganisms and the diluent comprises a cryoprotectant resulting in a final concentration of cryoprotectant in the faecal composition below 8% (v/v) cryoprotectant.

Description

NOVEL FAECAL COMPOSITION
Technical field of the invention
The present invention relates to a faecal composition for gut microbiota transplantation. In particular the present invention relates to a faecal composition for gut microbiota transplantation wherein the side effects has been reduced.
Background of the invention
The human body contains trillions of bacteria cells, protozoa and viruses, known as the microbiome. The microbiome has been linked to many aspects of human health, from gastrointestinal related diseases to obesity.
Focus on gut microbiota transplantation (GMT) for the prevention and treatment of intestinal disorders has been increasing in the human medicine. Increased clinical applications of GMT have provided convincing proofs that modification of the intestinal microbiota is an effective therapy for intestinal dysbiosis-related diseases
GMT refers to the process of implanting of faecal suspension from a healthy donor with the right gut microbiota composition into the recipient's intestinal tract to restore the community and function of intestinal microbiota and/or intestinal functionality.
Gut Microbiota Transplantation (GMT), also called faecal microbiota transplantation (FMT), has shown - in randomized, controlled clinical trials - resolved 80-90% of infections caused by recurrent C. difficile that does not respond to antibiotics.
During GMT, a faecal preparation from a carefully screened healthy stool-donor, is transplanted into the colon of the patient. There are multiple routes of administration (e.g., via colonoscopy, naso-enteric tube), each of which has unique risks and benefits.
Though the mechanism of gut microbiota transplantation yet has to be precisely determined. It is believed that GMT, in the treatment of C. difficile infection, works by repopulating the patient's microbiome with diverse microorganisms that competitively exclude C. difficile. In a healthy gut community, C. difficile is outcompeted or outgrown by many different bacterial species. However, receiving antibiotic treatment disrupts this ecosystem by killing those protective bacteria. C. difficile forms spores that are resistant to antibiotics. No longer outcompeted, antibiotic resistant C. difficile outgrowing in the gut and produces toxins, which causes severe diarrhoea, abdominal pain, and, often fever. With an infusion of GMT from a healthy donor's stool, the C. difficile may be outgrown.
Gut microbiome plays an important role in human and animal health by helping digestion and benefiting the immune system and many other aspects of health. An imbalance of gut microbiota/dysbiosis might be restored by gut microbiota transplantation from a healthy gut , which may contribute to the treatment of various diseases, such as ulcerative colitis and other gastrointestinal conditions. Jun Hu et al. describes the preparation of faecal compositions, where fresh feces are obtained from a healthy donor and diluted with sterile saline and homogenized in a standard blender. The slurry is then filtered three times through gauze, strainer, or 0.25 mm stainless steel sieves to eliminate the undigested and small particulate matter in the faecal suspension. The faecal suspension may be centrifuged at 6,000 x g for 15 min. The precipitate, without the supernatant, is re-suspended in fresh sterile saline. The resulting suspension should be transferred to the recipients immediately if a fresh faecal composition is to be used to ensuring faecal microbial viability.
Furthermore, Jun Hu et al. describes, preparation of faecal compositions used for GMT, that may preferably be provided as a frozen faecal suspension, which ensures durability, uniformity and viability of the faecal composition and the faecal suspension may be saved for later use. Comparative studies have demonstrated that frozen faecal material does not only simplify the practical steps of clinical human GMT, but also has the similar efficacy to fresh faecal material.
To improve the faecal microbial survival rates during the cryopreservation, fresh stool samples should be diluted with sterile saline homogenized and filtered using the protocol used in the preparation of the fresh faecal material above. Subsequently, the resulting suspension should be added to glycerol (a cryoprotectant) to get a final concentration of 10%. Finally, the faecal suspensions are labeled accurately and then stored, as soon as possible, at -80°C to ensure the faecal microbial survival. When there is the need to perform a GMT, the frozen faecal suspension should be thawed at and mixed with saline solution to obtain the required concentration and the infusion of faecal suspension should be implemented as soon as possible at room temperature. The disadvantage of the prior art, exemplified by Jun Hu et al. as described above, is that high amount of glycerol (cryoprotectant) is added to the faecal composition and that other beneficial products in the product, such as fungi, protozoa, viruses, vitamins and minerals produced by the healthy gut microbiota and the nutrition suitable for survival of the healthy gut microbiota in the diseased gut, are discharged together with the supernatant. However, the process is performed under aerobic condition and using standard blender adds more air to the product, which means there are no-anaerobic bacteria in the final product.
Despite the positive effect cryoprotectant, such as glycerol, has on survival rate of cells during cryopreservation, cryoprotectant, such as glycerol, has several disadvantages side effects like being harmful to the mucous membranes by increasing tissue osmolarity; or causing headaches, dizziness, bloating, nausea, vomiting, thirst, or diarrhoea, in particular the negative effects on the mucous membranes has been found to occur frequently and is undesirable.
Hence, an improved faecal composition would be advantageous, and in particular an efficient, a durable, uniform and viable faecal composition for GMT with no or reduced number of side effects would be advantageous.
Summary of the invention
Thus, an object of the present invention relates to a faecal composition for gut microbiota transplantation.
In particular, it is an object of the present invention to provide a faecal composition for GMT, that solves the above mentioned problems of the prior art with being harmful to the mucous membranes; or causing headaches, dizziness, bloating, nausea, vomiting, thirst, and/or diarrhoea.
Thus, one aspect of the invention relates to a faecal composition comprising a stool fraction and a diluent, wherein the stool fraction comprises one or more non-pathogenic microorganisms and the diluent comprises a cryoprotectant resulting in a final concentration of cryoprotectant in the faecal composition below 8% (v/v) cryoprotectant.
Another aspect of the present invention relates to a method for providing a faecal composition, the method comprises the steps of: (i) obtaining at least one sample of stool from a donor,
(ii) placing said sample obtained in step (i) in a sterile, preferably an anaerobic sterile, collection device
(iii) mixing (preferably gently mixing) the sample obtained in step (ii) with at least one diluent comprises a cryoprotectant resulting in a final concentration of cryoprotectant in the faecal composition below 8% (v/v) cryoprotectant
(iv) filtering the mixture obtained in step (iii), providing the faecal composition comprising a stool fraction and a diluent, wherein the diluent comprises a cryoprotectant resulting in a final concentration of cryoprotectant in the faecal composition below 8% (v/v) cryoprotectant.
Yet another aspect of the present invention relates to a device comprising a flexible, preferably anaerobic, container and a syringe, wherein the flexible container comprises a faecal composition according to the present invention. Still another aspect of the present invention relates to an animal feed product comprising the faecal composition according to the present invention.
A further aspect of the present invention relates to a faecal composition according to the present invention for use as a medicament.
An even further aspect of the present invention relates to a faecal composition according to the present invention for use in the treatment and/or prophylaxis of intestinal imbalance, such as chronic Inflammatory Bowel Disease; irritable bowel syndrome; dysbiosis; Clostridium difficile infections; diarrhea; idiopathic constipation; celiac disease; chronic urinary tract infection; infertility; type II diabetes; food allergies; cancer; refractory GvHD; obesity and morbid obesity; autism, sclerosis; constipation; chronic urinary tract infection or vaginal infection (including cystitis, mycoses); bone and joint infections; Parkinson's disease; Alzheimer's disease; schizophrenia and bipolar disorders; and gut dysbiosis associated with anti-cancer chemotherapy or immunotherapy. Detailed description of the invention
The inventor of the present invention surprising found that a faecal composition may be provided which has a significant reduction in the side effects and without compromising the efficiency, durability, uniformity and viability of the faecal composition.
A preferred embodiment of the present invention relates to a faecal composition comprising a stool fraction and a diluent, wherein the stool fraction comprises one or more non-pathogenic microorganisms and the diluent comprises a cryoprotectant resulting in a final concentration of cryoprotectant in the faecal composition below 8% (v/v) cryoprotectant.
The faecal composition according to the present invention relates to a composition obtained from a human or animal donor, preferably, from a healthy human donor. In the present context the term "donor" relates to a healthy human or animal-with the suitable gut microbiota delivering a faecal material. In the context of the present invention the term "recipient" relates to a sick human or animal receiving the healthy gut microbiota composition, vitamins and mineral produced by healthy gut microbiota, antimicrobial peptides, bacteria, protozoa, viruses, nutrition necessary for the healthy gut bacteria to survive in the disease prepared from faecal material delivered.
In an embodiment of the present invention the faecal material may be a healthy human or animal faeces or stool, preferably healthy human faeces or stool with the suitable gut microbiota. In the present context the terms "stool" and "faeces" may be used interchangeably.
The stool fraction may comprise one or more non-pathogenic microorganisms. In an embodiment of the present invention the one or more non-pathogenic microorganisms relates to one or more healthy gut bacteria, fungi, protozoa, viruses, present in the faecal material. In an embodiment of the present invention the faecal composition comprises vitamins and mineral produced by healthy gut microbiota, antimicrobial peptides, and nutrition necessary for the healthy gut bacteria to survive in a gut.
In the context of the present invention the term "non-pathogenic microorganism" relates to a microorganism that do not cause disease, harm or death to another organism, such as a human or an animal. In an embodiment of the present invention at least 50% of the non-pathogenic microorganisms in the faecal composition may be viable non-pathogenic microorganisms relative to the total number of non-pathogenic microorganisms in the faecal composition, such as at least 75%, e.g. at least 90%, such as at least 95%.
In a further embodiment of the present invention the faecal composition may comprise in the range of 104-1012 viable non-pathogenic facultative anaerobe, anaerobe and aerobe microorganisms, such as in the range of 104-1012, e.g. in the range of 106-1010, such as in the range of 108-109. Preferably, the viable non-pathogenic microorganisms may be viable non-pathogenic facultative anaerobe, anaerobe and aerobe microorganisms.
The faecal composition according to the present invention may be subjected to removal of undigested food. Furthermore, the faecal composition according to the present invention may preferably comprise vitamins and mineral produced by healthy gut microbiota, antimicrobial peptides, and nutrition necessary for the healthy gut bacteria to survive in a gut.
In an embodiment of the present invention a 50-gram stool fraction results in 12-28 gram faecal composition after filtration from undigested food, depending on the diet during the donation time.
In an embodiment according to the present invention the faecal composition does not comprises antibiotics.
In a further embodiment of the present invention the faecal composition according to the present invention comprises no worms; no pathogenic parasites; and/or no pathogenic microorganisms.
Preferably, the faecal composition may not comprise one or more of Clostridium difficile,· Clostridium Perfringens, Vibrio, Pleisiomonas, Aeromonas, enteric pathogens, including Salmonella, Shigella,· Campylobacter, Escherichia coli (0157 H7, Intimin-producing Escherichia coli, STEC, EPEC, ETEC, EIEC); Carbapenemase-producing enterobacteriae (CPE), Yersinia,· vancomycin-resistant enterococci (VRE); methicillin-resistant Staphylococcus aureus (MRSA); Helicobacter, Gram/negative multidrug-resistant bacteria; Enterobacteriaceae, such as Spectrum beta-lactamase (ESBL) producing Enterobacteriaceae, or carbapenem-resistant Enterobacteriaceae (CRE); Norovirus; Antigens and/or acid-fast staining for Giardia lamblia and Criptosporidium parvum; Protozoa (including Blastocystis hominis) and helminths; and/or faecal occult blood, Entamoeba histolytica, Sapovirus, Rotavirus, Astrovirus, Adenovirus, Coronavirus, worms, worm eggs and cysts, Bacillus cereus .
Preferably, the faecal composition may not comprises above mentioned Clostridium species,· enteric pathogens, including Salmonella, Shigella,· Campylobacter, above mentioned pathogenic Escherichia co//spp.; Yersinia,· vancomycin-resistant enterococci (VRE); methicillin-resistant Staphylococcus aureus (MRSA); Gram/negative multidrug- resistant bacteria; Enterobacteriaceae, such as Spectrum beta-lactamase (ESBL) producing Enterobacteriaceae, or carbapenem-resistant Enterobacteriaceae (CRE); Norovirus; Antigens and/or acid-fast staining for Giardia lamblia and Criptosporidium parvum;
Protozoa (including Blastocystis hominis) and helminths; and faecal occult blood, Entamoeba histolytica, Sapovirus, Rotavirus, Astrovirus, Adenovirus, Coronavirus, worms, worm eggs and cysts, Bacillus cereus.
In an embodiment of the present invention the faecal composition may be a fresh faecal composition.
In a preferred embodiment of the present invention the faecal may be a fresh or a frozen faecal composition.
In an embodiment of the present invention the faecal composition may comprise a diluent which is used to make the stool fraction more viscus. In an embodiment of the present invention the diluent may comprise an aqueous solution and/or a cryoprotectant.
The aqueous solution may comprise a salt, a mineral, and acid or an alkaline. Preferably, the aqueous solution comprises a salt, such as sodium chloride providing a saline solution. The saline solution comprises sodium chloride, preferably, the concentration of sodium chloride in the saline solution may be 2-30 g/l; such as 4-20 g/l; e.g. 7-19 g/l; such as about 9 g/l sodium chloride (0.9%).
The aqueous solution may preferably be a sterile aqueous solution.
Preferably the diluent, may be a saline containing diluent, thus resulting in a diluent that in addition to the cryoprotectant, preferably glycerol, comprises a saline solution.
In an embodiment of the present invention the faecal composition may be provided in dosage forms comprising 50 gram stool product and in the range of 50-150 ml saline comprising at most 10% (w/w) glycerol, such as at most 8% (w/w) glycerol, e.g. at most 5% (w/w) glycerol; such as at most 4% (w/w) glycerol, e.g. at most 3% (w/w) glycerol; such as at most 2% (w/w) glycerol, e.g. at most 1% (w/w) glycerol, relative to the saline solution; such as in the range of 60-125 ml, e.g. in the range of 65-100 ml, such as in the range of 70-80 ml, e.g. about 75 ml.
Preferably, the faecal composition according to the present invention may be provided in dosage forms consisting essentially of:
50 gram stool product; and in the range of 50-150 ml saline comprising at most 10% (w/w) glycerol, such as at most 8% (w/w) glycerol, e.g. at most 5% (w/w) glycerol; such as at most 4% (w/w) glycerol, e.g. at most 3% (w/w) glycerol; such as at most 2% (w/w) glycerol, e.g. at most 1% (w/w) glycerol, relative to the saline solution; such as in the range of 60-125 ml, e.g. in the range of 65-100 ml, such as in the range of 70- 80 ml, e.g. about 75 ml.
In the context of the present invention the term "stool product" relates to stool present in the faecal composition where undigested and small particulate material have been reduced or removed.
In an embodiment of the present invention the diluent may be added to the stool in several sequences, such as addition in 2 sequences, e.g. addition in 3 sequences, such as addition in 4 sequences.
In a further embodiment of the present invention addition of diluent to the stool fraction may be done by an addition of a saline solution, preferably a sterile saline solution, to increase viscosity followed by addition of a cryoprotectant before freezing. The addition of saline solution may involve several additions of saline solutions.
In yet an embodiment of the present invention the sterile cryoprotectant may be added to the stool fraction in combination with an sterile aqueous solution, such as a silane solution.
In a preferred embodiment of the present invention the diluent comprises a cryoprotectant resulting in a final concentration of cryoprotectant in the faecal composition below 8%
(v/v) cryoprotectant; such as 7% (v/v) cryoprotectant or below; e.g. 6% (v/v) cryoprotectant or below; such as 5% (v/v) cryoprotectant or below; e.g. 4% (v/v) cryoprotectant or below; such as 3% (v/v) cryoprotectant or below; e.g. 2% (v/v) cryoprotectant or below; such as 1% (v/v) cryoprotectant or below; e.g. 0.5% (v/v) cryoprotectant or below; such as 0.1% (v/v) cryoprotectant or below; e.g. 0.05% (v/v) cryoprotectant or below; such as 0.001% (v/v) cryoprotectant or below; e.g. 0.0001% (v/v) cryoprotectant or below; such as in the range of 0-8% (v/v) cryoprotectant; e.g. in the range of 0.0001-7% (v/v) cryoprotectant; such as in the range of 0.001-6% (v/v) cryoprotectant; e.g. in the range of 0.01-5% (v/v) cryoprotectant; such as in the range of 0.1-4% (v/v) cryoprotectant; e.g. in the range of 1-3% (v/v) cryoprotectant; such as in the range of 1.5-2.5% (v/v) cryoprotectant; e.g. in the range of 0.1-2% (v/v) cryoprotectant.
In an embodiment of the present invention the content of the diluent may constitute less than 5 times the content of the stool fraction on a (w/w) basis; such as less than 4 times the content of the stool fraction on a (w/w) basis; e.g. less than 3 times content of the content of the stool fraction on a (w/w) basis; such as less than 2.5 times the content of the stool fraction on a (w/w) basis; e.g. less than 2 times content of the content of the stool fraction on a (w/w) basis; such as less than 1.8 times the content of the stool fraction on a (w/w) basis; e.g. less than 1.6 times content of the content of the stool fraction on a (w/w) basis; such as less than 1.4 times the content of the stool fraction on a (w/w) basis; e.g. less than 1.2 times content of the content of the stool fraction on a (w/w) basis.
In a further embodiment of the present invention the cryoprotectant may be selected from the group consisting of glycerol; mannitol; sorbitol; propylene glycol; ethylene glycol; trehalose and its analogues; saccharose; galactose-lactose and mixtures hereof.
Preferably, the cryoprotectant may be glycerol.
The stool fraction and/or the faecal composition may comprise a particulate material as beneficial bacteria such as: Faecalbacterium, Prevotella bacteria, Lactobacilli species, Weissella spp., Bifidobacterium spp., Streptococcus thermophilus. The stool fraction also contains antimicrobial peptides, healthy gut microbiota composition, short chain fatty acid, which serves as nutrition for host cell functionality (only produced by healthy gut microbiota), Vitamins and mineral produced by healthy gut microbiota, bacteria, protozoa, viruses, nutrition necessary for the healthy gut bacteria to survive in the disease.
The particulate material may comprise a particle size of the stool fraction and/or the faecal composition is below 0.4 mm; such as 0.3 mm or below; e.g. 0.25 mm or below; such as 0.20 mm or below.
In an embodiment of the present invention the faecal composition may be a liquid faecal composition.
The faecal composition may be formulated into an enema formulation, a probe formulation or a colonoscopy formulation. In an embodiment of the present invention the stool fraction may be provided from a stool obtained from a mammal (donor). The mammal may be a human; a dog; a cat; a horse; cattle; sheep; goat; a chicken; a duck; and pig.
The selection of a suitable and healthy donor may follow a very strict scheme for collecting and inspecting donor faecal sample to ensure quality of the stool. The process of donor selection may be described below and may be demonstrated in the figure 1. Initially the donor should pass the preliminary donor health questionnaire regarding various health questions.
The donor stool may be obtained and tested. If approved (e.g. not comprising pathogenic microorganisms, antibiotics, presence of medicine, etc.) the donor may be accepted.
In an embodiment the donor stool may be placed in quarantine for 3 months at 5-8 degrees to investigate if the donor intestinal bacteria produce any toxins in the stool. If the donor is approved, the donor may be supplied with a stool collection kit, e.g. comprising a zip-bag for collecting the stool until processed.
Preferably the donor may deliver stool 3 times a week. Once the donor is accepted, they will be supplied with the collection kit and they must regularly donate up to 3 times a week. The quality of the stool may be checked frequently, such as 4 times a year. E.g. once a year, such as each second month, e.g. each month, such as each 14 days, e.g. each week, e.g. every time a stool donation is made.
A preferred embodiment of the present invention relates to a method for providing a faecal composition, the method comprises the steps of:
(i) obtaining at least one sample of stool from a donor,
(ii) placing said sample obtained in step (i) in a sterile collection device
(iii) mixing the sample obtained in step (ii) with at least one diluent comprises a cryoprotectant resulting in a final concentration of cryoprotectant in the faecal composition below 8% (v/v) cryoprotectant (iv) filtering the mixture obtained in step (iii), providing the faecal composition comprising a stool fraction and a diluent, wherein the diluent comprises a cryoprotectant resulting in a final concentration of cryoprotectant in the faecal composition below 8% (v/v) cryoprotectant.
The donor, healthy donor, may be analyzed and determined to be an acceptable healthy donor as described herein.
The liquid phase obtained from separating the solid and liquid phase obtained from the filtering performed in step (iv) may be included as part of the faecal composition.
In an embodiment of the present invention the faecal composition is not added a water immiscible compound. Preferably, the water immiscible compound is an oil compound.
Polyols, like glycerol, may be a water miscible compound.
In an embodiment of the present invention the faecal composition is not added a carbohydrate, an oligosaccharide or a polysaccharide compound.
In an embodiment of the present invention the faecal composition comprises:
(1) liquid phase obtained from the filtering performed in step (iv); antimicrobial peptides, short chain fatty acid, vitamins and/or minerals;
(2) microbiota, fungi, protozoa, viruses, and/or nutrition;
(3) glycerol;
(4) saline compound; and the faecal composition does not comprise
(5) added oil: and
(6) added carbohydrate, oligosaccharide or polysaccharide compound
The faecal composition according to the present invention may be used immediately or it may be frozen before use (in particular if not used immediately. Thus to improve viability and durability of the faecal composition the faecal composition may be stored for at least 6 hours at a temperature of -80°C or less, such as for at least 12 hours, e.g. for at least 24 hours, such as for at least 3 days, e.g. for at least 5 days, such as for at least 7 days, e.g. for at least 10 days, such as for at least 15 days, e.g. for at least 25 days, such as for at least 1 month, e.g. for at least 3 months, such as for at least 6 months, e.g. for at least 9 months, such as for at least 12 months or 2 till 3 years.
The faecal composition may be immediately stored at a temperature of -80°C or less. In order to improve viability of the faecal composition or for increasing storage of the faecal composition according to the present invention, the faecal composition may be stored (preferably immediately) at a temperature of -80°C or less, such as a temperature of -100°C or less, e.g. a temperature of -120°C or less, such as a temperature of -150°C or less.
In an embodiment of the present invention the process of freezing the faecal composition to a temperature of -100°C or less, e.g. a temperature of -120°C or less, such as a temperature of -150°C or less, may be performed in a stepwise procedure comprising a first freezing step to about -80C followed by a second freezing step to a temperature of - 120°C or less, such as a temperature of -150°C or less.
The faecal composition may be stored at the final storage temperature for a period of at least 40 minutes, e.g. at least 1 hour, e.g. at least 24 hours, e.g. at least 1 week, e.g. at least 2 weeks, e.g. at least 1 month, e.g. at least 3 months, e.g. at least 5 months, e.g. at least 1 year, e.g. at least 2 years, e.g. at least 3 years, preferably about 1 year.
In an embodiment of the present invention the stool desired for the purpose of the present invention may be characterised as a type 2-5 on the Bristol Stool Chart; such as a type 2- 5; e.g. a type 2-4; such as a type 3-4; e.g. a type 4.
In order to ensure a high-quality stool sample provided in step (i) a donor suitable for delivering gut microbiota transplantation may be tested (preferably from blood samples) for one or more, preferably all, of: · Cytomegalovirus, IgM, IgG
• Epstein-Barr virus IgM
• Treponema Pallidum IgG, IgM
• Hepatitis A
• HBV, Hepatitis B virus · HCV, Hepatitis C virus
• Hepatitis E virus
• Syphilis
• HIV-1 and HIV-2
• Corona virus · Entamoeba histolytica
• Complete blood cell count with differential: o Hematocrito, Hemoglobin, VCM, HCM, CHCM, Platelets,
Leukocyte, Differential Leukocyte count
• C-reactive protein and erythrocyte sedimentation rate • Coagulation factor, APTT
• IgG, IgA, IgM
• Albumin
• Creatinine and electrolytes (Calcium, K, Na, Cl, AST, ALT) · Aminotransferases, bilirubin, gamma-glutamyltransferase, alkaline phosphatase
Glucose, Cholesterol, Triglyceride, HDL, LDL, TSH
• Human T-lymphotropic virus types I and II antibodies
• Strongyloides stercoralis In order to further ensuring a high-quality stool sample provided in step (i) the stool from a donor suitable for delivering the stool sample may be tested for one or more, preferably all, of:
• Detection of Clostridium difficile, Clostridium Perfringens
• Detection of enteric pathogens, including Salmonella, Shigella · Campylobacter, Escherichia coli (0157 H7, Intimin-producing Escherichia coli,
STEC, EPEC, ETEC, EIEC), Yersinia, vancomycin-resistant enterococci (VRE), methicillin-resistant Staphylococcus aureus (MRSA, nasal or peri-rectal swabs is also ok if not stool), Gram/negative multidrug-resistant bacteria, Spectrum beta- lactamase (ESBL) producing Enterobacteriaceae, carbapenem-resistant Enterobacteriaceae (CRE), Carbapenemase-producing enterobacteriae (CPE)
• Norovirus
• Coronavirus in the blood and in feces
• Antigens and/or acid-fast staining for Giardia lamblia and Criptosporidium parvum
• Protozoa (including Blastocystis hominis) and helminths · Faecal occult blood testing
• Detection of Vibrio Cholera and Listeria monocytogens
• Antigens and/or acid-fast staining for Isospora and Microsporidia
• Calprotectin
• Helicobacter pylori faecal antigen · Rotavirus
• Pleisiomonas
• Aeromonas
• Entamoeba histolytica
• Sapovirus, · Astrovirus,
• Adenovirus,
• worms, worm eggs and cysts,
• Bacillus cereus, Preferably stool from the donor should be tested on a regularly basis, such as each 2nd months, preferably, for a period of at least 6 months after stool donation, e.g. at least 8 months, such as at least 10 months, e.g. at least 12 months, such as at least 18 months.
In an embodiment of the present invention the filtering performed in step (iv) may be performed using a filter having pores of diameter less than or equal to 0.4 mm; such as less than or equal to 0.3 mm; e.g. less than or equal to 0.25 mm; such as less than or equal to 0.20 mm.
Preferably, the filtering performed in step (iv) may be performed in a two-step filtering procedure.
The two-step filtering procedure may involve a first filtering process using a filter with a pore size in the range of 0.5-1 mm and/or a second filtering process using a filter with a pore size in the range of 0.01-0.25 mm.
In an embodiment of the present invention the method, after the sample obtained in (i) has been places in the sterile collection device (step (iii), may be performed anaerobically.
The sterile collection device may be a zip bag or a container. Preferably the zip bag or the container is a plastic zip bag or a plastic container.
In an embodiment of the present invention the mixing of the sample obtained in (ii) with at least one diluent (step (iii)) may be a gentle mixing. Preferably mixing may be performed manually or mechanically. Mechanically mixing may be performed using a grinder or a blender, preferably by grinding.
The faecal composition according to the present invention may be formulated into different products to make is easily used.
A preferred embodiment of the present invention relates to a device comprising a flexible container and a syringe wherein the flexible container comprises a faecal composition according to the present invention. This device may be provided for medical use, either by the doctor, or by a recipient himself.
Preferably, the device may be a rectal bulb syringe, rectal enema syringe or the like. Preferably, the device may be a disposable device. Another preferred embodiment of the present invention relates to an animal feed product comprising the faecal composition according to the present invention. By using the faecal composition according to the present invention, the use of antibiotics in a heard of pigs, cattle, sheep; and/or poultry may be reduced or even avoided.
An even further preferred embodiment of the present invention relates to a faecal composition according to the present invention for use as a medicament.
Yet a preferred embodiment of the present invention relates to a faecal composition according to the present invention for use in the treatment and/or prophylaxis of intestinal imbalance, such as chronic Inflammatory Bowel Disease; irritable bowel syndrome; dysbiosis; Clostridium difficile infections; diarrhea; idiopathic constipation; celiac disease; chronic urinary tract infection; infertility; type II diabetes; food allergies; cancer; refractory GvHD; obesity and morbid obesity; autism, sclerosis; constipation; chronic urinary tract infection or vaginal infection (including cystitis, mycoses); bone and joint infections; Parkinson's disease; Alzheimer's disease; schizophrenia and bipolar disorders; and gut dysbiosis associated with anti-cancer chemotherapy or immunotherapy.
In the context of the present invention the term "intestinal induced disease" relates to an imbalance in the gastrointestinal track, such as in the intestine caused by an inflammation in the gastrointestinal track, such as in the intestine, or in one or more parts of the intestine.
The disease caused by an imbalance in the intestine, e.g. intestinal induced disease, may be selected from the group consisting of chronic Inflammatory Bowel Disease; irritable bowel syndrome; dysbiosis; Clostridium difficile infections; diarrhea; idiopathic constipation; celiac disease; chronic urinary tract infection; infertility; type II diabetes; food allergies; cancer; refractory GvHD; obesity and morbid obesity; autism, sclerosis; constipation; chronic urinary tract infection or vaginal infection (including cystitis, mycoses); bone and joint infections; Parkinson's disease; Alzheimer's disease; schizophrenia and bipolar disorders; and gut dysbiosis associated with anti-cancer chemotherapy or immunotherapy.
This imbalance may be caused by an inflammation in the intestine or in one or more parts of the intestine.
Preferably, the faecal composition is an enema and is suitable for the treatment of chronic urinary tract infection or vaginal infection (including cystitis, mycoses) In an embodiment of the present invention an inflammation in the descending colon of a mammal, such as a human, may be treated and/or suppressed by administering an effective amount of the faecal composition by rectal administration of the faecal composition to the mammal. Preferably, the faecal composition for the oral administration may be provided in the form the rectal administration may be provided in the form of enema.
In a further embodiment of the present invention an inflammation in the sigmoid colon of a mammal, such as a human, may be treated and/or suppressed by administering an effective amount of the faecal composition by rectal administration of the faecal composition to the mammal. Preferably, the faecal composition for the rectal administration may be provided in the form of enema.
In yet an embodiment of the present invention an inflammation in the rectum of a mammal, such as a human, may be treated and/or suppressed by administering to the mammal an effective amount of the faecal composition mainly by rectal administration. Preferably, the faecal composition may be provided in the form of enema.
In another embodiment of the present invention an inflammation in the anal canal of a mammal, such as a human, may be treated and/or suppressed by administering to the mammal an effective amount of the faecal composition mainly by rectal administration. Preferably, the faecal composition may be provided in the form of enema.
The faecal composition according to the present invention may be suitable for the treatment of infertility. Without being bound by theory the inventor of the present invention believe that the treatment of infertility may be provided by stimulating female egg production. The faecal composition may be particular suitable for the treatment of infertility when:
The stool donor may preferably be a woman of childbearing potential (between 20- 25 years old), who is not using any birth control medication; and/or
- The stool donor may be taking a daily dosage of 100-500, preferably about 400 pg folic acid.
- The recipient should preferably also receive folic acid for a period before the treatment, preferably, for a couple week before the treatment; and/or the donor may preferably donate the stool, at her ovulation days; For the treatment of infertility, a rectal administration, preferably using enema, may be used. Preferably, the enema is used every night before bedtime, preferably for a period of at least 1 day before the ovulation period, even more preferably 2 days before the ovulation period; and during the ovulation days.
It should be noted that embodiments and features described in the context of one of the aspects of the present invention also apply to the other aspects of the invention. The invention will now be described in further details in the following non-limiting examples.
Examples Example 1
A fresh faecal composition was prepared following the below procedure:
1. A healthy donor with suitable intestinal microbiota was selected, and tested according to the present invention, and a fresh stool sample was obtained from an approved donor and subjected to a first microbial analysis,
2. 75 ML sterile anaerobic saline solution containing below 4% glycerol was added to a zip bag/ Bagpage XRCk containing 50-gram fresh stool and treated anaerobically.
3. The fresh stool sample (comprising the saline solution) was mixed manually for 5 minutes, but a BagMixer could also have been used 4. The mixture was gradually filtered down to <0.2 millimeter resulting in a fresh faecal composition.
5. After filtering the fresh faecal composition may be used immediately or stored in a plastic bottle at a temperature about 6-8°C if used within 6 hours after preparation. If stored for longer periods the faecal composition may be frozen to a temperature of -80°C or below.
6. The fresh faecal composition was subjected to a second microbial analysis and compared with the first microbial analysis performed and more than 95% viability was observed.
Example 2
A frozen faecal composition was prepared following the below procedure: 1. A suitable healthy donor was selected, and tested and approved according to the present invention, and a fresh stool sample was obtained and subjected to a first microbial analysis,
2. 75 ML sterile saline solution was added to a zip bag/ Bagpage XRCk containing 50- gram fresh stool and treated anaerobically.
3. The fresh stool sample (comprising the saline solution) was mixed manually for 5 minutes, but a BagMixer could also have been used
4. The mixture was gradually filtered down to <0.2 millimeter resulting in a fresh faecal composition. 5. After filtering the fresh faecal composition was mixed with glycerol resulting in a concentration of 4% (v/v) glycerol in the faecal composition.
6. The faecal composition comprising 4% (v/v) glycerol was immediately stored at - 80°C and stored for 1 year.
7. The frozen faecal composition was thawed in a warm water bath (0 °C). 8. The thawed faecal composition was subjected to a second microbial analysis and compared with the first microbial analysis performed and about 90% viability was observed.
References
Jun Hu et al. Front Microbiol. 2018; 9: 1328 (https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6018536/)

Claims

Claims
1. A faecal composition comprising a stool fraction and a diluent, wherein the stool fraction comprises one or more non-pathogenic microorganisms and the diluent comprises a cryoprotectant resulting in a final concentration of cryoprotectant in the faecal composition below 8% (v/v) cryoprotectant.
2. The faecal composition according to claim 1, wherein the cryoprotectant is selected from the group consisting of glycerol; mannitol; sorbitol; propylene glycol; ethylene glycol; trehalose and its analogues; saccharose; galactose-lactose and mixtures hereof.
Preferably, the cryoprotectant is glycerol.
3. The faecal composition according to anyone of claims 1-2, wherein the faecal composition is a fresh or frozen faecal composition.
4. A method for providing a faecal composition, the method comprises the steps of:
(i) obtaining at least one sample of stool from a donor,
(ii) placing said sample obtained in (i) in a sterile anaerobic collection device
(iii) mixing the sample obtained in (ii) with at least one diluent comprises a cryoprotectant resulting in a final concentration of cryoprotectant in the faecal composition below 8% (v/v) cryoprotectant
(iv) filtering the mixture obtained in (iii), providing the faecal composition comprising a stool fraction and a diluent, wherein the diluent comprises a cryoprotectant resulting in a final concentration of cryoprotectant in the faecal composition below 8% (v/v) cryoprotectant.
5. The method according to claim 4, wherein the freezing of the faecal composition is performed immediately to a temperature of -80°C or less.
6. The method according to claim 4, wherein the faecal composition is a fresh faecal composition.
7. The method according to claim 6, wherein the faecal composition is used for gut microbiota transplantation within 24 hours or less; such as within 20 hours or less; e.g. within 15 hours or less; such as within 10 hours or less; e.g. within 6 hours or less; such as within 5 hours or less; e.g. within 4 hours or less; such as within 3 hours or less; e.g. within 2 hours or less.
8. A device comprising a flexible container and a syringe wherein the flexible container comprises a faecal composition according to anyone of claims 1-3.
9. The device according to claim 8, wherein the device is a rectal bulb syringe, rectal enema syringe.
10. An animal feed product comprising the faecal composition according to anyone of claims 1-3.
11. A faecal composition according to anyone of claims 1-3 for use as a medicament.
12. A faecal composition according to anyone of claims 1-3 for use in the treatment and/or prophylaxis of intestinal imbalance, such as chronic Inflammatory Bowel Disease; irritable bowel syndrome; dysbiosis; Clostridium difficile infections; diarrhea; idiopathic constipation; celiac disease; chronic urinary tract infection; infertility; type II diabetes; food allergies; cancer; refractory GvHD; obesity and morbid obesity; autism, sclerosis; constipation; chronic urinary tract infection or vaginal infection (including cystitis, mycoses); bone and joint infections; Parkinson's disease; Alzheimer's disease; schizophrenia and bipolar disorders; and gut dysbiosis associated with anti-cancer chemotherapy or immunotherapy.
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Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20140363398A1 (en) * 2013-06-05 2014-12-11 Rebiotix, Inc. Microbiota restoration therapy (mrt), compositions and methods of manufacture
WO2018140931A1 (en) * 2017-01-30 2018-08-02 The Board Of Regents Of The University Of Texas System Compositions and methods for fecal microbiota transplantation
US20180289750A1 (en) * 2015-06-09 2018-10-11 Rebiotix, Inc. Microbiota restoration therapy (mrt) compositions and methods of manufacture
US20190247445A1 (en) * 2016-07-01 2019-08-15 Regents Of The University Of Minnesota Compositions and methods for c. difficile treatment
WO2019241523A1 (en) * 2018-06-14 2019-12-19 Rebiotix, Inc. Microbiota restoration therapy (mrt) compositions and methods of manufacture

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20140363398A1 (en) * 2013-06-05 2014-12-11 Rebiotix, Inc. Microbiota restoration therapy (mrt), compositions and methods of manufacture
US20180289750A1 (en) * 2015-06-09 2018-10-11 Rebiotix, Inc. Microbiota restoration therapy (mrt) compositions and methods of manufacture
US20190247445A1 (en) * 2016-07-01 2019-08-15 Regents Of The University Of Minnesota Compositions and methods for c. difficile treatment
WO2018140931A1 (en) * 2017-01-30 2018-08-02 The Board Of Regents Of The University Of Texas System Compositions and methods for fecal microbiota transplantation
WO2019241523A1 (en) * 2018-06-14 2019-12-19 Rebiotix, Inc. Microbiota restoration therapy (mrt) compositions and methods of manufacture

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
HAMILTON MATTHEW J ET AL: "Standardized Frozen Preparation for Transplantation of Fecal Microbiota for Recurrent Clostridium difficile Infection", AMERICAN JOURNAL OF GASTROENTEROLOGY, ELSEVIER SCIENCE INC, US, vol. 107, no. 5, 1 May 2012 (2012-05-01), pages 761 - 767, XP009161359, ISSN: 0002-9270, DOI: 10.1038/AJG.2011.482 *
JUN HU ET AL., FRONT MICROBIOL, vol. 9, 2018, pages 1328, Retrieved from the Internet <URL:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6018536>

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